Conditions: Avian Influenza H1N1 Influenza Influenza
Keywords: A/H5N1 A/H7N9 ASO3 Adjuvant influenza monovalent pH1N1 vaccines
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 30
Healthy volunteers: yes

Brief summary:
This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, inclusive, designed to evaluate and compare the immunogenicity between an intramuscular monovalent inactivated influenza A/H7N9 virus vaccine given with and without AS03 adjuvant, and an intramuscular unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine. The primary objectives are (1) assessing the serum anti-HA hemagglutination-inhibition (HAI) response to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v), and (2) identifying differentially expressed genes in human immune cells on Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine with or without AS03) and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 vaccine with or without AS03), compared to baseline assessments performed prior to each study vaccination (Days -7, 1, and 29).

Detailed description:
This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, inclusive, designed to evaluate and compare the immunogenicity between an intramuscular monovalent inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur given with and without AS03 adjuvant manufactured by GlaxoSmithKline, and an intramuscular unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi Pasteur. The primary objectives are (1) assessing the serum anti-HA hemagglutination-inhibition (HAI) response to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v), and (2) identifying differentially expressed genes in human immune cells on Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine with or without AS03) and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 vaccine with or without AS03), compared to baseline assessments performed prior to each study vaccination (Days -7, 1, and 29). The secondary objectives are: (1) compare plasma cytokine and chemokine profiles at specific time points and between treatment arms at post vaccination points, (2) assess the neutralizing antibody responses to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v vaccine), and (3) identify differentially expressed genes in human immune cells on Days 2, 4, and 8 following one intramuscular dose of influenza A/H3N2v vaccine compared to baseline assessments performed prior to study vaccination (Day -7 and Day 1).

Arms:
arm 1: 10 Subjects: one dose of 15 µg of A/H3N2v, at Day 1 arm 2: 10 Subjects: two doses of 3.75 µg of AH7N9 AS03,at Day 1 and at Day 29 arm 3: 10 Subjects: two doses of 3.75 µg of A/H7N9, at Day 1 and Day 29

Interventions: intervention 1: AS03 intervention 2: Influenza Virus Vaccine, Monovalent A/H3N2v A/Minnesota/11/2010 NYMC X-203 intervention 3: Monovalent influenza A/H7N9 virus vaccine
Intervention descriptions: intervention 1: AS03 oil-in-water emulsion adjuvant. intervention 2: Inactivated monovalent subvirion H3N2v vaccine containing hemagglutinin (HA) of A/Minnesota/11/2010 NYMC X-203 virus. intervention 3: Monovalent influenza A/H7N9 virus vaccine.

Protocol document:
Prot_000.pdf: 
DMID Protocol 14-0015 
Version 3.0 
H7N9 Systems Biology 
17 July 2019 
 ______________________________________________________________________________________________________  
 
 
A Phase II Study to Evaluate and Compare the Immunogenicity of 
Monovalent Inactivated Influenza A/H7N9 Virus Vaccine Administered with 
and without AS03 Adjuvant and Monovalent Inactivated Influenza A/H3N2v 
Virus Vaccine Administered without Adjuvant in Healthy Adults through 
Standard and Systems Biology Analyses 
 
DMID Protocol Number: 14-0015 
 
DMID Funding Mechanism: Vaccine and Treatment Evaluation Units 
 
Pharmaceutical Support Provided by: 
Sanofi Pasteur 
GlaxoSmithKline Biologicals 
 
IND Sponsor: Division of Microbiology and Infectious Diseases, National Institute of Allergy 
and Infectious Diseases, National Institutes of Health 
 
Principal Investigator: C. Buddy Creech, MD, MPH  
DMID Clinical Project Manager: Wendy Buchanan, BSN, MS 
DMID Medical Monitor: Mohamed Elsafy, MD 
DMID Medical Officer: Francisco Leyva, MD 
DMID Scientific Lead: Chris Roberts, PhD 
DMID Contracting Officer: Kathy Ormanoski, MS, CCRA 
 
Version Number: 3.0 
17 July 2019 


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STATEMENT OF COMPLIANCE 
This study will be carried out in accordance with Good Clinical Practice (GCP) as required by the 
following:  
• United States (US) Code of Federal Regulations (CFR) applicable to clinical studies (45 
CFR Part 46; 21 CFR Part 50, 21 CFR Part 54, 21 CFR Part 56, and 21 CFR Part 312); 
• International Conference on Harmonisation (ICH) E6; 62 Federal Register 25691 (1997); 
• The Health Insurance Portability and Accountability Act (HIPAA) Privacy Rule-Final 
Modification (45 CFR Parts 160 and 164); 
• National Institutes of Health (NIH) Clinical Terms of Award, as applicable. 
Compliance with these standards provides public assurance that the rights, safety and well-being 
of study subjects are protected, consistent with the principles that have their origin in the 
Declaration of Helsinki. 
All key personnel (all individuals responsible for the design and conduct of this study) have 
completed Human Subjects Protections Training. 


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SIGNATURE PAGE 
The signature below constitutes the approval of the protocol and the attachments, and provides 
the necessary assurances that this study will be conducted according to all stipulations of the 
protocol, including all statements regarding confidentiality, and according to local legal and 
regulatory requirements and applicable US federal regulations and ICH guidelines. 
 
Site Principal Investigator: 
 
Signed: _____________________________________________ 
Date: _____________ 
 
C. Buddy Creech, MD, MPH 
Associate Professor, Pediatric Infectious Diseases 
Director, Vanderbilt Vaccine Research Program 
 
 
 
 
 
 
 
 


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TABLE OF CONTENTS 
 
 
 
STATEMENT OF COMPLIANCE .......................................................................................................... I 
SIGNATURE PAGE .................................................................................................................................. II 
TABLE OF CONTENTS ........................................................................................................................ III 
LIST OF ABBREVIATIONS ................................................................................................................ VII 
PROTOCOL SUMMARY ........................................................................................................................ X 
1 
KEY ROLES ....................................................................................................................................... 1 
2 
BACKGROUND INFORMATION AND SCIENTIFIC RATIONALE........................................ 5 
2.1 
BACKGROUND INFORMATION ....................................................................................................... 5 
2.1.1 Public Readiness and Emergency Preparedness Act .................................................... 6 
2.2 
SCIENTIFIC RATIONALE ................................................................................................................. 7 
2.3 
POTENTIAL RISKS AND BENEFITS ................................................................................................. 8 
2.3.1 Potential Risks .............................................................................................................. 8 
2.3.2 Known Potential Benefits ........................................................................................... 16 
3 
STUDY OBJECTIVES AND OUTCOME MEASURES .............................................................. 17 
4 
STUDY DESIGN ............................................................................................................................... 22 
5 
STUDY ENROLLMENT AND WITHDRAWAL ......................................................................... 24 
5.1 
ELIGIBILITY CRITERIA................................................................................................................. 24 
5.1.1 Subject Inclusion Criteria ........................................................................................... 24 
5.1.2 Subject Exclusion Criteria .......................................................................................... 25 
5.2 
TREATMENT ASSIGNMENT PROCEDURES.................................................................................... 28 
5.2.1 Enrollment and Randomization Procedures ............................................................... 28 
5.2.2 Masking Procedures ................................................................................................... 30 
5.2.3 Reasons for Withdrawals and Discontinuation of Treatment ..................................... 31 
5.2.4 Handling of Withdrawals and Discontinuation of Treatment .................................... 33 
5.2.5 Termination of Study .................................................................................................. 33 
6 
STUDY INTERVENTION/INVESTIGATIONAL PRODUCT ................................................... 35 
6.1 
STUDY PRODUCT DESCRIPTION .................................................................................................. 35 
6.1.1 Acquisition.................................................................................................................. 36 
6.1.2 Formulation, Storage, Packaging, and Labeling ......................................................... 37 
6.1.3 Study Product Storage and Stability Procedures ........................................................ 37 
6.2 
DOSAGE, PREPARATION, AND ADMINISTRATION OF STUDY INTERVENTION/INVESTIGATIONAL 
PRODUCT ................................................................................................................................................. 38 


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6.3 
MODIFICATION OF STUDY INTERVENTION/INVESTIGATIONAL PRODUCT FOR A SUBJECT ......... 40 
6.4 
ACCOUNTABILITY PROCEDURES FOR THE STUDY INTERVENTION/ INVESTIGATIONAL 
PRODUCT(S) ............................................................................................................................................ 40 
6.5 
ASSESSMENT OF SUBJECT COMPLIANCE WITH STUDY INTERVENTION/INVESTIGATIONAL 
PRODUCT ................................................................................................................................................. 41 
6.6 
CONCOMITANT MEDICATIONS/TREATMENTS ............................................................................. 41 
7 
STUDY SCHEDULE ........................................................................................................................ 43 
7.1 
SCREENING AND ENROLLMENT VISITS ....................................................................................... 43 
7.1.1 Visit 00, Day -7, Screening, Clinic Visit, All Subjects .............................................. 43 
7.1.2 Visit 01, Day 1, Enrollment and First Study Vaccination (Dose 1), Clinic Visit, All 
Subjects .................................................................................................................................. 44 
7.2 
FOLLOW-UP VISITS...................................................................................................................... 46 
7.2.1 Visit 02, Day 2, Clinic Visit, All Subjects.................................................................. 46 
7.2.2 Visit 03, Day 4, Clinic Visit, All Subjects.................................................................. 47 
7.2.3 Visit 04, Day 8, Clinic Visit, A/H3N2v-vaccinated subjects only ............................. 47 
7.2.4 Visit 04, Day 8, Phone Call/Electronic Communication, A/H7N9-vaccinated subjects 
only 48 
7.2.5 Visit 05, Day 29, *Second Study Vaccination (Dose 2) for A/H7N9-vaccinated 
subjects only, Clinic Visit, All Subjects ................................................................................ 48 
7.2.6 Visit 06, Day 30, Clinic Visit, A/H7N9-vaccinated subjects only ............................. 50 
7.2.7 Visit 07, Day 32, Clinic Visit, A/H7N9-vaccinated subjects only ............................. 51 
7.2.8 Visit 08, Day 36, Clinic Visit, A/H7N9-vaccinated subjects only ............................. 52 
7.2.9 Visit 09, Day 57, Clinic Visit, A/H7N9-vaccinated subjects only ............................. 53 
7.2.10 Visit 10, Day 209, Phone Call/Electronic Communication, A/H7N9-vaccinated 
subjects only .......................................................................................................................... 53 
7.2.11 Visit 11, Day 394, Phone Call/Electronic Communication, A/H7N9-vaccinated 
subjects only .......................................................................................................................... 53 
7.3 
EARLY TERMINATION VISIT (IF NEEDED), ALL SUBJECTS .......................................................... 54 
7.4 
UNSCHEDULED VISIT (IF NEEDED), ALL SUBJECTS..................................................................... 55 
8 
STUDY PROCEDURES AND EVALUATIONS ........................................................................... 57 
8.1 
CLINICAL EVALUATIONS............................................................................................................. 57 
8.2 
LABORATORY EVALUATIONS ..................................................................................................... 58 
8.2.1 Clinical Laboratory Evaluations ................................................................................. 59 
8.2.2 Special Assays or Procedures ..................................................................................... 60 
8.2.2.1 
HAI and Neut Antibody Assays – Serologic Immunogenicity ....................................... 60 
8.2.2.2 
Systems Biology Studies – Cellular Immunogenicity .................................................... 60 
8.2.3 Specimen Preparation, Handling, and Shipping ......................................................... 65 
8.2.3.1 
Instructions for Specimen Preparation, Handling, and Storage ...................................... 65 
8.2.3.2 
Specimen Shipment......................................................................................................... 65 


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ASSESSMENT OF SAFETY ........................................................................................................... 66 
9.1 
SPECIFICATION OF SAFETY PARAMETERS ................................................................................... 66 
9.2 
METHODS AND TIMING FOR ASSESSING, RECORDING, AND ANALYZING SAFETY PARAMETERS
 
66 
9.2.1 Adverse Events ........................................................................................................... 66 
9.2.2 Reactogenicity ............................................................................................................ 68 
9.2.3 Additional Adverse Event Severity Grading .............................................................. 71 
9.2.4 Serious Adverse Events .............................................................................................. 71 
9.2.5 Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings .................................................................................................. 72 
9.3 
REPORTING PROCEDURES ........................................................................................................... 72 
9.3.1 Serious Adverse Events .............................................................................................. 73 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND ............ 73 
9.3.3 Reporting of Pregnancy .............................................................................................. 74 
9.4 
TYPE AND DURATION OF FOLLOW-UP OF SUBJECTS AFTER ADVERSE EVENTS ......................... 74 
9.5 
HALTING RULES .......................................................................................................................... 75 
9.6 
SAFETY OVERSIGHT .................................................................................................................... 76 
9.6.1 Independent Safety Monitor (ISM) ............................................................................ 76 
9.6.2 Safety Monitoring Committee (SMC) ........................................................................ 76 
10 CLINICAL MONITORING ............................................................................................................ 78 
10.1 
SITE MONITORING PLAN ............................................................................................................. 78 
11 STATISTICAL CONSIDERATIONS ............................................................................................ 79 
11.1 
INTRODUCTION ............................................................................................................................ 79 
11.2 
STUDY HYPOTHESES ................................................................................................................... 79 
11.3 
STUDY OUTCOME MEASURES ..................................................................................................... 80 
REFER TO SECTION 3 FOR STUDY OUTCOME MEASURES. ........................................................................ 80 
11.4 
SAMPLE SIZE CONSIDERATIONS .................................................................................................. 80 
11.4.1 Study Design............................................................................................................... 80 
11.4.2 Study Population......................................................................................................... 81 
11.4.3 Sample Size ................................................................................................................ 81 
11.4.4 Planned Interim Analyses ........................................................................................... 82 
11.4.5 Interim Safety Review ................................................................................................ 82 
11.4.6 Interim Immunogenicity Review ................................................................................ 82 
11.5 
FINAL ANALYSIS PLAN ............................................................................................................... 83 
11.5.1 Systems Biology Analyses ......................................................................................... 83 
11.5.1.1 Transcriptomics (RNA-seq) ............................................................................................ 83 
11.5.1.2 Proteomics ....................................................................................................................... 84 
11.5.2 Serologic Immunogenicity Data (HAI and Neut Antibody Assays) .......................... 85 
11.5.3 Cytokine/Chemokines ................................................................................................ 85 


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11.5.4 Primary Clinical Study Report (CSR) Database Lock ............................................... 85 
11.5.5 Analysis Populations .................................................................................................. 86 
11.5.6 Safety Data ................................................................................................................. 87 
11.5.7 Missing Values and Outliers ....................................................................................... 87 
12 DATA COLLECTION FORMS AND ACCESS TO SOURCE DATA/DOCUMENTS ............ 88 
13 QUALITY CONTROL AND QUALITY ASSURANCE .............................................................. 89 
14 ETHICS/PROTECTION OF HUMAN SUBJECTS ..................................................................... 90 
14.1 
ETHICAL STANDARD ................................................................................................................... 90 
14.2 
INSTITUTIONAL REVIEW BOARD ................................................................................................. 90 
14.3 
INFORMED CONSENT PROCESS.................................................................................................... 90 
14.3.1 Informed Consent ....................................................................................................... 90 
14.4 
EXCLUSION OF WOMEN, MINORITIES, AND CHILDREN (SPECIAL POPULATIONS) ...................... 92 
14.5 
SUBJECT CONFIDENTIALITY ........................................................................................................ 92 
14.6 
STUDY DISCONTINUATION .......................................................................................................... 93 
14.7 
COSTS, SUBJECT COMPENSATION, AND RESEARCH RELATED INJURIES .................................... 93 
14.8 
FUTURE USE OF STORED SPECIMENS .......................................................................................... 94 
15 DATA HANDLING AND RECORD KEEPING ........................................................................... 95 
15.1 
DATA MANAGEMENT RESPONSIBILITIES .................................................................................... 95 
15.2 
DATA CAPTURE METHODS.......................................................................................................... 95 
15.3 
TYPES OF DATA ........................................................................................................................... 96 
15.4 
TIMING/REPORTS ........................................................................................................................ 96 
15.5 
STUDY RECORDS RETENTION ..................................................................................................... 97 
15.6 
PROTOCOL DEVIATIONS .............................................................................................................. 97 
16 PUBLICATION POLICY ................................................................................................................ 99 
17 LITERATURE REFERENCES .................................................................................................... 101 
APPENDIX A: LIST OF POTENTIALLY IMMUNE-MEDIATED MEDICAL CONDITIONS . 106 
APPENDIX B: SCHEDULE OF EVENTS .......................................................................................... 109 
 
 


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LIST OF ABBREVIATIONS 
A/H1N1 
Influenza A Virus of the H1N1 Subtype 
A/H3N2v 
Influenza A Virus of the H3N2 Variant Subtype 
A/H5N1 
Influenza A Virus of the H5N1 Subtype 
A/H7N7 
Influenza A Virus of the H7N7 Subtype 
A/H7N9 
Influenza A Virus of the H7N9 Subtype 
AdvantageEDCSM 
Electronic Data Capture System 
AE 
Adverse Event/Adverse Experience 
AESIs 
Adverse Events of Special Interest 
AS03 
Adjuvant System (03) 
α 
Alpha 
β 
Beta 
BARDA 
Biomedical Advanced Research and Development Authority 
BMI 
Body Mass Index 
BP 
Blood Pressure 
BPM 
Beats Per Minute 
CAR 
Clinical Agents Repository 
CBA 
Cytometric Bead Arrays 
CDC 
Centers for Disease Control and Prevention 
CFR 
Code of Federal Regulations 
CHMP 
Committee for Medicinal Products for Human Use 
CI 
Confidence Interval 
CMI 
Cell-Mediated Immunity 
CSR 
Clinical Study Report 
°C 
Degrees Celsius 
°F 
Degrees Fahrenheit 
D 
Day(s) 
DC 
Dendritic Cell 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH 
eCRF 
Electronic Case Report Form 
ESR 
Erythrocyte Sedimentation Rate 
FACS 
Fluorescence-Activated Cell Sorting 
FDA 
Food and Drug Administration 
FDAAA 
Food and Drug Administration Amendments Act 
FWA 
Federalwide Assurance 
GBS 
Guillain-Barré Syndrome 


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GCP 
Good Clinical Practice 
GMT 
Geometric Mean Titer 
GSK 
GlaxoSmithKline Biologicals 
HA 
Hemagglutinin 
HAI 
Hemagglutination Inhibition 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HRSA 
Health Resources and Services Administration 
IATA 
International Air Transport Association 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
IFN 
Interferon 
IL 
Interleukin 
IM 
Intramuscular 
IND 
Investigational New Drug Application 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
iTRAQ 
Isobaric Tags for Relative and Absolute Quantitation 
ITT 
Intent-to-Treat 
IUD 
Intrauterine Device 
MAAEs 
Medically-Attended Adverse Events 
MACS 
Magnetic Cell Separation 
mcg or µg 
Microgram(s) 
MedDRA 
Medical Dictionary for Regulatory Activities 
mL 
Milliliter(s) 
mm 
Millimeter(s) 
mmHg 
Millimeters of Mercury 
MOP 
Manual of Procedures 
N 
Number of Subjects 
Neut 
Neutralizing or Neutralization 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, DHHS 
NK 
Natural Killer 
NIH 
National Institutes of Health 
NSAIDs 
Non-Steroidal Anti-Inflammatory Drugs 
NOCMCs 
New-Onset Chronic Medical Conditions 
OHRP 
Office for Human Research Protections 
PBS 
Phosphate-Buffered Saline 
PHI 
Protected Health Information 
PK 
Pharmacokinetics 


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PIMMCs 
Potentially Immune-Mediated Medical Conditions 
PMN 
Polymorphonuclear Leukocyte 
PP 
Per Protocol 
PREP Act 
Public Readiness and Emergency Preparedness Act 
QA 
Quality Assurance 
QC 
Quality Control 
RNA 
Ribonucleic Acid 
RP-HPLC 
Reversed-Phase High-Performance Liquid Chromatography 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SDCC 
Statistical and Data Coordinating Center 
SMC 
Safety Monitoring Committee 
SOC 
System Organ Class 
SOP 
Standard Operating Procedure 
SRID 
Single Radial Immunodiffusion 
TMT 
Tandem Mass Tags 
TNF 
Tumor Necrosis Factor 
US 
United States 
V 
Visit(s) 
VTEU 
Vaccine and Treatment Evaluation Unit 
WHO 
World Health Organization 


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PROTOCOL SUMMARY 
Title: 
A Phase II Study to Evaluate and Compare the Immunogenicity 
of Monovalent Inactivated Influenza A/H7N9 Virus Vaccine 
Administered with and without AS03 Adjuvant and Monovalent 
Inactivated Influenza A/H3N2v Virus Vaccine Administered 
without AS03 Adjuvant in Healthy Adults through Standard and 
Systems Biology Analyses 
Phase: 
II 
Population: 
Approximately 30 healthy male and non-pregnant female 
subjects aged 18 to 49 years old, inclusive, in the US 
Number of Sites: 
One Vaccine and Treatment Evaluation Unit (VTEU) site: 
Vanderbilt University 
Study Duration: 
Approximately 36 months 
Subject Participation 
Duration: 
Approximately 14 months 
Estimated Time to 
Complete 
Enrollment: 
Description of Agent 
or Intervention: 
Approximately 6 months 
 
Two doses (3.75 micrograms [mcg] of hemagglutinin [HA]) of a 
monovalent inactivated influenza A/H7N9 virus vaccine 
(A/Shanghai/2/2013) manufactured by Sanofi Pasteur 
administered intramuscularly approximately 28 days apart given 
with and without AS03 adjuvant manufactured by 
GlaxoSmithKline Biologicals (GSK) 
One dose (15 mcg HA) of a monovalent inactivated influenza 
A/H3N2v virus vaccine (A/Minnesota/11/2010) manufactured 
by Sanofi Pasteur administered intramuscularly 


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Study Objectives and 
Outcome Measures: 
 
 
Primary Objectives 
 
1. To assess the serum anti-HA hemagglutination-inhibition 
(HAI) response to influenza A/H7N9 antigen (with and 
without adjuvant) at Day 57 (approximately one month after 
the second study vaccination with A/H7N9 vaccine ± AS03) 
and influenza A/H3N2v antigen at Day 29 (approximately 
one month after the study vaccination with A/H3N2v 
vaccine). 
 
2. To identify differentially expressed genes in human immune 
cells on Days 2, 4, and 29 (following the first study 
vaccination with A/H7N9 vaccine ± AS03) and on Days 30, 
32, and 36 (following the second study vaccination with 
A/H7N9 vaccine ± AS03), compared to baseline assessments 
performed prior to each study vaccination (Days -7, 1, and 
29). 
 
Primary Outcome Measures 
 
1. Percentage of subjects achieving seroconversion (defined as 
either a pre-vaccination HAI titer <1:10 and a post-
vaccination HAI titer ≥1:40 or a pre- vaccination HAI titer 
≥1:10 and a minimum four-fold rise in post-vaccination HAI 
antibody titer) at approximately 28 days after the second 
study vaccination (A/H7N9 vaccine, with and without 
adjuvant). 
 
2. Percentage of subjects achieving seroconversion at 
approximately 28 days after the study vaccination with 
A/H3N2v vaccine. 
 
3. Number of differentially expressed genes based on RNA 
expression as determined by RNA-Seq analysis, in human 
immune cells at Days 2, 4, 29, 30, 32, and 36 after study 
vaccination with AS03-adjuvanted and unadjuvanted 
A/H7N9 vaccine, as compared to baseline assessments 
performed prior to vaccination (Days -7, 1, and 29). 


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Secondary Objectives 
 
1. To compare plasma cytokine and chemokine profiles at 
specific time points and between treatment arms: 
a. Post-vaccination time points [(Days 2 and 4; Day 8 
(A/H3N2v arm only)], and Days 30, 32, and 36 
(A/H7N9 arms only) with pre-vaccination time points 
(Days -7 and 1, prior to first study vaccination and 
Day 29, prior to second study vaccination (A/H7N9 
arms only). 
 
2. To assess the neutralizing antibody responses to influenza 
A/H7N9 antigen (with and without adjuvant) at Day 57 
(approximately one month after the second study vaccination 
with A/H7N9 vaccine ± AS03) and influenza A/H3N2v 
antigen at Day 29 (approximately one month after the study 
vaccination with A/H3N2v vaccine). 
 
3. To identify differentially expressed genes in human immune 
cells on Days 2, 4, and 8 following one intramuscular dose of 
influenza A/H3N2v vaccine compared to baseline 
assessments performed prior to study vaccination (Day -7 
and Day 1). 
 
Secondary Outcome Measures 
 
1. 
Plasma measurement of cytokines and chemokines at each 
study visit, comparing: 
a. 
Post-vaccination time points with pre-vaccination 
time points by treatment arms. 
b. 
AS03-adjuvanted influenza A/H7N9 responses to 
unadjuvanted influenza A/H7N9 responses. 
c. 
Influenza A/H3N2v vaccine to unadjuvanted 
influenza A/H7N9 vaccine. 
d. 
Influenza A/H3N2v vaccine to adjuvanted A/H7N9 
vaccine. 
 
2. 
Serologic response to influenza hemagglutinin: 


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a. 
Percentage of subjects achieving seroconversion 
(defined as either a pre-vaccination Neut titer <1:10 
and a post-vaccination Neut titer ≥1:40 or a pre-
vaccination Neut titer ≥1:10 and a minimum four-fold 
rise in post-vaccination Neut titer) at approximately 
28 days after the second study vaccination (A/H7N9 
vaccine, with and without adjuvant). 
b. 
Percentage of subjects achieving seroconversion at 
approximately 28 days after the study vaccination 
with influenza A/H3N2v vaccine. 
c. 
Geometric Mean Titers of serum HAI and Neut 
antibody at baseline (Day -7 and 1) and 28 days after 
influenza A/H3N2v vaccination. 
d. 
Geometric Mean Titers of serum HAI and Neut 
antibody at baseline (Day -7 and 1), and 28 days after 
both the first and the second dose of influenza 
A/H7N9 vaccine (with and without adjuvant). 
 
3. Number of differentially expressed genes based on RNA-Seq 
analysis in human immune cells at Days 2, 4, and 8 after 
study vaccination, as compared to baseline assessments 
(Days -7 and 1) after influenza A/H3N2v vaccine. 
 
Tertiary Objectives 
 
1. To assess the frequency of adverse events. 
 
2. To identify differentially abundant cellular proteins in 
human immune cells on Days 2, 4, and 8 following one 
intramuscular dose of influenza A/H3N2v vaccine compared 
to baseline assessments performed prior to study vaccination 
(Day -7 and Day 1). 
 
3. To identify differentially abundant cellular proteins in 
human immune cells on Days 2, 4, and 29 (following first 
study vaccination with A/H7N9 vaccine ± AS03) and Days 
30, 32, and 36 (following second study vaccination with 
A/H7N9 vaccine ± AS03), compared to baseline 


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assessments performed prior to study vaccination (Days -7, 
1, and 29). 
 
Tertiary Outcome Measures 
 
1. Occurrence of solicited local reactogenicity events within 7 
days after each study vaccination. 
 
2. Occurrence of solicited systemic reactogenicity events within 
7 days after each study vaccination. 
 
3. Occurrence of unsolicited AEs collected for approximately 
28 days after last study vaccination. 
 
4. Occurrence of SAEs and MAAEs including new-onset 
chronic medical conditions and potentially immune-mediated 
medical conditions for approximately 1 month (A/H3N2v 
arm) and 12 months (A/H7N9 arms) after last study 
vaccination. 
 
5. Number of differentially abundant proteins based on protein 
abundance as determined by quantitative proteomics in 
human immune cells at Days 2, 4, and 8 after study 
vaccination, as compared to baseline assessments (Days -7 
and 1) after influenza A/H3N2v vaccine. 
 
6. Number of differentially abundant proteins based on protein 
abundance as determined by quantitative proteomics analysis, 
in human immune cells at Days 2, 4, 29, 30, 32, and 36 after 
study vaccination with AS03-adjuvanted and unadjuvanted 
A/H7N9 vaccine comparing changes in protein abundance 
between treatment arms (A/H7N9 vaccine vs. A/H7N9 
vaccine + AS03). 
 
Exploratory Objectives 
 
1. To identify and characterize differentially expressed 
genes in human immune cells on Days 2, 4, and 29 
(following the first study vaccination with A/H7N9 


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vaccine ± AS03) and on Days 30, 32, and 36 (following 
the second study vaccination with A/H7N9 vaccine ± 
AS03) that differ between treatments (A/H7N9 vaccine 
vs. A/H7N9 vaccine + AS03). 
 
2. To identify and characterize differentially abundant 
cellular proteins in human immune cells on Days 2, 4, 
and 29 (following the first study vaccination with 
A/H7N9 vaccine ± AS03) and on Days 30, 32, and 36 
(following the second study vaccination with A/H7N9 
vaccine ± AS03) that differ between treatments (A/H7N9 
vaccine vs. A/H7N9 vaccine + AS03). 
 
3. To identify and characterize differentially expressed 
genes in human immune cells on Days 2 and 4 (following 
the first study vaccination) that differ in baseline 
responses (Days -7, and 1) between treatments (A/H3N2v 
vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. 
A/H7N9 vaccine + AS03). 
 
4. To identify and characterize differentially abundant 
cellular proteins in human immune cells on Days 2 and 4 
(following the first study vaccination) that differ in 
baseline responses (Days -7, and 1) between treatments 
(A/H3N2v vaccine vs. A/H7N9 vaccine and A/H3N2v 
vaccine vs. A/H7N9 vaccine + AS03). 
 
5. To identify transcriptomic and proteomic signatures in 
human immune cells that correlate with influenza 
seroconversion and with peak HAI titers. 
 
6. To correlate plasma cytokine profiles with transcriptomic 
and proteomic profiles in subjects receiving influenza 
A/H3N2v vaccine or influenza A/H7N9 ± AS03 vaccine. 
 
7. Exploratory inspection of transcriptomics\proteomics 
response signatures for subjects with unexpected or 
severe safety profiles that are related to the study 
products. 


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Exploratory Outcome Measures 
 
1. RNA expression, clusters of co-expressed genes, 
enriched pathways, and other functional modules as 
determined by RNA-Seq analysis, in human immune 
cells at Days 2, 4, 29, 30, 32, and 36 after study 
vaccination with AS03-adjuvanted and unadjuvanted 
A/H7N9 vaccine comparing changes in RNA expression 
between treatment arms (A/H7N9 vaccine vs. A/H7N9 
vaccine + AS03). 
 
2. Protein abundance, clusters of co-abundant proteins, 
enriched pathways, and other functional modules as 
determined by quantitative proteomics analysis, in human 
immune cells at Days 2, 4, 29, 30, 32, and 36 after study 
vaccination with AS03-adjuvanted and unadjuvanted 
A/H7N9 vaccine comparing changes in protein 
abundance between treatment arms (A/H7N9 vaccine vs. 
A/H7N9 vaccine + AS03). 
 
3. RNA expression, clusters of co-expressed genes, 
enriched pathways, and other functional modules as 
determined by RNA-Seq analysis, in human immune 
cells at Days 2, 4, and 29 after study vaccination 
comparing baseline changes in RNA expression between 
treatment arms (A/H3N2v vaccine vs. A/H7N9 vaccine 
and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
 
4. Protein abundance, clusters of co-abundant proteins, 
enriched pathways, and other functional modules as 
determined by quantitative proteomics analysis, in human 
immune cells at Days 2, 4, and 29 after first study 
vaccination comparing baseline changes (Days -7, and 1) 
in protein abundance between treatment arms (A/H3N2v 
vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. 
A/H7N9 vaccine + AS03). 
 


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5. Changes from baseline (Days -7, 1, and 29) in RNA 
expression or protein abundance that best correlate with 
influenza A/H7N9 seroconversion and with peak HAI 
titers at Days 2 and 4 (following first study vaccination) 
as well as Days 30, 32, and 36 (following second study 
vaccination). 
 
6. Changes from baseline (Days -7, 1, and 29) in RNA 
expression or protein abundance that best correlate with 
baseline changes in cytokine concentrations at Days 2, 4, 
8 (A/H3N2v arm only) and 29 (following first study 
vaccination) as well as Days 30, 32, and 36 (following 
second study vaccination). 
 
7. Exploratory summaries of transcriptomics/proteomics 
response signatures for subjects with unexpected or 
severe safety profiles. 
 
8. Lists of differential genes and proteins identified as part 
of the primary and secondary objectives. 
 
Description of Study 
Design: 
This is a single center, randomized, partially-blinded, Phase II, 
small, targeted, prospective study in approximately 30 healthy 
male and non-pregnant female subjects aged 18 to 49 years old, 
inclusive, designed to evaluate and compare the immunogenicity 
between an intramuscular monovalent inactivated influenza 
A/H7N9 virus vaccine manufactured by Sanofi Pasteur given 
with and without AS03 adjuvant manufactured by 
GlaxoSmithKline, and an intramuscular unadjuvanted 
monovalent inactivated influenza A/H3N2v virus vaccine 
manufactured by Sanofi Pasteur. 
This study will use a standard and systems biology approach to 
assess the human early gene and protein signatures expressed at 
baseline (approximately Days -7 and 1), and at approximately 
Days 2, 4, and 8 (A/H3N2v-vaccinated subjects only) after the 
first study vaccination in each treatment arm as well as at 
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subjects only.  Cellular immunogenicity (systems biology 
studies) data will be integrated with serologic immunogenicity 
(HAI and Neut antibody assays) and reactogenicity data to 
develop a systems model of the human immune response to 
unadjuvanted monovalent inactivated influenza A/H3N2v virus 
vaccine and monovalent inactivated influenza A/H7N9 virus 
vaccine given with and without AS03 adjuvant. 
This study will use venous blood samples and subject data 
collected from a total of thirty vaccinated subjects randomly 
divided into three equal treatment arms.  The first treatment arm 
(n=10) will be vaccinated with one dose of 15 µg of A/H3N2v 
HA.  The second treatment arm (n=10) will be vaccinated with 
two doses of 3.75 µg of A/H7N9 HA given with AS03 
approximately 28 days apart.  The third treatment arm (n=10) 
will be vaccinated with two doses of 3.75 µg of A/H7N9 HA 
given without AS03 adjuvant approximately 28 days apart.  All 
study vaccinations will be administered intramuscularly. 
Reactogenicity will be measured by the occurrence of solicited 
injection site and systemic reactions from the time of each study 
vaccination through 8 days after each study vaccination.  
Unsolicited non-serious adverse events (AEs) will be collected 
from the time of each study vaccination through approximately 
28 days after each study vaccination.  Serious adverse events 
(SAEs) and medically-attended adverse events (MAAEs) 
including new-onset chronic medical conditions (NOCMCs) and 
potentially immune-mediated medical conditions (PIMMCs) will 
be collected from the time of the first study vaccination through 
approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 
arms) after the last study vaccination. 
Venous blood samples (approximately 90 mL) will be collected 
from subjects at approximately Days -7 and 1 (immediately prior 
to the first study vaccination), and at approximately Days 2, 4; 
and 8 (A/H3N2v-vaccinated subjects only) after the first study 
vaccination for systems biology studies (cytokine and chemokine 
levels, immune cell activation status, and whole transcriptome 
and proteome profiles of the major blood immune cells).  


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Additionally, subjects in either treatment arm receiving A/H7N9 
vaccine will have additional blood samples collected at 
approximately Days 29 (this will be collected immediately prior 
to the second study vaccination for subjects in either group 
receiving A/H7N9), 30, 32, and 36 after the first study 
vaccination for systems biology studies. 
Serological assessment (hemagglutination inhibition (HAI) and 
neutralizing (Neut) antibody assays) will also be conducted on 
venous blood samples (approximately 10 mL) collected from 
subjects at Day 1 (immediately prior to the first study 
vaccination), and at approximately Days 29 (for A/H7N9-
vaccinated subjects this will be collected immediately prior to 
the second study vaccination) and 57 (A/H7N9-vaccinated 
subjects only) after the first study vaccination. 
We will quantify and characterize serum cytokine/chemokine 
levels, as well as transcriptomic and proteomic profiles from 
individual immune cell compartments, comparing this with 
standard serologic assessment to vaccine. 
Urine samples (~20 mL) will also be collected from subjects on 
Days -7, 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only), 29, 30, 
32, and 36 for future research.  Note: Days 29, 30, 32, and 36 
will apply only to A/H7N9-vaccinated subjects. 
 


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Overview of Study Design: 
Objective 
Study Day 
Assays 
Baseline pre-vaccination (1) 
Day -7 
RNA-seq/proteomics from Immune cell 
compartments (ICC) 
Urine samples 
Baseline pre-vaccination (2) and First Study Vaccination 
Day 1 
RNA-seq/proteomics from ICC 
Urine samples 
HAI and Neut Assays 
Early innate immune response to first study vaccination 
Day 2 
RNA-seq/proteomics from ICC 
Urine samples 
Intermediate innate immune response to first study 
vaccination 
Day 4 
RNA-seq/proteomics from ICC 
Urine samples 
Late innate immune response and early adaptive immune 
response to first study vaccination  
(A/H3N2v-vaccinated subjects only) 
Day 8 
RNA-seq/proteomics from ICC 
Urine samples 
Mature adaptive immune response to first study vaccination 
and new baseline for Second Study Vaccination 
Day 29 
RNA-seq/proteomics from ICC (A/H7N9-
vaccinated subjects only) 
Urine samples (A/H7N9 vaccinated-subjects 
only) 
HAI and Neut Assays 
Early innate immune response to second study vaccination 
(A/H7N9-vaccinated subjects only) 
Day 30 
RNA-seq/proteomics from ICC 
Urine samples 
Intermediate innate immune response to second study 
vaccination 
(A/H7N9-vaccinated subjects only) 
Day 32 
RNA-seq/proteomics from ICC 
Urine samples 
Late innate immune response and early adaptive immune 
response to second study vaccination  
(A/H7N9-vaccinated subjects only) 
Day 36 
RNA-seq/proteomics from ICC 
Urine samples 
Final serologic assessment 
(A/H7N9-vaccinated subjects only) 
Day 57 
HAI and Neut Assays 
 
Treatment Arms: 
Treatment Arms 
(n=10 vaccinated subjects per 
Treatment Arm) 
First Study Vaccination 
(Day 1) 
Second Study Vaccination 
(Day 29±2 days) 
1 
15 µg A/H3N2v* 
N/A 
2 
3.75 µg A/H7N9 + AS03 
3.75 µg A/H7N9 + AS03 
3 
3.75 µg A/H7N9* 
3.75 µg A/H7N9*  
Total N=30 vaccinated subjects 
* unadjuvanted 
 










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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
Since its emergence in Southeast Asia in 2013, influenza A/H7N9/Shanghai continues to infect 
humans and is associated with a high case fatality rate1.  The virus holds significant pandemic 
potential, since the avian population is rarely symptomatic (making avian disease surveillance 
difficult), and there is little preexisting immunity to the virus in the human population2; however, 
its current inability to sustain efficient human-to-human transmission has limited its spread3. 
Unfortunately, as with other avian influenza strains, the immunogenicity of inactivated vaccines 
is poor.  Higher doses of hemagglutinin (HA), a major target of the protective immune response, 
generate more frequent and higher antibody titers4; however clinical trials of influenza A/H5N1, 
influenza A/H7N7, and influenza A/H7N9 vaccines have clearly demonstrated that avian HA’s 
are substantially less immunogenic than seen with seasonal vaccines, even when given in very 
large doses5-7.  For example, in clinical studies of a subvirion inactivated influenza 
A/H5N1/Vietnam vaccine in healthy adults, two 90 mcg doses of vaccine were required to 
stimulate an antibody response in just over half (57%) of the recipients8.  A NIAID sponsored 
Phase I/II trial studying the safety and immunogenicity of an inactivated influenza A/H7N9 
vaccine revealed that even two doses of 90 mcg each were insufficient to generate measurable 
antibody responses in healthy adults9.  Moreover, adjuvant was required to induce protective 
responses against inactivated A/H7N9 vaccine in two NIAID sponsored Phase I clinical trials.  
While 59% of subjects seroconverted when vaccinated with 3.75 mcg of A/H7N9 vaccine in the 
presence of MF59 adjuvant, only 5% of subjects seroconverted when vaccinated with 45 mcg of 
A/H7N9 vaccine in the absence of MF59 adjuvant6,7. 
The use of adjuvants is a promising alternative to increasing HA content or administering 
additional doses of vaccine.  Adjuvants have the potential to increase both the cellular and 
humoral responses at a given dose of antigen, to decrease the amount of antigen needed in the 
vaccine, and to improve immunogenicity among those that generally respond poorly to influenza 
antigens (e.g., the elderly or immunocompromised).  While the addition of adjuvants has 
sometimes been associated with increased injection site reactogenicity and occasionally systemic 
reactogenicity, the overall safety of novel oil-in water adjuvants such as MF59 (Novartis 
Vaccines and Diagnostics) and AS03 (GSK) is reassuring10-14. 
Currently, one AS03-adjuvanted influenza A/H5N1 vaccine and one MF59-adjuvanted influenza 
A/H5N1 vaccine (FLUAD) are licensed in the US, and both adjuvants are licensed in Europe as 
part of multiple influenza vaccines.  When used as a component of influenza A/H5N1 vaccine, 


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AS03 leads to substantially improved HA responses when compared to non-adjuvanted 
vaccine15-21.  In an early study, two doses of A/H5N1 vaccine, with and without AS03 were 
given, with dosages ranging from 3.75 to 30 mcg of HA.  After two doses, only the 30 mcg non-
adjuvanted formulation met the Committee for Medicinal Products for Human Use (CHMP) 
criteria for seroconversion (>40%) while CHMP criteria were met by all adjuvanted 
formulations, even down to 3.75 mcg of HA.  Moreover, seroconversion criteria were met by HA 
doses of 7.5 mcg or greater in the adjuvanted group after only the first dose.  These data, as well 
as data from the 2009 pandemic A/H1N1 experience, demonstrate the potency and safety of 
AS03 as an influenza vaccine adjuvant. 
2.1.1 
Public Readiness and Emergency Preparedness Act 
The protocol and the study products (monovalent inactivated influenza A/H7N9 virus vaccine 
and monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi Pasteur as 
well as AS03 adjuvant manufactured by GlaxoSmithKline Biologicals) tested are covered under 
the Public Readiness and Emergency Preparedness Act (PREP Act).  Under the PREP Act, 
covered persons are immune from liability actions brought from the administration or use of a 
covered countermeasure that is the subject of a declaration.  The PREP Act provides immunity 
for covered persons (such as manufacturers, distributers, program planners and other qualified 
persons who prescribe, administer or dispense the study vaccines with or without AS03 adjuvant) 
from tort liability, unless the injury was caused by willful misconduct. 
The PREP Act also authorized a “Covered Countermeasures Process Fund” to provide 
compensation to eligible individuals who suffer specified injuries from administration or use of a 
countermeasure pursuant to the declaration.  Any requests for compensation must be filed within 
one year of administration or use of the countermeasure.  Requests would go to the Health 
Resources and Services Administration (HRSA) Preparedness Countermeasures Injury 
Compensation Program (http://www.hrsa.gov/cicp/).  Compensation may then be available for 
medical benefits, lost wages and death benefits to eligible individuals for specified injuries in 
accordance with regulations published by the Secretary of HRSA.  Eligibility for compensation 
and the injuries for which compensation may be available are further defined by regulation. 
An individual who suffers a serious physical injury or death from administration and use of the 
study vaccines with or without AS03 adjuvant must first seek compensation from the Covered 
Countermeasures Process Fund.  A serious physical injury means an injury that is life 
threatening, results in, or requires medical or surgical intervention to prevent, permanent 
impairment of body function or permanent damage to body structure.  Any compensation will be 
reduced by public or private insurance or worker’s compensation available to the injured 
individual. 


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If no funds have been appropriated to the compensation program, the Secretary of HRSA does 
not make a final determination on the individual’s request within 240 days, or if the individual 
decides not to accept the compensation, the injured individual or his representative may pursue a 
tort claim in the US District Court for the District of Columbia, but only if the claim involves 
willful misconduct, is pled with particularity required under the PREP Act, verified, and 
accompanied by an affidavit by a physician who did not treat the individual and certified medical 
records.  Any award is reduced by any public or private insurance or worker’s compensation 
available to the injured individual.  Awards for non-economic damages, such as pain, suffering, 
physical impairment, mental anguish, and loss of consortium are also limited.  If the individual 
accepts compensation, or if there is no willful misconduct, then the individual does not have a 
tort claim that can be filed in a United States Federal or a State court. 
2.2 
Scientific Rationale 
The cellular and molecular mechanisms of action by which novel oil-in-water emulsion 
adjuvants enhance the immune response to vaccination are not completely understood22,23.  
Recently published data from mouse models suggest that AS03 works locally to stimulate 
immune effectors24.  AS03 also induces the local production of monocyte-, dendritic cell (DC)-, 
and neutrophil-recruiting chemokines in mice, as well as the pro-inflammatory cytokines 
interleukin (IL)-6, IL-1β, IL-1α, tumor necrosis factor (TNF), and interferon (IFN)-gamma, and 
promotes recruitment of antigen-loaded monocytes to draining lymph nodes.  AS03 is unique 
among adjuvants in that it includes α- tocopherol, a highly bio-available form of Vitamin E.  The 
presence of α-tocopherol appears to be necessary to induce local cytokine responses and 
stimulate antigen-specific adaptive responses.  The mechanism of action in humans is not well 
defined.  Elucidating the mechanisms by which AS03 augments the immune response to 
influenza vaccine in humans is important not only in developing more efficient vaccine 
regimens, but also in determining the safety and the potential for adverse events after adjuvanted 
vaccine. 
Recently, an increased incidence of narcolepsy was noted in patients aged 4-19 after receiving 
Pandemrix™, a monovalent H1N1 vaccine adjuvanted with AS03 licensed for use in Europe 
during the 2009-2010 influenza season25-33.  In Finland, the risk of developing narcolepsy was 
nine times higher in patients aged 4-19 years who had received the pandemic vaccine than in 
those who had not.  All of these patients had the human leukocyte antigen (HLA) DQB1*0602 
genotype, a genotype which has been strongly linked to narcolepsy34.  The pathophysiology of 
narcolepsy has been postulated to result from low levels of the hypothalamic peptide hypocretin 
(also known as orexin) in the central nervous system, which is the result of selective cell death of 
hypocretin-containing neurons.  Many have hypothesized an immune-mediated phenomenon is 
responsible for hypocretin cell death since the disease is tightly associated with HLA haplotypes.  


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A recent report from Ahmed et al suggests that antibodies to influenza nucleoprotein cross-react 
with human hypocretin receptor 235.  The authors postulate that differences in vaccine NP 
content may explain the previously appreciated narcolepsy association with Pandemrix™, since 
NP content is higher than in some other influenza vaccines and the addition of AS03 may 
enhance the immune response to NP. 
This study will use a standard and systems biology approach to assess the human early gene and 
protein signatures expressed in healthy adults before and after each of two doses of a monovalent 
inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur administered 
intramuscularly approximately 28 days apart at 3.75 mcg of HA (A/Shanghai/2/2013) per 0.5 mL 
[adjuvanted] or 0.25 mL [unadjuvanted] dose given with or without AS03 adjuvant 
manufactured by GSK.  We will also compare these responses to one dose of unadjuvanted 
monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi Pasteur 
administered intramuscularly at 15 mcg of HA (A/Minnesota/11/2010) per 0.5 mL dose.  The 
addition of an interpandemic (seasonal) strain of influenza A/H3N2v may allow us to compare 
the innate and adaptive immune response between avian and seasonal influenza vaccines as well. 
2.3 
Potential Risks and Benefits 
2.3.1 
Potential Risks 
The potential risks of this study are those associated with having blood drawn, intramuscular 
(IM) injection of and possible reactions to the monovalent inactivated influenza A/H7N9 virus 
vaccine with or without AS03 adjuvant and monovalent inactivated influenza A/H3N2v virus 
vaccine as well as breach of confidentiality. 
Drawing blood may cause transient discomfort and fainting.  Fainting is usually transient and 
managed by having the subject lie down and elevate his/her legs.  Bruising at the blood draw site 
may occur, but can be prevented or lessened by applying pressure to the blood draw site for a 
few minutes after the blood is taken.  Intramuscular injection may also cause transient discomfort 
and fainting.  Drawing blood and IM injection may also cause infection.  The use of aseptic 
(sterile) technique will make infection at the site where blood will be drawn or where the study 
vaccination will be given extremely unlikely. 
The monovalent inactivated influenza A/H7N9 virus vaccine to be used in this study has never 
been tested for safety in animals.  However, the Division of Microbiology and Infectious 
Diseases (DMID), National Institute of Allergy and Infectious Diseases (NIAID) is sponsoring 
three ongoing Phase II clinical trials: two randomized, double-blinded, controlled clinical trials 
in healthy adults, 19 to 64 years old, inclusive, to assess the safety, reactogenicity, and 


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immunogenicity following receipt of two intramuscular doses, approximately 21 days apart, of a 
monovalent inactivated influenza A/H7N9 virus vaccine (produced by Sanofi Pasteur, 
Swiftwater, PA under contract to DHHS) administered admixed with or without adjuvants MF59 
(Novartis) or AS03 (GlaxoSmithKline) (DMID Protocol 13-0032; NCT019387426 and DMID 
Protocol 13-0033; NCT019422657); and one randomized, partially-blinded, controlled clinical 
trial in healthy adults, aged 65 years and older, to assess the safety, reactogenicity, and 
immunogenicity following receipt of three intramuscular doses of a monovalent inactivated 
influenza A/H7N9 virus vaccine (produced by Sanofi Pasteur, Swiftwater, PA under contract to 
DHHS) administered admixed with MF59 adjuvant (Novartis) at different intervals and dosages 
(DMID Protocol 13-0034; NCT02213354). 
For DMID Protocol 13-0032 and DMID Protocol 13-0033, the dosages of the inactivated 
influenza A/H7N9 vaccine evaluated were:  3.75, 7.5, 15, and 45 mcg of HA per dose.  
Enrollment in both clinical trials is complete with a total of 700 (DMID Protocol 13-0032) and 
980 (DMID Protocol 13-0033) study subjects enrolled, all doses of study products were 
administered, and safety follow-up of study subjects is complete.  Overall, the study products 
administered in both clinical trials were generally safe and well-tolerated.  For DMID Protocol 
13-0032, nine SAEs were reported.  All were assessed as being not related to study product.  
Two Adverse Events of Special Interest (AESIs) were reported in this clinical trial:  Hashimoto’s 
disease (assessed as not related to study product) and Hashimoto’s thyroiditis (assessed as related 
to study product).  For DMID Protocol 13-0033, sixteen SAEs were reported.  Fifteen of these 
SAEs were assessed as being not related to study product.  One SAE was considered to be 
related:  acute inferior myocardial infarction.  Two AESIs were reported in this clinical trial:  
psoriasiform dermatitis (assessed as not related to study product) and celiac disease (assessed as 
not related to study product). 
The manufacturing process used to produce the monovalent inactivated influenza A/H7N9 virus 
vaccine to be used in this study is based on Sanofi Pasteur’s current process for the production of 
their inactivated licensed seasonal influenza virus vaccine Fluzone®, except for a minor 
modification in the Phosphate-Buffered Saline (PBS) diluent in the formulation step that was 
made according to previous experiences of manufacturing of monovalent pandemic vaccines.  
This monovalent inactivated split influenza virus vaccine was derived from the influenza 
A/Shanghai/2/2013 virus.  Licensed release testing specifications were maintained, with a 
modification to the potency test.  
 
 
 
 
 


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We plan to use a 3.75 mcg of HA (A/Shanghai/2/2013) per 0.5 mL [adjuvanted] or 0.25 mL 
[unadjuvanted] dose in this study.  Consequently, the potential risks to subjects are anticipated to 
be similar to those observed for Sanofi Pasteur’s unadjuvanted licensed inter-pandemic 
(seasonal) influenza virus vaccines (Fluzone® and Fluzone® High-Dose), and their unadjuvanted 
licensed 2009 A/H1N1 and A/H5N1 monovalent vaccines. 
The manufacturing process used to produce the monovalent inactivated influenza A/H3N2v virus 
vaccine to be used in this study is based on Sanofi Pasteur’s current process for the production of 
their inactivated licensed seasonal influenza virus vaccine Fluzone®, except for a minor 
modification in the PBS diluent in the formulation step that was made to accommodate the 
nature of the diluted virus.  This monovalent inactivated split influenza virus vaccine was 
derived from the influenza A/Minnesota/11/2010 virus.  Consequently, the potential risks to 
subjects are anticipated to be similar to those observed for Sanofi Pasteur’s unadjuvanted 
licensed inter-pandemic (seasonal) influenza virus vaccines (Fluzone® and Fluzone® High-Dose), 
and their unadjuvanted licensed 2009 A/H1N1 and A/H5N1 monovalent vaccines.  The Division 
of Microbiology and Infectious Diseases (DMID), National Institute of Allergy and Infectious 
Diseases (NIAID) sponsored a Phase II open-label study in healthy adult and elderly populations 
to assess the safety, reactogenicity, and immunogenicity of the unadjuvanted A/H3N2v vaccine 
following receipt of two intramuscular doses (15mcg of HA/0.5mL) administered approximately 
21 days apart (DMID Protocol 12-0011; NCT0174608241).  Overall, the unadjuvanted A/H3N2v 
vaccine was generally safe and well-tolerated among the 210 vaccinated subjects.  Ten SAEs 
were reported.  All were assessed as being not related to study product. 
Occasionally, adult recipients of unadjuvanted licensed, inactivated influenza virus vaccines may 
develop influenza-like reactions, such as fever, feverishness (chills/shivering/sweating), fatigue 
(tiredness), malaise (general unwell feeling), myalgia (body aches/muscular pain), arthralgia 
(joint pain), headache, and/or nausea.  Some subjects may develop reactions at the injection site, 
including pruritus (itching), ecchymosis (bruising), erythema (redness), induration 
(hardness)/swelling, pain, and/or tenderness.  Most of these reactions peak in intensity in the first 
24 hours after vaccination and disappear without treatment within 1 or 2 days.  Analgesics (e.g., 


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acetaminophen, or ibuprofen or similar non-steroidal anti-inflammatory drugs (NSAIDs)) and 
rest may generally relieve or lessen these reactions.  Bruising can sometimes occur due to the 
vaccination procedure. 
In addition, post-marketing surveillance indicates AESIs (autoimmune, auto-inflammatory and 
immune-mediated diseases) as potential risks for pandemic vaccines based on those identified for 
the seasonal influenza vaccines including, but not limited to, neuritis, convulsions, severe 
allergic reactions, syncope, encephalitis, thrombocytopenia, vasculitis, and Guillain-Barré 
syndrome.  Reports of these reactions were rare; however, exact incidence rates cannot be 
precisely calculated. 
Acute and potentially life-threatening allergic reactions (i.e., anaphylaxis) are also possible.  
These reactions occur in about 1 in 4 million people given a vaccination.  These reactions can 
manifest as skin rash (hives), swelling around the mouth, throat or eyes (angioedema), difficulty 
breathing (bronchospasm), a fast pulse (tachycardia), or decrease in blood pressure 
(hypotension).  If these reactions occur, then they can usually be stopped by the administration of 
emergency medications by the study personnel.  As with any vaccine or medication, there is a 
very small chance of death, although researchers do not expect this to occur. 
During the swine influenza (H1N1) vaccine campaign of 1976, some recipients developed a 
paralytic illness called Guillain-Barré syndrome (GBS).  GBS is an acute inflammatory 
neuropathy characterized by weakness, hyporeflexia or areflexia, and elevated protein 
concentrations in cerebrospinal fluid.  The rate of GBS was significantly increased in individuals 
receiving the 1976 swine influenza (H1N1) vaccine at about 1 per 100,000 vaccine recipients.  
This syndrome has not been seen consistently with other influenza vaccines.  Most persons who 
develop GBS recover completely, although the recovery period may be as little as a few weeks 
or as long as a few years.  About 30% of those with GBS still have residual weakness after 3 
years and about 3% may suffer a relapse of muscle weakness and tingling sensations many years 
after the initial attack.  Intensive surveillance of GBS after administration of inactivated 
influenza virus vaccines since 1976 has shown a slight increase in risk over background cases 
(more than one additional case of GBS per million persons) following vaccination, typically with 
onset within 6 weeks after vaccination42.  Interestingly, although vaccination rates have increased 
in the last 10 years, the numbers of reported cases of vaccine-associated GBS have declined43.  A 
recent study in Canada showed that the 2009 H1N1 vaccine was associated with a small but 
significant risk of GBS in persons 50 years and older44.  An active, population-based surveillance 
study conducted in the United States during the 2009-2010 influenza season found less than 1 
excess GBS case per million doses of 2009 H1N1 vaccine administered – a rate similar to that 
associated with some previously administered annual influenza vaccines45-47.  Another study 
using the Medicare system showed an elevated risk of GBS with 2009 monovalent H1N1 


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vaccination (incidence rate ratio = 2.41, 95% confidence interval: 1.14, 5.11; attributable risk = 
2.84 per million doses administered, 95% confidence interval: 0.21, 5.48)48.  An international 
collaboration study also supported a conclusion of an association between 2009 H1N1 
vaccination and GBS49.  It is unknown if the administration of the monovalent inactivated 
influenza A/H7N9 virus vaccine or monovalent inactivated influenza A/H3N2v virus vaccine to 
be used in this study will result in an increased incidence of GBS as the mechanism leading to 
this adverse event has not been completely elucidated. 
As of November 22, 2015 (per the most current version of the manufacturer’s IB), data are 
available for 56 GSK-sponsored clinical trials of AS03-adjuvanted monovalent pandemic 
vaccines manufactured by GSK.  More than 18,000 adult (age ≥18 years) and 6,900 pediatric (6 
months to 17 years old) clinical trial participants have received at least one dose of a GSK-
manufactured, AS03-adjuvanted monovalent pandemic influenza vaccine.  Clinical data 
collected by GSK as of November 22, 2015 (per the most current version of the manufacturer’s 
IB), suggest that inactivated monovalent (pre)pandemic influenza virus antigens adjuvanted with 
AS03 have generally acceptable safety and benefit/risk profiles, though the incidence rates of 
solicited local and systemic adverse events are higher with AS03-adjuvanted antigens than with 
antigen alone, a licensed trivalent influenza vaccine, or placebo.  Some unsolicited adverse 
events (e.g., insomnia, dizziness, cystitis) were associated with a higher relative risk among 
H5N1/AS03 recipients in contrast to Fluarix® or placebo recipients. 
The information and guidance that follow are based on pre-clinical and clinical study results for 
GSK-manufactured AS03-adjuvanted monovalent pandemic vaccines, post- marketing safety 
surveillance data seen with unadjuvanted, inactivated trivalent seasonal influenza vaccines and (in 
the case of the H1N1 vaccines) post-marketing safety surveillance data seen to date for both 
Pandemrix™ and Arepanrix™ H1N1 vaccines. 
The reactogenicity profile in humans of GSK-manufactured AS03-adjuvanted vaccines is 
primarily conditioned by the presence of the adjuvant.  The incidence and severity of local 
redness, swelling, and pain at the injection site in recipients of AS03-adjuvanted vaccines are 
increased relative to monovalent pandemic influenza antigen alone, a licensed trivalent influenza 
vaccine, or placebo.  There is no increase in local and systemic reactogenicity events in 
recipients of AS03-adjuvanted vaccines after a second dose of vaccine relative to the first when 
given 21 days apart.  In young children (6 months to 6 years old), increased frequency of fever 
has been observed following a heterologous booster dose of adjuvanted vaccine administered 6 
months after the primary series. 
As of November 22, 2015 (per the most current version of the manufacturer’s IB), there has been 
no evidence collected by GSK in clinical trials to support a conclusion that any potential immune-
mediated disease or group of diseases was causally related to an AS03-adjuvanted vaccine.  There 


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have been no deaths in GSK clinical trials of AS03-adjuvanted pandemic influenza vaccines 
assessed as related to study vaccine.  A total of 1,428 non-fatal SAEs have been reported for adult 
subjects as of November 22, 2015. Fifteen of these events were deemed related to vaccination by 
the Investigator or GSK.  Of these, six occurred in recipients of an adjuvanted H1N1 vaccine: 
asthma, herpes zoster, hepatic enzyme increased, pain in extremity, polymyalgia rheumatic, and 
thrombocytopenia.  Three occurred in recipients of unadjuvanted H1N1 vaccine: alanine 
aminotransferase increased, hypersensitivity, and multiple sclerosis.  One SAE classified as 
related (myalgia) occurred in a subject who received a control product.  Four SAEs classified as 
related occurred in recipients of an adjuvanted H5N1 vaccine: autoimmune hepatitis, angina 
pectoris, pulmonary embolism, and non-Hodgkin’s lymphoma.  One SAE classified as related 
(lobar pneumonia) occurred in a recipient of an unadjuvanted H5N1 vaccine.  Overall, the 
reactogenicity and safety profile of AS03-adjuvanted pandemic vaccines is acceptable and no 
safety concerns have been identified in clinical trials. 
Narcolepsy is a chronic sleep disorder with a background incidence rate, based on US data, of 
approximately 1.37 per 100,000 per year, with a peak onset between 10 and 19 years of age in 
some datasets.  Narcolepsy, when associated with cataplexy is seen almost exclusively in 
individuals who are HLA DQB1*0602 allele carriers.  An autoimmune etiology has been 
proposed.  In the post-marketing period for adjuvanted H1N1 pandemic vaccines, several 
epidemiological studies conducted in several countries independently of GSK reported an 
increased risk of narcolepsy with or without cataplexy in subjects who were vaccinated with 
Dresden-manufactured H1N1 (Pandemrix™ H1N1) vaccine during the 2009-2010 season.  
These studies have described an absolute risk increase of narcolepsy of approximately 1.4 to 8 
additional cases per 100,000 vaccinated children/adolescents, and approximately one additional 
case per 100,000 vaccinated adults compared to background rates of 0.12 to 0.79 per 100,000 
children/adolescents per year and 0.67 to 1.10 per 100,000 adults per year.  The observed 
temporal association between narcolepsy and vaccination with Pandemrix™ H1N1 is not fully 
understood, and further research to evaluate the association between narcolepsy and 
Pandemrix™ H1N1, and other possible contributory factors to the development of narcolepsy 
during the 2009-2010 pandemic, such as genetic and environmental factors, is being conducted.  
A GSK-supported study was conducted in Quebec, Canada, to assess the risk of narcolepsy 
associated with Arepanrix™ H1N1, using various index dates, risk periods, observation periods, 
and epidemiological designs.  Overall, GSK considers that there is no strong evidence of an 
association between Q-Pan-H1N1 and narcolepsy in Quebec. 
No post-marketing data are available for AS03 administered in combination with any GSK-
manufactured H5N1, H7N1, H7N9 or H9N2 antigen.  However, millions of doses of GSK-
manufactured H1N1 antigens, combined with AS03, were administered in the context of the 
2009/10 pandemic response.  In addition to the adverse reactions reported in clinical trials, the 


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following have been reported during post-marketing experience with Pandemrix™ (H1N1) and 
Arepanrix™ (H1N1): 
• Immune system disorders 
o Rare: anaphylaxis, allergic reactions 
• Nervous system disorders 
o Rare: febrile convulsions (in subjects below 20 years of age), somnolence**, 
Guillain-Barré syndrome* 
*Spontaneous reports of Guillain-Barré syndrome have been received following 
vaccination with Arepanrix™ (H1N1); however, a causal association between 
vaccination and Guillain-Barré syndrome has not been established.  Data from a 
post-marketing epidemiological study in Canada indicate a small but significant 
increased relative risk of Guillain-Barré syndrome of 1.80 (95% CI, 1.63-4.62) in the 
56-day period following vaccination with Arepanrix™ (H1N1, in persons 50 years of 
age and older).  The number of GBS cases attributable to vaccination was 
approximately 2 per 1 million doses. 
**reported in patients with narcolepsy and as a temporary event following 
vaccination 
o Very rare1: narcolepsy with or without cataplexy 
1Frequency based on estimated attributable risk from epidemiological studies in 
several European countries. 
• Skin and subcutaneous tissue disorders 
o Rare: angioedema, generalized skin reactions, urticaria 
• General disorders and administration site conditions 
o Rare: injection site reactions (such as inflammation, mass, ecchymosis) 
 
From post-marketing surveillance with interpandemic (seasonal) trivalent vaccines, the following 
additional adverse events have been reported: 
• Blood and lymphatic system disorders 
o Transient thrombocytopenia 
• Nervous system disorders 
o Neuralgia, convulsions 
o Neurological disorders, such as encephalomyelitis, neuritis and Guillain-Barré 
syndrome 
• Vascular disorders 
o Vasculitis with transient renal involvement 
It is unknown if the monovalent inactivated influenza A/H7N9 virus vaccine with or without 
AS03 adjuvant or unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine poses 


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any risks to an unborn child.  As of November 22, 2015 (per the most current version of the 
manufacturer’s IB), the available data for women who become pregnant during clinical trials of 
AS03-adjuvanted (pre) pandemic influenza vaccines do not suggest any causal relationship 
between adverse pregnancy outcomes and receipt of an AS03-adjuvanted vaccine.  Women of 
childbearing potential who are not surgically sterile via tubal sterilization, bilateral 
oophorectomy or hysterectomy, or who are not postmenopausal for ≥1 year must agree to 
practice highly effective contraception that may include, but is not limited to, non-male sexual 
relationships, abstinence from sexual intercourse with a male partner, monogamous relationship 
with a vasectomized partner, male condoms that contain spermicide or with the use of applied 
spermicide, intrauterine devices, and licensed hormonal methods, with use of a highly effective 
method of contraception for a minimum of 30 days prior to study product exposure and agree to 
practice highly effective contraception for the duration of study product exposure, including 60 
days after their last study vaccination.  A highly effective method of contraception is defined as 
one that results in a low failure rate (i.e., less than 1% per year) when used consistently and 
correctly.  In addition to contraceptive use, all women of childbearing potential will be required 
to have a negative urine or serum pregnancy test within 24 hours prior to each study vaccination.  
If a female subject becomes pregnant while participating in this study, we will ask her 
permission to follow-up with her about her health and the health of her baby through pregnancy 
outcome. 
Subjects will be asked to provide personal health information (PHI).  All attempts will be made 
to keep this PHI confidential within the limits of the law.  However, there is a chance that 
unauthorized persons will see the subjects’ PHI.  All study records will be kept in a locked file 
cabinet or maintained in a locked room at the participating VTEU site.  Electronic files will be 
password-protected.  Only people who are involved in the conduct, oversight, monitoring, or 
auditing of this study will be allowed access to the PHI that is collected.  Any publications from 
this study will not use information that will identify subjects by name.  Organizations that may 
inspect and/or copy research records maintained at the participating VTEU site for quality 
assurance and data analysis include groups, such as the local Institutional Review Board (IRB), 
NIAID and Food and Drug Administration (FDA). 
A description of this clinical study will be available on http://www.ClinicalTrials.gov, as 
required by US Law.  This web site will not include information that can identify subjects.  At 
most, this web site will include a summary of the results. 
There may be other risks, discomforts, or side effects that are unknown at this time. 


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2.3.2 
Known Potential Benefits 
There are no known benefits attributable to the receipt of the monovalent inactivated influenza 
A/H7N9 virus vaccine with or without AS03 adjuvant, and there is no prospect of benefit at this 
time.  It is possible that vaccination using the monovalent inactivated influenza A/H7N9 virus 
vaccine with or with AS03 adjuvant will result in some protection against infection caused by the 
influenza A/H7N9 virus.  Vaccination using the monovalent inactivated influenza A/H7N9 virus 
vaccine with or without AS03 adjuvant may or may not provide protection against a serious 
infection with the influenza A/H7N9 virus, should the participant be exposed.  The duration of 
any such protection is currently unknown.  The monovalent inactivated influenza A/H7N9 virus 
vaccine with or without AS03 adjuvant is not expected to offer protection against circulating 
seasonal influenza viruses.  There may be pandemic preparedness benefits to society in the future 
if the monovalent inactivated influenza A/H7N9 virus vaccine and AS03 adjuvant being 
evaluated here prove to be sufficiently safe and immunogenic and can be employed if a need for 
widespread influenza A/H7N9 vaccination occurs. 
Similarly, there are no known benefits attributable to receipt of the unadjuvanted monovalent 
inactivated influenza A/H3N2v virus vaccine, but there is a prospect of benefit.  A/H3N2v has 
been seen in the United States as a variant of the seasonal influenza A/H3N2; the virus has been 
seen primarily in those with pig exposures.  As there may be cross-protection against other 
A/H3N2 viruses, and since the future circulation of A/H3N2v is unknown, vaccination may 
provide some benefit. 


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3 
STUDY OBJECTIVES AND OUTCOME MEASURES 
Primary Objectives 
 
1. To assess the serum anti-HA hemagglutination-inhibition (HAI) response to influenza 
A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the 
second study vaccination with A/H7N9 vaccine ± AS03) and influenza A/H3N2v antigen at 
Day 29 (approximately one month after the study vaccination with A/H3N2v vaccine). 
 
2. To identify differentially expressed genes in human immune cells on Days 2, 4, and 29 
(following the first study vaccination with A/H7N9 vaccine ± AS03) and on Days 30, 32, and 
36 (following the second study vaccination with A/H7N9 vaccine ± AS03), compared to 
baseline assessments performed prior to each study vaccination (Days -7, 1, and 29). 
 
Primary Outcome Measures 
 
1. Percentage of subjects achieving seroconversion (defined as either a pre-vaccination HAI 
titer <1:10 and a post-vaccination HAI titer ≥1:40 or a pre- vaccination HAI titer ≥1:10 and a 
minimum four-fold rise in post-vaccination HAI antibody titer) at approximately 28 days 
after the second study vaccination (A/H7N9 vaccine, with and without adjuvant). 
 
2. Percentage of subjects achieving seroconversion at approximately 28 days after the study 
vaccination with A/H3N2v vaccine. 
 
3. Number of differentially expressed genes based on RNA expression as determined by RNA-
Seq analysis, in human immune cells at Days 2, 4, 29, 30, 32, and 36 after study vaccination 
with AS03-adjuvanted and unadjuvanted A/H7N9 vaccine, as compared to baseline 
assessments performed prior to vaccination (Days -7, 1, and 29). 
 
Secondary Objectives 
 
1. To compare plasma cytokine and chemokine profiles at specific time points and between 
treatment arms: 
a. Post-vaccination time points [(Days 2 and 4; Day 8 (A/H3N2v arm only)], and Days 
30, 32, and 36 (A/H7N9 arms only) with pre-vaccination time points (Days -7 and 1, 
prior to first study vaccination and Day 29, prior to second study vaccination 
(A/H7N9 arms only). 
 


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2. To assess the neutralizing antibody responses to influenza A/H7N9 antigen (with and without 
adjuvant) at Day 57 (approximately one month after the second study vaccination with 
A/H7N9 vaccine ± AS03) and influenza A/H3N2v antigen at Day 29 (approximately one 
month after the study vaccination with A/H3N2v vaccine). 
 
3. To identify differentially expressed genes in human immune cells on Days 2, 4, and 8 
following one intramuscular dose of influenza A/H3N2v vaccine compared to baseline 
assessments performed prior to study vaccination (Day -7 and Day 1). 
 
Secondary Outcome Measures 
 
1. Plasma measurement of cytokines and chemokines at each study visit, comparing: 
a. 
Post-vaccination time points with pre-vaccination time points by treatment arms. 
b. 
AS03-adjuvanted influenza A/H7N9 responses to unadjuvanted influenza A/H7N9 
responses. 
c. 
Influenza A/H3N2v vaccine to unadjuvanted influenza A/H7N9 vaccine. 
d. 
Influenza A/H3N2v vaccine to adjuvanted A/H7N9 vaccine. 
 
2. 
Serologic response to influenza hemagglutinin: 
a. 
Percentage of subjects achieving seroconversion (defined as either a pre-vaccination 
Neut titer <1:10 and a post-vaccination Neut titer ≥1:40 or a pre-vaccination Neut titer 
≥1:10 and a minimum four-fold rise in post-vaccination Neut titer) at approximately 
28 days after the second study vaccination (A/H7N9 vaccine, with and without 
adjuvant). 
b. 
Percentage of subjects achieving seroconversion at approximately 28 days after the 
study vaccination with influenza A/H3N2v vaccine. 
c. 
Geometric Mean Titers of serum HAI and Neut antibody at baseline (Day -7 and 1) 
and 28 days after influenza A/H3N2v vaccination. 
d. 
Geometric Mean Titers of serum HAI and Neut antibody at baseline (Day -7 and 1), 
and 28 days after both the first and the second dose of influenza A/H7N9 vaccine 
(with and without adjuvant). 
 
3. Number of differentially expressed genes based on RNA-Seq analysis in human immune 
cells at Days 2, 4, and 8 after study vaccination, as compared to baseline assessments (Days -
7 and 1) after influenza A/H3N2v vaccine. 
 
Tertiary Objectives 
 
1. To assess the frequency of adverse events. 


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2. To identify differentially abundant cellular proteins in human immune cells on Days 2, 4, 
and 8 following one intramuscular dose of influenza A/H3N2v vaccine compared to 
baseline assessments performed prior to study vaccination (Day -7 and Day 1). 
 
3. To identify differentially abundant cellular proteins in human immune cells on Days 2, 4, 
and 29 (following first study vaccination with A/H7N9 vaccine ± AS03) and Days 30, 32, 
and 36 (following second study vaccination with A/H7N9 vaccine ± AS03), compared to 
baseline assessments performed prior to study vaccination (Days -7, 1, and 29). 
 
Tertiary Outcome Measures 
 
1. Occurrence of solicited local reactogenicity events within 7 days after each study 
vaccination. 
 
2. Occurrence of solicited systemic reactogenicity events within 7 days after each study 
vaccination. 
 
3. Occurrence of unsolicited AEs collected for approximately 28 days after last study 
vaccination. 
 
4. Occurrence of SAEs and MAAEs including new-onset chronic medical conditions and 
potentially immune-mediated medical conditions for approximately 1 month (A/H3N2v 
arm) and 12 months (A/H7N9 arms) after last study vaccination. 
 
5. Number of differentially abundant proteins based on protein abundance as determined by 
quantitative proteomics in human immune cells at Days 2, 4, and 8 after study 
vaccination, as compared to baseline assessments (Days -7 and 1) after influenza 
A/H3N2v vaccine. 
 
6. Number of differentially abundant proteins based on protein abundance as determined by 
quantitative proteomics analysis, in human immune cells at Days 2, 4, 29, 30, 32, and 36 
after study vaccination with AS03-adjuvanted and unadjuvanted A/H7N9 vaccine 
comparing changes in protein abundance between treatment arms (A/H7N9 vaccine vs. 
A/H7N9 vaccine + AS03). 
 
Exploratory Objectives 
 


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1. To identify and characterize differentially expressed genes in human immune cells on 
Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine ± AS03) 
and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 
vaccine ± AS03) that differ between treatments (A/H7N9 vaccine vs. A/H7N9 vaccine + 
AS03). 
 
2. To identify and characterize differentially abundant cellular proteins in human immune 
cells on Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine ± 
AS03) and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 
vaccine ± AS03) that differ between treatments (A/H7N9 vaccine vs. A/H7N9 vaccine + 
AS03). 
 
3. To identify and characterize differentially expressed genes in human immune cells on 
Days 2 and 4 (following the first study vaccination) that differ in baseline responses 
(Days -7, and 1) between treatments (A/H3N2v vaccine vs. A/H7N9 vaccine and 
A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
 
4. To identify and characterize differentially abundant cellular proteins in human immune 
cells on Days 2 and 4 (following the first study vaccination) that differ in baseline 
responses (Days -7, and 1) between treatments (A/H3N2v vaccine vs. A/H7N9 vaccine 
and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
 
5. To identify transcriptomic and proteomic signatures in human immune cells that correlate 
with influenza seroconversion and with peak HAI titers. 
 
6. To correlate plasma cytokine profiles with transcriptomic and proteomic profiles in 
subjects receiving influenza A/H3N2v vaccine or influenza A/H7N9 ± AS03 vaccine. 
 
7. Exploratory inspection of transcriptomics\proteomics response signatures for subjects 
with unexpected or severe safety profiles that are related to the study products. 
 
Exploratory Outcome Measures 
 
1. RNA expression, clusters of co-expressed genes, enriched pathways, and other functional 
modules as determined by RNA-Seq analysis, in human immune cells at Days 2, 4, 29, 
30, 32, and 36 after study vaccination with AS03-adjuvanted and unadjuvanted A/H7N9 
vaccine comparing changes in RNA expression between treatment arms (A/H7N9 
vaccine vs. A/H7N9 vaccine + AS03). 
 


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2. Protein abundance, clusters of co-abundant proteins, enriched pathways, and other 
functional modules as determined by quantitative proteomics analysis, in human immune 
cells at Days 2, 4, 29, 30, 32, and 36 after study vaccination with AS03-adjuvanted and 
unadjuvanted A/H7N9 vaccine comparing changes in protein abundance between 
treatment arms (A/H7N9 vaccine vs. A/H7N9 vaccine + AS03). 
 
3. RNA expression, clusters of co-expressed genes, enriched pathways, and other functional 
modules as determined by RNA-Seq analysis, in human immune cells at Days 2, 4, and 
29 after study vaccination comparing baseline changes in RNA expression between 
treatment arms (A/H3N2v vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. 
A/H7N9 vaccine + AS03). 
 
4. Protein abundance, clusters of co-abundant proteins, enriched pathways, and other 
functional modules as determined by quantitative proteomics analysis, in human immune 
cells at Days 2, 4, and 29 after first study vaccination comparing baseline changes (Days 
-7, and 1) in protein abundance between treatment arms (A/H3N2v vaccine vs. A/H7N9 
vaccine and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
 
5. Changes from baseline (Days -7, 1, and 29) in RNA expression or protein abundance that 
best correlate with influenza A/H7N9 seroconversion and with peak HAI titers at Days 2 
and 4 (following first study vaccination) as well as Days 30, 32, and 36 (following 
second study vaccination). 
 
6. Changes from baseline (Days -7, 1, and 29) in RNA expression or protein abundance that 
best correlate with baseline changes in cytokine concentrations at Days 2, 4, 8 (A/H3N2v 
arm only) and 29 (following first study vaccination) as well as Days 30, 32, and 36 
(following second study vaccination). 
 
7. Exploratory summaries of transcriptomics/proteomics response signatures for subjects 
with unexpected or severe safety profiles. 
 
8. Lists of differential genes and proteins identified as part of the primary and secondary 
objectives. 


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4 
STUDY DESIGN 
This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study 
in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, 
inclusive, designed to evaluate and compare the immunogenicity between an intramuscular 
monovalent inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur given 
with and without AS03 adjuvant manufactured by GlaxoSmithKline, and an intramuscular 
unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi 
Pasteur. 
This study will use a standard and systems biology approach to assess the human early gene and 
protein signatures expressed at baseline (approximately Days -7 and 1), and at approximately 
Days 2, 4, and 8 (A/H3N2v-vaccinated subjects only) after the first study vaccination in each 
treatment arm as well as at approximately Days 29, 30, 32, and 36 in A/H7N9-vaccinated 
subjects only.  Cellular immunogenicity (systems biology studies) data will be integrated with 
serologic immunogenicity (HAI and Neut antibody assays) and reactogenicity data to develop a 
systems model of the human immune response to unadjuvanted monovalent inactivated influenza 
A/H3N2v virus vaccine and monovalent inactivated influenza A/H7N9 virus vaccine given with 
and without AS03 adjuvant. 
This study will use venous blood samples and subject data collected from a total of thirty 
vaccinated subjects randomly divided into three equal treatment arms.  The first treatment arm 
(n=10) will be vaccinated with one dose of 15 µg of A/H3N2v HA.  The second treatment arm 
(n=10) will be vaccinated with two doses of 3.75 µg of A/H7N9 HA given with AS03 
approximately 28 days apart.  The third treatment arm (n=10) will be vaccinated with two doses 
of 3.75 µg of A/H7N9 HA given without AS03 adjuvant approximately 28 days apart.  All study 
vaccinations will be administered intramuscularly. 
Reactogenicity will be measured by the occurrence of solicited injection site and systemic 
reactions from the time of each study vaccination through 8 days after each study vaccination.  
Unsolicited non-serious adverse events (AEs) will be collected from the time of each study 
vaccination through approximately 28 days after each study vaccination.  Serious adverse events 
(SAEs) and medically-attended adverse events (MAAEs) including new-onset chronic medical 
conditions (NOCMCs) and potentially immune-mediated medical conditions (PIMMCs) will be 
collected from the time of the first study vaccination through approximately 1 month (A/H3N2v 
arm) and 12 months (A/H7N9 arms) after the last study vaccination. 
Venous blood samples (approximately 90 mL) will be collected from subjects at approximately 
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and 8 (A/H3N2v-vaccinated subjects only) after the first study vaccination for systems biology 
studies (cytokine and chemokine levels, immune cell activation status, and whole transcriptome 
and proteome profiles of the major blood immune cells).  Additionally, subjects in either 
treatment arm receiving A/H7N9 vaccine will have additional blood samples collected at 
approximately Days 29 (this will be collected immediately prior to the second study vaccination 
for subjects in either group receiving A/H7N9), 30, 32, and 36 after the first study vaccination 
for systems biology studies. 
Serological assessment (hemagglutination inhibition (HAI) and neutralizing (Neut) antibody 
assays) will also be conducted on venous blood samples (approximately 10 mL) collected from 
subjects at Day 1 (immediately prior to the first study vaccination), and at approximately Days 
29 (for A/H7N9-vaccinated subjects this will be collected immediately prior to the second study 
vaccination) and 57 (A/H7N9-vaccinated subjects only) after the first study vaccination. 
We will quantify and characterize serum cytokine/chemokine levels, as well as transcriptomic and 
proteomic profiles from individual immune cell compartments, comparing this with standard 
serologic assessment to vaccine. 
Urine samples (approximately 20 mL) will also be collected from subjects at approximately 
Days -7, 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only), 29, 30, 32, and 36 for future research.  
Note: Days 29, 30, 32, and 36 will apply only to A/H7N9-vaccinated subjects. 


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5 
STUDY ENROLLMENT AND WITHDRAWAL 
Approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, 
inclusive, who meet all eligibility criteria will be enrolled in this study from one participating 
VTEU site.  Enrollment will occur over a 6-month period, and the target population will reflect 
the community at large.  Information regarding this study may be mailed or emailed to potential 
subjects who have previously participated in vaccine trials conducted at the participating VTEU 
site.  Other forms and/or mechanisms of recruitment may also be used.  The local IRB will 
approve all materials prior to use. 
Subject Inclusion and Exclusion Criteria must be assessed by a study clinician licensed to make 
medical diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
No exemptions are granted on Subject Inclusion or Exclusion Criteria in DMID-sponsored 
studies.  Questions about eligibility should be directed toward the DMID Medical Officer. 
5.1 
Eligibility Criteria 
5.1.1 
Subject Inclusion Criteria 
Subjects eligible to participate in this study must meet all of the following inclusion criteria: 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Are able to understand and comply with planned study procedures and be available for all 
study visits. 
3. Are males or non-pregnant females, 18 to 49 years old, inclusive. 
4. Are in good health1. 
1As determined by medical history and targeted physical examination, if indicated based on medical history, to 
evaluate acute or currently ongoing chronic medical diagnoses or conditions, defined as those that have been present 
for at least 90 days, that would affect the assessment of the safety of subjects or the immunogenicity of study 
vaccinations.  Chronic medical diagnoses or conditions should be stable for the last 60 days.  This includes no change 
in chronic prescription medication, dose, or frequency as a result of deterioration of the chronic medical diagnosis or 
condition in the 60 days prior to enrollment.  Any prescription change that is due to change of health care provider, 
insurance company, etc., or that is done for financial reasons, as long as in the same class of medication, will not be 
considered a deviation of this inclusion criterion.  Any change in prescription medication due to improvement of a 
disease outcome, as determined by the site principal investigator or appropriate sub-investigator, will not be 
considered a deviation of this inclusion criterion.  Subjects may be on chronic or as needed (prn) medications if, in the 
opinion of the site principal investigator or appropriate sub-investigator, they pose no additional risk to subject safety 
or assessment of reactogenicity and immunogenicity and do not indicate a worsening of medical diagnosis or 


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condition.  Similarly, medication changes subsequent to enrollment and study vaccination are acceptable provided 
there was no deterioration in the subject’s chronic medical condition that necessitated a medication change, and there 
is no additional risk to the subject or interference with the evaluation of responses to study vaccination.  Note: Topical, 
nasal, and inhaled medications (with the exception of inhaled corticosteroids as outlined in the Subject Exclusion 
Criteria (see Section 5.1.2)), herbals, vitamins, and supplements are permitted. 
5. Oral temperature is less than 100.4°F. 
6. Pulse is 50 to 115 bpm, inclusive. 
7. Systolic blood pressure is 85 to 150 mm Hg, inclusive. 
8. Diastolic blood pressure is 55 to 95 mm Hg, inclusive. 
9. Erythrocyte sedimentation rate (ESR) is less than 30 mm per hour. 
10. Women of childbearing potential2 must use an acceptable contraception method3 from 30 
days before first study vaccination until 60 days after last study vaccination. 
2Not sterilized via tubal ligation, bilateral oophorectomy, hysterectomy or successful Essure® placement (permanent, 
non-surgical, non-hormonal sterilization) with documented radiological confirmation test at least 90 days after the 
procedure, and still menstruating or <1 year of the last menses if menopausal. 
3Includes, but is not limited to, non-male sexual relationships, abstinence from sexual intercourse with a male partner, 
monogamous relationship with vasectomized partner who has been vasectomized for 180 days or more prior to the 
subject receiving the first study vaccination, barrier methods such as condoms or diaphragms with spermicide or foam, 
effective intrauterine devices, NuvaRing®, and licensed hormonal methods such as implants, injectables or oral 
contraceptives (“the pill”). 
11. Women of childbearing potential must have a negative urine or serum pregnancy test 
within 24 hours prior to study vaccination. 
5.1.2 
Subject Exclusion Criteria 
Subjects eligible to participate in this study must not meet any of the following exclusion 
criteria: 
1. Have an acute illness4, as determined by the site principal investigator or appropriate 
sub-investigator, within 72 hours prior to study vaccination. 
4An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion 
of the site principal investigator or appropriate sub-investigator, the residual symptoms will not interfere with the 
ability to assess safety parameters as required by the protocol. 
2. Have any medical disease or condition that, in the opinion of the site principal 
investigator or appropriate sub-investigator, is a contraindication to study participation5. 


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5Including acute or chronic medical disease or condition, defined as persisting for at least 90 days that would place 
the subject at an unacceptable risk of injury, render the subject unable to meet the requirements of the protocol, or 
may interfere with the evaluation of responses or the subject’s successful completion of this study. 
3. Have immunosuppression as a result of an underlying illness or treatment, or use of 
anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study 
vaccination. 
4. Have known active neoplastic disease (excluding non-melanoma skin cancer) or a 
history of any hematologic malignancy. 
5. Have known human immunodeficiency virus (HIV), hepatitis B, or hepatitis C 
infection. 
6. Have known hypersensitivity or allergy to eggs, egg or chicken protein, squalene-based 
adjuvants, or other components of the study vaccines. 
7. Have a history of severe reactions following previous immunization with licensed or 
unlicensed influenza virus vaccines. 
8. Have a personal or family history of narcolepsy. 
9. Have a history of Guillain-Barré syndrome. 
10. Have a history of convulsions or encephalomyelitis within 90 days prior to study 
vaccination. 
11. Have a history of a potentially immune-mediated medical condition6. 
6Refer to Appendix A: List of Potentially Immune-Mediated Medical Conditions. 
12. Have a history of alcohol or drug abuse within 5 years prior to study vaccination. 
13. Have any diagnosis, current or past, of schizophrenia, bipolar disease, or other 
psychiatric diagnosis that may interfere with subject compliance or safety evaluations. 
14. Have been hospitalized for psychiatric illness, history of suicide attempt, or 
confinement for danger to self or others within 10 years prior to study vaccination. 
15. Have taken oral or parenteral (including intra-articular) corticosteroids of any dose 
within 30 days prior to study vaccination. 


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16. Have taken high-dose inhaled corticosteroids within 30 days prior to study vaccination.  
High-dose defined as >840 mcg/day of beclomethasone dipropionate CFC or 
equivalent. 
17. Received licensed live vaccine within 30 days prior to the first study vaccination, or 
plans to receive licensed live vaccine within 30 days before or after each study 
vaccination. 
18. Received licensed inactivated vaccine within 14 days prior to the first study 
vaccination, or plans to receive licensed inactivated vaccine within 14 days before or 
after each study vaccination. 
19. Received immunoglobulin or other blood products (with exception of Rho D 
immunoglobulin) within 90 days prior to study vaccination. 
20. Received an experimental agent7 within 30 days prior to the first study vaccination, or 
expects to receive an experimental agent8 during the 13-month study-reporting period. 
7Including vaccine, drug, biologic, device, blood product, or medication. 
8Other than from participation in this study. 
21. Are participating or plan to participate in another clinical trial with an interventional 
agent9 that will be received during the 13-month study-reporting period. 
9Including licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication. 
22. Prior participation in a clinical trial of influenza A/H7 vaccine10 or have a history of 
influenza A/H7 virus actual or potential exposure or infection prior to the first study 
vaccination. 
10And assigned to a group receiving influenza A/H7 vaccine, does not apply to documented placebo recipients. 
23. Prior participation in a clinical trial of influenza A/H3N2v vaccine11 or have a history 
of influenza A/H3N2v virus actual or potential exposure or infection prior to the first 
study vaccination. 
11And assigned to a group receiving influenza A/H3N2v vaccine, does not apply to documented placebo recipients. 
24. Occupational exposure to or substantial direct physical contact12 with birds in the past 
year or during the 28 days after each study vaccination. 
12Casual contact with birds at petting zoos or county or state fairs or having pet birds does not exclude subjects from 
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25. Occupational exposure to or substantial direct physical contact13 with pigs in the past 
year or during the 28 days after each study vaccination. 
13Casual contact with pigs at petting zoos or county or state fairs does not exclude subjects from study participation. 
26. Female subjects who are breastfeeding or plan to breastfeed at any given time from the 
first study vaccination until 30 days after the last study vaccination. 
27. Plan to travel outside the US (continental US, Hawaii, and Alaska) within 28 days after 
each study vaccination. 
28. Blood donation or planned blood donation within 30 days before enrollment until 30 
days after the last blood draw for this study. 
5.2 
Treatment Assignment Procedures 
5.2.1 
Enrollment and Randomization Procedures 
Per International Conference on Harmonisation (ICH) guideline E6: Good Clinical Practice 
(GCP), screening records will be kept at the participating VTEU site to document the reason why 
an individual was screened, but failed study entry criteria.  The reasons why individuals failed 
screening will be recorded in the Statistical and Data Coordinating Center’s (SDCC) 
AdvantageEDCSM (Electronic Data Capture System). 
Once consented and upon entry of demographic data and confirmation of eligibility for this 
study, the subject will be enrolled.  Subjects will be assigned randomly to 1 of 3 treatment arms.  
The first treatment arm (n=10) will be vaccinated with one dose of 15 µg of A/H3N2v HA.  The 
second treatment arm (n=10) will be vaccinated with two doses of 3.75 µg of A/H7N9 HA given 
with AS03 approximately 28 days apart.  The third treatment arm (n=10) will be vaccinated with 
two doses of 3.75 µg of A/H7N9 HA given without AS03 adjuvant approximately 28 days apart.  
All study vaccinations will be administered intramuscularly. 
Enrollment of subjects will be done online using the enrollment module of AdvantageEDCSM.  
The randomization code will be prepared by statisticians at the SDCC and included in the 
enrollment module for this study.  AdvantageEDCSM will assign each subject to a treatment arm 
after the demographic and eligibility data have been entered into the system.  A designated 
individual at the participating VTEU site will be provided with a code list for emergency 
unblinding purposes, which will be kept in a secure place. 
Instructions for use of the enrollment module are included in the AdvantageEDCSM User’s 
Guide.  Manual back-up procedures and instructions are provided for use in the event that the 


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participating VTEU site temporarily loses access to the Internet or the online enrollment system 
is unavailable. 
Subjects who sign the informed consent form and are randomized but do not receive study 
vaccine may be replaced.  Subjects who sign the informed consent form, and are randomized and 
vaccinated, and subsequently withdraw, or are withdrawn or terminated from this study, or are 
lost to follow-up may be replaced.  The randomization scheme for this study is presented in the 
table below. 
 
 


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Treatment Arms 
Treatment Arms 
(n=10 vaccinated subjects per 
Treatment Arm) 
First Study Vaccination 
(Day 1) 
Second Study Vaccination 
(Day 29±2 days) 
1 
15 µg A/H3N2v* 
N/A 
2 
3.75 µg A/H7N9 + AS03 
3.75 µg A/H7N9 + AS03 
3 
3.75 µg A/H7N9* 
3.75 µg A/H7N9*  
Total N=30 vaccinated subjects 
* unadjuvanted 
5.2.2 
Masking Procedures 
This is a partially-blind clinical study as the unadjuvanted A/H3N2v vaccine treatment arm will 
be open-label to avoid the need for placebo administration at approximately Day 29.  
Investigators and study personnel will remain blinded to allocation of A/H7N9 with or without 
AS03. 
The randomization scheme will be generated by the SDCC and provided to unblinded study 
personnel (i.e., research pharmacists performing study vaccination preparations and unblinded 
study vaccine administrators) at the participating VTEU site. 
The unblinded study vaccine administrator is a study personnel member credentialed to 
administer vaccines and may also participate in dose preparation, but will not be involved in 
study-related assessments or have subject contact for data collection following study vaccine 
administration. 
Laboratory personnel performing antibody, cytokine\chemokine, and RNA-Seq assays will be 
masked to A/H7N9 and A/H3N2v treatment groups, subject ID, and study visit for all study 
specimens.  For proteomics assays, A/H7N9 treatment groups and study visit will be masked. 
Subject IDs will be replaced by random IDs generated by the SDCC to allow proteomics batch-
processing of specimens obtained from the same subject.  In all cases, the SDCC will generate 
random specimen picklists for each laboratory to determine which specimens will be tested. 
After all subjects have completed Day 57, the database will be locked to facilitate data analysis. 
At the discretion of DMID, unmasking of laboratories may occur after all data have been locked, 
analyzed and interpreted and the final CSR has been completed. 
The Safety Monitoring Committee (SMC) may receive data in aggregate and presented by 
treatment arm.  The SMC may also be provided with expected and observed rates of the expected 
AEs in an unblinded fashion, and may request the treatment arm be unblinded for an individual 


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subject if required for safety assessment.  The SMC will review grouped and unblinded data in 
the closed session only. 
5.2.3 
Reasons for Withdrawals and Discontinuation of Treatment 
Subjects may voluntarily withdraw their consent for study participation at any time and for any 
reason, without penalty. 
A subject may withdraw or be withdrawn from this study for any of the following reasons: 
• Medical disease or condition, or any new clinical findings for which continued 
participation, in the opinion of the site principal investigator or appropriate sub-
investigator, would compromise the safety of the subject, or would interfere with the 
subject's successful completion of this study, or would interfere with the evaluation of 
responses. 
• Subject no longer meets eligibility criteria (see Section 5.1).  Note: Medication changes 
within 60 days, as specified in Subject Inclusion Criterion #4, are exclusionary for receipt 
of the first study vaccination only. 
• As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
• Subject withdrawal of consent. 
• Subject lost to follow-up. 
• Termination of this study. 
• New information becomes available that makes further participation unsafe. 
The second study vaccination will not be administered to an A/H7N9-vaccinated subject if any 
of the following criteria are met: 
• Medical condition or medication change for which continued participation, in the opinion 
of the site principal investigator or appropriate sub-investigator, would pose a risk to the 
subject or would be likely to confound interpretation of the results. 
• Presence of signs or symptoms that could confound or confuse assessment of study 
vaccine reactogenicity.  For subjects with injection site or systemic signs or symptoms, or 
with an acute illness, including an oral temperature greater than or equal to 100.4°F, the 
second study vaccination should be postponed/deferred until signs, symptoms, or acute 
illness have resolved, or are improving as further specified below, and if within the 
acceptable protocol-specified window for that visit (Day 29±2 days post first study 


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vaccination).  No exceptions to the protocol-specified window will be made.  Note for 
afebrile, acute illness only: If a subject is afebrile, his/her acute illness is nearly resolved 
with only minor residual symptoms remaining, this occurs within the acceptable protocol-
specified window for that visit (Day 29±2 days post first study vaccination), and, in the 
opinion of the site principal investigator or appropriate sub-investigator, the residual 
symptoms will not interfere with the ability to assess safety parameters as required by the 
protocol, the subject may receive the second study vaccination without further approval 
from the DMID Medical Officer.  No exceptions to the protocol-specified window will be 
made. 
• Grade 2 or 3 solicited or unsolicited adverse event that is ongoing, whether or not it is 
improved or resolving.  An unresolved or continuing Grade 1 adverse event is 
permissible following the documented determination by the site principal investigator or 
appropriate sub-investigator, that it would not render study vaccination unsafe or interfere 
with the evaluation of responses. 
• Grade 3 solicited or unsolicited adverse event that occurs without alternative etiology in 
the 8 days following the first study vaccination. 
• Severe or sustained reaction or disability related to the first study vaccination. 
• New onset of illness or condition that meets the Subject Exclusion Criteria (see Section 
5.1.2). 
• Subject no longer meets eligibility criteria (see Section 5.1).  Note: Medication changes 
subsequent to the first study vaccination are not exclusionary for receipt of the second 
study vaccination provided there was no deterioration in the subject’s chronic medical 
condition that necessitated a medication change, and there is no additional risk to the 
subject or interference with the evaluation of responses to study vaccination. 
• As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
• Subject refusal of further study vaccination. 
• Subject withdrawal of consent. 
• Subject lost to follow-up. 
• Termination of this study. 
• New information becomes available that makes further participation unsafe. 


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5.2.4 
Handling of Withdrawals and Discontinuation of Treatment 
The primary reason for withdrawal from this study will be recorded on the Study Status data 
collection form.  Subjects will be encouraged to complete the Early Termination Visit.  The 
Early Termination Visit procedures are listed in Section 7.3. 
Although subjects are free to withdraw at any time or may be withdrawn by the site principal 
investigator or appropriate sub-investigator at any time (see Section 5.2.3), those subjects who 
receive only one dose of study vaccine will be encouraged to remain in this study for follow-up 
safety assessments (may be conducted by phone call/electronic communication (e.g., email, text 
message) rather than in person) continuing through approximately 12 months after their last 
study vaccination.  These subjects will also be encouraged to provide a venous blood sample for 
serological assessment (HAI and Neut antibody assays) at approximately 28 days after their last 
study vaccination.  Collection of venous blood samples for systems biology studies will be 
obtained at serial time points indicated per the study schedule, if possible.  See the protocol-
specific Manual of Procedures (MOP) for alternate follow-up requirements. 
Every attempt will be made to follow all adverse events, including solicited injection site and 
systemic reactions, unsolicited non-serious adverse events, serious adverse events, and 
medically-attended adverse events including new-onset chronic medical conditions and 
potentially immune-medicated medical conditions, ongoing at the time of early withdrawal 
through resolution as per applicable collection times defined for the specific type of adverse 
event. 
In the case of subjects who fail to appear for a follow-up safety assessment, extensive effort (i.e., 
three documented contact attempts via phone calls, e-mails, etc., made on separate occasions and 
followed by a certified letter) will be made to locate or recall them, or at least to determine their 
health status.  These efforts will be documented in the subject’s study records. 
Subjects who withdraw, or are withdrawn or terminated from this study, or are lost to follow-up 
after signing the informed consent form and randomization but before receipt of study vaccine 
may be replaced.  Similarly, subjects in the A/H7N9 study arms who do not receive their second 
vaccination will be replaced to ensure that at least 10 evaluable subjects per A/H7N9 study arm 
are included in the analyses.  
5.2.5 
Termination of Study 
Although the sponsor has every intention of completing this study, it reserves the right to 
terminate this study at any time for clinical or administrative reasons.  Reasons for termination 


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include, but are not limited to, study closure due to SMC review and recommendation and at the 
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6 
STUDY INTERVENTION/INVESTIGATIONAL PRODUCT 
6.1 
Study Product Description 
A/H7N9 Vaccine 
Sanofi Pasteur has developed a monovalent inactivated influenza A/H7N9 virus vaccine for 
intramuscular use.  The manufacturing process for the production of this monovalent 
inactivated influenza A/H7N9 virus vaccine is similar to the manufacturing process for the 
production of its licensed Influenza Virus Vaccine Fluzone® family of products, except for a 
minor modification in the PBS diluent in the formulation step that was made according to 
previous experiences of manufacturing of monovalent pandemic vaccines.  This monovalent 
inactivated split influenza virus vaccine was derived from the influenza A/Shanghai/2/2013 
virus.  Licensed release testing specifications were maintained, with a modification to the 
potency test.  
 
 
 
 
 
 
 
 
 
 
  We 
plan to use a 3.75 mcg of HA (A/Shanghai/2/2013) per 0.5 mL [adjuvanted] or 0.25 mL 
[unadjuvanted] dose in this study. 
AS03 Adjuvant [Adjuvant System (03)] 
 
 
A/H3N2v Vaccine 
Sanofi Pasteur has developed a monovalent inactivated influenza A/H3N2v. The resulting 
reassortant is designated A/Minnesota/11/2010 NYMC X-203 virus vaccine for 
intramuscular use.  The manufacturing process for the production of this monovalent 




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6.1.2 
Formulation, Storage, Packaging, and Labeling 
A/H7N9 Vaccine 
The monovalent inactivated influenza A/H7N9 virus vaccine is supplied as a sterile, clear, 
and slightly opalescent suspension in single-dose vials containing 7.5 mcg of HA per 0.5 mL.  
Each vial contains a fill volume of 0.7 mL.  It contains no preservative (i.e., non-thimerosal) 
or antibiotics.  The vials containing study product must be stored at 2°C to 8°C (35.6°F to 
46.4°F).  Do not freeze.  Vials will be provided with latex-free stoppers. 
AS03 Adjuvant 
The AS03 adjuvant is supplied as a preservative-free, oil-in-water, whitish to yellowish 
homogenous milky liquid emulsion in single-use vials containing a fill volume of 3.15 mL.  
The vials containing study product must be stored at 2°C to 8°C (35.6°F to 46.4°F).  Do not 
freeze.  Vials will be provided with latex-free stoppers. 
A/H3N2v Vaccine 
The monovalent inactivated influenza A/H3N2v virus vaccine is supplied as a sterile, clear, 
and slightly opalescent suspension. A minor modification in the PBS diluent in the 
formulation step was made to accommodate the nature of the diluted virus in single-dose, 
prefilled syringes, without needles, containing 15 mcg of HA per 0.5mL.  It contains no 
preservative (i.e., non-thimerosal), antibiotics, or latex.  The vaccine includes porcine gelatin 
(0.05%) as a stabilizer.  The prefilled syringes containing study product must be stored at 
2°C to 8°C (35°F to 46°F).  Do not freeze. 
Each of these study products will be labeled according to manufacturer specifications and 
include the statement “Caution: New Drug – Limited by Federal Law to Investigational Use.” 
Further details are included in the respective, applicable Investigator’s Brochures and 
Supplemental Information for the Investigator’s Brochures for the A/H7N9 vaccine, AS03 
adjuvant, and A/H3N2v vaccine as well as protocol-specific MOP. 
Sterile empty vials will be provided with latex-free stoppers. 
6.1.3 
Study Product Storage and Stability Procedures 
The temperature of the storage unit must be manually recorded daily (excluding non-business 
days and holidays as applicable), continuously monitored and recorded during the duration of 
this study per the participating VTEU site’s standard operating procedures, and documentation 


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will be maintained.  If the temperature fluctuates outside of the required range, the affected study 
product(s) must be quarantined at the correct storage temperature and labeled as ‘Do Not Use’ 
(until further notice).  The research pharmacist must alert the site principal investigator and study 
coordinator, if the temperature fluctuates outside of the required range.  In the event the 
temperature fluctuates outside of the required range, including accidental deep-freezing or 
disruption of the cold chain, the affected study product(s) must not be administered.  The site 
principal investigator or responsible person should immediately contact the DMID Product 
Support Team at DMIDProductSupportTeam@niaid.nih.gov for further instructions before any 
additional study vaccinations are administered.  Based on the information collected, DMID 
and/or the manufacturer will determine whether the affected study product(s) can be used.  If it 
cannot be used, the site will receive specific instructions on how to return the affected study 
product(s) to the DMID CAR or destroy it on site.  Additional instructions for quarantine are 
provided in the protocol-specific MOP. 
6.2 
Dosage, Preparation, and Administration of Study 
Intervention/Investigational Product 
See the protocol-specific MOP Appendices B and C for detailed information on the preparation, 
labeling, storage, and administration of A/H7N9 vaccine given with or without AS03 adjuvant.  
Study vaccine preparation will be performed by the participating VTEU site’s research 
pharmacist on the same day of study vaccine administration.  No study vaccine preparation is 
required for the A/H3N2v vaccine as it is contained in a prefilled syringe and the entire contents 
will be administered to subjects per treatment assignment. 
Visually inspect the A/H7N9 vaccine, AS03 adjuvant, and A/H3N2v vaccine upon receipt and 
prior to use.  If the study product(s) appear(s) to have been damaged, contaminated or discolored, 
contain visible particulate matter, or if there are any concerns regarding its integrity, do NOT use 
the affected study product(s).  The affected study product(s) must be quarantined at 2°C to 8°C 
(35.6°F to 46.4°F) and labeled as ‘Do Not Use’ (until further notice).  The site principal 
investigator or responsible person should immediately contact the DMID Product Support Team 
at DMIDProductSupportTeam@niaid.nih.gov and DMID Clinical Project Manager for further 
instructions before any additional study vaccinations are administered.  Based on the information 
collected, DMID and/or the manufacturer will determine whether the affected study product(s) 
can be used.  If it cannot be used, the site will receive specific instructions on how to return the 
affected study product(s) to the DMID CAR or destroy it on site.  If the A/H7N9 vaccine, AS03 
adjuvant, or A/H3N2v vaccine is unusable, study personnel will use another vial or prefilled 
syringe from the study supply.  Replacement vials or prefilled syringes may be requested by 


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contacting DMID.  Additional instructions for quarantine and DMID contact information are 
provided in the protocol-specific MOP. 
Also, visually inspect the A/H7N9 vaccine plus AS03 adjuvant admixture prior to use.  The 
admixture will be milky (whitish to yellowish) in appearance.  If it appears to have been 
damaged, contaminated or discolored, contain visible particulate matter, or if there are any 
concerns regarding its integrity, do NOT use it.  The affected A/H7N9 vaccine plus AS03 
adjuvant admixture must be quarantined at room temperature and labeled as ‘Do Not Use’ (until 
further notice).  The site principal investigator or responsible person should immediately contact 
the DMID Product Support Team at DMIDProductSupportTeam@niaid.nih.gov and DMID 
Clinical Project Manager for further instructions before any additional study vaccinations are 
administered.  Based on the information collected, DMID and/or the manufacturer will determine 
whether the A/H7N9 vaccine plus AS03 adjuvant admixture can be used.  If it cannot be used, 
the site will receive specific instructions on how to send the A/H7N9 vaccine plus AS03 
adjuvant admixture to the DMID CAR or destroy it on site.  If the A/H7N9 vaccine plus AS03 
adjuvant admixture is unusable, the participating VTEU site’s research pharmacist will prepare 
another vial.  Additional instructions for quarantine and DMID contact information are provided 
in the protocol-specific MOP. 
Once mixed, the A/H7N9 vaccine plus AS03 adjuvant admixture must be stored at room 
temperature in an upright position and must be used within 8 hours. 
Study vaccine administration will be performed by an unblinded study team member who is 
credentialed to administer vaccines and may also participate in dose preparation, but will not be 
involved in study-related assessments or have subject contact for data collection following study 
vaccine administration.  Each dose of study vaccine will be administered to subjects via a single 
IM injection into the deltoid muscle of the preferred arm on the day of each study vaccination.  
Study vaccinations subsequent to the first study vaccination may be given in the same preferred 
arm as long as there is no interference with the reactogenicity assessment.  The site of injection 
(right or left arm) will be recorded on the appropriate data collection form.  Aseptic technique 
will be used for the withdrawal and administration of each dose of study vaccine using a 
disposable, sterile needle appropriate in length for each subject and a 1-mL disposable, sterile 
syringe (except for the A/H3N2v vaccine as it is contained in a prefilled syringe).  See the 
protocol-specific MOP for information on how to administer IM injections.  For the A/H7N9 
vaccine and A/H7N9 vaccine plus AS03 adjuvant admixture, each dose of study vaccine must be 
administered within 30 minutes of drawing into the syringe, and the prepared syringe must 
remain at room temperature until administered.  Note: Each 0.5 mL dose of AS03-adjuvanted 
A/H7N9 vaccine contains one- 0.25 mL dose of AS03 adjuvant. 


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6.3 
Modification of Study Intervention/Investigational Product for a 
Subject 
There will be no dose modifications.  If a subject’s second study vaccination is deferred, it 
should be rescheduled to occur within the acceptable protocol-specified window for that visit 
(Day 29 ± 2 days post first study vaccination).  No exceptions to the protocol-specified window 
will be made. 
Subjects who do not receive the second study vaccination will continue with follow-up safety 
assessments (may be conducted by phone call/electronic communication (e.g., email, text 
message) rather than in person) continuing through approximately 12 months after their first 
study vaccination.  These subjects will also be encouraged to provide a venous blood sample for 
serological assessment (HAI and Neut antibody assays) at approximately 28 days after their first 
study vaccination.  See Sections 5.2.3 and 5.2.4 for reasons for and handling of withdrawals and 
discontinuation of treatment.  See the protocol-specific MOP for alternate follow-up 
requirements. 
6.4 
Accountability Procedures for the Study Intervention/ 
Investigational Product(s) 
After receipt of the A/H7N9 vaccine, AS03 adjuvant, A/H3N2v vaccine, and sterile empty vials, 
the site principal investigator is responsible for study product distribution and disposition, and 
has ultimate responsibility for study product accountability.  The site principal investigator may 
delegate to the participating VTEU site’s research pharmacist responsibility for study product 
accountability.  The participating VTEU site’s research pharmacist will be responsible for 
maintaining complete records and documentation of study product receipt, accountability, 
dispensation, storage conditions, and final disposition of the study product(s).  The study product 
accountability records and dispensing logs will also capture, as appropriate for type of study 
product, vial numbers, including final mixed vial number, date of study vaccine 
preparation/administration, time of study vaccine preparation, expiration of study vaccine 
preparation, time study vaccine is drawn into the syringe, expiration time of study vaccine drawn 
into the syringe, and amount of study vaccine withdrawn for administration.  Time of study 
vaccine administration to the subject will be captured on the appropriate data collection form.  
All study product(s), including the amount of A/H7N9 vaccine, AS03 adjuvant, A/H3N2v 
vaccine, and A/H7N9 vaccine plus AS03 adjuvant admixture, whether administered or not, must 
be documented on the appropriate study product accountability record or dispensing log.  The 
sponsor's monitoring staff will verify the participating VTEU site’s study product accountability 
records and dispensing logs per the site monitoring plan. 


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Used and unused vials of A/H7N9 vaccine, AS03 adjuvant, and A/H7N9 vaccine plus AS03 
adjuvant admixture and unused syringes of A/H3N2v vaccine will be retained until monitored 
and released for disposition as applicable.  This can occur on an ongoing basis for used vials of 
A/H7N9 vaccine, AS03 adjuvant, and A/H7N9 vaccine plus AS03 adjuvant admixture.  Used 
syringes of A/H3N2v vaccine will be discarded immediately after administration.  Final 
disposition of the unused A/H7N9 vaccine, AS03 adjuvant, A/H3N2v vaccine, and sterile empty 
vials will be determined by DMID and communicated to the participating VTEU sites by the 
DMID Clinical Project Manager. 
6.5 
Assessment of Subject Compliance with Study 
Intervention/Investigational Product 
Study product will be administered to subjects by an unblinded study vaccine administrator via 
IM injection at all dosing times according to subject treatment assignment and as described in 
Section 6.2.  Thus, subject compliance is not anticipated to be an issue.  Deviations from the 
dose schedule may only occur as described in Section 6.3. 
6.6 
Concomitant Medications/Treatments 
Administration of any medications, therapies, or vaccines will be recorded on the appropriate 
data collection form.  Concomitant medications will include all current medications and 
medications taken within 60 days prior to signing the informed consent form through 
approximately 28 days after the last study vaccination or through the early termination visit (if 
prior to 28 days after the last study vaccination), whichever occurs first.  Medications reported in 
the electronic case report form (eCRF) are limited to those taken within 30 days prior to the first 
study vaccination through approximately 28 days after the last study vaccination.  Prescription 
and over-the-counter drugs will be included as well as herbals, vitamins and supplements.  In 
addition, receipt of non-study influenza vaccines will be solicited through approximately 28 days 
after the last study vaccination, and reported in the eCRF. 
Use of new medication should prompt evaluation for the presence of a new diagnosis of chronic 
medical disease or condition. 
Medications that might interfere with the evaluation of the investigational product(s) should not 
be used during the study-reporting period (approximately 12 months after the last study 
vaccination) unless clinically indicated as part of the subject’s health care.  Medications in this 
category include the prohibited medications per the Subject Exclusion Criteria (see Section 
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other medications that should not be used due to a risk to subject safety or assessment of 
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7 
STUDY SCHEDULE 
Complete study schedule details listed by type of visit are described below.  Refer also to 
Sections 4 and 8 and Appendix B. 
7.1 
Screening and Enrollment Visits 
7.1.1 
Visit 00, Day -7, Screening, Clinic Visit, All Subjects 
(Window: Day -14 to Day -5 prior to first study vaccination) 
Potential subjects will be screened for eligibility up to 14 days, but no less than 5 days prior to 
the administration of the first study vaccination.  The following activities will be performed: 
• Subjects will be provided with a description of this study (purpose and study procedures) 
and asked to read and sign the informed consent form.  The informed consent form will 
be signed prior to performing any study procedures, including any screening procedures. 
• Demographic information will be obtained by interview of subjects. 
• Eligibility criteria will be reviewed with subjects. 
• Complete medical history will be obtained by interview of subjects to ensure eligibility. 
• All concomitant medications taken within 60 days prior to signing the informed consent 
form will be reviewed with subjects.  Medications reported in the eCRF are limited to 
those taken within 30 days prior to the first study vaccination. 
• Subject receipt of licensed seasonal influenza vaccines over the previous two seasons, 
what type (inactivated or live attenuated), and approximate date of vaccination will be 
recorded on the appropriate data collection form, if known.  Prior receipt of licensed 
seasonal influenza vaccine is not exclusionary, as long as it has been administered within 
the allowable window (see Section 5.1.2). 
• Vital signs, including oral temperature, pulse, and blood pressure, will be obtained to 
ensure eligibility.  Subjects must not eat or drink anything hot or cold, or smoke within 
10 minutes prior to taking oral temperature. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 


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review of complete medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• A urine or serum pregnancy test will be performed on all women of childbearing 
potential and must be negative to ensure eligibility. 
• Approximately 4 mL of venous blood will be collected for ESR, and performed locally 
by the site.  The ESR value must be confirmed as less than 30 mm per hour prior to 
randomization and first study vaccination. The ESR, if initially elevated, may be repeated 
once to determine eligibility. 
• Approximately 90 mL of venous blood will be collected for baseline cytokine and 
chemokine levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
7.1.2 
Visit 01, Day 1, Enrollment and First Study Vaccination (Dose 1), Clinic Visit, 
All Subjects 
• Subject’s willingness to participate will be reconfirmed and documented in the subject’s 
study records prior to performing any further study procedures, including administration 
of the first study vaccination. 
• Eligibility criteria, including results of the ESR evaluation, will be reviewed with subjects 
prior to the first study vaccination to ensure continued eligibility. 
• Complete medical history and any updates obtained by interview of subjects since the 
screening visit will be reviewed with subjects prior to the first study vaccination to ensure 
continued eligibility. 
• All concomitant medications will be reviewed with subjects prior to the first study 
vaccination for accuracy and completeness.  Any new concomitant medications taken 
since the screening visit will be reviewed with subjects and assessed for continued 
eligibility prior to the first study vaccination.  Medications reported in the eCRF are 
limited to those taken within 30 days prior to the first study vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, will be obtained prior 
to the first study vaccination.  Vital signs assessed on Day 1 prior to the first study 


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vaccination will be considered as baseline.  Subjects must not eat or drink anything hot or 
cold, or smoke within 10 minutes prior to taking oral temperature. 
• Height and weight will be collected prior to the first study vaccination for the calculation 
of Body Mass Index (BMI). 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed prior to the first 
study vaccination, if indicated based on review of complete medical history and any 
updates obtained by interview of subjects since the screening visit, by a study clinician 
licensed to make medical diagnoses and listed on the Form FDA 1572 as the site 
principal investigator or sub-investigator. 
• A urine or serum pregnancy test will be performed within 24 hours prior to the first study 
vaccination on all women of childbearing potential.  Results must be negative and known 
prior to randomization and first study vaccination. 
• Approximately 10 mL of venous blood will be collected immediately prior to the first 
study vaccination for baseline HAI and Neut antibody assays. 
• Approximately 90 mL of venous blood will be collected immediately prior to the first 
study vaccination for baseline cytokine and chemokine levels and systems biology 
studies. 
• Approximately 20 mL of urine will be collected prior to the first study vaccination for 
future research. 
• Subjects will be enrolled in AdvantageEDCSM and assigned randomly to a treatment arm 
prior to the first study vaccination. 
• Pre-administration reactogenicity assessments will be performed prior to the first study 
vaccination to establish baseline.  Subjects will then receive a single dose of study 
vaccine via IM injection into the deltoid muscle of the preferred arm.  The site of 
injection (right or left arm) and time of administration will be recorded on the appropriate 
data collection form.  Subjects will be observed in the clinic for at least 20 minutes after 
the first study vaccination.  The first study vaccination site will be examined, post-
administration reactogenicity assessments will be performed, and any AE/SAEs will be 
recorded on the appropriate data collection form prior to discharge from the clinic. 


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• Subjects will be provided with a memory aid and other study-related materials to record 
daily oral temperature, solicited injection site and systemic reactions, any unsolicited 
AEs, and concomitant medications.  Subjects will be encouraged to take their oral 
temperature around the same time each day.  Subjects must not eat or drink anything hot 
or cold, or smoke within 10 minutes prior to taking oral temperature.  Subjects will be 
instructed on how to use their memory aid and how to measure and record AEs prior to 
discharge from the clinic.  Subjects will be instructed to notify the study center if they 
develop any severe reactions after the first study vaccination.  If the site principal 
investigator or appropriate sub-investigator deems the reaction severe enough, further 
instructions will be given to the subject on the proper course of action, including a return 
to the clinic for immediate evaluation if appropriate. 
7.2 
Follow-up Visits 
Follow-up visits are scheduled in reference to study vaccination dates as indicated for each visit 
window. 
7.2.1 
Visit 02, Day 2, Clinic Visit, All Subjects 
(Window: Day 2+1 day post first study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The first study vaccination site will be examined. 


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• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
7.2.2 
Visit 03, Day 4, Clinic Visit, All Subjects 
(Window: Day 4+1 day post first study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The first study vaccination site will be examined. 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
7.2.3 
Visit 04, Day 8, Clinic Visit, A/H3N2v-vaccinated subjects only 
(Window: Day 8+2 days post first study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 


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• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The first study vaccination site will be examined. 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
7.2.4 
Visit 04, Day 8, Phone Call/Electronic Communication, A/H7N9-vaccinated 
subjects only 
(Window: Day 8+2 days post first study vaccination) 
Study personnel will contact subjects by phone or electronic communication (e.g., email, text 
message) to solicit any AE/SAE and concomitant medication information (including solicitation 
for receipt of any non-study influenza vaccines) and review information on their memory aid. 
7.2.5 
Visit 05, Day 29, *Second Study Vaccination (Dose 2) for A/H7N9-vaccinated 
subjects only, Clinic Visit, All Subjects 
(Window: Day 29±2 days post first study vaccination) 
*This visit is for all subjects; however only A/H7N9-vaccinated subjects will receive the 
second study vaccination (Dose 2). 
• A/H7N9-vaccinated subjects only – Eligibility criteria, will be reviewed with subjects 
prior to the second study vaccination* to ensure continued eligibility. 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 


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obtained by interview of subjects (prior to the second study vaccination*) and note any 
changes since the previous clinic visit or contact will be noted. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form (prior to the second 
study vaccination*). 
• All AE/SAEs will be recorded on the appropriate data collection form (prior to the 
second study vaccination*). 
• Vital signs, including oral temperature, pulse, and blood pressure, will be obtained prior 
to the second study vaccination*.  Subjects must not eat or drink anything hot or cold, or 
smoke within 10 minutes prior to taking oral temperature.  Note: Vital signs are not 
required for A/H3N2v-vaccinated subjects or A/H7N9-vaccinated subjects who are 
discontinued from receipt of the second study vaccination and are being followed for 
safety. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed (prior to the second 
study vaccination*), if indicated based on review of interim medical history, by a study 
clinician licensed to make medical diagnoses and listed on the Form FDA 1572 as the site 
principal investigator or sub-investigator. 
• A/H7N9-vaccinated subjects only – A urine or serum pregnancy test will be performed 
within 24 hours prior to the second study vaccination* on all women of childbearing 
potential.  Results must be negative and known prior the second study vaccination*. 
• Approximately 10 mL of venous blood will be collected for HAI and Neut antibody 
assays on all subjects.  A/H7N9-vaccinated subjects only – Blood should be collected 
immediately prior to the second study vaccination*. 
• A/H7N9-vaccinated subjects only – Approximately 90 mL of venous blood will be 
collected immediately prior to the second study vaccination* for cytokine and chemokine 
levels and systems biology studies. 
• A/H7N9-vaccinated subjects only - Approximately 20 mL of urine will be collected 
(prior to the second study vaccination*) for future research. 
• A/H7N9-vaccinated subjects only – Pre-administration reactogenicity assessments will be 
performed prior to the second study vaccination* to establish baseline.  Subjects will then 


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receive a single dose of study vaccine via IM injection into the deltoid muscle of the 
preferred arm.  The second study vaccination may be given in the same preferred arm as 
long as there is no interference with the reactogenicity assessment.  The site of injection 
(right or left arm) and time of administration will be recorded on the appropriate data 
collection form.  Subjects will be observed in the clinic for at least 20 minutes after the 
second study vaccination.  The second study vaccination site will be examined, post-
administration reactogenicity assessments will be performed, and any AE/SAEs will be 
recorded on the appropriate data collection form prior to discharge from the clinic. 
• A/H7N9-vaccinated subjects only – Subjects will be provided with a memory aid and 
other study-related materials to record daily oral temperature, solicited injection site and 
systemic reactions, any unsolicited AEs, and concomitant medications.  Subjects will be 
encouraged to take their oral temperature around the same time each day.  Subjects must 
not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral 
temperature.  Subjects will be instructed on how to use their memory aid and how to 
measure and record AEs prior to discharge from the clinic.  Subjects will be instructed to 
notify the study center if they develop any severe reactions after the second study 
vaccination.  If the site principal investigator or appropriate sub-investigator deems the 
reaction severe enough, further instructions will be given to the subject on the proper 
course of action, including a return to the clinic for immediate evaluation if appropriate. 
Note: This will be the last visit for those randomized to the A/H3N2v arm. 
7.2.6 
Visit 06, Day 30, Clinic Visit, A/H7N9-vaccinated subjects only 
(Window: Day 2+1 day post second study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 


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review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The second study vaccination site will be examined. 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
7.2.7 
Visit 07, Day 32, Clinic Visit, A/H7N9-vaccinated subjects only 
(Window: Day 4+1 day post second study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The second study vaccination site will be examined. 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 


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7.2.8 
Visit 08, Day 36, Clinic Visit, A/H7N9-vaccinated subjects only 
(Window: Day 8+2 days post second study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The second study vaccination site will be examined. 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies. 
• Approximately 20 mL of urine will be collected for future research. 
 
 


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7.2.9 
Visit 09, Day 57, Clinic Visit, A/H7N9-vaccinated subjects only 
(Window: Day 29±2 days post second study vaccination) 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• Approximately 10 mL of venous blood will be collected for HAI and Neut antibody 
assays. 
7.2.10 
Visit 10, Day 209, Phone Call/Electronic Communication, A/H7N9-vaccinated 
subjects only 
(Window: Day 181±14 days post second study vaccination) 
Subjects will be contacted by phone or electronic communication (e.g., email, text message) to 
query for safety events.  Adverse events limited to SAEs and MAAEs including new-onset 
chronic medical conditions and potentially immune-mediated medical conditions that have 
occurred since the previous clinic visit or contact will be solicited.  Based on the information, 
subjects may be asked to return to the clinic for evaluation. 
7.2.11 
Visit 11, Day 394, Phone Call/Electronic Communication, A/H7N9-vaccinated 
subjects only 
(Window: Day 366±14 days post second study vaccination) 
Subjects will be contacted by phone or electronic communication (e.g., email, text message) to 
query for safety events.  Adverse events limited to SAEs and MAAEs including new-onset 
chronic medical conditions and potentially immune-mediated medical conditions that have 


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occurred since the previous clinic visit or contact will be solicited.  Based on the information, 
subjects may be asked to return to the clinic for evaluation. 
7.3 
Early Termination Visit (if needed), All Subjects 
The following activities will be performed at the early termination visit for subjects who 
withdraw, or are withdrawn or terminated from this study: 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects (if within 8 days after the last 
study vaccination). 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form (if prior to 28 days 
after the last study vaccination). 
• All AE/SAEs will be recorded on the appropriate data collection form.  AEs will be 
limited to SAEs and MAAEs including new-onset chronic medical conditions and 
potentially immune-mediated medical conditions if after 28 days following the last study 
vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated.  Subjects must not eat or drink anything hot or cold, or smoke within 10 
minutes prior to taking oral temperature. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• The study vaccination site will be examined (if within 8 days after the last study 
vaccination). 
• Post-administration reactogenicity assessments will be performed (if within 8 days after 
the last study vaccination). 


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• Approximately 10 mL of venous blood will be collected for HAI and Neut antibody 
assays (if prior to 28 days after the last study vaccination). 
• Approximately 90 mL of venous blood will be collected for cytokine and chemokine 
levels and systems biology studies (if prior to 28 days after the last study vaccination). 
• Approximately 20 mL of urine will be collected for future research (if prior to 28 days 
after the last study vaccination). 
7.4 
Unscheduled Visit (if needed), All Subjects 
Unscheduled visits may occur at any time during this study.  Any of the following activities may 
be performed: 
• Interim medical history, including an assessment for new medical conditions and 
symptoms suggestive of a potentially immune-mediated medical condition, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted (if indicated). 
• Memory aid information will be reviewed with subjects (if within 8 days after the last 
study vaccination). 
• All concomitant medications (including solicitation for receipt of any non-study influenza 
vaccines) will be recorded on the appropriate data collection form (if prior to 28 days 
after the last study vaccination). 
• All AE/SAEs will be recorded on the appropriate data collection form.  AEs will be 
limited to SAEs and MAAEs including new-onset chronic medical conditions and 
potentially immune-mediated medical conditions if after 28 days following the last study 
vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated.  Subjects must not eat or drink anything hot or cold, or smoke within 10 
minutes prior to taking oral temperature. 
• A targeted physical examination, including an assessment for signs suggestive of a 
potentially immune-mediated medical condition, may be performed, if indicated based on 
review of interim medical history, by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
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• The study vaccination site will be examined (if within 8 days after the last study 
vaccination). 
• Post-administration reactogenicity assessments will be performed (if within 8 days after 
the last study vaccination). 


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8 
STUDY PROCEDURES AND EVALUATIONS 
8.1 
Clinical Evaluations 
Complete medical history will be obtained by interview of subjects at the screening visit and will 
be updated on Day 1 prior to the first study vaccination.  Subjects will be queried regarding a 
history of significant medical disorders of the head, eyes, ears, nose, throat, mouth, 
cardiovascular system, lungs, gastrointestinal tract, liver, pancreas, kidney, urologic system, 
nervous system, blood, lymph nodes, endocrine system, musculoskeletal system, skin, and 
genital/reproductive tract.  A history of any allergies, cancer, immunodeficiency, psychiatric 
illness, substance abuse, and autoimmune disease will be solicited.  At follow-up visits after the 
first study vaccination, an interim medical history will be obtained by interview of subjects 
noting any changes since the previous clinic visit or contact.  The interim medical history should 
include an assessment for new medical conditions and symptoms suggestive of a potentially 
immune-mediated medical condition. 
Medications history (concomitant medications) will include a review of all current medications 
and medications taken within 60 days prior to signing the informed consent form through 
approximately 28 days after the last study vaccination or through the early termination visit (if 
prior to 28 days after the last study vaccination), whichever occurs first.  Medications reported in 
the eCRF are limited to those taken within 30 days prior to the first study vaccination through 
approximately 28 days after the last study vaccination.  Prescription and over-the-counter drugs 
will be included as well as herbals, vitamins and supplements.  In addition, receipt of non-study 
influenza vaccines will be solicited through approximately 28 days after the last study 
vaccination, and reported in the eCRF.  Use of new medication should prompt evaluation for the 
presence of a new diagnosis of chronic medical disease or condition.  Assessment of eligibility 
will include a review of all permitted and prohibited medications per the Subject Inclusion and 
Exclusion Criteria (see Sections 5.1.1 and 5.1.2).  In addition, the site principal investigator or 
appropriate sub-investigator may identify other medications that should not be used due to a risk 
to subject safety or assessment of reactogenicity and immunogenicity. 
At the screening visit, a targeted physical examination may be performed, if indicated based on 
subject’s complete medical history, by a study clinician licensed to make medical diagnoses and 
listed on the Form FDA 1572 as the site principal investigator or sub-investigator.  On Day 1 
prior to the first study vaccination and at follow-up visits after the first study vaccination, a 
targeted physical examination may be performed, if indicated based on subject’s interim medical 
history, by a study clinician licensed to make medical diagnoses and listed on the Form FDA 
1572 as the site principal investigator or sub-investigator.  Targeted physical examinations 


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should also include an assessment for signs suggestive of a potentially immune-mediated 
medical condition. 
Vital signs (oral temperature, pulse, and blood pressure) will be collected at the screening visit 
and prior to each study vaccination.  Vital signs assessed on Day 1 prior to the first study 
vaccination will be considered as baseline.  Subjects must not eat or drink anything hot or cold, 
or smoke within 10 minutes prior to taking oral temperature. 
Height and weight will be collected on Day 1 prior to the first study vaccination for the 
calculation of BMI. 
Reactogenicity assessments will include an assessment of solicited adverse events occurring 
from the time of each study vaccination through 8 days after each study vaccination, which 
includes an assessment of injection site reactions including pruritus (itching), ecchymosis 
(bruising), erythema (redness), induration (hardness)/swelling, pain, and tenderness as well as 
systemic reactions including fever, feverishness (chills/shivering/sweating), fatigue (tiredness), 
malaise (general unwell feeling), myalgia (body aches/muscular pain exclusive of the injection 
site), arthralgia (joint pain exclusive of the injection site), headache, and nausea.  Pre-
administration reactogenicity assessments will be performed prior to each study vaccination to 
establish baseline, then the study vaccination will be given. 
Subjects will be observed in the clinic for at least 20 minutes after each study vaccination.  The 
study vaccination site will be examined, post-administration reactogenicity assessments will be 
performed, and any AE/SAEs will be recorded on the appropriate data collection form prior to 
discharge from the clinic.  The study vaccination site will also be examined at follow-up visits 
occurring through approximately 8 days after each study vaccination. 
All subjects will complete a subject memory aid from the time of each study vaccination through 
8 days after each study vaccination.  Subject memory aids will be reviewed with the subjects for 
adverse events (solicited injection site and systemic reactions and unsolicited AEs) at follow-up 
visits occurring through approximately 8 days after each study vaccination. 
8.2 
Laboratory Evaluations 
The volume of venous blood to be collected for ESR, HAI and Neut antibody assays, cytokine 
and chemokine levels, and systems biology studies is presented in the table below. 
 
 




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evaluation will be performed locally by the site.  A venous blood sample (approximately 4 mL) 
will be collected from each subject at the screening visit. 
8.2.2 
Special Assays or Procedures 
8.2.2.1 HAI and Neut Antibody Assays – Serologic Immunogenicity 
Assays to determine serum levels of HAI and Neut antibodies will be performed at Southern 
Research.  Venous blood samples (approximately 10 mL) for HAI and Neut antibody assays 
will be collected from subjects at Day 1 (immediately prior to the first study vaccination), 
and at approximately Days 29 (for A/H7N9-vaccinated subjects this will be collected 
immediately prior to the second study vaccination) and 57 (A/H7N9-vaccinated subjects 
only) after the first study vaccination.  Subjects who withdraw early will have HAI and Neut 
antibody assays run on available sera, including sera from the early termination visit, if 
available. 
The Vanderbilt VTEU site will remain blinded to the HAI and Neut antibody assay results 
performed at Southern Research until after all subjects have completed Day 57 visits and all 
serologic laboratory results are complete. 
8.2.2.2 Systems Biology Studies – Cellular Immunogenicity 
Venous blood samples collected from subjects at time points and amounts described below 
will be used to measure changes in cytokine and chemokine levels, immune cell activation 
status, and whole transcriptome and proteome profiles of major blood immune cells to 
identify and quantify changes in gene expression.  We will use mathematical modeling of the 
accumulated data to identify ribonucleic acid (RNA) or protein expression signatures that 
correlate significantly with the observed serum HAI and Neut antibody response, cytokine 
and chemokine response, and immune cell activation response.  Urine samples 
(approximately 20 mL) will also be collected from subjects at approximately Days -7 1, 2, 4, 
8 (A/H3N2v-vaccinated subjects only), 29, 30, 32, and 36 for future research.  Note: Days 
29, 30, 32, and 36 will apply only to A/H7N9-vaccinated subjects.  By comparing subjects 
vaccinated with A/H7N9 vaccine given with and without AS03 adjuvant, our study may 
identify biomarkers that indicate that the AS03 adjuvant has successfully enhanced the 
immunological response. We may also identify differences between receipt of A/H7N9 
vaccine given with and without AS03 adjuvant and unadjuvanted A/H3N2v vaccine. 
For systems biology analysis of the subject’s immune and blood cells responses to 
unadjuvanted A/H3N2v vaccine and A/H7N9 vaccine given with and without AS03 
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transcriptome and proteome profiles of major blood immune cells will be determined in 
venous blood samples collected at approximately Days -7, 1, 2, 4, and 8 (A/H3N2v-
vaccinated subjects only).  These will also be performed on venous blood samples collected 
at approximately Days 29, 30, 32, and 36 from A/H7N9-vaccinated subjects only.  RNA 
transcript abundance in blood immune cells collected at approximately Days -7, 1, 2, 4, and 8 
(A/H3N2v-vaccinated subjects only) will be determined by RNA-Seq and gene expression 
quantifications will be obtained by counting mapped reads.  RNA-Seq will also be performed 
on blood immune cells collected at approximately Days 29, 30, 32, and 36 from A/H7N9-
vaccinated subjects only.  Relative cellular protein changes in subject’s blood immune cells 
collected at approximately Days -7, 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only) will be 
identified by shotgun proteomics and quantified using either Isobaric Tags for Relative and 
Absolute Quantitation (iTRAQ) or Tandem Mass Tags (TMT)53.  These will also be 
performed on blood immune cells collected at approximately Days 29, 30, 32, and 36 from 
A/H7N9-vaccinated subjects only.  Similar comparisons will also be made between those 
subjects receiving A/H7N9 vaccine given with and without AS03 adjuvant and those subjects 
receiving unadjuvanted A/H3N2v vaccine. 
(a) Venous blood samples will be collected prior to the first study vaccination at 
approximately Days -7 and 1.  The pre-study vaccination time points permit setting 
the baseline immune activity for each subject enrolled in this study.  This is a 
significant advantage, because it will correct for baseline variations between subjects 
that are caused by uncontrollable parameters such as genomic and/or metagenomic 
variations, asymptomatic infections, personal habits (e.g., tobacco use), or even over-
the-counter medications. 
(b) Additional venous blood samples will be collected after the first study vaccination at 
approximately Days 2, 4, 8 (A/H3N2v-vaccinated subjects only), 29 and 57 
(A/H7N9-vaccinated subjects only) to monitor the serologic and cellular response to 
study vaccination.  On Day 29, for A/H3N2v-vaccinated subjects, only serologic 
assays will be performed. 
(c) For the approximately Days -7, 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only), and 29 
(A/H7N9-vaccinated subjects only) approximately 90 to 100 mL of venous blood will 
be collected for serological assessment (HAI and Neut antibody assays, as 
applicable), cytokine and chemokine levels, and systems biology studies.  On Day 29, 
for A/H3N2v-vaccinated subjects, only serologic assays will be performed.  For the 
final serological assessment at approximately Day 57, approximately 10 mL of 
venous blood will be collected from A/H7N9-vaccinated subjects only.  The venous 
blood samples will be immediately processed to avoid possible sample variation 


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caused by freezing and thawing cells. 
Urine samples (approximately 20 mL) will also be collected from subjects at 
approximately Days -7, 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only), 29, 30, 32, 
and 36 for future research.  Note: Days 29, 30, 32, and 36 will apply only to A/H7N9-
vaccinated subjects. 
Day -7: Baseline pre-study vaccination time point #1 – approximately 10 mL for 
cytokine and chemokine levels + approximately 80 mL for systems 
biology studies = 90 mL of venous blood 
Day 1: 
Baseline pre-study vaccination time point #2 (immediately prior to the 
first study vaccination) – approximately 10 mL for cytokine and 
chemokine levels + approximately 10 mL for HAI and Neut antibody 
assays + approximately 80 mL for systems biology studies = 100 mL of 
venous blood 
Day 2: 
To monitor early innate immune response – approximately 10 mL for 
cytokine and chemokine levels + approximately 80 mL for systems 
biology studies = 90 mL of venous blood 
Day 4: 
To monitor intermediate innate immune response – approximately 10 
mL for cytokine and chemokine levels + approximately 80 mL for 
systems biology studies = 90 mL of venous blood 
Day 8: 
A/H3N2v-vaccinated subjects only.  To monitor late innate immune 
response & early adaptive immune response – approximately 10 mL for 
cytokine and chemokine levels + approximately 80 mL for systems 
biology studies = 90 mL of venous blood 
Day 29: A/H3N2v-vaccinated subjects – approximately 10 mL for HAI and Neut 
antibody assays 
A/H7N9-vaccinated subjects.  To monitor the mature adaptive immune 
response (this will be collected immediately prior to the second study 
vaccination) – approximately 10 mL for cytokine and chemokine levels 
+ approximately 10 mL for HAI and Neut antibody assays + 80 mL for 
systems biology studies = 100 mL of venous blood 
Day 30: A/H7N9-vaccinated subjects only.  To monitor early innate immune 
response to second study vaccination – approximately 10 mL for 


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cytokine and chemokine levels + approximately 80 mL for systems 
biology studies = 90 mL of venous blood 
Day 32: A/H7N9-vaccinated subjects only.  To monitor late-intermediate innate 
immune response to second study vaccination – approximately 10 mL 
for cytokine and chemokine levels + approximately 80 mL for systems 
biology studies = 90 mL of venous blood 
Day 36: A/H7N9-vaccinated subjects only.  To monitor late innate immune 
response & early adaptive immune response to second study vaccination 
– approximately 10 mL for cytokine and chemokine levels + 
approximately 80 mL for systems biology studies = 90 mL of venous 
blood 
Day 57: A/H7N9-vaccinated subjects only.  Final serological assessment (HAI 
and Neut antibody assays) – approximately 10 mL for HAI and Neut 
antibody assays 
(d) A fraction of the collected venous blood samples will be analyzed as follows in e-k to 
measure the immunological response signature for each subject in response to study 
vaccination.  The emerging immunologic signatures will be used in systems biology 
modeling. 
(e) HAI- and Neut-specific Antibody Response: HAI- and Neut-specific antibody 
response will be measured at specified time points (see above for HAI and Neut 
antibody assays) to confirm the effective influenza immunity and to correlate with the 
systems biology studies data.  HAI and Neut antibody assays will be performed by 
Southern Research. 
(f) Cytokine and Chemokine Response: Using the Vanderbilt Immunology Core, 
cytokine and chemokine levels in venous blood samples will be quantitatively 
measured at approximately Days -7, 1, 2, 4, and 8 (A/H3N2v-vaccinated subjects 
only), as listed above in (c).  This will also be performed on venous blood samples 
collected at approximately Days 29, 30, 32, and 36 from A/H7N9-vaccinated subjects 
only. 
The majority of the collected venous blood samples will be used to isolate immune 
cell subsets to profile the transcriptome and proteome profiles in response to study 
vaccination.  The gene expression signatures along with the immunologic signatures 
will be used in the systems biology approach to identify biomarkers or gene 


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expression signatures that predict an immunological response. 
(g) Fractionation of Peripheral Blood into Immune Cell Subsets: Magnetic cell 
separation (MACS) using antibody-coated magnetic beads is a fast and efficient way 
to isolate specific immune cell populations directly from venous blood samples.  
Using Ficoll separation and immune subset marker-specific magnetic beads on the 
fresh venous blood samples, we will isolate 6 major immune cell subsets: 
1. Neutrophils (Polymorphonuclear Leukocyte [PMN]) 
2. Monocytes  
3. Natural Kill (NK) cells 
4. B-cells 
5. T-cells 
6. Dendritic cells (DC) 
All isolated immune cell subsets will be stored at -80°C.  The subjects’ samples will 
be used for in-depth systems biology analysis to optimize the discovery of gene 
expression signatures that predicts an early immunological response and protection.  
This will be performed by the Vanderbilt Immunology Core. 
(h) RNA-Seq Transcriptome Sequencing: Transcriptome profiling using RNA-Seq will 
be performed on immune cell subsets collected at approximately Days -7, 1, 2, 4, and 
8 (A/H3N2v-vaccinated subjects only), as listed above in (c) and (g) to identify and 
quantify the RNA transcript response after study vaccination.  This will also be 
performed on immune cell subsets collected at approximately Days 29, 30, 32, and 36 
from A/H7N9-vaccinated subjects only. Total RNA will be isolated using standard 
methods and subjected to ultra-high-throughput parallel next generation sequencing.  
This will be performed by the Vanderbilt Immunology Core and Hudson Alpha. 
(i) Proteomics Profiling: To identify and quantify changes in the immune cell’s 
proteome after study vaccination, quantitative and shotgun proteomics using 
multidimensional liquid chromatography coupled with tandem mass spectrometry 
will be performed on immune cell samples collected at approximately Days -7, 1, 2, 
4, and 8 (A/H3N2v-vaccinated subjects only), as listed above in (c) and (g).  These 
will also be performed on immune cell subsets collected at approximately Days 29, 
30, 32, and 36 from A/H7N9-vaccinated subjects only. This will be performed by the 
Vanderbilt Proteomics Core. 
(j) Systems Biology Analysis: Our goal is to identify the unique RNA and protein 
biomarkers that correlate with the immunological responses from A/H7N9 vaccine 
given with and without AS03 adjuvant.  We may also identify differences between 


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receipt of A/H7N9 vaccine given with and without AS03 adjuvant and unadjuvanted 
A/H3N2v vaccine. 
8.2.3 
Specimen Preparation, Handling, and Shipping 
8.2.3.1 Instructions for Specimen Preparation, Handling, and Storage 
Instructions for specimen preparation, handling, and storage are included in the 
protocol-specific MOP. 
8.2.3.2 Specimen Shipment 
Specimen shipment will occur at intervals during the course of this study following 
all applicable International Air Transport Association (IATA) requirements and 
according to the specifics for storage temperature and documentation as detailed in 
the protocol-specific MOP. 
Specimens for HAI and Neut antibody assays will be shipped from the participating 
VTEU site to the DMID CAR, and then provided by the DMID CAR to Southern 
Research in a blinded manner once they all become available to the DMID CAR. 
Specimens for systems biology studies will remain at the Vanderbilt VTEU site as 
experiments are being conducted.  Once RNA is extracted from subjects’ immune cell 
subsets, these RNA samples will be sent directly to HudsonAlpha for RNA-
sequencing given the susceptible for sample degradation during multiple shipments. 
Any specimens remaining once the protocol-defined systems biology studies are 
completed will be shipped to the DMID CAR. 
Further instructions for specimen shipment are included in the protocol-specific 
MOP. 


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9 
ASSESSMENT OF SAFETY 
9.1 
Specification of Safety Parameters 
Safety will be assessed by the frequency and severity of: 
1. Study vaccine-related serious adverse events occurring from the time of the first study 
vaccination through approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 
arms) after the last study vaccination. 
2. Solicited Adverse Events – reactogenicity events occurring from the time of each study 
vaccination through 8 days after each study vaccination: 
a) Injection site reactions including pruritus (itching), ecchymosis (bruising), erythema 
(redness), induration (hardness)/swelling, pain, and tenderness. 
b) Systemic reactions including fever, feverishness (chills/shivering/sweating), fatigue 
(tiredness), malaise (general unwell feeling), myalgia (body aches/muscular pain 
exclusive of the injection site), arthralgia (joint pain exclusive of the injection site), 
headache, and nausea. 
3. Unsolicited Adverse Events – study vaccine-related non-serious adverse events occurring 
from the time of each study vaccination through approximately 28 days after each study 
vaccination. 
4. Medically-Attended Adverse Events including new-onset chronic medical conditions and 
potentially immune-mediated medical conditions occurring from the time of the first study 
vaccination through approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 arms) 
after the last study vaccination. 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
9.2.1 
Adverse Events 
Adverse Event (AE):  International Conference on Harmonisation (ICH) E6 defines an AE as 
any untoward medical occurrence in a patient or clinical investigation subject administered a 
pharmaceutical product regardless of its causal relationship to the study treatment.  An AE can 
therefore be any unfavorable and unintended sign (including an abnormal laboratory finding), 
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The occurrence of an AE may come to the attention of study personnel during study visits and 
interviews of a study recipient presenting for medical care, or upon review by a study monitor. 
AEs, including solicited local (injection site) and systemic (subjective and quantitative) 
reactions, not meeting the protocol-defined criteria for SAEs will be captured on the appropriate 
data collection form and eCRF.  Information to be collected for unsolicited non-serious AEs 
includes event description, date of onset, licensed study physician’s assessment of severity and 
relationship to study product and alternate etiology (if not related to study product) (assessed 
only by those with the training and authority to make a diagnosis and listed on the Form FDA 
1572 as the site principal investigator or sub-investigator), date of resolution of the event, 
seriousness and outcome.  AEs occurring during the collection and reporting period will be 
documented appropriately regardless of relationship.  AEs will be followed through resolution. 
Any medical condition that is present at the time that the subject is screened will be considered 
as baseline and not reported as an AE.  However, if the severity of any pre-existing medical 
condition increases, it will be recorded as an AE. 
AEs must be graded for severity and assessed for relationship to study product (see definitions 
below).  Adverse events characterized as intermittent require documentation of onset and 
duration of each episode.  The start and stop date of each reported AE will be recorded on the 
appropriate data collection form and eCRF. 
FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether or not considered drug related. 
Medically-Attended Adverse Events (MAAEs):  For each unsolicited AE experienced, the 
subject will be asked if he/she had received medical attention, defined as hospitalization, an 
emergency room visit, or an otherwise unscheduled visit to or from medical personnel for any 
reason.  AEs characterized by such unscheduled medical care will be designated as medically-
attended adverse events.  MAAEs can also include new-onset chronic medical conditions, or 
potentially immune-mediated medical conditions (also known as Adverse Events of Special 
Interest [AESIs]). 
New-Onset Chronic Medical Conditions (NOCMCs):  New-onset chronic medical conditions are 
defined as any new ICD-10 diagnosis that is applied to the subject during the duration of the 
study, after receipt of the study agent, that is expected to continue for at least 3 months and 
requires continued health care intervention. 
Potentially Immune-Mediated Medical Conditions (PIMMCs):  Potentially immune-mediated 
medical conditions constitute a group of AEs that includes diseases which are clearly 


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autoimmune in etiology and other inflammatory and/or neurologic disorders which may or may 
not have autoimmune etiologies.  PIMMCs currently in effect are presented in Appendix A: List 
of Potentially Immune-Mediated Medical Conditions. 
Severity of Event:  AEs will be assessed by a licensed study physician listed on the Form FDA 
1572 as the site principal investigator or appropriate sub-investigator using a protocol-defined 
grading system (see Sections 9.2.2 and 9.2.3).  For events not included in the protocol-defined 
grading system, the following guidelines will be used to quantify severity: 
• 
Mild (Grade 1):  Events require minimal or no treatment and do not interfere with the 
subject’s daily activities. 
• 
Moderate (Grade 2):  Events result in a low level of inconvenience or concern with 
therapeutic measures.  Moderate events may cause some interference with functioning 
and daily activities. 
• 
Severe (Grade 3):  Events interrupt the subject’s daily activities and may require systemic 
drug therapy or other treatment.  Severe events are usually incapacitating. 
Relationship to Study Product:  The licensed study physician’s assessment of an AE's 
relationship to study product is part of the documentation process, but it is not a factor in 
determining what is or is not reported in this study.  If there is any doubt as to whether a clinical 
observation is an AE, the event should be reported.  The relationship to study product must be 
assessed for AEs using the terms: related or not related.  In a clinical trial, the study product must 
always be suspect.  To help assess, the following guidelines are used: 
• Related – There is a reasonable possibility that the study product caused the adverse 
event.  Reasonable possibility means that there is evidence to suggest a causal 
relationship between the study product and the adverse event. 
• Not Related – There is not a reasonable possibility that the administration of the study 
product caused the event. 
9.2.2 
Reactogenicity 
Reactogenicity events are AEs that are common and known to occur following administration of 
these types of study vaccines.  The following Toxicity Grading Scales will be used to grade 
solicited local (injection site) and systemic (subjective and quantitative) reactions: 
 
 


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Local (Injection Site) Reactogenicity Grading 
Local (Injection Site) 
Reaction 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Pain – experienced 
without touching the 
injection site 
(spontaneous discomfort) 
Subject is aware of pain, 
but it does not interfere 
with daily activity, and 
no pain medication is 
taken 
There is interference with 
daily activity or it 
requires repeated use of a 
non-narcotic pain reliever 
for >24 hours 
Pain prevents daily activity 
or requires any use of a 
narcotic pain reliever 
Tenderness – hurts only 
when injection site is 
touched or the arm is 
moved 
The area immediately 
surrounding the injection 
site hurts only when 
touched or with arm 
motion, and it does not 
interfere with daily 
activity 
The area immediately 
surrounding the injection 
site hurts when touched or 
with arm motion, and it 
interferes with daily 
activity 
The area immediately 
surrounding the injection 
site hurts when touched or 
with arm motion, and it 
prevents daily activity 
Pruritus (Itching) 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Ecchymosis (Bruising)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Erythema (Redness)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Induration 
(Hardness)/Swelling* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
* Will also be measured in mm but size will not be used as halting criteria. 
 
Ecchymosis (bruising), erythema (redness), and induration (hardness)/swelling as analyzed by 
measurement will be graded as follows: 
Local (Injection Site) Reactogenicity Measurements 
Local (Injection Site) Reaction 
Small 
Medium 
Large 
Ecchymosis (Bruising)* 
<20 mm 
20 mm – 50 mm 
>50 mm 
Erythema (Redness)* 
<20 mm 
20 mm – 50 mm 
>50 mm 
Induration (Hardness)/Swelling* 
<20 mm 
20 mm – 50 mm 
>50 mm 
* Will not be used as halting criteria. 
 
 


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Subjective Systemic Reactogenicity Grading 
Systemic (Subjective) 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Feverishness 
(Chills/Shivering/Sweating) 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Fatigue (Tiredness) 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Malaise (General Unwell 
Feeling) 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Myalgia (Body 
Aches/Muscular Pain)* 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Arthralgia (Joint Pain)* 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Headache 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
Nausea 
No interference 
with daily activity 
Some interference with 
daily activity 
Significant interference, 
prevents daily activity 
* Not at injection site. 
 
Oral temperature# will be graded as follows: 
Quantitative Systemic Reactogenicity Grading 
Systemic (Quantitative) 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Fever* - oral† 
38.0°C – 38.4°C 
100.4°F – 101.1°F 
38.5°C – 38.9°C 
101.2°F – 102.0°F 
>38.9°C 
>102.0°F 
# Oral temperature assessed on Day 1 prior to the first study vaccination will be considered as baseline. 
* A fever can be considered not related to the study product if an alternative etiology can be documented. 
† Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral temperature. 
 
 


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9.2.3 
Additional Adverse Event Severity Grading 
Pulse and blood pressure# will be graded as follows: 
Physiologic Parameter 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Bradycardia - beats per minute 
45 – 49 
40 – 44 
<40 
Tachycardia - beats per minute 
116 – 130 
131 – 155 
>155 
Hypotension (systolic) mm Hg 
80 – 84 
75 – 79 
<75 
Hypotension (diastolic) mm Hg 
50 – 54 
45 – 49 
<45 
Hypertension (systolic) mm Hg 
151 – 155 
156 – 160 
>160 
Hypertension (diastolic) mm Hg 
96 – 100 
101 – 105 
>105 
# Pulse and blood pressure assessed on Day 1 prior to the first study vaccination will be considered as baseline. 
9.2.4 
Serious Adverse Events 
Serious Adverse Event (SAE):  An adverse event or suspected adverse reaction is considered 
“serious” if, in the view of either the site principal investigator or sponsor, it results in any of the 
following outcomes: 
• Death, 
• a life-threatening adverse event*, 
• inpatient hospitalization or prolongation of existing hospitalization, 
• a persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions, or 
• a congenital anomaly/birth defect. 
• Important medical events that may not result in death, be life-threatening, or require 
hospitalizations may be considered serious when, based upon appropriate medical 
judgment they may jeopardize the patient or subject and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition.  Examples of such 
medical events include allergic bronchospasm requiring intensive treatment in an 
emergency room or at home, blood dyscrasias or convulsions that do not result in 
inpatient hospitalization, or the development of drug dependency or drug abuse. 


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• All events described as Guillain-Barré syndrome will also be considered SAEs. 
* Life-threatening adverse event.  An adverse event is considered “life-threatening” if, in the 
view of either the site principal investigator or sponsor, its occurrence places the patient or 
subject at immediate risk of death.  It does not include an adverse event that, had it occurred in a 
more severe form, might have caused death. 
SAEs will be: 
• 
Assessed for severity and relationship to study product and alternate etiology (if not 
related to study product) by a licensed study physician listed on the Form FDA 1572 as 
the site principal investigator or sub-investigator. 
• 
Recorded on the appropriate SAE form and eCRF. 
• 
Followed through resolution by a licensed study physician listed on the Form FDA 1572 
as the site principal investigator or sub-investigator. 
• 
Reviewed and evaluated by an Independent Safety Monitor (ISM), the SMC (periodic 
review unless related), DMID, and the IRB. 
9.2.5 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values 
or Abnormal Clinical Findings 
The site principal investigator or appropriate sub-investigator is responsible for recording all 
AE/SAEs that are observed or reported during this study, regardless of the relationship to study 
product.  AE/SAEs, abnormal clinical laboratory values, or abnormal clinical findings will be 
collected, assessed, documented, reported, and followed appropriately. 
9.3 
Reporting Procedures 
Solicited injection site and systemic reactogenicity events will be documented and reported from 
the time of each study vaccination through 8 days after each study vaccination. 
Unsolicited non-serious AEs will be documented and reported from the time of each study 
vaccination through approximately 28 days after each study vaccination. 
SAEs and MAAEs including new-onset chronic medical conditions and potentially immune-
mediated medical conditions will be documented and reported from the time of the first study 
vaccination through approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 arms) after 
the last study vaccination. 


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9.3.1 
Serious Adverse Events 
Any AE that meets a protocol-defined serious criterion must be submitted immediately (within 
24 hours of site awareness) on an SAE form to the DMID Pharmacovigilance Group at the 
following address: 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support (CROMS) 
6500 Rock Spring Dr., Suite 650 
Bethesda, MD 20814, USA 
SAE Hot Line: 1-800-537-9979 (US) or 1-301-897-1709 (outside US) 
SAE FAX: 1-800-275-7619 (US) or 1-301-897-1710 (outside US) 
SAE Email Address: PVG@dmidcroms.com 
In addition to the SAE form, selected SAE data fields must also be entered into 
AdvantageEDCSM.  Please see the protocol-specific MOP for details regarding this procedure. 
Other supporting documentation of the event may be requested by the DMID Pharmacovigilance 
Group and should be provided as soon as possible. 
The site will send a copy of the SAE report(s) to the ISM when they are provided to the DMID 
Pharmacovigilance Group.  The DMID Medical Monitor and DMID Clinical Project Manager 
will be notified of the SAE by the DMID Pharmacovigilance Group.  The DMID Medical 
Monitor will review and assess the SAE for regulatory reporting and potential impact on study 
subject safety and protocol conduct. 
 
At any time after completion of this study, if the site principal investigator or appropriate sub-
investigator becomes aware of an SAE that is suspected to be related to study product, the site 
principal investigator or appropriate sub-investigator will report the event to the DMID 
Pharmacovigilance Group. 
9.3.2 
Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND 
Following notification from the site principal investigator or appropriate sub-investigator, 
DMID, the Investigational New Drug (IND) sponsor, will report any suspected adverse reaction 
that is both serious and unexpected.  DMID will report an adverse event as a suspected adverse 
reaction only if there is evidence to suggest a causal relationship between the drug and the 
adverse event.  DMID will notify FDA and all participating site principal investigators (i.e., all 
principal investigators to whom the sponsor is providing drug under its IND(s) or under any 
principal investigator’s IND(s)) in an IND safety report of potential serious risks from clinical 
trials or any other source, as soon as possible, but in no case later than 15 calendar days after the 


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sponsor determines that the information qualifies for reporting as specified in 21 CFR Part 
312.32.  DMID will also notify FDA of any unexpected fatal or life-threatening suspected 
adverse reaction as soon as possible, but in no case later than 7 calendar days after the sponsor’s 
initial receipt of the information.  Relevant follow up information to an IND safety report will be 
submitted as soon as the information is available.  Upon request from FDA, DMID will submit to 
FDA any additional data or information that the agency deems necessary, as soon as possible, 
but in no case later than 15 calendar days after receiving the request. 
All serious events designated as “not related” to study product(s), will be reported to the FDA at 
least annually in a summary format. 
9.3.3 
Reporting of Pregnancy 
Pregnancies occurring in study subjects will be reported via AdvantageEDCSM on the Pregnancy 
Report form.  No further study vaccinations will be administered to pregnant subjects, but with 
the subject’s permission all protocol-required venous blood samples will be obtained and the 
subject will continue to be followed for safety for the duration of this study.  Efforts will be 
made to follow all pregnancies reported during the course of this study to pregnancy outcome 
pending the subject’s permission. 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events 
AEs will be collected, assessed, and followed through resolution from the time of each study 
vaccination through approximately 28 days after each study vaccination. 
SAEs and MAAEs including new-onset chronic medical conditions and potentially immune-
mediated medical conditions will be collected, assessed, and followed from the time of the first 
study vaccination through resolution even if this extends beyond the study-reporting period 
(approximately Day 29 for A/H3N2v-vaccinated subjects and approximately 12 months after the 
last study vaccination for A/H7N9 + AS03). 
Resolution of an AE/SAE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic. 
If the site principal investigator or appropriate sub-investigator becomes aware of an acute 
febrile illness and the site principal investigator or appropriate sub-investigator decides to bring 
the subject in for an evaluation to determine etiology, then the site principal investigator or 
appropriate sub-investigator, at their own discretion, can determine if specific viral testing should 
be performed. 


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Follow-up procedures, evaluations, and outcomes will be recorded on the appropriate data 
collection form. 
9.5 
Halting Rules 
Further enrollment and study vaccinations will be halted for SMC review/recommendation if any 
of the following are reported: 
• Any subject experiences ulceration, abscess, or necrosis at the injection site related to 
study product administration. 
• Any subject experiences laryngospasm, bronchospasm, or anaphylaxis within 1 day after 
administration of study product that is considered related to study product. 
• Two or more subjects experience generalized urticaria within 3 days after administration 
of study product that is considered related to study product. 
• Any subject experiences a study vaccine-related SAE from the time of the first study 
vaccination through the subject’s last study visit. 
• Any subject experiences acute weakness of limbs and/or cranial nerve innervated muscles 
(description of potential signal of GBS) after administration of study product. 
• Any subject develops a potentially immune-mediated medical condition after 
administration of study product. 
This study will also be halted for SMC review/recommendation if, within 8 days after 
administration of each study vaccination, any of the following occurs: 
• 10% or more of subjects (with a minimum of 2 subjects) who received at least one dose 
of study vaccine to date experience the same severe (Grade 3) study vaccine-related 
injection site reaction.  Ecchymosis (bruising), erythema (redness), and induration 
(hardness)/swelling will also be measured in mm but size will not be used as halting 
criteria. 
• 10% or more of subjects (with a minimum of 2 subjects) who received at least one dose 
of study vaccine to date experience the same severe (Grade 3) study vaccine-related 
subjective systemic reaction, for which the severity (grade) is corroborated by study 
personnel. 


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• 10% or more of subjects (with a minimum of 2 subjects) who received at least one dose 
of study vaccine to date experience the same severe (Grade 3) study vaccine-related 
quantitative systemic reaction. 
Grading scales for solicited local (injection site) and systemic (subjective and quantitative) 
reactions are included in Section 9.2.2. 
If any of the halting rules are met following any subject receipt of any study vaccination, then 
this study will not continue with the remaining enrollments or study vaccinations without a 
review by and recommendation from the SMC to proceed. 
DMID retains the authority to suspend additional enrollment and study interventions/ 
administration of study product during the entire study, as applicable. 
9.6 
Safety Oversight 
9.6.1 
Independent Safety Monitor (ISM) 
The ISM is a physician with relevant expertise whose primary responsibility is to provide 
independent safety monitoring in a timely fashion.  The ISM will review SAEs in real time and 
other AEs as needed and provide an independent medical assessment and recommendation to 
DMID.  The participating VTEU site will have an ISM with experience in infectious diseases or 
internal medicine, in close proximity to the participating VTEU site, and have the authority to 
readily access study participant records. 
9.6.2 
Safety Monitoring Committee (SMC) 
Safety oversight will be conducted by a SMC that is an independent group of experts that 
monitors subject safety and advises DMID.  The SMC members will be separate and 
independent of study personnel participating in this study and should not have scientific, 
financial or other conflict of interest related to this study.  The SMC will consist of members 
with appropriate expertise to contribute to the interpretation of the data from this study. 
The SMC will review study progress and participant, clinical, safety, and reactogenicity data at 
the following time points: 
• Data review for safety at study specific time frames; at least annually 
• After all 28-day post first study vaccination safety data are available. 


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• Ad hoc when a halting rule is met, for immediate concerns regarding observations during 
this study, or as needed. 
The SMC will operate under the rules of a DMID-approved charter that will be approved at the 
organizational meeting of the SMC.  At this time, each data element that the SMC needs to 
assess will be clearly defined.  Procedures for SMC reviews/meetings will be defined in the 
charter.  The SMC will review applicable data to include, but not limited to, study progress and 
participant, clinical, safety, and reactogenicity data.  Reports may include enrollment and 
demographic information, medical history, concomitant medications, physical assessments, and 
solicited and unsolicited AE/SAEs.  Additional data may be requested by the SMC, and interim 
statistical reports may be generated as deemed necessary and appropriate by DMID.  The SMC 
may receive data in aggregate and presented by treatment arm.  The SMC may also be provided 
with expected and observed rates of the expected AEs in an unblinded fashion, and may request 
the treatment assignment be unblinded for an individual subject if required for safety assessment.  
The SMC will review grouped and unblinded data in the closed session only.  The SMC will 
meet and review this data at scheduled time points or ad hoc as needed during this study as 
defined in the SMC charter.  As an outcome of each review/meeting, the SMC will make a 
recommendation as to the advisability of proceeding with study vaccinations (as applicable), and 
to continue, modify, or terminate this study. 
DMID or the SMC chair may convene the SMC on an ad hoc basis according to protocol criteria 
or if there are immediate concerns regarding observations during the course of the study.  The 
DMID Medical Monitor is empowered to stop enrollment and study vaccinations if adverse 
events that meet the halting criteria are reported.  The DMID Medical Monitor and the ISM will 
be responsible for reviewing SAEs in real time.  The SMC will review SAEs on a regular basis 
and ad hoc during the study. 


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10 
CLINICAL MONITORING 
10.1 Site Monitoring Plan 
Site monitoring will be conducted to ensure that human subjects protections, study and 
laboratory procedures, study intervention administration, and data collection processes are of 
high quality and meet sponsor, ICH/GCP guidelines and applicable regulations, and that this 
study is conducted in accordance with the protocol, protocol-specific MOP and applicable 
sponsor standard operating procedures.  DMID, the sponsoring agency, or its designee will 
conduct site-monitoring visits as detailed in the clinical monitoring plan. 
Site visits will be made at standard intervals as defined by DMID and may be made more 
frequently as directed by DMID.  Monitoring visits will include, but are not limited to, review of 
regulatory files, accountability records, eCRFs, informed consent forms, medical and laboratory 
reports, and protocol compliance.  Site monitors will have reasonable access to the participating 
VTEU site, study personnel, and all study documentation according to the DMID-approved site 
monitoring plan.  Study monitors will meet with the site principal investigators to discuss any 
problems and actions to be taken and document visit findings and discussions. 


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11 
STATISTICAL CONSIDERATIONS 
11.1 Introduction 
This study will use a systems biology approach to get a better understanding of how human 
immune cells respond to influenza A/H7N9 vaccination on the molecular level and how the 
addition of AS03 adjuvant modulates this response. 
Specifically, this study will assess and characterize changes in gene expression and protein 
abundance in immune cells obtained from healthy adults after each of two doses of a monovalent 
inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur administered 
intramuscularly approximately 28 days apart at 3.75 mcg of HA (A/Shanghai/2/2013) per 0.5 mL 
[adjuvanted] or 0.25 mL [unadjuvanted] dose given with or without AS03 adjuvant 
manufactured by GSK.  We will also compare these responses to one dose of unadjuvanted 
monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi Pasteur 
administered intramuscularly at 15 mcg of HA (A/Minnesota/11/2010) per 0.5 mL dose. 
To elucidate the higher order organization of genes and proteins that differ between the three 
study arms, clusters with correlated response profiles, enriched functional modules and pathways 
as well as their placement within molecular networks will be determined.  In addition, 
cytokine/chemokine concentrations well as serum anti-HA hemagglutination-inhibition (HAI) 
and neutralizing antibody concentrations will be measured.  The goal is to correlate these 
measurements with responses on the molecular level to identify groups of genes and proteins that 
are associated with cytokine/chemokine production, seroconversion, or peak titer. 
11.2 Study Hypotheses 
This Phase II clinical study is not designed to test a specific hypothesis.  Rather it is intended as 
an exploratory hypothesis-generating study to reveal new insights into how human immune cells 
respond to influenza A/H7N9 or A/H3N2 vaccination and how AS03 modulates responses to 
A/H7N9. 


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11.3 Study Outcome Measures 
Refer to Section 3 for study outcome measures. 
11.4 Sample Size Considerations 
11.4.1 
Study Design 
This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study 
in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, 
inclusive, designed to evaluate and compare the immunogenicity between an intramuscular 
monovalent inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur given 
with and without AS03 adjuvant manufactured by GlaxoSmithKline, and an intramuscular 
unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi 
Pasteur. 
This study will use a standard and systems biology approach to assess the human early gene and 
protein signatures expressed at baseline (approximately Days -7 and 1), and at approximately 
Days 2, 4, and 8 (A/H3N2v-vaccinated subjects only) after the first study vaccination in each 
treatment arm as well as at approximately Days 29, 30, 32, and 36 in A/H7N9-vaccinated 
subjects only.  Cellular immunogenicity (systems biology studies) data will be integrated with 
serologic immunogenicity (HAI and Neut antibody assays) and reactogenicity data to develop a 
systems model of the human immune response to unadjuvanted monovalent inactivated influenza 
A/H3N2v virus vaccine and monovalent inactivated influenza A/H7N9 virus vaccine given with 
and without AS03 adjuvant. 
This study will use venous blood samples and subject data collected from a total of thirty 
vaccinated subjects randomly divided into three equal treatment arms.  The first treatment arm 
(n=10) will be vaccinated with one dose of 15 µg of A/H3N2v HA.  The second treatment arm 
(n=10) will be vaccinated with two doses of 3.75 µg of A/H7N9 HA given with AS03 
approximately 28 days apart.  The third treatment arm (n=10) will be vaccinated with two doses 
of 3.75 µg of A/H7N9 HA given without AS03 adjuvant approximately 28 days apart.  All study 
vaccinations will be administered intramuscularly. 
Reactogenicity will be measured by the occurrence of solicited injection site and systemic 
reactions from the time of each study vaccination through 8 days after each study vaccination.  
Unsolicited non-serious adverse events (AEs) will be collected from the time of each study 
vaccination through approximately 28 days after each study vaccination.  Serious adverse events 
(SAEs) and medically-attended adverse events (MAAEs) including new-onset chronic medical 


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conditions (NOCMCs) and potentially immune-mediated medical conditions (PIMMCs) will be 
collected from the time of the first study vaccination through approximately 1 month (A/H3N2v 
arm) and 12 months A/(H7N9 arms) after the last study vaccination. 
Venous blood samples (approximately 90 mL) will be collected from subjects at approximately 
Days -7 and 1 (immediately prior to the first study vaccination), and at approximately Days 2, 4, 
and 8 (A/H3N2v-vaccinated subjects only), after the first study vaccination for systems biology 
studies (cytokine and chemokine levels, immune cell activation status, and whole transcriptome 
and proteome profiles of the major blood immune cells).  Additionally, subjects in either 
treatment arm receiving A/H7N9 vaccine will have additional blood samples collected at 
approximately Days 29 (this will be collected immediately prior to the second study vaccination 
for subjects in either group receiving A/H7N9), 30, 32, and 36 after the first study vaccination 
for systems biology studies. 
Serological assessment (hemagglutination inhibition (HAI) and neutralizing (Neut) antibody 
assays) will also be conducted on venous blood samples (approximately 10 mL) collected from 
subjects at Day 1 (immediately prior to the first study vaccination), and at approximately Days 
29 (for A/H7N9-vaccinated subjects this will be collected immediately prior to the second study 
vaccination) and 57 (A/H7N9-vaccinated subjects only) after the first study vaccination. 
We will quantify and characterize serum cytokine/chemokine levels, as well as transcriptomic 
and proteomic profiles from individual immune cell compartments, comparing this with standard 
serologic assessment to vaccine. 
Urine samples (approximately 20 mL) will also be collected from subjects at approximately 
Days -7 1, 2, 4, 8 (A/H3N2v-vaccinated subjects only), 29, 30, 32, and 36 for future research.  
Note: Days 29, 30, 32, and 36 will apply only to A/H7N9-vaccinated subjects. 
11.4.2 
Study Population 
The study population for this clinical study is approximately 30 healthy male and non-pregnant 
female subjects aged 18 to 49 years old, inclusive, who meet all eligibility criteria.  The subjects 
will be recruited from the general population at the participating VTEU site that has substantial 
experience conducting influenza vaccine studies. 
11.4.3 
Sample Size 
The sample size is not based on a formal hypothesis.  Rather it is based on practical 
considerations with the goal of gathering enough information to learn more about immune 
system responses to the vaccine/adjuvant on the molecular level.  DMID Protocol 10-0074, 


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entitled:  “A Randomized, Double-Blinded, Controlled, Phase I Study in Healthy Adults to Assess 
the Safety, Reactogenicity, and Immunogenicity of Intramuscular Subvirion Inactivated 
Monovalent Influenza A/H5N1 Virus Vaccine Administered With and Without AS03 Adjuvant: 
Standard & Systems Biology Analyses”, showed that sample sizes in this range are sufficient to 
detect key differential signals and carry out systems biology analyses. 
11.4.4 
 Planned Interim Analyses 
Interim analyses would only be used to terminate this study in the event that unanticipated safety 
events deemed to be of sufficient concern require such action by the sponsor.  These assessments 
will not be made on the basis of testing a formal statistical hypothesis; therefore, p-value 
adjustment will not be made to any analyses.  A SMC will be convened by DMID to review 
study progress and participant, clinical, safety, and reactogenicity data. 
11.4.5 
Interim Safety Review 
An interim safety review may include enrollment and demographic information, medical history, 
concomitant medications, physical assessments, and solicited and unsolicited AE/SAEs.  
Additional data may be requested by the SMC, and interim statistical reports may be generated 
as deemed necessary and appropriate by DMID.  The SMC may receive data in aggregate and 
presented by treatment arm.  The SMC may also be provided with expected and observed rates 
of the expected AEs in an unblinded fashion, and may request the treatment arm be unblinded for 
an individual subject if required for safety assessment.  The SMC will review grouped and 
unblinded data in the closed session only.  The SMC will meet and review this data at scheduled 
time points or ad hoc as needed during this study as defined in the SMC charter.  As an outcome 
of each review/meeting, the SMC will make a recommendation as to the advisability of 
proceeding with study vaccinations (as applicable), and to continue, modify, or terminate this 
study. 
Additionally, this study will be monitored to determine if any of the halting rules described in 
Section 9.5 are met. 
11.4.6 
Interim Immunogenicity Review 
An interim immunogenicity review is not planned. 


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11.5 Final Analysis Plan 
Prior to database lock, a Statistical Analysis Plan that will outline in detail the planned statistical 
analyses for this study will be developed and finalized.  In the following section an abbreviated 
version is provided. 
11.5.1 
Systems Biology Analyses 
11.5.1.1 
Transcriptomics (RNA-seq) 
RNA-Seq data will be pre-processed by removing adapters and low-quality reads and mapping 
sequences to the latest human reference genome using splice-aware alignment software such as 
HISAT2..  Gene expression quantification will be carried out by using Subread against a 
common gene model reference.  Systematic differences between samples will be normalized 
using the TMM method as implemented in the edgeR R package.  Post normalization, genes that 
are known ribosomal, transfer, and mitochondrial RNA genes or genes located on the X or Y 
chromosomes will be excluded to avoid gender-specific effects Moderated log counts per million 
(LCPM) for the remaining genes will be determined using edgeR. LCPM will be used to identify 
lowly expressed genes, technical bias (e.g. batch effects or GC-content bias), and outlying 
samples.  More specifically, Principal Component Analysis, non-metric multidimensional 
scaling, and correlation heatmaps based on LCPM will be carried out to identify potential global 
outliers and batch effects.  Negative binomial models as implemented in edgeR will be used to 
determine differentially expressed genes for each endpoint and time point after removal of lowly 
expressed genes and outliers.  If technical bias is detected and the effect is balanced across 
comparison groups, it will be accounted for by including a covariate as part of the negative 
binomial models.  To compensate for testing multiple genes, p-values will be adjusted for by 
calculating false-discovery rates (FDR) based on the Benjamini-Hochberg procedure which 
controls the false positive rate among significantly differentially expressed genes.  Genes with a 
FDR-value < 0.05 and a fold change of greater or equal to 1.5-fold (up or down regulation) will 
be deemed to be significantly differentially expressed (DE).  Baseline responses (fold change in 
LCPM) of DE genes will be visualized using heatmaps.  Unsupervised multiscale bootstrapping 
will be carried out to identify robust gene clusters with correlated responses (pvclust R package).  
Visualizations of fold change time trends for robust gene cluster will be provided. In addition, 
higher order organization of DE genes will be investigated using gene set enrichment analysis 
based on the latest MSigDB and KEGG databases accounting for gene length bias (goseq R 
package).  Pathway maps color-coded by treatment effect will be provided for significantly 
enriched KEGG pathways.  Experimentally determined protein-protein interaction networks 
including known Influenza A-host interactions centered around DE genes will be generated using 
data from the IntAct database and visualized using Cytoscape. Regularized canonical correlation 


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(mixOmics R package), linear and logistic regression models (glmnet R package) based on 
baseline fold change in LCPM will be carried out to determine gene responses that are correlated 
with cytokine/chemokine concentrations, peak titer, or seroconversion status, respectively. 
11.5.1.2 
Proteomics 
A suitable global minimum reporter ion intensity threshold for peptide quantifications will be 
determined by exploring overall intensity distributions.  Reported intensities below this threshold 
will be treated as missing.  Next, peptide intensities will be divided by the reference channel ion 
intensities followed by log2 transformation of the ratio.  Systematic differences between 
experiments will be accounted for by scaling channel ion intensities so that the channel medians 
of the log2 ratios are the same for all samples within a cell type.  Log2 protein reference ratios 
(LPR) are then calculated as the median of the normalized peptide log2 reference ratios. 
Global patterns in LPR will be investigated using principal component analysis, hierarchical 
clustering, and multidimensional scaling to identify potential outlying samples and systematic 
effects.  For these multivariate analyses, missing data will be imputed for each immune cell type 
using the k-nearest neighbors algorithm implemented in the impute R package.  To accommodate 
smaller sample sizes, linear models as implemented in the limma R package  will be used to 
identify differentially abundant proteins (DA) for each endpoint and time point. The advantage 
of the empirical Bayes approach as implemented in limma is that it leverages information across 
all proteins during the inference process thereby increasing statistical power. Proteins with a fold 
change that differs by at least 1.2 fold (up or down regulation) and individual p-value of <0.05 
will be deemed to be significantly differentially abundant. 
Baseline responses of DA proteins will be visualized using heatmaps.  Higher order organization 
of DA proteins will be investigated using gene set enrichment analysis based on the latest 
MSigDB and KEGG databases using the Fisher’s exact test implementation provided by the 
goseq R package.  Pathway maps color-coded by treatment effect will be provided for 
significantly enriched KEGG pathways.  Experimentally determined protein-protein interaction 
networks including known Influenza A-host interactions centered around DA proteins will be 
generated using data from the IntAct database and visualized using Cytoscape.  Regularized 
canonical correlation (mixOmics R package), linear and logistic regression models (glmnet R 
package) based on baseline fold changes in LPR will be carried out to determine protein 
responses that are correlated with cytokine/chemokine concentrations, peak titer, or 
seroconversion status, respectively.  The CD-HIT software will be used to determine protein 
families within the protein sequence database.  This information will be integrated when 
presenting lists of DA proteins, heatmaps, and protein-interaction networks. 


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11.5.2 
Serologic Immunogenicity Data (HAI and Neut Antibody Assays) 
The percentage of subjects achieving seroconversion (defined as either a pre-vaccination titer 
<1:10 and a post-vaccination titer ≥1:40 or a pre-vaccination titer ≥1:10 and a minimum four-
fold rise in post-vaccination antibody titer) approximately 28 days following study vaccination 
will be summarized for each assay (HAI and Neut) and study arm using exact 95% confidence 
intervals.  Comparisons between study arms will be carried out using a Fisher’s exact test.  
Geometric Mean Titers (GMT) of serum HAI and Neut antibody and their 95% confidence 
intervals will be computed for Day -7 and 1 (baseline) and approximately 28 days following last 
study vaccination by transforming results to a logarithmic scale, assuming asymptotic normality 
conditions were satisfied on this scale, computing the mean and then converting back to the 
original scale.  Differences between the treatments will be evaluated for Day 29 using two-sided 
t-test adjusting for unequal variance if necessary. 
11.5.3 
Cytokine/Chemokines 
Cytokine/chemokine concentrations as well as their baseline fold changes will be summarized 
using minimum, Q1, median, Q3, maximum, and 95% CI of the median.  The 95% CI of the 
median concentration and median fold change from baseline will be determined by using the 
bootstrap method and visualized using time trend plots.  To identify cytokines/chemokines that 
show a differential response from baseline, a Wilcoxon signed-rank test will be carried out for 
each study arm and post-vaccination time point.  Significant differences in fold changes between 
study arms for shared post-vaccination time points will be evaluated using a Wilcoxon rank-sum 
test. 
11.5.4 
Primary Clinical Study Report (CSR) Database Lock 
Clinical, safety, and reactogenicity data through approximately 28 days after the last study 
vaccination will represent the primary clinical database for this study.  Once the last subject 
completes the visit that occurs approximately 28 days after the last study vaccination, the 
primary clinical database will be cleaned, monitored and locked.  Unblinded analyses of safety, 
reactogenicity, and available immunogenicity data may be performed by the SDCC after the 
primary clinical database is locked.  These analyses will be made available to the sponsor for 
planning subsequent studies and to the clinical investigators for publication.  These analyses will 
not be used to make any decisions concerning the conduct of this study.  As it is anticipated 
subjects will remain in the long-term, tertiary safety follow-up at the time of these analyses, the 
presentation of these data will be in a format that prevents inadvertent unblinding of the sponsor 
or clinical investigators to any individual subject’s treatment assignment or immune response 
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clinical investigators and sponsor, such as the presence of extreme values or noteworthy AEs, a 
presentation by treatment arm will not be provided until after the long-term, tertiary safety 
follow-up is complete. 
Serologic immunogenicity data from HAI and Neut antibody assays and processed cellular 
immunogenicity data from systems biology studies may be analyzed as results are available once 
the primary clinical database is locked.  These analyses will be considered the final results for 
this study and included as part of the clinical study report (CSR). 
The primary CSR will be completed when all primary and secondary endpoint data are available.  
Any available data from the long-term, tertiary safety follow-up of applicable subjects may also 
be included.  Additional data from the long-term, tertiary safety follow-up of applicable subjects, 
exploratory correlation analyses, and data from the proteomics portion of the study will be 
included in one or more addenda to the CSR. 
A formal statistical analysis plan will be developed and finalized prior to database lock. 
11.5.5 
Analysis Populations 
The Safety Analysis population includes all subjects who received at least one dose of study 
vaccine. 
The intent-to-treat (ITT) population includes all subjects who received at least one dose of study 
vaccine and contributed both pre- and at least one post-study vaccination blood samples for 
serological assessment (HAI or Neut antibody assays) for which valid results were reported. 
The per protocol (PP) population includes all subjects in the ITT subset with the following 
exclusions: 
• Data from all available visits for subjects found to be ineligible at baseline. 
• Data from all visits subsequent to major protocol deviations, such as: 
• Second study vaccination not received, 
• Receipt of non-study licensed live vaccine within 30 days before or after each 
study vaccination, 
• Receipt of non-study licensed inactivated vaccine within 14 days before or after 
each study vaccination, 
• Receipt of immunosuppressive therapy (e.g., corticosteroids) within 30 days 
before or after each study vaccination. 


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• Data from any visit that occurs substantially out of window. 
11.5.6 
Safety Data 
Summaries and analysis of safety data will be presented for the Safety Analysis Population. 
These include reactogenicity following each study vaccination, unsolicited adverse events, and 
SAEs/MAAEs (including new-onset chronic medical conditions and potentially immune-
mediated medical conditions.  These safety data will be analyzed, in an exploratory manner, in 
the context of transcriptomic/proteomic changes. 
11.5.7 
Missing Values and Outliers 
All attempts will be made to collect all data per protocol.  As missing data are expected to be 
minimal, no imputation will be performed for missing values.  Any data point that appears to be 
erroneous or inexplicable based on clinical judgment will be investigated as a possible outlier.  If 
data points are identified as outliers, sensitivity analyses will be performed to examine the 
impact of including or excluding the outliers.  Any substantive differences in these analyses will 
be reported. 


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12 
DATA COLLECTION FORMS AND ACCESS TO SOURCE 
DATA/DOCUMENTS 
The participating VTEU site will maintain appropriate medical and research records for this 
study, in compliance with ICH E6, Section 4.9, and regulatory and institutional requirements for 
the protection of confidentiality of subjects.  The participating VTEU site will permit authorized 
representatives of the DMID, its designees, and appropriate regulatory agencies to examine (and 
when required by applicable law, to copy) clinical study records for the purposes of quality 
assurance reviews, audits, monitoring and evaluation of the study safety and progress.  These 
representatives will be permitted access to all source data, which include, but are not limited to, 
hospital records, clinical and office charts, laboratory notes, memoranda, evaluation checklists, 
pharmacy dispensing records, recorded data from automated instruments, copies or transcriptions 
certified after verification as being accurate and complete, microfiches, photographic negatives, 
microfilm or magnetic media, x-rays, and subject files and records kept at the pharmacy, at the 
laboratories, and medico-technical departments involved in the study.  Source data are all 
information, original records of clinical findings, observations, or other activities in a clinical 
study necessary for the reconstruction and evaluation of the study. 
Interview of subjects is sufficient for obtaining medical history.  Solicitation of medical records 
from the subject’s primary care provider is not required unless needed. 


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13 
QUALITY CONTROL AND QUALITY ASSURANCE 
Following a written DMID-accepted site quality management plan, the participating VTEU site 
is responsible for conducting routine quality assurance (QA) and quality control (QC) activities 
to internally monitor study progress and protocol compliance.  The site principal investigators 
will provide direct access to all study-related sites, source data/data collection forms, and reports 
for the purpose of monitoring and auditing by the sponsor, and inspection by local and regulatory 
authorities.  The site principal investigators will ensure all study personnel are appropriately 
trained and applicable documentations are maintained on site. 
The SDCC will implement quality control procedures beginning with the data entry system and 
generate data quality control checks that will be run on the database.  Any missing data or data 
anomalies will be communicated to the participating VTEU site for clarification and resolution. 


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14 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 Ethical Standard 
The site principal investigators will ensure that this study is conducted in full conformity with 
the principles of the Belmont Report:  Ethical Principles and Guidelines for the Protection of 
Human Subjects of Research of the National Commission for the Protection of Human Subjects 
of Biomedical and Behavioral Research (April 18, 1979) and codified in 45 CFR 46, 21 CFR 50 
and 56, and ICH E6; 62 Federal Regulations 25691 (1997), if applicable.  The site principal 
investigators’ institution will hold a current Federalwide Assurance (FWA) issued by Office of 
Human Research Protection (OHRP) for federally funded research. 
14.2 Institutional Review Board 
Prior to enrollment of subjects into this study, the approved protocol and informed consent form 
will be reviewed and approved by the appropriate IRB listed on its FWA. 
The responsible official for the IRB will sign the IRB letter of approval of the protocol prior to 
the start of this study and a copy will be provided to DMID.  The IRB FWA number will be 
provided to DMID. 
Should amendments to the protocol be required, the amendments will be written by the sponsor 
and provided to the site principal investigator for submission to the IRB. 
14.3 Informed Consent Process 
14.3.1 
Informed Consent 
The site principal investigators will choose subjects in accordance with the eligibility criteria 
detailed in Section 5.1.  Before any study procedures are performed, subjects must sign an 
informed consent form that complies with the requirements of 21 CFR Part 50 and 45 CFR 46 
and the local IRB.  Study personnel may employ recruitment efforts prior to obtaining study 
consent if a patient-specific screening consent is on record or if the IRB has agreed that chart 
review is allowed without a fully executed screening consent.  In cases where there is not a 
patient-specific screening consent on record, site clinical staff may pre-screen via chart review 
and refer potential subjects to the research staff.  Research staff would obtain written consent per 
the standard informed consent process before conducting protocol-specific screening activities. 


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Informed consent is a process that is initiated prior to an individual agreeing to participate in a 
study and continuing throughout the individual’s study participation.  Before any study 
procedures are performed, including pre-screening of subjects for eligibility, subjects will 
receive a comprehensive explanation of the proposed study procedures and study 
interventions/products.  This will include the nature, risks and possible benefits of this study, 
alternate therapies, any known AEs, the investigational status of the study interventions/products, 
and the other elements that are part of obtaining proper informed consent.  Subjects will also 
receive a detailed explanation of the proposed use and disclosure of their protected health 
information, including specifically their serum and urine samples.  Subjects will be allowed 
sufficient time to consider participation in this research study, after having the nature, risks and 
possible benefits of this study explained to them, and have the opportunity to discuss this study 
with their family, friends or legally authorized representative or think about it prior to agreeing to 
participate. 
Informed consent forms describing in detail the study interventions/products, study procedures, 
risks and possible benefits will be given to subjects.  The informed consent form must not 
include any exculpatory statements.  Informed consent forms will be IRB-approved and subjects 
will be asked to read and review the appropriate document.  Upon reviewing the appropriate 
document, the site principal investigator (or designee) will explain this research study to subjects 
and answer any questions that may arise.  Subjects must sign the informed consent form, and 
written documentation of the informed consent process is required prior to starting any study 
procedures specifically for this study, including determining eligibility and administering study 
product. 
By signing the informed consent form, subjects agree to complete all study procedures required 
by this study, unless the subject withdraws voluntarily, or is withdrawn or terminated from this 
study for any reason.  The rights and welfare of subjects will be protected by emphasizing to 
subjects that the quality of their medical care will not be adversely affected if they decline to 
participate in or withdraw from this study. 
DMID will provide the site principal investigators, in writing, any new information that 
significantly impacts the subjects' risk of receiving the investigational products.  This new 
information will be communicated by the site principal investigators to subjects who consent to 
participate in this study in accordance with IRB requirements.  The informed consent document 
will be updated and subjects will be re-consented per IRB requirements, if necessary.  Subjects 
will be given a copy of all informed consent forms that they sign. 


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14.4 Exclusion of Women, Minorities, and Children (Special Populations) 
This study will be inclusive of all adults who meet the Subject Inclusion Criteria (see Section 
5.1.1) and do not meet the Subject Exclusion Criteria (see Section 5.1.2), regardless of religion, 
sex, or ethnic background.  Should the outcome of this study be deemed acceptable, additional 
studies may be initiated including those in other populations. 
It is unknown if the monovalent inactivated influenza A/H7N9 virus vaccine with or without 
AS03 adjuvant or unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine poses 
any risks to an unborn child.  As of November 22, 2015 (per the most current version of the 
manufacturer’s IB), the available data for women who become pregnant during clinical trials of 
AS03-adjuvanted (pre) pandemic influenza vaccines do not suggest any causal relationship 
between adverse pregnancy outcomes and receipt of an AS03-adjuvanted vaccine.  Women of 
childbearing potential who are not surgically sterile via tubal sterilization, bilateral 
oophorectomy or hysterectomy, or who are not postmenopausal for ≥1 year must agree to 
practice highly effective contraception that may include, but is not limited to, non-male sexual 
relationships, abstinence from sexual intercourse with a male partner, monogamous relationship 
with a vasectomized partner, male condoms that contain spermicide or with the use of applied 
spermicide, intrauterine devices, and licensed hormonal methods, with use of a highly effective 
method of contraception for a minimum of 30 days prior to study product exposure and agree to 
practice highly effective contraception for the duration of study product exposure, including 60 
days after their last study vaccination.  A highly effective method of contraception is defined as 
one that results in a low failure rate (i.e., less than 1% per year) when used consistently and 
correctly.  In addition to contraceptive use, all women of childbearing potential will be required 
to have a negative urine or serum pregnancy test within 24 hours prior to each study vaccination.  
If a female subject becomes pregnant while participating in this study, we will ask her 
permission to follow-up with her about her health and the health of her baby through pregnancy 
outcome. 
Children will not be included in this study. 
14.5 Subject Confidentiality 
Subject confidentiality is strictly held in trust by the site principal investigators, other study 
personnel, the sponsor, and their agents.  This confidentiality is extended to cover testing of 
biological samples, in addition to the clinical information relating to participating subjects.  
Subjects will have code numbers and will not be identified by name. 


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The study protocol, documentation, data, and all other information generated will be held in 
strict confidence.  No information concerning this study or the data will be released to any 
unauthorized third party without prior written approval of the sponsor. 
All information provided by the sponsor and all data and information generated by the 
participating VTEU site as part of this study (other than a subject’s medical records) will be kept 
confidential by the site principal investigators and other study personnel to the extent permitted 
by law.  This information and data will not be used by the site principal investigator or other 
study personnel for any purpose other than conducting this study.  These restrictions do not apply 
to: (1) information which becomes publicly available through no fault of the site principal 
investigator or other study personnel; (2) information which is necessary to disclose in 
confidence to an IRB solely for the evaluation of this study; (3) information which is necessary 
to disclose in order to provide appropriate medical care to a study subject; or (4) study results 
which may be published as described in Section 16. 
The study monitor, applicable regulatory authorities, such as the FDA, or other authorized 
representatives of the sponsor may inspect all documents and records required to be maintained 
by the site principal investigators.  This includes, but is not limited to, medical records (office, 
clinic, or hospital) and pharmacy records for the subjects in this study.  The participating VTEU 
site will permit access to such records. 
14.6 Study Discontinuation 
If this study is discontinued, subjects who have signed the informed consent form and are 
randomized and vaccinated will continue to be followed for safety for the duration of this study.  
No further study vaccinations will be administered. 
14.7 Costs, Subject Compensation, and Research Related Injuries 
There is no cost to subjects for taking part in this study. 
Subjects may be compensated for their participation in this study.  Compensation will be in 
accordance with the local IRB’s policies and procedures, and subject to IRB approval. 
If it is determined by the participating VTEU site and the site principal investigators that an 
injury occurred to a subject as a direct result of the tests or treatments that are done for this study, 
then referrals to appropriate health care facilities will be provided to the subject.  Study 
personnel will try to reduce, control, and treat any complications from this study.  Immediate 
medical treatment may be provided by the participating VTEU site, such as giving emergency 
medications to stop immediate allergic reactions to the study vaccines.  No financial 


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compensation will be provided to the subject by the participating VTEU site for any injury 
suffered due to participation in this study. 
14.8 Future Use of Stored Specimens 
Subjects will be asked for permission to keep any remaining serum from venous blood samples 
for possible use in future research studies, such as examining additional immunological 
assessments or testing for antibodies against other viruses or bacteria.  Urine samples 
(approximately 20 mL) will also be collected from subjects for future research. 
Samples will be stored indefinitely at a central clinical storage facility and may be shared with 
investigators at the participating VTEU site and with other investigators at other institutions.  
The samples will not be sold or used directly for production of any commercial product.  No 
human genetic tests will be performed on the samples.  Each sample will be encoded (labeled) 
only with a barcode and a unique tracking number to protect subject confidentiality. 
There are no benefits to subjects in the collection, storage and subsequent use of their 
specimens for future research.  Reports about future research done with subjects’ samples will 
NOT be kept in their health records. 
Subjects may be given the option to decide if they want their samples to be used for future 
research or have their samples destroyed at the end of this study.  The subject’s decision can be 
changed at any time by notifying the study doctors or nurses in writing; however, if a subject 
originally consents to future use and subsequently changes his/her decision, any data from a 
previously collected sample may still be used for this research. 


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15 
DATA HANDLING AND RECORD KEEPING 
The site principal investigator is responsible to ensure the accuracy, completeness, legibility, and 
timeliness of the data reported. 
Data collection forms will be derived from the eCRF and provided by the SDCC to record and 
maintain data for each subject enrolled in this study.  All data collection forms should be 
completed in a neat, legible manner to ensure accurate interpretation of data.  Black or blue ink is 
required to ensure clarity of reproduced copies.  When making a change or correction, cross out 
the original entry with a single line and initial and date the change.  Do not erase, overwrite, or 
use correction fluid or tape on the original. 
Data reported in the eCRF derived from the data collection forms should be consistent with the 
data collection forms or the discrepancies should be explained. 
The sponsor and/or its designee will provide guidance to the site principal investigators and other 
study personnel on making corrections to the data collection forms and eCRF. 
15.1 Data Management Responsibilities 
All data collection forms and laboratory reports must be reviewed by the clinical team and data 
entry personnel, who will ensure that they are accurate and complete.  Adverse events must be 
recorded on the appropriate data collection form, assessed for severity and relationship, and 
reviewed by the site principal investigator or appropriate sub-investigator. 
Data collection is the responsibility of the study personnel at the participating VTEU site under 
the supervision of the respective site principal investigators.  During this study, the site principal 
investigators must maintain complete and accurate documentation for this study. 
The SDCC for this study will be responsible for data management, quality review, analysis, and 
reporting of the study data. 
15.2 Data Capture Methods 
Clinical (including, but not limited to, AE/SAEs, concomitant medications, medical history, and 
physical assessments), reactogenicity, serologic immunogenicity (HAI and Neut antibody 
assays), and processed cellular immunogenicity (systems biology studies) data will be entered 
into a 21 CFR 11-compliant Internet Data Entry System provided by the SDCC.  The data 
system includes password protection and internal quality checks, such as automatic range checks, 


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to identify data that appear inconsistent, incomplete, or inaccurate.  Clinical and reactogenicity 
data will be entered directly from the data collection forms completed by the study personnel. 
15.3 Types of Data 
Data for this study will include clinical, safety, and outcome measures (e.g., reactogenicity, 
serologic immunogenicity (HAI and Neut antibody assays), and processed cellular 
immunogenicity (systems biology studies) data). 
15.4 Timing/Reports 
Clinical, safety, and reactogenicity data through approximately 28 days after the last study 
vaccination will represent the primary clinical database for this study.  Once the last subject 
completes the visit that occurs approximately 28 days after the last study vaccination, the 
primary clinical database will be cleaned, monitored and locked.  Unblinded analyses of safety, 
reactogenicity, and available immunogenicity data may be performed by the SDCC after the 
primary clinical database is locked.  These analyses will be made available to the sponsor for 
planning subsequent studies and to the clinical investigators for publication.  These analyses will 
not be used to make any decisions concerning the conduct of this study.  As it is anticipated 
subjects will remain in the long-term, tertiary safety follow-up at the time of these analyses, the 
presentation of these data will be in a format that prevents inadvertent unblinding of the sponsor 
or clinical investigators to any individual subject’s treatment assignment or immune response 
data.  When it is not possible to display the data by treatment arm and maintain the blind for the 
clinical investigators and sponsor, such as the presence of extreme values or noteworthy AEs, a 
presentation by treatment arm will not be provided until after the long-term, tertiary safety 
follow-up is complete. 
Serologic immunogenicity data from HAI and Neut antibody assays and processed cellular 
immunogenicity data from systems biology studies may be analyzed as results are available once 
the primary clinical database is locked.  These analyses will be considered the final results for 
this study and included as part of the clinical study report (CSR). 
The primary CSR will be completed when all primary and secondary endpoint data are available.  
Any available data from the long-term, tertiary safety follow-up of applicable subjects may also 
be included.  Additional data from the long-term, tertiary safety follow-up of applicable subjects, 
exploratory correlation analyses, and data from the proteomics portion of the study will be 
included in one or more addenda to the CSR. 


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Interim statistical reports may be generated as deemed necessary and appropriate by DMID.  
Safety summary reports may be generated for the SMC. 
After full analysis and final reporting is complete, and upon request and DMID approval, the 
SDCC will provide the participating VTEU site with a summary of results by treatment arm 
and/or subject treatment assignments.  In this regard, the participating VTEU site requesting such 
information to share with study subjects must do so in compliance with their respective IRB 
guidelines. 
15.5 Study Records Retention 
Study records and reports, including, but not limited to, eCRFs, source documents, informed 
consent forms, and study drug disposition records, shall be maintained for 2 years after a 
marketing application is approved for the drug for the indication for which it is being 
investigated; or, if no application is to be filed or if the application is not approved for the drug, 
until 2 years after the investigation is discontinued and FDA has been so notified.  The 
participating VTEU site must contact DMID for authorization prior to the destruction of any 
study records.  Informed consent forms for future use will be maintained as long as the sample 
exists. 
15.6 Protocol Deviations 
A protocol deviation is any noncompliance with the study protocol, GCP or protocol-specific 
MOP requirements.  The noncompliance may be either on the part of the subject, the site 
principal investigators, or other study personnel.  As a result of deviations, corrective actions are 
to be developed by the site and implemented promptly. 
These practices are consistent with ICH E6: 
 
4.5 
Compliance with Protocol, Sections 4.5.1, 4.5.2, and 4.5.3 
5.1 
Quality Assurance and Quality Control, Section 5.1.1 
5.20 
Noncompliance, Sections 5.20.1, and 5.20.2. 
It is the responsibility of the site principal investigators and other study personnel to use 
continuous vigilance to identify and report deviations within five working days of identification 
of the protocol deviation, or within five working days of the scheduled protocol-required 
activity.  All deviations must be promptly reported to DMID, via the SDCC’s AdvantageEDCSM. 


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All protocol deviations, as defined above, must be addressed in study subject data collection 
forms.  A completed copy of the DMID Protocol Deviation Form must be maintained in the 
Regulatory File, as well as in the subject's chart.  Protocol deviations must be sent to the local 
IRB/IEC per its guidelines.  The site principal investigator and other study personnel are 
responsible for knowing and adhering to their IRB requirements. 


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16 
PUBLICATION POLICY 
All investigators funded by the NIH must submit or have submitted for them to the National 
Library of Medicine’s PubMed Central (http://www.ncbi.nlm.nih.gov/pmc/) an electronic 
version of their final, peer-reviewed manuscripts upon acceptance for publication, to be made 
publicly available no later than 12 months after the official date of publication.  The NIH Public 
Access Policy ensures the public has access to the published results of NIH funded research.  It 
requires investigators to submit final peer-reviewed journal manuscripts that arise from NIH 
funds to the digital archive PubMed Central upon acceptance for publication.  Further, the policy 
stipulates that these papers must be accessible to the public on PubMed Central no later than 12 
months after publication. 
Refer to: 
• 
NIH Public Access Policy, http://publicaccess.nih.gov/ 
• 
NIH Office of Extramural Research (OER) Grants and Funding, 
http://grants.nih.gov/grants/oer.htm 
Following completion of this clinical trial, the lead principal investigator is expected to publish 
the results of this research in a scientific journal.  The International Committee of Medical 
Journal Editors (ICMJE) member journals have adopted a trials-registration policy as a condition 
for publication.  This policy requires that all clinical trials be registered in a public trials registry 
such as ClinicalTrials.gov* (http://clinicaltrials.gov/), which is sponsored by the National 
Library of Medicine.  Other biomedical journals are considering adopting similar policies. 
The ICMJE defines a clinical trial as any research project that prospectively assigns human 
subjects to intervention or comparison groups to study the cause-and-effect relationship between 
a medical intervention and a health outcome.  Studies designed for other purposes, such as to 
study pharmacokinetics or major toxicity (e.g., Phase I trials), would be exempt from this policy. 
It is the responsibility of DMID to register this clinical trial in an acceptable registry.  Any 
clinical trial starting enrollment after 01 July 2005 must be registered on or before subject 
enrollment.  For clinical trials that began enrollment prior to this date, the ICMJE member 
journals will require registration by 13 September 2005, before considering the results of the 
clinical trial for publication. 
For clinical trials in which DMID is not the IND/IDE sponsor, or there is no IND/IDE, and 
DMID does not provide data management services, it is the responsibility of the investigator to 
register the clinical trial and post results in compliance with Public Law 110-85, the Food and 
Drug Administration Amendments Act of 2007 (FDAAA). 


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Refer to: 
• 
Public Law 110-85, Section 801, Clinical Trial Databases 
*Journal Citation:  De Angelis C, Drazen JM, Frizelle FA, Haug C, Hoey J, Horton R, et al. 
Clinical trial registration: a statement from the International Committee of Medical Journal 
Editors. N Engl J Med. 2004; 351:1250-1. 


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17 
LITERATURE REFERENCES 
1. 
Zhang W, Wang L, Hu W, et al. Epidemiologic characteristics of cases for influenza 
A(H7N9) virus infections in China. Clin Infect Dis. 2013;57(4):619-620. 
2. 
Zhou H, Zheng Y, Wang L, et al. First serologic study for influenza A (H7N9) virus 
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Xie L, Ding H, Kao QJ, et al. Clinical and epidemiological survey and analysis of the 
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7. 
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Treanor JJ, Campbell JD, Zangwill KM, Rowe T, Wolff M. Safety and immunogenicity 
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of pandemic influenza A (H1N1) 2009 vaccine: systematic review and meta-analysis. 
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Durando P, Iudici R, Alicino C, et al. Adjuvants and alternative routes of administration 
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Atmar RL, Keitel WA. Adjuvants for pandemic influenza vaccines. Current topics in 
microbiology and immunology. 2009;333:323-344. 
15. 
Godeaux O, Izurieta P, Madariaga M, Drame M, Li P, Vaughn DW. Immunogenicity and 
safety of AS03A-adjuvanted H5N1 influenza vaccine prepared from bulk antigen after 
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Risi G, Frenette L, Langley JM, et al. Immunological priming induced by a two-dose 
series of H5N1 influenza antigen, administered alone or in combination with two 
different formulations of AS03 adjuvant in adults: Results of a randomised single 
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Yang PC, Yu CJ, Chang SC, et al. Safety and immunogenicity of a split-virion AS03A-
adjuvanted A/Indonesia/05/2005 (H5N1) vaccine in Taiwanese adults. Journal of the 
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18. 
Langley JM, Risi G, Caldwell M, et al. Dose-sparing H5N1 A/Indonesia/05/2005 pre-
pandemic influenza vaccine in adults and elderly adults: a phase III, placebo-controlled, 
randomized study. J Infect Dis. 2011;203(12):1729-1738. 
19. 
Moris P, van der Most R, Leroux-Roels I, et al. H5N1 influenza vaccine formulated with 
AS03 A induces strong cross-reactive and polyfunctional CD4 T-cell responses. Journal 
of clinical immunology. 2011;31(3):443-454. 
20. 
Nicholson KG, Abrams KR, Batham S, et al. Immunogenicity and safety of a two-dose 
schedule of whole-virion and AS03A-adjuvanted 2009 influenza A (H1N1) vaccines: a 
randomised, multicentre, age-stratified, head-to-head trial. Lancet Infect Dis. 
2011;11(2):91-101. 
21. 
Diez-Domingo J, Garces-Sanchez M, Baldo JM, et al. Immunogenicity and Safety of 
H5N1 A/Vietnam/1194/2004 (Clade 1) AS03-adjuvanted prepandemic candidate 
influenza vaccines in children aged 3 to 9 years: a phase ii, randomized, open, controlled 
study. Pediatr Infect Dis J. 2010;29(6):e35-46. 
22. 
Caproni E, Tritto E, Cortese M, et al. MF59 and Pam3CSK4 boost adaptive responses to 
influenza subunit vaccine through an IFN type I-independent mechanism of action. J 
Immunol. 2012;188(7):3088-3098. 
23. 
Seubert A, Monaci E, Pizza M, O'Hagan DT, Wack A. The adjuvants aluminum 
hydroxide and MF59 induce monocyte and granulocyte chemoattractants and enhance 
monocyte differentiation toward dendritic cells. J Immunol. 2008;180(8):5402-5412. 
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Morel S, Didierlaurent A, Bourguignon P, et al. Adjuvant System AS03 containing alpha-
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via modulation of Nrf2 the expression and turnover of hypocretin in vitro and its 
implication to the development of narcolepsy. Vaccine. 2014;32(25):2980-2988. 
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Ahmed SS, Schur PH, MacDonald NE, Steinman L. Narcolepsy, 2009 A(H1N1) 
pandemic influenza, and pandemic influenza vaccinations: what is known and unknown 
about the neurological disorder, the role for autoimmunity, and vaccine adjuvants. 
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29. 
van der Most R, Van Mechelen M, Destexhe E, Wettendorff M, Hanon E. Narcolepsy 
and A(H1N1)pdm09 vaccination: shaping the research on the observed signal. Hum 
Vaccin Immunother. 2014;10(3):572-576. 
30. 
Dauvilliers Y, Arnulf I, Lecendreux M, et al. Increased risk of narcolepsy in children and 
adults after pandemic H1N1 vaccination in France. Brain : a journal of neurology. 
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during, and after the influenza A(H1N1)pdm09 pandemic and vaccination campaigns. 
Vaccine. 2013;31(8):1246-1254. 
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Nohynek H, Jokinen J, Partinen M, et al. AS03 adjuvanted AH1N1 vaccine associated 
with an abrupt increase in the incidence of childhood narcolepsy in Finland. PLoS One. 
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Nishino S, Okuro M, Kotorii N, et al. Hypocretin/orexin and narcolepsy: new basic and 
clinical insights. Acta physiologica (Oxford, England). 2010;198(3):209-222. 
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Ahmed SS, Volkmuth W, Duca J, et al. Antibodies to influenza nucleoprotein cross-react 
with human hypocretin receptor 2. Sci Transl Med. 2015;7(294):294ra105. 
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virology. 1975;47(2):157-166. 
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Palache AM, Beyer WE, Sprenger MJ, et al. Antibody response after influenza 
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Keitel WA, Jackson LA, Edupuganti S, et al. Safety and Immunogenicity of a Subvirion 
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Lasky T, Terracciano GJ, Magder L, et al. The Guillain-Barre syndrome and the 1992-
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Haber P, DeStefano F, Angulo FJ, et al. Guillain-Barre syndrome following influenza 
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De Wals P, Deceuninck G, Toth E, et al. Risk of Guillain-Barre syndrome following 
H1N1 influenza vaccination in Quebec. Jama. 2012;308(2):175-181. 
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Wise ME, Viray M, Sejvar JJ, et al. Guillain-Barre syndrome during the 2009-2010 
H1N1 influenza vaccination campaign: population-based surveillance among 45 million 
Americans. Am J Epidemiol. 2012;175(11):1110-1119. 
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Juurlink DN, Stukel TA, Kwong J, et al. Guillain-Barre syndrome after influenza 
vaccination in adults: a population-based study. Arch Intern Med. 2006;166(20):2217-
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47. 
Salmon DA, Proschan M, Forshee R, et al. Association between Guillain-Barre syndrome 
and influenza A (H1N1) 2009 monovalent inactivated vaccines in the USA: a meta-
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48. 
Polakowski LL, Sandhu SK, Martin DB, et al. Chart-confirmed guillain-barre syndrome 
after 2009 H1N1 influenza vaccination among the Medicare population, 2009-2010. Am J 
Epidemiol. 2013;178(6):962-973. 
49. 
Dodd CN, Romio SA, Black S, et al. International collaboration to assess the risk of 
Guillain Barre Syndrome following Influenza A (H1N1) 2009 monovalent vaccines. 
Vaccine. 2013;31(40):4448-4458. 
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Bioinformatics (Oxford, England). 2009;25(9):1105-1111. 
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Wang H, Qian WJ, Mottaz HM, et al. Development and evaluation of a micro- and 
nanoscale proteomic sample preparation method. Journal of proteome research. 
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Di Silvestre D, Zoppis I, Brambilla F, Bellettato V, Mauri G, Mauri P. Availability of 
MudPIT data for classification of biological samples. Journal of clinical bioinformatics. 
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Ross PL, Huang YN, Marchese JN, et al. Multiplexed protein quantitation in 
Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Molecular & 
cellular proteomics : MCP. 2004;3(12):1154-1169. 
 


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APPENDICES 
APPENDIX A: LIST OF POTENTIALLY IMMUNE-MEDIATED MEDICAL CONDITIONS 
APPENDIX B: SCHEDULE OF EVENTS 
 
 


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APPENDIX A: LIST OF POTENTIALLY IMMUNE-MEDIATED 
MEDICAL CONDITIONS 
(also known as Adverse Events of Special Interest (AESIs)) 
Gastrointestinal disorders 
• Celiac disease 
• Crohn’s disease 
• Ulcerative colitis 
• Ulcerative proctitis 
 
Liver disorders 
• Autoimmune cholangitis 
• Autoimmune hepatitis 
• Primary biliary cirrhosis 
• Primary sclerosing cholangitis 
 
Metabolic diseases 
• Addison’s disease 
• Autoimmune thyroiditis (including Hashimoto thyroiditis) 
• Diabetes mellitus type I 
• Grave's or Basedow’s disease 
 
Musculoskeletal disorders 
• Antisynthetase syndrome 
• Dermatomyositis 
• Juvenile chronic arthritis (including Still’s disease) 
• Mixed connective tissue disorder 
• Polymyalgia rheumatic 
• Polymyositis 
• Psoriatic arthropathy 
• Relapsing polychondritis 
• Rheumatoid arthritis 
• Scleroderma, including diffuse systemic form and CREST syndrome 
• Spondyloarthritis, including ankylosing spondylitis, reactive arthritis (Reiter's Syndrome) and 
undifferentiated spondyloarthritis 
• Systemic lupus erythematosus 
• Systemic sclerosis 
 
Neuroinflammatory disorders 
• Acute disseminated encephalomyelitis, including site specific variants (e.g., non-infectious 
encephalitis, encephalomyelitis,myelitis, myeloradiculomyelitis) 
• Cranial nerve disorders, including paralyses/paresis (e.g., Bell’s palsy) 


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• Guillain-Barré syndrome, including Miller Fisher syndrome and other variants 
• Immune-mediated peripheral neuropathies and plexopathies, including chronic inflammatory 
demyelinating polyneuropathy, multifocal motor neuropathy and polyneuropathies associated 
with monoclonal gammopathy 
• Multiple sclerosis 
• Narcolepsy 
• Optic neuritis 
• Transverse myelitis 
• Myasthenia gravis, including Eaton-Lambert syndrome 
 
Skin disorders 
• Alopecia areata 
• Autoimmune bullous skin diseases, including pemphigus, pemphigoid and dermatitis 
herpetiformis 
• Cutaneous lupus erythematosus 
• Erythema nodosum 
• Morphoea 
• Lichen planus 
• Psoriasis 
• Sweet’s syndrome 
• Vitiligo 
 
Vasculitides 
• Large vessels vasculitis including: giant cell arteritis such as Takayasu's arteritis and temporal 
arteritis 
• Medium sized and/or small vessels vasculitis including: polyarteritis nodosa, Kawasaki's 
disease, microscopic polyangiitis,Wegener's granulomatosis, Churg–Strauss syndrome (allergic 
granulomatous angiitis), Buerger’s disease thromboangiitis obliterans, necrotizing vasculitis and 
anti-neutrophil cytoplasmic antibody (ANCA) positive vasculitis (type unspecified), Henoch- 
Schonlein purpura, Behcet's syndrome, leukocytoclastic vasculitis 
 
Others 
• Antiphospholipid syndrome 
• Autoimmune hemolytic anemia 
• Autoimmune glomerulonephritis (including IgA nephropathy, glomerulonephritis rapidly 
progressive, membranous glomerulonephritis, membranoproliferative glomerulonephritis, and 
mesangioproliferative glomerulonephritis) 
• Autoimmune myocarditis/cardiomyopathy 
• Autoimmune thrombocytopenia 
• Goodpasture syndrome 
• Idiopathic pulmonary fibrosis 
• Pernicious anemia 
• Raynaud’s phenomenon 
• Sarcoidosis 


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• Sjögren’s syndrome 
• Stevens-Johnson syndrome 
• Uveitis 
 








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@ Complete medical history will be obtained by interview of the subjects at the screening visit and will be updated on Day 1 prior to the first study vaccination and 
interim medical history will be obtained by interview of the subjects at follow-up visits after the first study vaccination. 
! Including solicitation through Visit 09 for receipt of non-study influenza vaccines. 
√ All current medications and medications taken within 60 days prior to signing the informed consent form. Medications reported in the eCRF are limited to those 
taken within 30 days prior to the first study vaccination. 
$ Vital signs assessed on Day 1 prior to the first study vaccination will be considered as baseline. 
% Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral temperature. 
() Targeted physical examination if indicated based on review of complete or interim medical history. 
^ Will be performed on all women of childbearing potential at screening and within 24 hours prior to each study vaccination and results must be negative and known 
prior to each study vaccination. 
1 Including cytokine and chemokine levels. 
~ A/H7N9-vaccinated subjects only. 
& Inclusive of reactogenicity assessments performed on the day of each study vaccination through 8 days after each study vaccination. 
# Assessment of AE/SAE limited to SAEs and MAAEs including new-onset chronic medical conditions and potentially immune-mediated medical conditions. 
** Assessment of AE/SAE limited to SAEs and MAAEs including new-onset chronic medical conditions and potentially immune-mediated medical conditionsif after 28 days 
following the last study vaccination. 
2 To be performed locally by the site. The ESR value must be confirmed as <30 mm/hr prior to randomization and first study vaccination. 
3 A/H3N2v-vaccinated subjects only. 
Ω Visit 04 is an in-clinic assessment for A/H3N2v-vaccinated subjects and a phone call/electronic communication (e.g., email, text message) assessment for A/H7N9-vaccinated 
subjects. 
 

SAP_001.pdf: 
CLINICAL RESEARCH IN INFECTIOUS DISEASES 
 
 
 
STATISTICAL ANALYSIS PLAN 
for 
DMID Protocol: 14-0015 
Study Title: 
 
A Phase II Study to Evaluate and Compare the Immunogenicity of Monovalent 
Inactivated Influenza A/H7N9 Virus Vaccine Administered with and without AS03 
Adjuvant and Monovalent Inactivated Influenza A/H3N2v Virus Vaccine 
Administered without Adjuvant in Healthy Adults through Standard and Systems 
Biology Analyses 
 
NCT02921997 
 
Version 1.0 
 
DATE: 21 NOVEMBER 2019 
THIS COMMUNICATION IS PRIVILEGED AND CONFIDENTIAL  
 


Statistical Analysis Plan - DMID Protocol:14-0015  
Version 1.0 
21 November 2019 
- ii - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
STUDY TITLE 
Protocol Number Code: 
DMID Protocol: 14-0015 
Development Phase: 
Phase II 
Products: 
Monovalent Inactivated Influenza A/Shanghai/02/2013 (H7N9) Virus 
Vaccine Administered with and without AS03 Adjuvant 
 
Monovalent Inactivated Influenza A/Minnesota/11/2010 (H3N2)v Virus 
Vaccine Administered without Adjuvant 
Form/Route: 
Intramuscular 
Indication Studied: 
Influenza 
Sponsor: 
Division of Microbiology and Infectious Diseases 
National Institute of Allergy and Infectious Diseases 
National Institutes of Health 
Clinical Trial Initiation Date: 
01 November 2016 
Clinical Trial Completion Date: 
14 February 2018 
Date of the Analysis Plan: 
21 November 2019 
Version Number: 
1.0 
This study was performed in compliance with Good Clinical Practice. 
Information contained in this publication is the property of Division of Microbiology and Infectious Diseases and is confidential. This 
information may not be disclosed to third parties without written authorization from Division of Microbiology and Infectious Diseases. This 
report may not be reproduced, stored in a retrieval system or transmitted in any form or by any means - electronic, mechanical, recording or 
otherwise - without the prior authorization from Division of Microbiology and Infectious Diseases. This document must be returned to Division 
of Microbiology and Infectious Diseases upon request. 
 


Statistical Analysis Plan - DMID Protocol:14-0015  
Version 1.0 
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TABLE OF CONTENTS 
STUDY TITLE ............................................................................................................................... II 
TABLE OF CONTENTS ...............................................................................................................III 
LIST OF ABBREVIATIONS ....................................................................................................... VI 
1. 
PREFACE .....................................................................................................................1 
2. 
INTRODUCTION ........................................................................................................2 
2.1. 
Purpose of the Analyses ................................................................................................2 
3. 
STUDY OBJECTIVES AND ENDPOINTS ................................................................3 
3.1. 
Study Objectives ...........................................................................................................3 
3.2. 
Endpoints ......................................................................................................................4 
3.3. 
Study Definitions and Derived Variables .....................................................................6 
4. 
INVESTIGATIONAL PLAN .......................................................................................7 
4.1. 
Overall Study Design and Plan .....................................................................................7 
4.2. 
Discussion of Study Design, Including the Choice of Control Groups ........................8 
4.3. 
Selection of Study Population ......................................................................................8 
4.4. 
Treatments ..................................................................................................................11 
4.4.1. 
Treatments Administered ............................................................................................11 
4.4.2. 
Identity of Investigational Product(s) .........................................................................12 
4.4.3. 
Method of Assigning Subjects to Treatment Groups (Randomization) .....................12 
4.4.4. 
Selection of Doses in the Study ..................................................................................13 
4.4.5. 
Selection and Timing of Dose for Each Subject .........................................................13 
4.4.6. 
Blinding ......................................................................................................................13 
4.4.7. 
Prior and Concomitant Therapy ..................................................................................13 
4.4.8. 
Treatment Compliance ................................................................................................14 
4.5. 
Immunogenicity and Safety Variables ........................................................................14 
4.5.1. 
Immunogenicity Variables ..........................................................................................14 
4.5.2. 
-Omics Variables ........................................................................................................14 
4.5.3. 
Safety Variables ..........................................................................................................15 
5. 
SAMPLE SIZE CONSIDERATIONS .......................................................................16 
6. 
GENERAL STATISTICAL CONSIDERATIONS ....................................................17 
6.1. 
General Principles .......................................................................................................17 


Statistical Analysis Plan - DMID Protocol:14-0015  
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6.2. 
Timing of Analyses .....................................................................................................17 
6.3. 
Analysis Populations ..................................................................................................17 
6.3.1. 
Safety Analysis Population .........................................................................................17 
6.3.2. 
Intention-to-Treat Analysis (ITT) Population ............................................................17 
6.3.3. 
Per protocol (PP) Population ......................................................................................17 
6.3.4. 
The -Omics Analysis Population ................................................................................18 
6.4. 
Covariates and Subgroups ..........................................................................................18 
6.5. 
Missing Data ...............................................................................................................18 
6.6. 
Interim Analyses and Data Monitoring ......................................................................18 
6.7. 
Multicenter Studies .....................................................................................................18 
6.8. 
Multiple Comparisons/Multiplicity ............................................................................18 
7. 
STUDY SUBJECTS ...................................................................................................19 
7.1. 
Disposition of Subjects ...............................................................................................19 
7.2. 
Protocol Deviations ....................................................................................................19 
8. 
IMMUNOGENICITY EVALUATION .....................................................................20 
8.1. 
Serum Antibody Analysis ...........................................................................................20 
8.2. 
Plasma Cytokines/Chemokine Analysis .....................................................................20 
9. 
TRANSCRIPTOMICS EVALUATIONS ..................................................................21 
9.1. 
RNA-Seq Data Processing and Qc .............................................................................21 
9.1.1. 
Human Reference Genome Alignment and Quality Assessment ...............................21 
9.1.2. 
Gene Expression Quantification and Data Normalization ..........................................21 
9.1.3. 
Global Outlier and Batch Effect Detection .................................................................21 
9.1.4. 
Pre-Vaccination Transcriptomics Activity .................................................................22 
9.1.5. 
Log Fold Change Calculations ...................................................................................22 
9.1.6. 
Determination of Cut Offs for Filtering Lowly Expressed Genes ..............................22 
9.2. 
Identification of Differentially Expressed Genes .......................................................22 
9.2.1. 
Identification of DE Genes Compared to Pre-First Vaccination ................................23 
9.2.2. 
Identification of DE Genes Compared to Pre-Second Vaccination ............................23 
9.2.3. 
Identification of Genes with Differential Responses Between Study Arms ...............23 
9.3. 
Determination of higher order functional and temporal organization of 
differentially expressed genes .....................................................................................24 
9.3.1. 
Subject-level Response Heatmaps ..............................................................................24 


Statistical Analysis Plan - DMID Protocol:14-0015  
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9.3.2. 
Identification of Co-Expressed Gene Clusters ...........................................................24 
9.3.3. 
Pathway Enrichment Analysis ....................................................................................24 
9.3.4. 
Protein-Protein Interactions Network Analysis ..........................................................25 
10. 
SAFETY EVALUATIONS ........................................................................................26 
10.1. 
Demographic and Other Baseline Characteristics ......................................................26 
10.2. 
Adverse Events ...........................................................................................................26 
10.2.1. 
Solicited Events and Symptoms .................................................................................26 
10.2.2. 
Unsolicited Adverse Events ........................................................................................26 
10.3. 
Deaths, Serious Adverse Events and other Significant Adverse Events ....................27 
10.4. 
Vital Signs and Physical Evaluations .........................................................................27 
11. 
TERTIARY AND EXPLORATORY PROTEOMICS ANALYSES ........................28 
12. 
EXPLORATORY CORRELATION ANALYSES ....................................................29 
13. 
REPORTING CONVENTIONS ................................................................................30 
14. 
REFERENCES ...........................................................................................................31 
15. 
LISTING OF TABLES, FIGURES, AND LISTINGS ..............................................32 
APPENDICES ...............................................................................................................................33 
APPENDIX 1. TABLE MOCK-UPS..........................................................................................34 
APPENDIX 2. FIGURE MOCK-UPS ........................................................................................92 
APPENDIX 3. LISTINGS MOCK-UPS ...................................................................................107 
 


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LIST OF ABBREVIATIONS 
A/H3N2v 
Influenza A Virus of the H3N2 Variant Subtype 
A/H5N1 
Influenza A Virus of the H5N1 Subtype 
A/H7N9 
Influenza A Virus of the H7N9 Subtype 
AdvantageEDCSM 
Electronic Data Capture System 
AE 
Adverse Event/Adverse Experience 
AS03 
Adjuvant System (03) 
α 
Alpha 
BMI 
Body Mass Index 
BPM 
Beats Per Minute 
CDC 
Centers for Disease Control and Prevention 
CDF 
Cumulative Distribution Function 
CI 
Confidence Interval 
CSR 
Clinical Study Report 
°C 
Degrees Celsius 
°F 
Degrees Fahrenheit 
D 
Day(s) 
DE 
Differentially Expressed 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH 
eCRF 
Electronic Case Report Form 
ESR 
Erythrocyte Sedimentation Rate 
FDA 
Food and Drug Administration 
FDR 
False-discovery Rate 
GCP 
Good Clinical Practice 
GMT 
Geometric Mean Titer 
GTF 
Gene Transfer Format 
HA 
Hemagglutinin 
HAI 
Hemagglutination Inhibition 
HIV 
Human Immunodeficiency Virus 
ICH 
International Conference on Harmonisation 
IFN 
Interferon 
IL 
Interleukin 
IM 
Intramuscular 
IQR 
Interquartile Range 
ITT 
Intent-to-Treat 
LCPM 
Log2 Counts per Million 
LFC 
Log2 Fold Change 
LOD 
Limit of Detection 
MAAEs 
Medically-Attended Adverse Events 
mcg or µg 
Microgram(s) 
MedDRA 
Medical Dictionary for Regulatory Activities 


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mL 
Milliliter(s) 
mm 
Millimeter(s) 
mmHg 
Millimeters of Mercury 
MOP 
Manual of Procedures 
N 
Number of Subjects 
Neut or NT 
Neutralizing or Neutralization 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, DHHS 
NK 
Natural Killer 
NIH 
National Institutes of Health 
NOCMCs 
New-Onset Chronic Medical Conditions 
PBS 
Phosphate-Buffered Saline 
PIMMCs 
Potentially Immune-Mediated Medical Conditions 
PP 
Per Protocol 
QC 
Quality Control 
RCDF 
Reverse Cumulative Distribution Function 
RNA 
Ribonucleic Acid 
RP-HPLC 
Reversed-Phase High-Performance Liquid Chromatography 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SAP 
Statistical Analysis Plan 
SDCC 
Statistical and Data Coordinating Center 
SOC 
System Organ Class 
SRID 
Single Radial Immunodiffusion 
TMM 
Trimmed Mean of M-values 
TNF 
Tumor Necrosis Factor 
US 
United States 
V 
Visit(s) 
VTEU 
Vaccine and Treatment Evaluation Unit 
WHO 
World Health Organization 
 


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1. 
PREFACE 
The Statistical Analysis Plan (SAP) for “A Phase II Study to Evaluate and Compare the 
Immunogenicity of Monovalent Inactivated Influenza A/H7N9 Virus Vaccine Administered with 
and without AS03 Adjuvant and Monovalent Inactivated Influenza A/H3N2v Virus Vaccine 
Administered without Adjuvant in Healthy Adults through Standard and Systems Biology 
Analyses” (DMID Protocol 14-0015) describes and expands upon the statistical information 
presented in the protocol. 
This document describes all planned analyses including sample tables, listings, and figures. 
Regarding the final analyses and Clinical Study Report (CSR), this SAP follows the International 
Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for 
Human Use (ICH) Guidelines, as indicated in Topic E3 (Structure and Content of Clinical Study 
Reports), and more generally is consistent with Topic E8 (General Considerations for Clinical 
Trials) and Topic E9 (Statistical Principles for Clinical Trials). The structure and content of the 
SAP provides sufficient detail to meet the requirements identified by the FDA and ICH, while all 
work planned and reported for this SAP will follow internationally accepted guidelines published 
by the American Statistical Association and the Royal Statistical Society for statistical practice.  
This document contains four sections: (1) a review of the study design, (2) general statistical 
considerations, (3) comprehensive statistical analysis methods for immunogenicity, 
transcriptomics, cytokine/chemokine, and safety outcomes, and (4) a list of proposed tables, 
figures, and listings. Within the table, figure, and listing mock-ups (Appendices 1, 2, and 3), 
references to CSR sections are included. Any deviation from this SAP will be described and 
justified in protocol amendments and/or in the CSR, as appropriate. The reader of this SAP is 
encouraged to also review the study protocol for details on conduct of the study and the 
operational aspects of clinical assessments. Tertiary and exploratory proteomics assessments and 
correlation analyses are presented in a separate addendum to this SAP. 
 


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2. 
INTRODUCTION 
2.1. 
Purpose of the Analyses 
The primary goal of these analyses is to characterize and compare gene expression responses from subjects 
receiving two doses of monovalent inactivated influenza A/Shanghai/02/2013 (H7N9) virus vaccine, hereafter 
referred to as A/H7N9 vaccine, administered with AS03 adjuvant with those subjects that received the same 
vaccine without AS03 post first and second dose. This direct comparison will potentially reveal gene 
expression signatures that are modulated by AS03 which in turn could lead to a better understanding of the 
immune response-enhancing capabilities of the AS03 adjuvant. As a co-primary endpoint, anti-HA 
hemagglutination-inhibition (HAI) antibody responses will be assessed. The secondary endpoint analysis 
focuses on serum neutralizing antibodies and plasma cytokine/chemokine profiles pre- and post-vaccination. 
As part of the tertiary analysis, A/H7N9 transcriptomics responses will be compared to a control arm 
receiving one dose of monovalent inactivated influenza A/Minnesota/11/2010 (H3N2)v virus vaccine, 
hereafter referred to as A/H3N2v vaccine. The goal is to compare the innate and adaptive immune responses 
observed between the two avian (A/H7N9) study arms, and between the unadjuvanted avian (A/H7N9) and a 
swine-origin influenza vaccine (A/H3N2v) that is more similar to seasonal influenza A/H3N2. 
Additional tertiary and exploratory endpoint analyses are aimed at characterizing and comparing proteomics 
signatures and assessing the correlation between omics and serum antibody and plasma cytokines/chemokines 
as well as safety outcomes. The goal of the correlation analysis is to integrate the different data to develop a 
systems model of the human immune response to unadjuvanted monovalent inactivated influenza A/H3N2v 
virus vaccine and monovalent inactivated influenza A/H7N9 virus vaccine given with and without AS03 
adjuvant.  
 


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3. 
STUDY OBJECTIVES AND ENDPOINTS 
3.1. 
Study Objectives 
Primary Objectives 
1. To assess the serum anti-HA hemagglutination-inhibition (HAI) response to influenza A/H7N9 antigen 
(with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with 
A/H7N9 vaccine ± AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the 
study vaccination with A/H3N2v vaccine). 
2. To identify differentially expressed genes in human immune cells on Days 2, 4, and 29 (following the first 
study vaccination with A/H7N9 vaccine ± AS03) and on Days 30, 32, and 36 (following the second study 
vaccination with A/H7N9 vaccine ± AS03), compared to baseline assessments performed prior to each 
study vaccination (Days -7, 1, and 29). 
Secondary Objectives 
1. To compare plasma cytokine and chemokine profiles at specific time points and between treatment arms: 
a. Post-vaccination time points [(Days 2 and 4; Day 8 (A/H3N2v arm only)], and Days 30, 32, and 36 
(A/H7N9 arms only) with pre-vaccination time points (Days -7 and 1, prior to first study 
vaccination and Day 29, prior to second study vaccination (A/H7N9 arms only). 
2. To assess the neutralizing antibody responses to influenza A/H7N9 antigen (with and without adjuvant) at 
Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine ± AS03) and 
influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with 
A/H3N2v vaccine). 
3. To identify differentially expressed genes in human immune cells on Days 2, 4, and 8 following one 
intramuscular dose of influenza A/H3N2v vaccine compared to baseline assessments performed prior to 
study vaccination (Day -7 and Day 1). 
Tertiary Objectives 
1. To assess the frequency of adverse events. 
2. To identify differentially abundant cellular proteins in human immune cells on Days 2, 4, and 8 following 
one intramuscular dose of influenza A/H3N2v vaccine compared to baseline assessments performed prior 
to study vaccination (Day -7 and Day 1). 
3. To identify differentially abundant cellular proteins in human immune cells on Days 2, 4, and 29 
(following first study vaccination with A/H7N9 vaccine ± AS03) and Days 30, 32, and 36 (following 
second study vaccination with A/H7N9 vaccine ± AS03), compared to baseline assessments performed 
prior to study vaccination (Days -7, 1, and 29). 
Exploratory Objectives 
1. To identify and characterize differentially expressed genes in human immune cells on Days 2, 4, and 29 
(following the first study vaccination with A/H7N9 vaccine ± AS03) and on Days 30, 32, and 36 
(following the second study vaccination with A/H7N9 vaccine ± AS03) that differ between treatments 
(A/H7N9 vaccine vs. A/H7N9 vaccine + AS03). 


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2. To identify and characterize differentially abundant cellular proteins in human immune cells on Days 2, 
4, and 29 (following the first study vaccination with A/H7N9 vaccine ± AS03) and on Days 30, 32, and 
36 (following the second study vaccination with A/H7N9 vaccine ± AS03) that differ between treatments 
(A/H7N9 vaccine vs. A/H7N9 vaccine + AS03). 
3. To identify and characterize differentially expressed genes in human immune cells on Days 2 and 4 
(following the first study vaccination) that differ in baseline responses (Days -7, and 1) between 
treatments (A/H3N2v vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
4. To identify and characterize differentially abundant cellular proteins in human immune cells on Days 2 
and 4 (following the first study vaccination) that differ in baseline responses (Days -7, and 1) between 
treatments (A/H3N2v vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
5. To identify transcriptomic and proteomic signatures in human immune cells that correlate with influenza 
seroconversion and with peak HAI titers. 
6. To correlate plasma cytokine profiles with transcriptomic and proteomic profiles in subjects receiving 
influenza A/H3N2v vaccine or influenza A/H7N9 ± AS03 vaccine. 
7. Exploratory inspection of transcriptomics\proteomics response signatures for subjects with unexpected or 
severe safety profiles that are related to the study products. 
3.2. 
Endpoints 
Primary Endpoints 
1. Percentage of subjects achieving seroconversion (defined as either a pre-vaccination HAI titer <1:10 and a 
post-vaccination HAI titer ≥1:40 or a pre- vaccination HAI titer ≥1:10 and a minimum four-fold rise in 
post-vaccination HAI antibody titer) at approximately 28 days after the second study vaccination 
(A/H7N9 vaccine, with and without adjuvant). 
2. Percentage of subjects achieving seroconversion at approximately 28 days after the study vaccination with 
A/H3N2v vaccine. 
3. Number of differentially expressed genes based on RNA expression as determined by RNA-Seq analysis, 
in human immune cells at Days 2, 4, 29, 30, 32, and 36 after study vaccination with AS03-adjuvanted and 
unadjuvanted A/H7N9 vaccine, as compared to baseline assessments performed prior to vaccination (Days 
-7, 1, and 29). 
Secondary Endpoints 
1. 
Plasma measurement of cytokines and chemokines at each study visit, comparing: 
a. 
Post-vaccination time points with pre-vaccination time points by treatment arms. 
b. 
AS03-adjuvanted influenza A/H7N9 responses to unadjuvanted influenza A/H7N9 responses. 
c. 
Influenza A/H3N2v vaccine to unadjuvanted influenza A/H7N9 vaccine. 
d. 
Influenza A/H3N2v vaccine to adjuvanted A/H7N9 vaccine. 
2. 
Serologic response to influenza hemagglutinin: 
a. 
Percentage of subjects achieving seroconversion (defined as either a pre-vaccination Neut titer 
<1:10 and a post-vaccination Neut titer ≥1:40 or a pre-vaccination Neut titer ≥1:10 and a minimum 
four-fold rise in post-vaccination Neut titer) at approximately 28 days after the second study 
vaccination (A/H7N9 vaccine, with and without adjuvant). 


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b. 
Percentage of subjects achieving seroconversion at approximately 28 days after the study 
vaccination with influenza A/H3N2v vaccine. 
c. 
Geometric Mean Titers of serum HAI and Neut antibody at baseline (Day -7 and 1) and 28 days 
after influenza A/H3N2v vaccination. 
d. 
Geometric Mean Titers of serum HAI and Neut antibody at baseline (Day -7 and 1), and 28 days 
after both the first and the second dose of influenza A/H7N9 vaccine (with and without adjuvant). 
3. Number of differentially expressed genes based on RNA-Seq analysis in human immune cells at Days 2, 
4, and 8 after study vaccination, as compared to baseline assessments (Days -7 and 1) after influenza 
A/H3N2v vaccine. 
Tertiary Endpoints 
1. Occurrence of solicited local reactogenicity events within 7 days after each study vaccination. 
2. Occurrence of solicited systemic reactogenicity events within 7 days after each study vaccination. 
3. Occurrence of unsolicited AEs collected for approximately 28 days after last study vaccination. 
4. Occurrence of SAEs and MAAEs including new-onset chronic medical conditions and potentially 
immune-mediated medical conditions for approximately 1 month (A/H3N2v arm) and 12 months 
(A/H7N9 arms) after last study vaccination. 
5. Number of differentially abundant proteins based on protein abundance as determined by quantitative 
proteomics in human immune cells at Days 2, 4, and 8 after study vaccination, as compared to baseline 
assessments (Days -7 and 1) after influenza A/H3N2v vaccine. 
6. Number of differentially abundant proteins based on protein abundance as determined by quantitative 
proteomics analysis, in human immune cells at Days 2, 4, 29, 30, 32, and 36 after study vaccination with 
AS03-adjuvanted and unadjuvanted A/H7N9 vaccine comparing changes in protein abundance between 
treatment arms (A/H7N9 vaccine vs. A/H7N9 vaccine + AS03). 
Exploratory Endpoints 
1. RNA expression, clusters of co-expressed genes, enriched pathways, and other functional modules as 
determined by RNA-Seq analysis, in human immune cells at Days 2, 4, 29, 30, 32, and 36 after study 
vaccination with AS03-adjuvanted and unadjuvanted A/H7N9 vaccine comparing changes in RNA 
expression between treatment arms (A/H7N9 vaccine vs. A/H7N9 vaccine + AS03). 
2. Protein abundance, clusters of co-abundant proteins, enriched pathways, and other functional modules 
as determined by quantitative proteomics analysis, in human immune cells at Days 2, 4, 29, 30, 32, 
and 36 after study vaccination with AS03-adjuvanted and unadjuvanted A/H7N9 vaccine comparing 
changes in protein abundance between treatment arms (A/H7N9 vaccine vs. A/H7N9 vaccine + 
AS03). 
3. RNA expression, clusters of co-expressed genes, enriched pathways, and other functional modules as 
determined by RNA-Seq analysis, in human immune cells at Days 2, 4, and 29 after study vaccination 
comparing baseline changes in RNA expression between treatment arms (A/H3N2v vaccine vs. 
A/H7N9 vaccine and A/H3N2v vaccine vs. A/H7N9 vaccine + AS03). 
4. Protein abundance, clusters of co-abundant proteins, enriched pathways, and other functional modules 
as determined by quantitative proteomics analysis, in human immune cells at Days 2, 4, and 29 after 
first study vaccination comparing baseline changes (Days -7, and 1) in protein abundance between 


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treatment arms (A/H3N2v vaccine vs. A/H7N9 vaccine and A/H3N2v vaccine vs. A/H7N9 vaccine + 
AS03). 
5. Changes from baseline (Days -7, 1, and 29) in RNA expression or protein abundance that best 
correlate with influenza A/H7N9 seroconversion and with peak HAI titers at Days 2 and 4 (following 
first study vaccination) as well as Days 30, 32, and 36 (following second study vaccination). 
 
6. Changes from baseline (Days -7, 1, and 29) in RNA expression or protein abundance that best 
correlate with baseline changes in cytokine concentrations at Days 2, 4, 8 (A/H3N2v arm only) and 29 
(following first study vaccination) as well as Days 30, 32, and 36 (following second study 
vaccination). 
7. Exploratory summaries of transcriptomics/proteomics response signatures for subjects with 
unexpected or severe safety profiles. 
8. Lists of differential genes and proteins identified as part of the primary and secondary objectives. 
3.3. 
Study Definitions and Derived Variables 
Derived safety variables are described in Table 3, Table 4, Table 5, Table 6, and Table 7 in this document. 
The following assay-specific criteria will be used to determine seroconversion for the hemagglutination 
inhibition (HAI) and neutralization (NT) titers: 
• a pre-vaccination titer <1:10 and a post-vaccination titer ≥1:40 or a pre-vaccination titer ≥1:10 and 
a minimum of a four-fold rise in post-vaccination antibody titer. 


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4. 
INVESTIGATIONAL PLAN 
4.1. 
Overall Study Design and Plan 
This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study in 
approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, inclusive, designed 
to evaluate and compare the immunogenicity between an intramuscular monovalent inactivated influenza 
A/H7N9 virus vaccine manufactured by Sanofi Pasteur given with and without AS03 adjuvant manufactured 
by GlaxoSmithKline, and an intramuscular unadjuvanted monovalent inactivated influenza A/H3N2v virus 
vaccine manufactured by Sanofi Pasteur. 
This study will use a standard and systems biology approach to assess the human early gene and protein 
signatures expressed at baseline (approximately Days -7 and 1), and at approximately Days 2, 4, and 8 
(A/H3N2v-vaccinated subjects only) after the first study vaccination in each treatment arm as well as at 
approximately Days 29, 30, 32, and 36 in A/H7N9-vaccinated subjects only. Cellular immunogenicity 
(systems biology studies) data will be integrated with serologic immunogenicity (HAI and Neut antibody 
assays) and reactogenicity data to develop a systems model of the human immune response to unadjuvanted 
monovalent inactivated influenza A/H3N2v virus vaccine and monovalent inactivated influenza A/H7N9 
virus vaccine given with and without AS03 adjuvant. 
This study will use venous blood samples and subject data collected from a total of thirty vaccinated subjects 
randomly divided into three equal treatment arms (Table 1).  The first treatment arm (n=10) will be vaccinated 
with one dose of 15 µg of A/H3N2v HA.  The second treatment arm (n=10) will be vaccinated with two doses 
of 3.75 µg of A/H7N9 HA given with AS03 approximately 28 days apart.  The third treatment arm (n=10) 
will be vaccinated with two doses of 3.75 µg of A/H7N9 HA given without AS03 adjuvant approximately 28 
days apart.  All study vaccinations will be administered intramuscularly. 
Reactogenicity will be measured by the occurrence of solicited injection site and systemic reactions from the 
time of each study vaccination through 8 days after each study vaccination.  Unsolicited non-serious adverse 
events (AEs) will be collected from the time of each study vaccination through approximately 28 days after 
each study vaccination.  Serious adverse events (SAEs) and medically-attended adverse events (MAAEs) 
including new-onset chronic medical conditions (NOCMCs) and potentially immune-mediated medical 
conditions (PIMMCs) will be collected from the time of the first study vaccination through approximately 1 
month (A/H3N2v arm) and 12 months (A/H7N9 arms) after the last study vaccination. 
Venous blood samples (approximately 90 mL) will be collected from subjects at approximately Days -7 and 1 
(immediately prior to the first study vaccination), and at approximately Days 2, 4; and 8 (A/H3N2v-
vaccinated subjects only) after the first study vaccination for systems biology studies (cytokine and 
chemokine levels, immune cell activation status, and whole transcriptome and proteome profiles of the major 
blood immune cells).  Additionally, subjects in either treatment arm receiving A/H7N9 vaccine will have 
additional blood samples collected at approximately Days 29 (this will be collected immediately prior to the 
second study vaccination for subjects in either group receiving A/H7N9), 30, 32, and 36 after the first study 
vaccination for systems biology studies. 
Serological assessment (hemagglutination inhibition (HAI) and neutralizing (Neut) antibody assays) will also 
be conducted on venous blood samples (approximately 10 mL) collected from subjects at Day 1 (immediately 
prior to the first study vaccination), and at approximately Days 29 (for A/H7N9-vaccinated subjects this will 
be collected immediately prior to the second study vaccination) and 57 (A/H7N9-vaccinated subjects only) 
after the first study vaccination. Plasma cytokine/chemokine levels, as well as transcriptomic and proteomic 


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profiles from individual immune cell compartments will be measured and integrated with standard serologic 
assessment to vaccine. 
Urine samples (approximately 20 mL) will also be collected from subjects at approximately Days -7, 1, 2, 4, 8 
(A/H3N2v-vaccinated subjects only), 29, 30, 32, and 36 for future research. 
4.2. 
Discussion of Study Design, Including the Choice of Control Groups 
The main study arms are the two arms receiving influenza A/H7N9 virus vaccine with or without AS03 
adjuvant (Study Arms 2 and 3). The goal is to better understand how the AS03 adjuvant modulates the 
immune response to influenza A/H7N9 virus on the molecular level. The addition of the interpandemic 
(seasonal) strain of influenza A/H3N2v was chosen to compare innate and adaptive immune response between 
avian (A/H7N9) and swine-origin influenza vaccines (A/H3N2v) more closely resembling seasonal strains of 
influenza A. 
4.3. 
Selection of Study Population 
Subject Inclusion Criteria 
Subjects eligible to participate in this study must meet all of the following inclusion criteria: 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Are able to understand and comply with planned study procedures and be available for all study visits. 
3. Are males or non-pregnant females, 18 to 49 years old, inclusive. 
4. Are in good health1. 
1As determined by medical history and targeted physical examination, if indicated based on medical history, to evaluate acute or 
currently ongoing chronic medical diagnoses or conditions, defined as those that have been present for at least 90 days, that would 
affect the assessment of the safety of subjects or the immunogenicity of study vaccinations.  Chronic medical diagnoses or conditions 
should be stable for the last 60 days.  This includes no change in chronic prescription medication, dose, or frequency as a result of 
deterioration of the chronic medical diagnosis or condition in the 60 days prior to enrollment.  Any prescription change that is due to 
change of health care provider, insurance company, etc., or that is done for financial reasons, as long as in the same class of 
medication, will not be considered a deviation of this inclusion criterion.  Any change in prescription medication due to improvement of 
a disease outcome, as determined by the site principal investigator or appropriate sub-investigator, will not be considered a deviation of 
this inclusion criterion.  Subjects may be on chronic or as needed (prn) medications if, in the opinion of the site principal investigator or 
appropriate sub-investigator, they pose no additional risk to subject safety or assessment of reactogenicity and immunogenicity and do 
not indicate a worsening of medical diagnosis or condition.  Similarly, medication changes subsequent to enrollment and study 
vaccination are acceptable provided there was no deterioration in the subject’s chronic medical condition that necessitated a medication 
change, and there is no additional risk to the subject or interference with the evaluation of responses to study vaccination.  Note: 
Topical, nasal, and inhaled medications (with the exception of inhaled corticosteroids as outlined in the Subject Exclusion Criteria (see 
Section 5.1.2)), herbals, vitamins, and supplements are permitted. 
5. Oral temperature is less than 100.4°F. 
6. Pulse is 50 to 115 bpm, inclusive. 
7. Systolic blood pressure is 85 to 150 mm Hg, inclusive. 
8. Diastolic blood pressure is 55 to 95 mm Hg, inclusive. 
9. Erythrocyte sedimentation rate (ESR) is less than 30 mm per hour. 


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10. Women of childbearing potential2 must use an acceptable contraception method3 from 30 days before 
first study vaccination until 60 days after last study vaccination. 
2Not sterilized via tubal ligation, bilateral oophorectomy, hysterectomy or successful Essure® placement (permanent, non-surgical, non-
hormonal sterilization) with documented radiological confirmation test at least 90 days after the procedure, and still menstruating or <1 
year of the last menses if menopausal. 
3Includes, but is not limited to, non-male sexual relationships, abstinence from sexual intercourse with a male partner, monogamous 
relationship with vasectomized partner who has been vasectomized for 180 days or more prior to the subject receiving the first study 
vaccination, barrier methods such as condoms or diaphragms with spermicide or foam, effective intrauterine devices, NuvaRing®, and 
licensed hormonal methods such as implants, injectables or oral contraceptives (“the pill”). 
11. Women of childbearing potential must have a negative urine or serum pregnancy test within 24 hours 
prior to study vaccination. 
Subject Exclusion Criteria 
Subjects eligible to participate in this study must not meet any of the following exclusion criteria: 
1. Have an acute illness4, as determined by the site principal investigator or appropriate sub-
investigator, within 72 hours prior to study vaccination. 
4An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion of the site 
principal investigator or appropriate sub-investigator, the residual symptoms will not interfere with the ability to assess safety 
parameters as required by the protocol. 
2. Have any medical disease or condition that, in the opinion of the site principal investigator or 
appropriate sub-investigator, is a contraindication to study participation5. 
5Including acute or chronic medical disease or condition, defined as persisting for at least 90 days that would place the subject at an 
unacceptable risk of injury, render the subject unable to meet the requirements of the protocol, or may interfere with the evaluation of 
responses or the subject’s successful completion of this study. 
3. Have immunosuppression as a result of an underlying illness or treatment, or use of anticancer 
chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study vaccination. 
4. Have known active neoplastic disease (excluding non-melanoma skin cancer) or a history of any 
hematologic malignancy. 
5. Have known human immunodeficiency virus (HIV), hepatitis B, or hepatitis C infection. 
6. Have known hypersensitivity or allergy to eggs, egg or chicken protein, squalene-based adjuvants, or 
other components of the study vaccines. 
7. Have a history of severe reactions following previous immunization with licensed or unlicensed 
influenza virus vaccines. 
8. Have a personal or family history of narcolepsy. 
9. Have a history of Guillain-Barré syndrome. 
10. Have a history of convulsions or encephalomyelitis within 90 days prior to study vaccination. 


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11. Have a history of a potentially immune-mediated medical condition6. 
6Refer to Appendix A: List of Potentially Immune-Mediated Medical Conditions. 
12. Have a history of alcohol or drug abuse within 5 years prior to study vaccination. 
13. Have any diagnosis, current or past, of schizophrenia, bipolar disease, or other psychiatric diagnosis 
that may interfere with subject compliance or safety evaluations. 
14. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for danger 
to self or others within 10 years prior to study vaccination. 
15. Have taken oral or parenteral (including intra-articular) corticosteroids of any dose within 30 days 
prior to study vaccination. 
16. Have taken high-dose inhaled corticosteroids within 30 days prior to study vaccination.  High-dose 
defined as >840 mcg/day of beclomethasone dipropionate CFC or equivalent. 
17. Received licensed live vaccine within 30 days prior to the first study vaccination, or plans to receive 
licensed live vaccine within 30 days before or after each study vaccination. 
18. Received licensed inactivated vaccine within 14 days prior to the first study vaccination, or plans to 
receive licensed inactivated vaccine within 14 days before or after each study vaccination. 
19. Received immunoglobulin or other blood products (with exception of Rho D immunoglobulin) 
within 90 days prior to study vaccination. 
20. Received an experimental agent7 within 30 days prior to the first study vaccination, or expects to 
receive an experimental agent8 during the 13-month study-reporting period. 
7Including vaccine, drug, biologic, device, blood product, or medication. 
8Other than from participation in this study. 
21. Are participating or plan to participate in another clinical trial with an interventional agent9 that will 
be received during the 13-month study-reporting period. 
9Including licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication. 
22. Prior participation in a clinical trial of influenza A/H7 vaccine10 or have a history of influenza A/H7 
virus actual or potential exposure or infection prior to the first study vaccination. 
10And assigned to a group receiving influenza A/H7 vaccine, does not apply to documented placebo recipients. 
23. Prior participation in a clinical trial of influenza A/H3N2v vaccine11 or have a history of influenza 
A/H3N2v virus actual or potential exposure or infection prior to the first study vaccination. 
11And assigned to a group receiving influenza A/H3N2v vaccine, does not apply to documented placebo recipients. 
24. Occupational exposure to or substantial direct physical contact12 with birds in the past year or during 
the 28 days after each study vaccination. 




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slight modifications in the formulation step, to the manufacturing process for the production of its licensed 
Influenza Virus Vaccine Fluzone® and the H1N1 influenza vaccine A/California/07/2009 NYMC X-179A.  
Using the Fluzone® production process, an A/H3N2v reassortant was derived from the A/Minnesota/11/2010 
strain produced by classical reassortant technology at New York Medical College (NYMC) and provided by 
the Centers for Disease Control and Prevention (CDC).  This monovalent inactivated split influenza virus 
vaccine is prepared from influenza virus propagated in embryonated chicken eggs.  Licensed release testing 
specifications were maintained 
4.4.2. 
Identity of Investigational Product(s) 
A/H7N9 Vaccine 
The monovalent inactivated influenza A/H7N9 virus vaccine was supplied as a sterile, clear, and slightly 
opalescent suspension in single-dose vials containing 7.5 mcg of HA per 0.5 mL.  Each vial contained a fill 
volume of 0.7 mL.  It contained no preservative (i.e., non-thimerosal) or antibiotics.  The vials containing 
study product had to be stored at 2°C to 8°C (35.6°F to 46.4°F) and not frozen.  Vials were provided with 
latex-free stoppers. 
AS03 Adjuvant 
The AS03 adjuvant was supplied as a preservative-free, oil-in-water, whitish to yellowish homogenous milky 
liquid emulsion in single-use vials containing a fill volume of 3.15 mL.  The vials containing study product 
had to be stored at 2°C to 8°C (35.6°F to 46.4°F) and not frozen.  Vials were provided with latex-free 
stoppers. 
A/H3N2v Vaccine 
The monovalent inactivated influenza A/H3N2v virus vaccine was supplied as a sterile, clear, and slightly 
opalescent suspension. A minor modification in the PBS diluent in the formulation step was made to 
accommodate the nature of the diluted virus in single-dose, prefilled syringes, without needles, containing 15 
mcg of HA per 0.5mL.  It contained no preservative (i.e., non-thimerosal), antibiotics, or latex.  The vaccine 
included porcine gelatin (0.05%) as a stabilizer.  The prefilled syringes containing study product had to be 
stored at 2°C to 8°C (35°F to 46°F) and not frozen. 
Each of these study products were labeled according to manufacturer specifications and included the 
statement “Caution: New Drug – Limited by Federal Law to Investigational Use.” 
Further details are included in the respective, applicable Investigator’s Brochures and Supplemental 
Information for the Investigator’s Brochures for the A/H7N9 vaccine, AS03 adjuvant, and A/H3N2v vaccine 
as well as protocol-specific MOP. 
4.4.3. 
 Method of Assigning Subjects to Treatment Groups (Randomization) 
Per International Conference on Harmonisation (ICH) guideline E6: Good Clinical Practice (GCP), screening 
records were kept at the participating VTEU site to document the reason why an individual was screened, but 
failed study entry criteria. The reasons why individuals failed screening were recorded in the Statistical and 
Data Coordinating Center’s (SDCC) AdvantageEDCSM (Electronic Data Capture System). 
Once consented and upon entry of demographic data and confirmation of eligibility for this study, the subject 
was enrolled.  Subjects were assigned randomly to 1 of 3 treatment arms (Table 1).  The first treatment arm 
(n=10) was vaccinated with one dose of 15 µg of A/H3N2v HA.  The second treatment arm (n=10) was 
vaccinated with two doses of 3.75 µg of A/H7N9 HA given with AS03 approximately 28 days apart.  The 


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third treatment arm (n=10) was vaccinated with two doses of 3.75 µg of A/H7N9 HA given without AS03 
adjuvant approximately 28 days apart.  All study vaccinations were administered intramuscularly. 
Enrollment of subjects was done online using the enrollment module of AdvantageEDCSM.  The 
randomization code was prepared by statisticians at the SDCC and included in the enrollment module for this 
study.  AdvantageEDCSM assigned each subject to a treatment arm after the demographic and eligibility data 
had been entered into the system.  A designated individual at the participating VTEU site was provided with a 
code list for emergency unblinding purposes, which was to be kept in a secure place. 
4.4.4. 
Selection of Doses in the Study 
As with other avian influenza strains, the immunogenicity of inactivated vaccines is poor.  Higher doses of 
hemagglutinin (HA), a major target of the protective immune response, has been shown to generate more 
frequent and higher antibody titers. Thus, two doses of 3.75 mcg of HA (A/Shanghai/2/2013) were 
administered for the A/H7N9 arms. Standard dosing of 15 mcg of HA (A/H3N2v) was chosen to more closely 
resemble the dosing administered as part of seasonal influenza vaccination. While 15 mcg represents a 4-fold 
increase in HA amount, the vaccine was unadjuvanted and matches what is formulated in seasonal standard 
dose inactivated influenza vaccines.   
4.4.5. 
Selection and Timing of Dose for Each Subject 
The dose of vaccine was chosen primarily to mirror the amount provided in DMID 10-0074 in which 3.75 
mcg of influenza A/H5N1 virus vaccine was given with or without AS03. This reduced amount represents the 
minimal amount of HA needed to generate an immune response in the presence of an oil-in-water adjuvant 
system, based on prior studies. The HA content was kept the same for the unadjuvanted dose in an effort to 
measure the adjuvant effect specifically. Further, in previous studies, two doses of avian-strain influenza 
vaccine were required to generate an adaptive response, and typically the second dose was given 3-4 weeks 
after the first dose.  
4.4.6. 
Blinding 
This was a partially-blind clinical study as the unadjuvanted A/H3N2v vaccine treatment arm was open-label 
to avoid the need for placebo administration at approximately Day 29.  Investigators and study personnel 
remained blinded to allocation of A/H7N9 with or without AS03. The randomization scheme was generated 
by the SDCC and provided to unblinded study personnel (i.e., research pharmacists performing study 
vaccination preparations and unblinded study vaccine administrators) at the participating VTEU site. The 
unblinded study vaccine administrator was a study personnel member credentialed to administer vaccines and 
may also have participated in dose preparation but was not involved in study-related assessments or had 
subject contact for data collection following study vaccine administration. 
4.4.7. 
Prior and Concomitant Therapy 
Administration of any medications, therapies, or vaccines was recorded on the appropriate data collection 
form.  Concomitant medications included all current medications and medications taken within 60 days prior 
to signing the informed consent form through approximately 28 days after the last study vaccination or 
through the early termination visit (if prior to 28 days after the last study vaccination), whichever occurred 
first. Medications reported in the electronic case report form (eCRF) were limited to those taken within 30 
days prior to the first study vaccination through approximately 28 days after the last study vaccination.  
Prescription and over-the-counter drugs were included as well as herbals, vitamins and supplements.  In 
addition, receipt of non-study influenza vaccines was solicited through approximately 28 days after the last 


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study vaccination and reported in the eCRF. Use of new medication should have prompted evaluation for the 
presence of a new diagnosis of chronic medical disease or condition. Medications that might interfere with the 
evaluation of the investigational product(s) should not be have been used during the study-reporting period 
(approximately 12 months after the last study vaccination) unless clinically indicated as part of the subject’s 
health care.  Medications in this category included the prohibited medications per the Subject Exclusion 
Criteria (see Section 4.3).  In addition, the site principal investigator or appropriate sub-investigator may 
identify other medications that should not have been used due to a risk to subject safety or assessment of 
reactogenicity and immunogenicity. 
4.4.8. 
Treatment Compliance 
Study product was administered to subjects by an unblinded study vaccine administrator via intramuscular 
(IM) injection at all dosing times according to subject treatment assignment and as described in Section 6.2 in 
the protocol. The number of administered vaccine doses was recorded. 
4.5. 
Immunogenicity and Safety Variables 
 
The following section describes the collection of safety and immunogenicity variables. For a detailed 
schedule of activities refer to Appendix B of the protocol.  
4.5.1. 
Immunogenicity Variables 
Venous blood samples (approximately 10 mL) for HAI and Neut antibody assays were collected from 
subjects at Day 1 (immediately prior to the first study vaccination), and at approximately Days 29 (for 
A/H7N9-vaccinated subjects this was collected immediately prior to the second study vaccination) and 57 
(A/H7N9-vaccinated subjects only) after the first study vaccination. Subjects who withdrew early had HAI 
and Neut antibody assays run on available sera, including sera from the early termination visit, if available.  
Individual HAI and Neut results will be reported by the central immunology laboratory for the 
A/Shanghai/02/2013xPR8 vaccine strain and the A/Minnesota/11/2010 vaccine strain for the A/H3N2v study 
arm. Assay results will be reported as a reciprocal titer with values of 10*2k, where k=0, 1, 2, etc. The lower 
limit of detection for the HAI and Neut assays is 1:10; values below the limit of detection will be imputed for 
analysis using one-half the limit of detection (10/2 = 5). For analysis, the geometric mean (calculated on 
natural log scale) of repeated results for each sample will be computed and used as the response for all 
subsequent calculations. The variation arising from multiple measurements for an individual at a single time 
point will not be used in subsequent calculations/statistics. See Section 3.3 for definitions of derived variables 
for the analysis of HAI and Neut data. 
Cytokine and chemokine levels in plasma obtained from venous blood samples were quantitatively measured 
at approximately Days -7, 1, 2, 4, and 8 (A/H3N2v-vaccinated subjects only) and at approximately Days 29, 
30, 32, and 36 for A/H7N9-vaccinated subjects only using the Luminex platform. Cytokine and chemokine 
levels will be reported as concentrations. Values below the plate-specific LOD for each cytokine and 
chemokine will be imputed using 1/2 LOD.  
4.5.2. 
-Omics Variables 
Transcriptome profiling using RNA-Seq was performed on immune cell subsets collected at approximately 
Days -7, 1, 2, 4, and 8 (A/H3N2v-vaccinated subjects only) and at approximately Days 29, 30, 32, and 36 for 
A/H7N9-vaccinated subjects only. Total RNA was isolated using standard methods and subjected to ultra-


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high-throughput parallel next generation sequencing. Gene expression levels for each gene were estimated 
based on the number of mapping sequencing reads. 
Proteomics profiling was based on shotgun proteomics using multidimensional liquid chromatography 
coupled with tandem mass spectrometry and performed on immune cell samples collected at approximately 
Days -7, 1, 2, 4, and 8 (A/H3N2v-vaccinated subjects only) and at approximately Days 29, 30, 32, and 36 for 
A/H7N9-vaccinated subjects only. Protein abundance levels for each protein were estimated based on the 
sample peak intensity ratio compared to the sample that was prepared using a pool of all samples. 
4.5.3. 
Safety Variables 
Safety assessments included: 
1. Study vaccine-related serious adverse events occurring from the time of the first study vaccination through 
approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 arms) after the last study vaccination. 
2. Solicited Adverse Events – reactogenicity events occurring from the time of each study vaccination 
through 7 days after each study vaccination: 
a) Injection site reactions including pruritus (itching), ecchymosis (bruising), erythema (redness), 
induration (hardness)/swelling, pain, and tenderness. 
b) Systemic reactions including fever, feverishness (chills/shivering/sweating), fatigue (tiredness), 
malaise (general unwell feeling), myalgia (body aches/muscular pain exclusive of the injection 
site), arthralgia (joint pain exclusive of the injection site), headache, and nausea. 
3. Unsolicited Adverse Events – study vaccine-related non-serious adverse events occurring from the time of 
each study vaccination through approximately 28 days after each study vaccination. 
4. Medically-Attended Adverse Events including new-onset chronic medical conditions and potentially 
immune-mediated medical conditions occurring from the time of the first study vaccination through 
approximately 1 month (A/H3N2v arm) and 12 months (A/H7N9 arms) after the last study vaccination. 


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5. 
SAMPLE SIZE CONSIDERATIONS 
The sample size is not based on a formal hypothesis. Rather it is based on practical considerations with the 
goal of gathering enough information to learn more about immune system responses to the vaccine/adjuvant 
on the molecular level.  DMID Protocol 10-0074, entitled:  “A Randomized, Double-Blinded, Controlled, 
Phase I Study in Healthy Adults to Assess the Safety, Reactogenicity, and Immunogenicity of Intramuscular 
Subvirion Inactivated Monovalent Influenza A/H5N1 Virus Vaccine Administered With and Without AS03 
Adjuvant: Standard & Systems Biology Analyses”, showed that sample sizes in this range are sufficient to 
detect key differential signals and carry out systems biology analyses. 


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6. 
GENERAL STATISTICAL CONSIDERATIONS 
6.1. 
General Principles 
Continuous variables will be summarized using descriptive statistics (n, mean, standard deviation, median, 
interquartile range (IQR), minimum, and maximum) unless otherwise specified. The frequency and 
percentages (based on the analysis population size) of observed levels will be reported for all categorical 
measures. In general, all data will be listed and sorted by study arm and subject, and when appropriate, by 
study visit number within subject. All summary tables will be structured with a column for each study arm in 
the following order: Study Arm 1, Study Arm 2, and Study Arm 3. 
6.2. 
Timing of Analyses 
The primary CSR will be completed when all primary and secondary endpoint data are available and the data 
have been cleaned and locked. Any available data from the long-term, tertiary safety follow-up of applicable 
subjects may also be included.  Additional data from the long-term, tertiary safety follow-up of applicable 
subjects, exploratory correlation analyses, and data from the proteomics portion of the study will be included 
in one or more addenda to the CSR. 
6.3. 
Analysis Populations 
6.3.1. 
Safety Analysis Population 
The Safety Analysis population includes all subjects who received at least one dose of study vaccine. 
6.3.2. 
Intention-to-Treat Analysis (ITT) Population 
The intent-to-treat (ITT) population includes all subjects who received at least one dose of study vaccine and 
contributed both pre- and at least one post-study vaccination blood samples for serological assessment (HAI 
or Neut antibody assays) for which valid results were reported. 
6.3.3. 
Per protocol (PP) Population 
The per protocol (PP) population includes all subjects in the ITT subset with the following exclusions: 
• Data from all available visits for subjects found to be ineligible at baseline. 
• Data from all visits subsequent to major protocol deviations, such as: 
− Second study vaccination not received, 
− Receipt of non-study licensed live vaccine within 30 days before or after each study 
vaccination, 
− Receipt of non-study licensed inactivated vaccine within 14 days before or after each study 
vaccination, 
− Receipt of immunosuppressive therapy (e.g., corticosteroids) within 30 days before or after 
each study vaccination. 
− Data from any visit that occurs substantially out of window. Substantially out of window is 
defined as any visit that occurred outside of the pre-defined protocol window except for the 
following visits for which windows will be redefined as: 


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o Day 36: +4/-4 
o Day 57: +7/-4 
6.3.4. 
The -Omics Analysis Population 
The ‘omics analysis population for the -omics (transcriptomics and proteomics) endpoint analyses will 
include all subjects who received at least one study vaccination and contributed at least one pre- and one post-
vaccination sample for ‘omics testing for which valid results were reported. In addition, the following 
exclusions will be applied: 
• post-second dose samples from subjects in Study Arms 2 and 3 that did not receive second vaccination 
will be excluded. 
• samples that were collected substantially out-of-window will be excluded.  Substantially out of window 
is defined as any visit that occurred outside of the pre-defined protocol window except for the following 
visits for which windows will be redefined as: 
o Day 36: +4/-4 
o Day 57: +7/-4 
6.4. 
Covariates and Subgroups 
There is no a priori plan to adjust the analyses of safety and immunogenicity for covariates or to report results 
by subgroups. 
6.5. 
Missing Data 
For proteomics data relative protein abundance values will be imputed using the k-nearest neighbors 
imputation algorithm as implemented in the impute R package (See SAP Addendum A for further details).  
Otherwise, no imputation will be performed for missing values.  
6.6. 
Interim Analyses and Data Monitoring 
No interim safety or immunogenicity review was conducted. 
6.7. 
Multicenter Studies 
This was a single site study. 
6.8. 
Multiple Comparisons/Multiplicity 
No multiple testing adjustments will be carried out for the safety, immunogenicity, and cytokine/chemokine 
endpoints. For -omics endpoints, the false-discovery rates (FDR) based on the Benjamini-Hochberg procedure 
as implemented in the R p.adjust function will be used to control the false positive rate among significantly 
differentially expressed genes, proteins, and enriched pathways. 


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7. 
STUDY SUBJECTS 
7.1. 
Disposition of Subjects 
A flowchart presenting the disposition of study subjects, adapted from the CONSORT statement will be 
included (Figure 1). The flowchart will summarize the number of subjects who were eligible, enrolled and 
randomized, lost to follow-up, and analyzed, by study arm. Screened subjects who were ineligible for 
enrollment in the study will be presented by inclusion and exclusion criteria (Table 10). A listing of all 
subjects that received study product is provided in Listing 1. A listing of subjects who discontinued or 
terminated early from the study will be tabulated in Listing 2. The disposition of subjects in the study will be 
tabulated by study arm (Table 8). This table will show the number of subjects who were screened, enrolled 
and received the first vaccination, received both vaccinations, completed -omics blood draws, completed 
follow-up, and were included in the per-protocol analysis population. For each analysis population, a 
summary of excluded subjects by study arm and time point will be presented as shown in Table 9. A tabular 
listing of all subjects, visits, and observations excluded from certain analysis populations will be provided as 
specified in Listing 5. 
7.2. 
Protocol Deviations 
A summary of protocol deviations will be presented by the deviation category, type, and study arm in Table 2. 
This table will provide both the number of subjects and the number of deviations for each category and study 
arm. All subject-specific protocol deviations and non-subject-specific protocol deviations will be included in 
Listing 3 and Listing 4, respectively. 


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8. 
IMMUNOGENICITY EVALUATION 
8.1. 
Serum Antibody Analysis 
All immunogenicity analyses will be conducted for both the ITT and PP analysis populations. For each 
analysis population (ITT and PP) and serum antibody assay (HAI and Neut), geometric mean titers (GMTs 
and two-sided 95% confidence interval (CI) based on t-distribution) and percent seroconversion ([n/N (%)] 
and an exact (Clopper-Pearson) two-sided 95% CI as defined in Section 3.3) will be summarized by study arm 
and post-vaccination time point (Table 14, Table 15, Table 16, Table 17). GMTs over time will be 
summarized using time trend plots (Figure 2). Subject-level serum antibody titer responses at Day 29 post-last 
vaccination for all three study arms will be contrasted using bar plots (Figure 3). Differences between the 
treatments will be evaluated at Day 29 post-last dose using two-sided t-tests adjusting for unequal variance if 
necessary using a standard t-test or Welch’s t-test (Table 18, Table 19, Table 20, Table 21). The decision on 
whether to adjust for unequal variance will be based on inspection of variances for the three study arms. 
Differences in seroconversion rates for Day 29 post-last dose will be assessed using a two-sided Fisher’s exact 
test (Table 22, Table 23, Table 24, Table 25). Data listings of HAI and Neut assay results will be provided in 
Listing 8 sorted by study group, subject ID, and visit. 
8.2. 
Plasma Cytokines/Chemokine Analysis 
Pre-first vaccination cytokine/chemokine concentration will be determined using the mean concentrations 
among pre-vaccination measurements at Day -7 to -1 and Day 1. Cytokine/chemokine concentrations as well 
as their baseline fold changes will be summarized using minimum, Q1, median, Q3, maximum, and 95% CI 
of the median (starting with Table 26 and ending with Table 121). The 95% CI of the median concentration 
and median fold change from baseline will be determined by using the bootstrap method using 1,000 
bootstrap replicates and will be visualized using time trend plots (Figure 4) and radar plots (Figure 5). To 
identify cytokines/chemokines that show a differential response from baseline, a two-sided Wilcoxon signed-
rank test will be carried out for each study arm and post-vaccination time point (Table 122, Table 123).  
Significant differences in fold changes between study arms for shared post-vaccination time points will be 
evaluated using a Wilcoxon rank-sum test (Table 124, Table 125). Data listings of cytokine/chemokine assay 
results will be provided as shown in Listing 9 sorted by study group, subject ID, and visit. 


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9. 
TRANSCRIPTOMICS EVALUATIONS 
The following sections describe the analysis of RNA-Seq endpoint data for the main Clinical Study Report. 
Analysis will be based on the PP analysis population. To the extent possible, transcriptomics analyses will be 
implemented using the R statistical programming language [1] and will utilize published R packages such as 
made available via the Bioconductor online resource [2]. At the start of the analysis, the most-up-to-date 
Bioconductor version, R statistical software, and other third-party software will be identified and used. 
9.1. 
RNA-Seq Data Processing and Qc 
9.1.1. 
Human Reference Genome Alignment and Quality Assessment 
The latest version of the human reference genome assembly, gene models, and associated gene and transcript 
annotation information in the form of a Gene Transfer Format (GTF) file will be obtained from the 
ENSEMBL database [3]. Reference dataset versions will be documented in the CSR. Adapter sequences and 
low-quality 5’ ending sequences will be removed from raw sequencing reads using the Trimmomatic software 
[4]. The impact of quality filtering will be assessed using FastQC software [5]. Quality-filtered reads will then 
be aligned to the reference transcriptome/genome using the latest version of HISAT2 splice-aware sequence 
aligner [6]. For each sample, the quality of reference alignments will be evaluated using the RSeQC software 
[7]. Quality measures for each laboratory will be summarized in tabular form, univariate boxplots and 
multivariate starplots (Figure 6). 
9.1.2. 
Gene Expression Quantification and Data Normalization 
Gene expression quantification will be carried out on the gene level using the featureCounts function of the 
Subread software [8]. Reads that overlap with multiple genes or map to multiple genomic locations on the 
reference genome will be excluded. Known ribosomal, transfer, and mitochondrial RNA genes will be 
removed from the final read count analysis dataset. In addition, genes located on the X and Y chromosomes 
will be excluded from the final read count analysis dataset to avoid gender-related biases. 
Systematic sample differences in sequencing coverage will be corrected for by calculating scaling factors for 
each sample using the trimmed mean of M-values (TMM) method [9] as implemented in the edgeR software 
[10]. TMM-normalized moderated log2 counts per million (LCPM) for each gene will be calculated using the 
edgeR software [10]. As part of this step, a sequencing coverage–scaled count of 0.5 reads will be added to 
each gene to avoid taking the log of zero. LCPM distributions will be visualized using univariate boxplots and 
cumulative distribution function (CDF) plots before and after TMM-normalization. 
9.1.3. 
Global Outlier and Batch Effect Detection 
TMM-normalized LCPM will be standardized (z-score: mean=0, variance=1) and LCPM distributions across 
samples will be inspected for outliers (potential sample mislabeling, experimental error, etc.) and systematic 
effects (sample ordering, batch processing, etc.) using hierarchical clustering, multidimensional scaling, and 
principal component analysis (Figure 7). If outlying samples are discovered, corresponding clinical and 
laboratory metadata will be inspected for abnormalities. Based on these findings, a decision will be made as to 
whether the respective RNA-Seq observation should be removed from downstream analyses. The decision 
will be recorded in the CSR. If a noticeable batch effect is observed, unless an imbalance across vaccine 
groups is observed, the batch effect may be accounted for by adding a batch blocking factor as part of the 
negative binomial models (see Section 9.2). 


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9.1.4. 
Pre-Vaccination Transcriptomics Activity 
The two pre-first vaccination time point RNA-Seq samples (Day -7 to -1 and Day 1) measure the baseline 
transcriptomics activity for each subject enrolled in this study. For each subject, baseline transcriptomic 
activity will be represented by averaging results for the two pre-first vaccination samples by using the sum of 
the read counts for each gene followed by TMM normalization as described in Section 9.1.2. Globally 
outlying Day -7 to -1 or Day 1 RNA-Seq samples will be excluded (see Section 9.1.3). The combined pre-first 
vaccination sample will be used as a reference to assess transcriptomics responses at Days 2, 4, 8, or 29 post-
first vaccination. The Day 29 time point RNA-Seq sample also measures the pre-second vaccination 
transcriptomics activity for each subject enrolled in this study. This sample will be used as a reference to 
assess post-second vaccination responses (Days 2, 4, and 8 post-second vaccination) for subjects that received 
a second vaccine dose (Study Arms 2 and 3). 
9.1.5. 
Log Fold Change Calculations 
Log2 fold changes (LFC) per gene for each subject and post-vaccination day will be calculated by subtracting 
pre-vaccination LCPM values from the respective post-vaccination day LCPM value. Two sets of LFC will be 
determined to separately assess post-first and post-second vaccination responses: 
1. Day 2, 4, 8, or 29 vs. pre-first vaccination (averaged Day -7 to -1 and Day 1 results) LCPM for Study 
Arms 1, 2, and 3. 
2. Day 30, 32, and 36 vs. pre-second vaccination (Day 29) LCPM for Study Arms 2 and 3 
9.1.6. 
Determination of Cut Offs for Filtering Lowly Expressed Genes 
Transcriptome-wide LCPM results will be used to identify a suitable minimum LCPM filter cut off to exclude 
lowly expressed genes excluding outlying samples. Genes with mean normalized expression levels below X 
log2 counts per million across all samples (all study arms and time points) will be considered to be lowly 
expressed and will be excluded from the differential expression analysis. To determine a suitable cut off value 
for X, reverse cumulative distribution function (RCDF) plots summarizing the percentage of genes that 
exceeded a certain LCPM cut off will be plotted on the y-axis (Figure 8). The goal is to choose a cut off that 
retains between 10,000 and 14,000 genes for differential analysis. A default cut off of 1 count per million will 
be used if the targeted gene coverage is met. The filtered set will be used as input for down-stream 
transcriptomics endpoint analysis described in subsequent sections. 
9.2. 
Identification of Differentially Expressed Genes 
Negative binomial generalized linear models as implemented in the edgeR software [10] will be applied to 
identify differentially expressed (DE) genes after exclusion of outlying samples and lowly expressed genes. 
TMM-adjusted total read counts per sample will be included in the models as an offset to account for 
systematic sample differences. If systematic technical bias is detected and the effect is balanced across study 
arms, it will be accounted for by including a covariate or factor as part of the negative binomial models.  
For all study arms, DE gene analysis will be carried out for each post-first vaccination study day (Day 2, 4, 8 
or 29) in relation to pre-first vaccination gene expression levels (Section 9.2.1). For Study Arms 2 and 3, DE 
gene analysis will be carried out for each post-second vaccination study day (Day 30, 32, and 36) in relation 
to pre-second vaccination (Day 29) gene expression levels (Section 9.2.2). In addition, differences in DE gene 
responses between Study Arms will be assessed for all possible pairwise study arm comparisons 
(Section 9.2.3). 


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To control for testing multiple genes, the false-discovery rate (FDR) based on the Benjamini-Hochberg 
procedure [12] as implemented in the p.adjust R function will be applied for each model. In all cases, genes 
with an absolute fold change (effect size) of ≥ 1.5 compared to pre-vaccination or between Study Arms and 
FDR-adjusted p-value < 0.05 will be considered significantly DE. Depending on observed magnitude of 
differential expression effects, the pre-specified fold change cut off of 1.5-fold may be revised. The number of 
DE genes will be tabulated by study arm and time point (Table 126 and Table 127). In addition, DE gene 
listings that include gene annotations, log2 fold change estimates, p-values, FDR-adjusted p-values, likelihood 
ratio statistic and average LCPMs will be provided for each post-vaccination day (Table 128). 
Treatment effects in terms of overall fold changes and FDR-adjusted p-values will be summarized for all 
genes that passed the low expression cut off using MA plots and Volcano plots (Figure 9). Overlap in DE 
genes between post-vaccination days and Study Arms will be summarized using UpSet plots as outlined in 
Figure 10 both in terms of overall numbers and separately for up/down- regulated DE genes. 
The following sections describe the negative binomial model parameterization and hypotheses that will be 
tested. 
9.2.1. 
Identification of DE Genes Compared to Pre-First Vaccination 
For each post-first vaccination day and study arm, a negative binomial model will be fit to pre- and post-first 
vaccination read counts. Each model will include coefficients to estimate subject and pre- vs. post-first 
vaccination effects. The subject effects for estimating subject-specific pre-first vaccination levels will be 
added to account for paired samples from the same subject. Effects will be parameterized in the design matrix 
using dummy variables with one variable for each subject and an additional variable for the vaccination 
effect. For vaccination effects, the value will be set to 1 for post-first vaccination and 0 for pre-first 
vaccination observations. For each gene, the statistical significance of the post- vs. pre-vaccination effect will 
be evaluated using a likelihood ratio testing the following hypothesis on the log2 scale: 
H0: µpost-vaccination - µpre-vaccination = 0, H1: µpost-vaccination - µpre-vaccination≠ 0 
Genes with an absolute fold change (effect size) of ≥ 1.5 compared to pre-first vaccination and FDR-adjusted 
p-value < 0.05 will be considered to be significantly DE. 
9.2.2. 
Identification of DE Genes Compared to Pre-Second Vaccination 
For subjects receiving a second vaccination, for each post-second vaccination day (Day 30, 32, and 36) and 
study arm (Study Arms 2 and 3), a negative binomial model will be fit to pre- and post-second vaccination 
read counts across subjects. The model will be parameterized and evaluated as described in Section 9.2.1, 
except using Day 29 as the pre-vaccination reference. 
9.2.3. 
Identification of Genes with Differential Responses Between Study Arms 
For each shared post-first vaccination day (Day 1, 2 and pairwise study arm comparison, a negative binomial 
model will be fit to pre- and post-first vaccination read counts across the respective study arm pair (Study 
Arms 2 vs. 3, Study Arms 2 vs. 1, Arms 3 vs. 1). Each model will include coefficients to estimate subject and 
pre- vs. post-first vaccination effects for each of the two study arms that are being compared. For each shared 
post-first vaccination day, genes that significantly differed in their post vs. pre-first vaccination response 
between study arms will then be identified using contrasts testing the following hypothesis for study arm pair 
X and Y: 
H0: (µpost-vaccination Study Arm X - µpre-vaccination Study Arm X) – (µpost-vaccination Study Arm Y - µpre-vaccination Study Arm Y]) = 0 
H1: (µpost-vaccination Study Arm X - µpre-vaccination Study Arm X) – (µpost-vaccination Study Arm Y - µpre-vaccination Study Arm Y]) ≠ 0 


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Similarly, for all post-second vaccination timepoints, models will be fit to identify DE genes between Study 
Arm 2 vs. 3 except using Day 29 as the pre-vaccination reference. 
Genes with an absolute fold change difference (effect size) of ≥ 1.5 in their response between study arms and 
FDR-adjusted p-value < 0.05 will be considered significantly DE between Study Arms. 
9.3. 
Determination of higher order functional and temporal organization of 
differentially expressed genes 
9.3.1. 
Subject-level Response Heatmaps 
Per-subject baseline log2 fold change patterns for DE genes will be visualized across Study Arms for each 
post-vaccination day and fold change set (post-first vaccination and post-second vaccination) using heatmaps 
(Figure 11, left panel). Genes identified as DE for any of the comparisons described in Section 9.2 will be 
included as part of this analysis. Heatmaps will include subject and gene dendrograms based on uncentered 
Pearson correlation distance and complete linkage hierarchical clustering. 
9.3.2. 
Identification of Co-Expressed Gene Clusters 
Unsupervised multiscale bootstrapping as implemented in the pvclust R package [12] will be carried out to 
identify robust clusters of genes with correlated log2 fold change responses for each fold change set (post-first 
vaccination and post-second vaccination). Bootstrap resampling will be based on uncentered Pearson 
correlation distances between log2 fold change responses in combination with complete linkage clustering. 
Genes identified as DE for any of the comparisons described in Section 9.2 will be included as part of this 
analysis. Clusters with a bootstrap probability > 0.95 and maximum distance between cluster members of 0.5 
(equivalent to minimum uncentered Pearson correlation of 0.5 among all members) will be considered robust 
gene clusters. Resulting robust gene cluster dendrograms will be visualized (Figure 11, middle panel) and 
cluster information will be tabulated (Table 129). In addition, mean log2 fold change for robust gene clusters 
(mean across all member genes in a cluster) and associated 95% bootstrap CIs (1000 replicates each) will be 
visualized by study arm across the respective post-vaccination days (Figure 11, right panel). 
9.3.3. 
Pathway Enrichment Analysis 
Pathway enrichment analysis will be carried out separately for each DE gene set identified (post vs. pre-
vaccination as well as between study arm comparisons) using published pathway information obtained from 
KEGG [13] (KEGG Pathways), MSigDB [14] (MSigDB Reactome Pathways, MSigDB GO Biological 
Process, and MSigDB Immunologic Signatures), and Blood Transcription Modules [15]. Pathway enrichment 
analysis will be conducted using the goseq algorithm [16] to adjust for RNA-Seq-specific gene length bias1. 
Pathways with a FDR-adjusted p-value < 0.05 will be considered significantly enriched. Due to the relatively 
increased number of pathways in the MSigDB Immunologic Signatures set, a more stringent FDR-adjusted  
                                                 
 
 
 
 
1 For RNA-Seq, longer genes tend to obtain higher read counts compared to shorter genes as they occupy a higher proportion of the transcriptome/genome space. As 
such gene expression based on read counts is not only proportional to the actual expression but a combination of read length and actual expression. This results in 
greater statistical power for detecting differential genes if they are long and highly expressed. The effect needs to be addressed in gene set enrichment analysis as 
comparisons are made between sets of genes. Otherwise gene sets that on average contain larger genes will more likely be identified as significantly enriched 
compared to gene sets with on average smaller genes. 


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p-value < 0.01 will be used. Pathway enrichment results including gene set annotation, gene set size, number 
of DE genes in a gene set by up and down-regulation, Jaccard similarity index, p-value, and FDR-adjusted  
p-value will be listed (Table 130). For enriched KEGG pathways, log2 fold change from baseline will be 
overlaid on top of pathway maps and significantly up/and down-regulated genes (DE genes) will be 
highlighted to facilitate visual interpretation (Figure 12, left panel). Overlap between sets of identified 
enriched pathways will be summarized using UpSet plots (similar to Figure 10). Gene set enrichment trends 
across study arms and post-vaccination days will be visualized using heatmaps color-coded by enrichment 
score (-1 x log10(enrichment p-value)) (Figure 12, middle panel) as well as time trend plots of median 
pathway responses (based on median average log2 fold changes of all pathway members) and associated 95% 
bootstrap confidence intervals (CIs) (similar to Figure 11, right panel). 
9.3.4. 
Protein-Protein Interactions Network Analysis 
To further characterize DE genes, experimentally determined protein-protein interaction networks including 
known Influenza A-host interactions centered around DE genes will be generated using data from the IntAct 
database [17]. Networks will be visualized using Cytoscape [18] (Figure 12, right panel). This analysis will be 
performed for DE genes that differed between the H7N9 arms. 
 


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10. 
SAFETY EVALUATIONS 
Summaries and analysis of safety data will be presented for the Safety Analysis Population. These include 
reactogenicity following each study vaccination, unsolicited adverse events, and adverse events of special 
interest including SAEs/MAAEs, new-onset chronic medical conditions, and potentially immune-mediated 
medical conditions. The goal of the safety summaries is to provide information to guide the exploratory 
correlation analysis to identify transcriptomic/proteomic changes that are predictive of certain safety profiles. 
The exploratory correlation analysis will be described in an addendum to this SAP. The following sections 
describe planned baseline and safety summaries. 
10.1. 
Demographic and Other Baseline Characteristics 
Summaries of age, sex, ethnicity, and race will be presented by study arm and overall for all enrolled subjects 
(Table 11 and Table 12). Ethnicity will be categorized as Hispanic or Latino, or not Hispanic and not Latino. 
In accordance with NIH reporting policy, subjects may self-designate as belonging to more than one race or 
may refuse to identify a race, the latter reflected in the CRF as “No” to each racial option. Individual subject 
listings will be presented for all demographics (Listing 6); pre-existing medical conditions (Listing 7); and 
concomitant medications (Listing 15). The number of subjects with pre-existing medical conditions by 
MedDRA System Organ Class and study arm will we provided as shown in Table 13. Summaries of 
medications that were started prior to dosing and continuing at the time of dosing will be presented by WHO 
Drug Level 1 and Level 2 codes and study arm for subjects in the safety population (Table 153). The number 
of doses of study product administered to subjects will be presented by study arm as part of the subject 
disposition table (Table 8). 
10.2. 
Adverse Events 
10.2.1. 
Solicited Events and Symptoms 
Systemic and upper respiratory (local) solicited adverse events were collected pre-vaccination, and systemic 
and local solicited adverse events were collected 30 minutes post-vaccination and then daily for 7 days after 
first vaccination and graded on a scale of 0 (absent), 1 (mild), 2 (moderate) and 3 (severe). The grading scales 
for solicited systemic and local (administration site) adverse events are provided in Table 3, Table 4, Table 5, 
and Table 6. The percentage of subjects and associated exact Clopper–Pearson 95% CI reporting at least one 
solicited adverse event will be summarized for each solicited adverse event, any systemic symptom, any local 
symptom, and any symptoms (Table 132, Table 133, Table 134). In addition, for each systemic and local 
event, any systemic event, any local event, and any solicited event, the maximum severity over 7 days after 
each vaccination will be summarized for the Safety population. For each event the denominator is the number 
of subjects with non-missing data for the specific event (Table 135, Table 136, Table 137). The number of 
subjects reporting a solicited adverse event will be summarized for each day post vaccination in a summary 
table (Table 138, Table 139, Table 140, Table 141, Table 142) and graphically in a bar chart (Figure 13). 
Solicited adverse events by subject will be presented in Listing 10 and Listing 11. 
10.2.2. 
Unsolicited Adverse Events 
When calculating the incidence of unsolicited AEs (i.e., on a per subject basis), each subject will only be 
counted once at the highest severity and/or relationship, and any repetitions of the same AEs within a subject 
will be ignored; the denominator will be the total number of subjects in the safety population. The number and 
percentage and associated 95% CI of subjects reporting at least one unsolicited AE will be summarized by 
MedDRA system organ class and preferred term (Table 143, Table 144, Table 145). Denominators for 


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percentages are the number of subjects who received the study vaccination summarized. In addition, a 
summary of the percentage of subjects by MedDRA system organ class, preferred term, severity, and 
relationship to study product will be provided (Table 146, Table 147, Table 148). Bar charts of the incidence 
of non-serious adverse events by severity and MedDRA system organ class will be generated as shown in 
Figure 14. A listing of all reported AEs by subject sorted by study group, subject ID, and AE Number will be 
presented as specified in  
Listing 12.  
10.3. 
Deaths, Serious Adverse Events and other Significant Adverse Events 
The number and percentage of subjects experiencing adverse events of special interest will be tabulated by 
study arm as presented in Table 131. The following listings will be presented including Subject ID, AE 
Description, AE Onset Date/End Date, Last Vaccination Received/Days Post Vaccination, Reason Reported 
as an SAE, Relationship to Treatment, Alternate Etiology if not Related, Outcome, and Duration of Event 
(days): 
• Deaths and Serious Adverse Events (Table 149) 
• Non-Serious, Unsolicited, Moderate or Severe Adverse Events (Table 150) 
• Adverse Events of Special Interest (Table 151) 
10.4. 
Vital Signs and Physical Evaluations 
Vital signs will be tabulated by study arm in Table 152. A listing of vital signs will be presented (Listing 13). 
Targeted physical examinations will be performed, if indicated, based on a subject’s medical history. A listing 
of physical exam findings will be presented (Listing 14). 


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11. 
TERTIARY AND EXPLORATORY PROTEOMICS ANALYSES 
Analysis of tertiary and exploratory endpoints will be described in separate addenda to this SAP which will be 
finalized prior to analysis of the proteomics data. 


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12. 
EXPLORATORY CORRELATION ANALYSES 
Correlation analysis between the -omics data (transcriptomics and proteomics) and serum antibody/plasma 
cytokines as well as correlated with certain safety profiles will be described in separate addenda to this SAP 
which will be finalized prior to the proteomics data lock. 


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13. 
REPORTING CONVENTIONS 
P-values ≥0.0001 and ≤0.9999 will be reported to 4 decimal places; p-values less than 0.0001 will be reported 
as “<0.0001”; p-values greater than 0.9999 will be reported as “> 0.9999”. The mean, standard deviation, and 
any other statistics other than quantiles, will be reported to one decimal place greater than the original data. 
Quantiles, such as median, Q1, Q3, minimum and maximum will use the same number of decimal places as 
the original data. 


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14. 
REFERENCES 
1. Statistical Package, R. "R: A language and environment for statistical computing." Vienna, Austria: R 
Foundation for Statistical Computing (2009). 
2. Gentleman, Robert C., et al. "Bioconductor: open software development for computational biology 
and bioinformatics." Genome biology 5.10 (2004): R80. 
3. Hubbard, T., et al. "The Ensembl genome database project." Nucleic acids research 30.1 (2002): 38-
41. 
4. Bolger, Anthony M., Marc Lohse, and Bjoern Usadel. "Trimmomatic: a flexible trimmer for Illumina 
sequence data." Bioinformatics 30.15 (2014): 2114-2120. 
5. http://www.bioinformatics.babraham.ac.uk/projects/fastq 
6. Kim, Daehwan, Ben Langmead, and Steven L. Salzberg. "HISAT: a fast spliced aligner with low 
memory requirements." Nature methods 12.4 (2015): 357. 
7. Wang, Liguo, Shengqin Wang, and Wei Li. "RSeQC: quality control of RNA-seq experiments." 
Bioinformatics 28.16 (2012): 2184-2185. 
8. Liao, Yang, Gordon K. Smyth, and Wei Shi. "featureCounts: an efficient general purpose program for 
assigning sequence reads to genomic features." Bioinformatics 30.7 (2014): 923-930. 
9. Robinson, Mark D., and Alicia Oshlack. "A scaling normalization method for differential expression 
analysis of RNA-seq data." Genome Biol 11.3 (2010): R25. 
10. Robinson, Mark D., Davis J. McCarthy, and Gordon K. Smyth. "edgeR: a Bioconductor package for 
differential expression analysis of digital gene expression data." Bioinformatics 26.1 (2010): 139-140. 
11. Benjamini, Yoav, and Yosef Hochberg. "Controlling the false discovery rate: a practical and powerful 
approach to multiple testing." Journal of the Royal Statistical Society. Series B (Methodological) 
(1995): 289-300. 
12. Suzuki, Ryota, and Hidetoshi Shimodaira. "Pvclust: an R package for assessing the uncertainty in 
hierarchical clustering." Bioinformatics 22.12 (2006): 1540-1542. 
13. Kanehisa, Minoru, and Susumu Goto. "KEGG: kyoto encyclopedia of genes and genomes." Nucleic 
acids research 28.1 (2000): 27-30. 
14. Liberzon, Arthur, et al. "Molecular signatures database (MSigDB) 3.0." Bioinformatics 27.12 (2011): 
1739-1740. 
15. Li, Shuzhao, et al. "Molecular signatures of antibody responses derived from a systems biology study 
of five human vaccines." Nature immunology 15.2 (2014): 195.  
16. Young, Matthew D., et al. "Method Gene ontology analysis for RNA-seq: accounting for selection 
bias." Genome Biol 11 (2010): R14. 
17. Kerrien, Samuel, et al. "IntAct—open source resource for molecular interaction data." Nucleic acids 
research 35.suppl_1 (2006): D561-D565. 
18. Shannon, Paul, et al. "Cytoscape: a software environment for integrated models of biomolecular 
interaction networks." Genome research 13.11 (2003): 2498-2504 


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15. 
LISTING OF TABLES, FIGURES, AND LISTINGS  
Table, figure, and listing shells are presented in Appendices 1, 2, and 3. 
 
 


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APPENDICES 


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APPENDIX 1. TABLE MOCK-UPS 
 
LIST OF TABLES 
Table 1: 
Study Design and Sample Size ...................................................................................43 
Table 2: 
Distribution of Protocol Deviations by Category, Type, and Treatment 
Group ..........................................................................................................................44 
Table 3: 
Local (Injection Site) Reactogenicity Grading ...........................................................46 
Table 4: 
Local (Injection Site) Reactogenicity Measurements .................................................46 
Table 5: 
Subjective Systemic Reactogenicity Grading .............................................................46 
Table 6: 
Quantitative Systemic Reactogenicity Grading ..........................................................47 
Table 7: 
Additional Adverse Event Severity Grading ..............................................................47 
Table 8: 
Subject Disposition by Treatment Group – All Enrolled Subjects .............................48 
Table 9: 
Analysis Populations by Treatment Group and Time Point – All Enrolled 
Subjects .......................................................................................................................49 
Table 10: Ineligibility Summary of Screen Failures ...................................................................50 
Table 11: Summary of Categorical Demographic and Baseline Characteristics  by 
Treatment Group - All Enrolled Subjects ...................................................................51 
Table 12: Summary of Continuous Demographic and Baseline Characteristics  by 
Treatment Group - All Enrolled Subjects ...................................................................52 
Table 13: Summary of Subjects with Pre-Existing Medical Conditions by MedDRA 
System Organ Class and Treatment Group ................................................................53 
Table 14: Summaries of Hemagglutination Inhibition Antibody Titer by Study Day 
and Treatment Group in the Intent-to-Treat Population .............................................54 
Table 15: Summaries of Hemagglutination Inhibition Antibody Titer by Study Day 
and Treatment Group in the Per-Protocol Population ................................................54 
Table 16: Summaries of Neutralizing Antibody by Study Day and Treatment Group in 
the Intent-to-Treat Population .....................................................................................54 
Table 17: Summaries of Neutralizing Antibody by Study Day and Treatment Group in 
the Per-Protocol Population ........................................................................................54 
Table 18: t-Test Results for Comparing Day 29 Post-Last Vaccination 
Hemagglutination Inhibition Antibody Titer between Treatment Groups in 
the Intent-to-Treat Population .....................................................................................55 
Table 19: t-Test Results for Comparing Day 29 Post-Last Vaccination 
Hemagglutination Inhibition Antibody Titer Between Treatment Groups in 
the Per-Protocol Population ........................................................................................55 
Table 20: t-Test Results for Comparing Day 29 Post-Last Vaccination Neutralizing 
Antibody Titer Between Treatment Groups in the Intent-to-Treat Population ..........55 


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Table 21: t-Test Results for Comparing Day 29 Post-Last Vaccination Neutralizing 
Antibody Titer Between Treatment Groups in the Per-Protocol Population ..............55 
Table 22: Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination 
Seroconversion based on Hemagglutination Inhibition Antibody Titer 
Between Treatment Groups in the Intent-to-Treat Population ...................................56 
Table 23: Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination 
Seroconversion based on Hemagglutination Inhibition Antibody Titer 
Between Treatment Groups in the Per-Protocol Population .......................................56 
Table 24: Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination 
Seroconversion based on Neutralizing Antibody between Treatment Groups 
in the Intent-to-Treat Population ................................................................................56 
Table 25: Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination 
Seroconversion based on Neutralizing Antibody between Treatment Groups 
in the Per-Protocol Population ....................................................................................56 
Table 26: Fractalkine Concentration by Study Visit Day and Treatment Group in the 
Intent-to-Treat Population ..........................................................................................57 
Table 27: Fractalkine Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................59 
Table 28: GM-CSF Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 29: GM-CSF Concentration by Study Visit Day and Treatment Group in the Per-
Protocol Population ....................................................................................................59 
Table 30: Interferon-gamma Concentration by Study Visit Day and Treatment Group 
in the Intent-To-Treat Population ...............................................................................59 
Table 31: Interferon-gamma Concentration by Study Visit Day and Treatment Group 
in the Per-Protocol Population ....................................................................................59 
Table 32: Interleukin 1 beta Concentration by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................59 
Table 33: Interleukin 1 beta Concentration by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................59 
Table 34: Interleukin 2 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 35: Interleukin 2 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................59 
Table 36: Interleukin 4 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 37: Interleukin 4 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................59 


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Table 38: Interleukin 5 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 39: Interleukin 5 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................59 
Table 40: Interleukin 6 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 41: Interleukin 6 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................59 
Table 42: Interleukin 7 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................59 
Table 43: Interleukin 7 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 44: Interleukin 8 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 
Table 45: Interleukin 8 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 46: Interleukin 10 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 
Table 47: Interleukin 10 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 48: Interleukin-12, p70 Concentration by Study Visit Day and Treatment Group 
in the Intent-To-Treat Population ...............................................................................60 
Table 49: Interleukin-12, p70 Concentration by Study Visit Day and Treatment Group 
in the Per-Protocol Population ....................................................................................60 
Table 50: Interleukin 13 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 
Table 51: Interleukin 13 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 52: IL-17A Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 
Table 53: IL-17A Concentration by Study Visit Day and Treatment Group in the Per-
Protocol Population ....................................................................................................60 
Table 54: Interleukin 21 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 
Table 55: Interleukin 21 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 56: Interleukin 23 Concentration by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................60 


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Table 57: Interleukin 23 Concentration by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................60 
Table 58: Interferon-inducible T Cell Alpha Chemoattractant Concentration by Study 
Visit Day and Treatment Group in the Intent-To-Treat Population ...........................60 
Table 59: Interferon-inducible T Cell Alpha Chemoattractant Concentration by Study 
Visit Day and Treatment Group in the Per-Protocol Population ................................60 
Table 60: Macrophage Inflammatory Protein 1 Alpha Concentration by Study Visit 
Day and Treatment Group in the Intent-To-Treat Population ....................................60 
Table 61: Macrophage Inflammatory Protein 1 Alpha Concentration by Study Visit 
Day and Treatment Group in the Per-Protocol Population .........................................60 
Table 62: Macrophage Inflammatory Protein-1 Beta Concentration by Study Visit Day 
and Treatment Group in the Intent-To-Treat Population ............................................61 
Table 63: Macrophage Inflammatory Protein-1 Beta Concentration by Study Visit Day 
and Treatment Group in the Per-Protocol Population ................................................61 
Table 64: Macrophage Inflammatory Protein-3 Alpha Concentration by Study Visit 
Day and Treatment Group in the Intent-To-Treat Population ....................................61 
Table 65: Macrophage Inflammatory Protein-3 Alpha Concentration by Study Visit 
Day and Treatment Group in the Per-Protocol Population .........................................61 
Table 66: Tumor Necrosis Factor Alpha Concentration by Study Visit Day and 
Treatment Group in the Intent-To-Treat Population ..................................................61 
Table 67: Tumor Necrosis Factor Alpha Concentration by Study Visit Day and 
Treatment Group in the Per-Protocol Population .......................................................61 
Table 68: Interferon Gamma-Induced Protein 10 Concentration by Study Visit Day 
and Treatment Group in the Intent-To-Treat Population ............................................61 
Table 69: Interferon Gamma-Induced Protein 10 Concentration by Study Visit Day 
and Treatment Group in the Per-Protocol Population ................................................61 
Table 70: Interferon-Alpha Concentration by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................61 
Table 71: Interferon-Alpha Concentration by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................61 
Table 72: Interferon-Beta Concentration by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................61 
Table 73: Interferon-Beta Concentration by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................61 
Table 74: Fractalkine Fold Change by Study Visit Day and Study Group in the Intent-
To-Treat Population ....................................................................................................62 
Table 75: Fractalkine Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 


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Table 76: GM-CSF Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 77: GM-CSF Fold Change by Study Visit Day and Treatment Group in the Per-
Protocol Population ....................................................................................................64 
Table 78: Interferon-gamma Fold Change by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................64 
Table 79: Interferon-gamma Fold Change by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................64 
Table 80: Interleukin 1 beta Fold Change by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................64 
Table 81: Interleukin 1 beta Fold Change by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................64 
Table 82: Interleukin 2 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 83: Interleukin 2 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 84: Interleukin 4 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 85: Interleukin 4 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 86: Interleukin 5 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 87: Interleukin 5 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 88: Interleukin 6 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 89: Interleukin 6 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 90: Interleukin 7 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 91: Interleukin 7 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 92: Interleukin 8 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 
Table 93: Interleukin 8 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 94: Interleukin 10 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................64 


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Table 95: Interleukin 10 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................64 
Table 96: Interleukin-12, p70 Fold Change by Study Visit Day and Treatment Group 
in the Intent-To-Treat Population ...............................................................................64 
Table 97: Interleukin-12, p70 Fold Change by Study Visit Day and Treatment Group 
in the Per-Protocol Population ....................................................................................64 
Table 98: Interleukin 13 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................65 
Table 99: Interleukin 13 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................65 
Table 100: IL-17A Fold Change by Study Visit Day and Treatment Group in the Intent-
To-Treat Population ....................................................................................................65 
Table 101: IL-17A Fold Change by Study Visit Day and Treatment Group in the Per-
Protocol Population ....................................................................................................65 
Table 102: Interleukin 21 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................65 
Table 103: Interleukin 21 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................65 
Table 104: Interleukin 23 Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................65 
Table 105: Interleukin 23 Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................65 
Table 106: Interferon-inducible T Cell Alpha Chemoattractant Fold Change by Study 
Visit Day and Treatment Group in the Intent-To-Treat Population ...........................65 
Table 107: Interferon-inducible T Cell Alpha Chemoattractant Fold Change by Study 
Visit Day and Treatment Group in the Per-Protocol Population ................................65 
Table 108: Macrophage Inflammatory Protein 1 Alpha Fold Change by Study Visit Day 
and Treatment Group in the Intent-To-Treat Population ............................................65 
Table 109: Macrophage Inflammatory Protein 1 Alpha Fold Change by Study Visit Day 
and Treatment Group in the Per-Protocol Population ................................................65 
Table 110: Macrophage Inflammatory Protein-1 Beta Fold Change by Study Visit Day 
and Treatment Group in the Intent-To-Treat Population ............................................65 
Table 111: Macrophage Inflammatory Protein-1 Beta Fold Change by Study Visit Day 
and Treatment Group in the Per-Protocol Population ................................................65 
Table 112: Macrophage Inflammatory Protein-3 Alpha Fold Change by Study Visit Day 
and Treatment Group in the Intent-To-Treat Population ............................................65 
Table 113: Macrophage Inflammatory Protein-3 Alpha Fold Change by Study Visit Day 
and Treatment Group in the Per-Protocol Population ................................................65 


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Table 114: Tumor Necrosis Factor Alpha Fold Change by Study Visit Day and 
Treatment Group in the Intent-To-Treat Population ..................................................65 
Table 115: Tumor Necrosis Factor Alpha Fold Change by Study Visit Day and 
Treatment Group in the Per-Protocol Population .......................................................65 
Table 116: Interferon Gamma-Induced Protein 10 Fold Change by Study Visit Day and 
Treatment Group in the Intent-To-Treat Population ..................................................65 
Table 117: Interferon Gamma-Induced Protein 10 Fold Change by Study Visit Day and 
Treatment Group in the Per-Protocol Population .......................................................66 
Table 118: Interferon-Alpha Fold Change by Study Visit Day and Treatment Group in 
the Intent-To-Treat Population ...................................................................................66 
Table 119: Interferon-Alpha Fold Change by Study Visit Day and Treatment Group in 
the Per-Protocol Population ........................................................................................66 
Table 120: Interferon-Beta Fold Change by Study Visit Day and Treatment Group in the 
Intent-To-Treat Population .........................................................................................66 
Table 121: Interferon-Beta Fold Change by Study Visit Day and Treatment Group in the 
Per-Protocol Population ..............................................................................................66 
Table 122: Wilcoxon Signed-Rank Test Results for Comparing Cytokine/Chemokine 
Concentrations between Pre- and Post-Vaccination Time Points in the 
Intent-To-Treat Population .........................................................................................67 
Table 123: Wilcoxon Signed-Rank Test Results for Comparing Cytokine/Chemokine 
Concentrations between Pre- and Post-Vaccination Time Points in the Per-
Protocol Population ....................................................................................................68 
Table 124: Wilcoxon Rank Sum Test Results for Comparing Fold Changes between 
Study Groups in the Intent-To-Treat Population ........................................................69 
Table 125: Wilcoxon Rank Sum Test Results for Comparing Fold Changes between 
Study Groups in the Per-Protocol Population .............................................................70 
Table 126: Genes Differentially Expressed Compared to Pre-Vaccination by Treatment 
Group, Cell Type, and Post-Vaccination Day ............................................................71 
Table 127: Genes Differentially Expressed Between Treatment Groups by Immune Cell 
Type, and Post-Vaccination Day ................................................................................73 
Table 128: Genes Differentially Expressed at Day 2 Compared to Pre-First Vaccination 
(RNA-Seq, Monocytes, Study Arm 1 A/H3N2v, D1) ................................................75 
Table 129: Clusters of Co-Expressed Differential Genes (Monocytes, Days 2-29) .....................76 
Table 130: KEGG Pathways Enriched in Differentially Expressed Genes at Day 2 
Compared to Pre-First Vaccination (RNA-Seq, Monocytes, Study Arm 1 
A/H3N2v, D1) ............................................................................................................77 
Table 131: Number and Percentage of Subjects Experiencing Adverse Events of Special 
Interest ........................................................................................................................78 


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Table 132: Number and Percentage of Subjects Experiencing Solicited Events with 95% 
Confidence Intervals by Symptom, and Treatment Group (Post-First 
Vaccination) ................................................................................................................79 
Table 133: Number and Percentage of Subjects Experiencing Solicited Events with 95% 
Confidence Intervals by Symptom, and Treatment Group (Post-Second 
Vaccination) ................................................................................................................79 
Table 134: Number and Percentage of Subjects Experiencing Solicited Events with 95% 
Confidence Intervals by Symptom, and Treatment Group (Post-Any 
Vaccination) ................................................................................................................79 
Table 135: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Maximum Severity, and Treatment Group (Post-First 
Vaccination) ................................................................................................................80 
Table 136: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Maximum Severity, and Treatment Group (Post-Second 
Vaccination) ................................................................................................................81 
Table 137: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Maximum Severity, and Treatment Group (Post-Any 
Vaccination) ................................................................................................................81 
Table 138: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Severity, and Post-Vaccination Day (Post-First Vaccination, 
Study Arm 1 A/H3N2v, D1) .......................................................................................82 
Table 139: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Severity, and Post-Vaccination Day (Post-First Vaccination, 
Study Arm 2 A/H7N9 + AS03, D1, 29) .....................................................................84 
Table 140: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Severity, and Post-Vaccination Day (Post-First Vaccination, 
Study Arm 3 A/H7N9 + PBS, D1, 29) .......................................................................84 
Table 141: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Severity, and Post-Vaccination Day (Post-Second Vaccination, 
Study Arm 2 A/H7N9 + AS03, D1, D29) ..................................................................84 
Table 142: Number and Percentage of Subjects Experiencing Solicited Events by 
Symptom, Severity, and Post-Vaccination Day (Post-Second Vaccination, 
Study Arm 3 A/H7N9 + PBS, D1, 29) .......................................................................84 
Table 143: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events with 95% Confidence Intervals by MedDRA® System Organ Class 
and Preferred Term, and Treatment Group (Post-First Vaccination) .........................85 
Table 144: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events with 95% Confidence Intervals by MedDRA® System Organ Class 
and Preferred Term, and Treatment Group (Post-Second Vaccination) .....................85 


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Table 145: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events with 95% Confidence Intervals by MedDRA® System Organ Class 
and Preferred Term, and Treatment Group (Post-Any Vaccination) .........................85 
Table 146: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA® System Organ Class and Preferred Term, Maximum 
Severity, and Relationship (Study Arm 1 A/H3N2v, D1) ..........................................86 
Table 147: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA® System Organ Class and Preferred Term, Maximum 
Severity, Relationship (Study Arm 2 A/H7N9 + AS03, D1, 29) ...............................86 
Table 148: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA® System Organ Class and Preferred Term, Maximum 
Severity, Relationship (Study Arm 3 A/H7N9 + PBS, D1, 29) .................................86 
Table 149: Listing of Serious Adverse Events .............................................................................87 
Table 150: Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events ...............87 
Table 151: Listing of Adverse Events of Special Interest ............................................................88 
Table 152: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group – Any Assessment ...........................................................................................89 
Table 153: Number and Percentage of Subjects with Prior and Concurrent Medications 
by WHO Drug Classification and Treatment Group ..................................................91 
 


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9.1 
Overall Study Design and Plan Description 
Table 1: 
Study Design and Sample Size 
Subjects 
First Study Vaccination 
(Day 1) 
Second Study Vaccination 
(Day 29±2 days) 
Study Arm Label 
10 
15 µg A/H3N2v* 
N/A 
Study Arm 1  
A/H3N2v, D1  
10 
3.75 µg A/H7N9 + AS03 
3.75 µg A/H7N9 + AS03 
Study Arm 2  
A/H7N9 + AS03 
D1, 29 
10 
3.75 µg A/H7N9+PBS 
3.75 µg A/H7N9 + PBS  
Study Arm 3  
A/H7N9 + PBS, D1, 29 
Total N=30 vaccinated subjects 
*Unadjuvanted


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10.2 
Protocol Deviations 
Table 2: 
Distribution of Protocol Deviations by Category, Type, and Treatment Group 
Category 
Deviation Type 
Study Arm 1 
A/H3N2v, D1 
(N=X) 
Study Arm 2 
A/H7N9 + AS03, 
D1, 29 (N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 
29 (N=X) 
All Subjects 
(N=X) 
No. 
of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. 
of 
Dev. 
Eligibility/enrollment 
Any type 
 
 
 
 
 
 
 
 
 
Did not meet inclusion criterion 
x 
x 
 
 
x 
x 
x 
x 
 
Met exclusion criterion 
 
 
 
 
 
 
 
 
 
ICF not signed prior to study 
procedures 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
Treatment 
administration 
schedule 
Any type 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
Missed treatment administration 
 
 
 
 
 
 
 
 
 
Delayed treatment 
administration 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
Follow-up visit 
schedule 
Any type 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
Protocol 
procedure/assessment 
Any type 
 
 
 
 
 
 
 
 
 
Incorrect version of ICF signed 
 
 
 
 
 
 
 
 
 
Blood not collected 
 
 
 
 
 
 
 
 
 
Urine not collected 
 
 
 
 
 
 
 
 
 
Other specimen not collected 
 
 
 
 
 
 
 
 
 
Too few aliquots obtained 
 
 
 
 
 
 
 
 
 
Specimen result not obtained 
 
 
 
 
 
 
 
 
 
Required procedure not 
conducted 
 
 
 
 
 
 
 
 
 
Required procedure done 
incorrectly 
 
 
 
 
 
 
 
 
 
Study product temperature 
excursion 
 
 
 
 
 
 
 
 


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Table 2: Distribution of Protocol Deviations by Category, Type, and Treatment Group (continued) 
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Category 
Deviation Type 
Study Arm 1 
A/H3N2v, D1 
(N=X) 
Study Arm 2 
A/H7N9 + AS03, 
D1, 29 (N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 
29 (N=X) 
All Subjects 
(N=X) 
No. 
of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. of 
Dev. 
No. of 
Subj. 
No. 
of 
Dev. 
 
Specimen temperature 
excursion 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
Treatment 
administration 
Any type 
 
 
 
 
 
 
 
 
 
Required procedure done 
incorrectly 
 
 
 
 
 
 
 
 
 
Study product temperature 
excursion 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
Blinding 
policy/procedure 
Any type 
 
 
 
 
 
 
 
 
 
Treatment unblinded 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
N=Number of enrolled subjects. 
 
 


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12.2.2 Displays of Adverse Events 
Solicited Adverse Event Grading Scale 
Table 3: 
Local (Injection Site) Reactogenicity Grading 
Local (Injection Site) 
Reaction  
Mild (Grade 1)  
Moderate (Grade 2)  
Severe (Grade 3) 
Pain – experienced 
without touching the 
injection site 
(spontaneous 
discomfort) 
Subject is aware of 
pain, but it does not 
interfere with daily 
activity, and no pain 
medication is taken 
There is interference 
with daily activity or it 
requires repeated use 
of a non-narcotic pain 
reliever for >24 hours 
Pain prevents daily 
activity or requires any 
use of a narcotic pain 
reliever 
Tenderness – hurts 
only when injection 
site is touched or the 
arm is moved 
The area immediately 
surrounding the 
injection site hurts only 
when touched or with 
arm motion, and it does 
not interfere with daily 
activity 
The area immediately 
surrounding the 
injection site hurts 
when touched or with 
arm motion, and it 
interferes with daily 
activity 
The area immediately 
surrounding the injection 
site hurts when touched 
or with arm motion, and it 
prevents daily activity 
Pruritus (Itching)  
Does not interfere with 
daily activity 
Interferes with daily 
activity  
Prevents daily activity 
Ecchymosis 
(Bruising)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity  
Prevents daily activity 
Erythema (Redness)*  
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Induration 
(Hardness)/Swelling* 
Does not interfere with 
daily activity 
Interferes with daily 
activity  
Prevents daily activity 
* Will also be measured in mm but size will not be used as halting criteria. 
 
Table 4: 
Local (Injection Site) Reactogenicity Measurements 
Local (Injection Site) Reaction  
Small  
Medium  
Large 
Ecchymosis (Bruising)*  
<20 mm  
20 mm – 50 mm  
>50 mm 
Erythema (Redness)*  
<20 mm  
20 mm – 50 mm  
>50 mm 
Induration (Hardness)/Swelling*  
<20 mm  
20 mm – 50 mm  
>50 mm 
* Will not be used as halting criteria. 
 
Table 5: 
Subjective Systemic Reactogenicity Grading 
Systemic (Subjective)  
Mild (Grade 1)  
Moderate (Grade 2)  
Severe (Grade 3) 
Feverishness 
(Chills/Shivering/Sweating) 
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
Fatigue (Tiredness)  
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
Malaise (General Unwell 
Feeling) 
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
Myalgia (Body 
Aches/Muscular Pain)* 
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 


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Systemic (Subjective)  
Mild (Grade 1)  
Moderate (Grade 2)  
Severe (Grade 3) 
Arthralgia (Joint Pain)*  
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
Headache  
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
Nausea  
No interference 
with daily activity 
Some interference 
with daily activity 
Significant interference, 
prevents daily activity 
* Not at injection site. 
 
Table 6: 
Quantitative Systemic Reactogenicity Grading 
Systemic (Quantitative)  
Mild (Grade 1)  
Moderate (Grade 2)  
Severe (Grade 3) 
Fever* - oral† 
38.0°C – 38.4°C 
100.4°F – 101.1°F 
38.5°C – 38.9°C 
101.2°F – 102.0°F 
>38.9°C 
>102.0°F 
Note: Oral temperature assessed on Day 1 prior to the first study vaccination will be considered as baseline. 
* A fever can be considered not related to the study product if an alternative etiology can be documented. 
† Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral 
temperature. 
 
Table 7: 
Additional Adverse Event Severity Grading 
Physiologic Parameter 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Bradycardia - beats per minute 
45 – 49 
40 – 44 
<40 
Tachycardia - beats per minute 
116 – 130 
131 – 155 
>155 
Hypotension (systolic) mm Hg 
80 – 84 
75 – 79 
<75 
Hypotension (diastolic) mm Hg 
50 – 54 
45 – 49 
<45 
Hypertension (systolic) mm Hg 
151 – 155 
156 – 160 
>160 
Hypertension (diastolic) mm Hg 
96 – 100 
101 – 105 
>105 


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14.1 
Description of Study Subjects 
14.1.1 Disposition of Subjects 
Table 8: 
Subject Disposition by Treatment Group – All Enrolled Subjects 
Subject 
Disposition  
Study Arm 1 A/H3N2v, 
D1 (N=X) 
Study Arm 2 A/H7N9 + AS03, 
D1, 29 (N=X) 
Study Arm 3 A/H7N9 + PBS, 
D1, 29 (N=X) 
n 
% 
n 
% 
n 
% 
Screened 
x 
-- 
x 
-- 
x 
-- 
Enrolled/Randomized 
x 
100 
x 
100 
x 
100 
Received One Treatment 
x 
xx 
x 
xx 
x 
xx 
Received All Scheduled Treatments a 
x 
xx 
x 
xx 
x 
xx 
Completed Day 2 Omics Blood Draw 
x 
xx 
x 
xx 
x 
xx 
Completed Day 4 Omics Blood Draw 
x 
xx 
x 
xx 
x 
xx 
Completed Day 8 Omics Blood Draw 
x 
xx 
n/a 
n/a 
n/a 
n/a 
Completed Day 29 Omics Blood Draw 
n/a 
n/a 
x 
xx 
x 
xx 
Completed Day 30 Omics Blood Draw 
n/a 
n/a 
x 
xx 
x 
xx 
Completed Day 32 Omics Blood Draw 
n/a 
n/a 
x 
xx 
x 
xx 
Completed Day 36 Omics Blood Draw 
n/a 
n/a 
x 
xx 
x 
xx 
Completed Follow-up (Study Day 29)a 
 
 
n/a 
n/a 
n/a 
n/a 
Completed Follow-up (Study Day 366)a 
n/a 
n/a 
x 
xx 
x 
xx 
Completed Per Protocolb 
x 
xx 
x 
xx 
x 
xx 
N=number of enrolled subjects 
a Refer to Listing of Early Terminations or Discontinued Subjects for reasons subjects discontinued or terminated early. 
b Refer to Listing of Analysis Populations Exclusions for reasons subjects are excluded from the Analysis populations. 
 
 


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Table 9: 
Analysis Populations by Treatment Group and Time Point – All Enrolled Subjects 
Analysis 
Populations 
Time Point 
 
Reason Subjects Excluded 
Study Arm 1 
(A/H3N2v, D1) 
(N=X) 
Study Arm 1 
(A/H3N2v, D1) 
(N=X) 
Group 3  
3.75 μg A/H7N9 
+ PBS 
(D1, 29) (N=X) 
 
n 
% 
n 
% 
% 
n 
Safety Analysis 
Population 
Any Time Point 
Any Reason 
x 
xx 
x 
xx 
x 
xx 
 
 
[Reason 1, for example: Did not meet 
eligibility criteria] 
 
 
 
 
 
 
 
 
[Reason 2] 
 
 
 
 
 
 
 
 
[Reason 3] 
 
 
 
 
 
 
 
 
[Reason 4] 
 
 
 
 
 
 
ITT Analysis 
Population 
Any Time Point 
Any Reason 
 
 
 
 
 
 
 
 
[Reason 1] 
 
 
 
 
 
 
 
 
[Reason 2] 
 
 
 
 
 
 
PP Analysis 
Population 
Any Time Point 
Any Reason 
 
 
 
 
 
 
 
 
[Reason 1] 
 
 
 
 
 
 
 
 
[Reason 2] 
 
 
 
 
 
 
 
Day 29 (Post-
Vaccination 1) 
Any Reason 
 
 
 
 
 
 
 
 
[Reason 1, for example: Did not meet 
eligibility criteria] 
 
 
 
 
 
 
 
 
[Reason 2] 
 
 
 
 
 
 
 
 
[Reason 3] 
 
 
 
 
 
 
N=Number of enrolled subjects. 
 


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Table 10: 
Ineligibility Summary of Screen Failures 
Inclusion/Exclusion Category 
Inclusion/Exclusion Criterion 
na 
%b 
Inclusion and Exclusion 
Number of subjects failing any eligibility criterion 
x 
xx 
Inclusion 
Any inclusion criterion 
x 
xx 
 
[inclusion criterion 1] 
x 
xx 
 
[inclusion criterion 2] 
x 
xx 
 
[inclusion criterion 3] 
x 
xx 
Exclusion 
Any exclusion criterion 
x 
xx 
 
[exclusion criterion 1] 
x 
xx 
 
[exclusion criterion 2] 
x 
xx 
 
[exclusion criterion 3] 
x 
xx 
Eligible but not enrolled 
 
x 
xx 
a More than one criterion may be marked per subject. 
b Denominator for percentages is the total number of screen failures. 
 
 


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14.1.2 Demographic Data by Study Group 
Table 11: 
Summary of Categorical Demographic and Baseline Characteristics  
by Treatment Group - All Enrolled Subjects 
Variable 
Characteristic 
Study Arm 1 
A/H3N2v, D1 (N=X) 
Study Arm 2 
A/H7N9 + AS03, 
D1, 29 (N=X) 
Study Arm 3 
A/H7N9 + 
PBS, D1, 29 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
Sex  
Male 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Female 
 
 
 
 
 
 
 
Ethnicity  
Not Hispanic or Latino 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Hispanic or Latino 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
Unknown 
 
 
 
 
 
 
 
Race  
American Indian or Alaska Native 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Asian 
 
 
 
 
 
 
 
 
Native Hawaiian or Other Pacific 
Islander 
 
 
 
 
 
 
 
 
Black or African American 
 
 
 
 
 
 
 
 
White 
 
 
 
 
 
 
 
 
Multi-Racial 
 
 
 
 
 
 
 
 
Unknown 
 
 
 
 
 
 
 
N=Number of enrolled subjects. 
 
 


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Table 12: 
Summary of Continuous Demographic and Baseline Characteristics  
by Treatment Group - All Enrolled Subjects 
Variable 
Statistic 
Study Arm 1 
A/H3N2v, D1 
(N=X) 
Study Arm 2 A/H7N9 
+ AS03, D1, 29 (N=X) 
Study Arm 3 
A/H7N9 + PBS, 
D1, 29 (N=X) 
All Subjects 
(N=X) 
Age (units) 
Mean 
x x 
x.x 
x.x 
x x 
 
Standard Deviation 
x x 
x.x 
x.x 
x x 
 
Median 
x 
x 
x 
x 
 
Minimum 
x 
x 
x 
x 
 
Maximum 
x 
x 
x 
x 
BMI 
Mean 
x x 
x.x 
x.x 
x x 
 
Standard Deviation 
x x 
x.x 
x.x 
x x 
 
Median 
x 
x 
x 
x 
 
Minimum 
x 
x 
x 
x 
 
Maximum 
x 
x 
x 
x 
N=Number of enrolled subjects. 
 
 


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14.1.3 Prior and Concurrent Medical Conditions 
Table 13: 
Summary of Subjects with Pre-Existing Medical Conditions by MedDRA System Organ Class and Treatment Group 
MedDRA System Organ Class 
Study Arm 1 
 A/H3N2v, D1 (N=X) 
Study Arm 2 
 A/H7N9 + AS03, D1, 29 
(N=X) 
Study Arm 3  
A/H7N9 + PBS, D1, 29 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
[SOC 1] 
 
 
 
 
 
 
 
[SOC 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N=Number of subjects in the Safety Population 
n = Number of subjects reporting medical history within the specified SOC. A subject is only counted once per SOC. 
 
 


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14.2 
Immunogenicity Data 
14.2.1 Serum Antibody Data 
Table 14: 
Summaries of Hemagglutination Inhibition Antibody Titer by Study Day and Treatment 
Group in the Intent-to-Treat Population 
Time Point 
Statistic 
Study Arm 1 
A/H3N2v, D1 (N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 
29 (N=X) 
Study Arm 3 A/H7N9 
+ PBS, D1, 29 (N=X) 
Day 1 (Pre-Vaccination 1) 
n 
x 
x 
x 
  
GMT (95% CI) 
x x (x.x,x x) 
x.x (x x,x.x) 
x x (x x,x.x) 
Day 29 (Post-Vaccination 
1) 
n 
x 
x 
x 
  
GMT (95% CI) 
x x (x.x,x x) 
x.x (x x,x.x) 
x x (x x,x.x) 
  
Seroconversion n/N (%) [95% 
CI] 
xx/xx (xx%) [xx, xx] 
xx/xx (xx%) [xx, xx] 
xx/xx (xx%) [xx, xx] 
Day 29 (Post-Vaccination 
2) 
n 
n/a 
x 
x 
  
GMT (95% CI) 
n/a 
x.x (x x,x.x) 
x x (x x,x.x) 
  
Seroconversion n/N (%) [95% 
CI] 
n/a 
xx/xx (xx%) [xx, xx] 
xx/xx (xx%) [xx, xx] 
N = Number of subjects in the Intent-to-Treat Population  
n = Number of subjects with available results 
GMT = Geometric mean titer 
Seroconversion is defined as having a pre -vaccination titer <1:10 and a post - vaccination titer ≥1:40 or a pre - vaccination titer ≥1:10 and a 
minimum four -fold rise in post-vaccination antibody titer 
 
Tables with similar format: 
Table 15: 
Summaries of Hemagglutination Inhibition Antibody Titer by Study Day and Treatment 
Group in the Per-Protocol Population 
Table 16: 
Summaries of Neutralizing Antibody by Study Day and Treatment Group in the Intent-
to-Treat Population 
Table 17: 
Summaries of Neutralizing Antibody by Study Day and Treatment Group in the Per-
Protocol Population 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 18: 
t-Test Results for Comparing Day 29 Post-Last Vaccination Hemagglutination Inhibition 
Antibody Titer between Treatment Groups in the Intent-to-Treat Population 
Statistic 
Study Arm 2  
A/H7N9 + AS03, D1, 29 (N=X) 
vs. Study Arm 3 A/H7N9 + 
PBS, D1, 29 (N=X) 
Study Arm 2   
A/H7N9 + AS03, D1, 29 (N=X) 
vs. Study Arm 1  
A/H3N2v, D1 (N=X) 
Study Arm 3 A/H7N9 + PBS, 
D1, 29 (N=X) vs. 
Study Arm 1  
A/H3N2v, D1 (N=X) 
n 
xx 
xx 
xx 
GMT Ratio (95% CI) 
x xx (x xx,x xx) 
x xx (x xx,x xx) 
x.xx (x.xx,x.xx) 
t-statistic 
x xxx 
x.xxx 
x xxx 
P-value 
x xxx 
x.xxx 
x xxx 
N = Number of subjects in the Intent-to-Treat Population 
n = Number of subjects with available results 
P-value =  two-sided Welch’s or standard t-test P-value 
 
Tables with similar format: 
Table 19: 
t-Test Results for Comparing Day 29 Post-Last Vaccination Hemagglutination Inhibition 
Antibody Titer Between Treatment Groups in the Per-Protocol Population 
Table 20: 
t-Test Results for Comparing Day 29 Post-Last Vaccination Neutralizing Antibody Titer 
Between Treatment Groups in the Intent-to-Treat Population 
Table 21: 
t-Test Results for Comparing Day 29 Post-Last Vaccination Neutralizing Antibody Titer 
Between Treatment Groups in the Per-Protocol Population 
 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 22: 
Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination Seroconversion based on Hemagglutination Inhibition 
Antibody Titer Between Treatment Groups in the Intent-to-Treat Population 
  
Study Arm 2  
A/H7N9 + AS03, D1, 29 (N=X) vs. Study 
Arm 3 A/H7N9 + PBS, D1, 29 (N=X) 
Study Arm 2   
A/H7N9 + AS03, D1, 29 (N=X) vs. Study 
Arm 1  
A/H3N2v, D1 (N=X) 
Study Arm 3 A/H7N9 + PBS, D1, 29 (N=X) 
vs. 
Study Arm 1  
A/H3N2v, D1 (N=X) 
n 
xx 
xx 
xx 
Seroconversion n/N (%) [95% CI] 
xx/xx (xx%) [xx, xx] 
xx/xx (xx%) [xx, xx] 
xx/xx (xx%) [xx, xx] 
Exact Odds Ratio(95% CI) 
x.xxx 
x xxx 
x.xxx 
p-value 
x.xxx 
x xxx 
x.xxx 
N = Number of subjects in the Intent-to-Treat Population 
n = Number of subjects with available results 
P-value = two-sided Fisher’s Exact test P-vale. 
 
Tables with similar format: 
Table 23: 
Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination Seroconversion based on Hemagglutination Inhibition 
Antibody Titer Between Treatment Groups in the Per-Protocol Population 
Table 24: 
Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination Seroconversion based on Neutralizing Antibody 
between Treatment Groups in the Intent-to-Treat Population 
Table 25: 
Fisher’s Exact Test Results for Comparing Day 29 Post-Last Vaccination Seroconversion based on Neutralizing Antibody 
between Treatment Groups in the Per-Protocol Population 


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14.2.2 Plasma Cytokine/Chemokine Data 
Table 26: 
Fractalkine Concentration by Study Visit Day and Treatment Group in the Intent-to-Treat Population 
Time Point 
Statistic 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Day 1 (Pre-Vaccination 1) 
n 
x 
x 
x 
 
Min 
x xx 
x xx 
x xx 
 
Q1 
x x 
x.x 
x x 
 
Median (95% CI) 
x x (x x - x.x) 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
x x 
x.x 
x x 
 
Max 
x xx 
x xx 
x xx 
Day 2 (Post-Vaccination 1) 
n 
x 
x 
x 
 
Min 
x xx 
x xx 
x xx 
 
Q1 
x x 
x.x 
x x 
 
Median (95% CI) 
x x (x x - x.x) 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
x x 
x.x 
x x 
 
Max 
x xx 
x xx 
x xx 
Day 4 (Post-Vaccination 1) 
n 
x 
x 
x 
 
Min 
x xx 
x xx 
x xx 
 
Q1 
x x 
x.x 
x x 
 
Median (95% CI) 
x x (x x - x.x) 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
x x 
x.x 
x x 
 
Max 
x xx 
x xx 
x xx 
Day 8 (Post-Vaccination 1) 
n 
x 
n/a 
n/a 
 
Min 
x xx 
n/a 
n/a 
 
Q1 
x x 
n/a 
n/a 
 
Median (95% CI) 
x x (x x - x.x) 
n/a 
n/a 
 
Q3 
x x 
n/a 
n/a 


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Time Point 
Statistic 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
 
Max 
x xx 
n/a 
n/a 
Day 29 (Post-Vaccination 1) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
Day 2 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
Day 4 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
Day 8 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Time Point 
Statistic 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
N= Number of subjects in the Intent-to-Treat analysis population 
n= Number of subjects with valid results 
Min=Minimum 
Q1= First quartile 
Q3=Third quartile 
Max=Maximum 
The 95% CI of the median is based on the bootstrap method. 
 
Tables with similar format: 
Table 27: 
Fractalkine Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 28: 
GM-CSF Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 29: 
GM-CSF Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 30: 
Interferon-gamma Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 31: 
Interferon-gamma Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 32: 
Interleukin 1 beta Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 33: 
Interleukin 1 beta Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 34: 
Interleukin 2 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 35: 
Interleukin 2 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 36: 
Interleukin 4 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 37: 
Interleukin 4 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 38: 
Interleukin 5 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 39: 
Interleukin 5 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 40: 
Interleukin 6 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 41: 
Interleukin 6 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 42: 
Interleukin 7 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 43: 
Interleukin 7 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 44: 
Interleukin 8 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 45: 
Interleukin 8 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 46: 
Interleukin 10 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 47: 
Interleukin 10 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 48: 
Interleukin-12, p70 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 49: 
Interleukin-12, p70 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 50: 
Interleukin 13 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 51: 
Interleukin 13 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 52: 
IL-17A Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 53: 
IL-17A Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 54: 
Interleukin 21 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 55: 
Interleukin 21 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 56: 
Interleukin 23 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 57: 
Interleukin 23 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 58: 
Interferon-inducible T Cell Alpha Chemoattractant Concentration by Study Visit Day and Treatment Group in the Intent-To-
Treat Population 
Table 59: 
Interferon-inducible T Cell Alpha Chemoattractant Concentration by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 60: 
Macrophage Inflammatory Protein 1 Alpha Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 61: 
Macrophage Inflammatory Protein 1 Alpha Concentration by Study Visit Day and Treatment Group in the Per-Protocol 
Population 


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Table 62: 
Macrophage Inflammatory Protein-1 Beta Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 63: 
Macrophage Inflammatory Protein-1 Beta Concentration by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 64: 
Macrophage Inflammatory Protein-3 Alpha Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 65: 
Macrophage Inflammatory Protein-3 Alpha Concentration by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 66: 
Tumor Necrosis Factor Alpha Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 67: 
Tumor Necrosis Factor Alpha Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 68: 
Interferon Gamma-Induced Protein 10 Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 69: 
Interferon Gamma-Induced Protein 10 Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 70: 
Interferon-Alpha Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 71: 
Interferon-Alpha Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 72: 
Interferon-Beta Concentration by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 73: 
Interferon-Beta Concentration by Study Visit Day and Treatment Group in the Per-Protocol Population 
 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 74: 
Fractalkine Fold Change by Study Visit Day and Study Group in the Intent-To-Treat Population 
Time Point 
Statistic 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Day 2 (Post-Vaccination 1) 
n 
x 
x 
x 
 
Min 
x xx 
x xx 
x xx 
 
Q1 
x x 
x.x 
x x 
 
Median (95% CI) 
x x (x x - x.x) 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
x x 
x.x 
x x 
 
Max 
x xx 
x xx 
x xx 
Day 4 (Post-Vaccination 1) 
n 
x 
x 
x 
 
Min 
x xx 
x xx 
x xx 
 
Q1 
x x 
x.x 
x x 
 
Median (95% CI) 
x x (x x - x.x) 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
x x 
x.x 
x x 
 
Max 
x xx 
x xx 
x xx 
Day 8 (Post-Vaccination 1) 
n 
x 
n/a 
n/a 
 
Min 
x xx 
n/a 
n/a 
 
Q1 
x x 
n/a 
n/a 
 
Median (95% CI) 
x x (x x - x.x) 
n/a 
n/a 
 
Q3 
x x 
n/a 
n/a 
 
Max 
x xx 
n/a 
n/a 
Day 29 (Post-Vaccination 1) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Time Point 
Statistic 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Day 2 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
Day 4 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
Day 8 (Post-Vaccination 2) 
n 
n/a 
x 
x 
 
Min 
n/a 
x xx 
x xx 
 
Q1 
n/a 
x.x 
x x 
 
Median (95% CI) 
n/a 
x.x (x x - x.x) 
x x (x x - x x) 
 
Q3 
n/a 
x.x 
x x 
 
Max 
n/a 
x xx 
x xx 
N= Number of subjects in the Intent-to-Treat population 
n= Number of subjects with valid results 
Min=Minimum 
Q1= First quartile 
Q3=Third quartile 
Max=Maximum 
The 95% CI of the median is based on the bootstrap method. 
 
 
 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Tables with similar format: 
Table 75: 
Fractalkine Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 76: 
GM-CSF Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 77: 
GM-CSF Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 78: 
Interferon-gamma Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 79: 
Interferon-gamma Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 80: 
Interleukin 1 beta Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 81: 
Interleukin 1 beta Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 82: 
Interleukin 2 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 83: 
Interleukin 2 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 84: 
Interleukin 4 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 85: 
Interleukin 4 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 86: 
Interleukin 5 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 87: 
Interleukin 5 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 88: 
Interleukin 6 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 89: 
Interleukin 6 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 90: 
Interleukin 7 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 91: 
Interleukin 7 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 92: 
Interleukin 8 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 93: 
Interleukin 8 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 94: 
Interleukin 10 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 95: 
Interleukin 10 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 96: 
Interleukin-12, p70 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 97: 
Interleukin-12, p70 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 98: 
Interleukin 13 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 99: 
Interleukin 13 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 100: 
IL-17A Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 101: 
IL-17A Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 102: 
Interleukin 21 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 103: 
Interleukin 21 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 104: 
Interleukin 23 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 105: 
Interleukin 23 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 106: 
Interferon-inducible T Cell Alpha Chemoattractant Fold Change by Study Visit Day and Treatment Group in the Intent-To-
Treat Population 
Table 107: 
Interferon-inducible T Cell Alpha Chemoattractant Fold Change by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 108: 
Macrophage Inflammatory Protein 1 Alpha Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 109: 
Macrophage Inflammatory Protein 1 Alpha Fold Change by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 110: 
Macrophage Inflammatory Protein-1 Beta Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 111: 
Macrophage Inflammatory Protein-1 Beta Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 112: 
Macrophage Inflammatory Protein-3 Alpha Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat 
Population 
Table 113: 
Macrophage Inflammatory Protein-3 Alpha Fold Change by Study Visit Day and Treatment Group in the Per-Protocol 
Population 
Table 114: 
Tumor Necrosis Factor Alpha Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 115: 
Tumor Necrosis Factor Alpha Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 116: 
Interferon Gamma-Induced Protein 10 Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 117: 
Interferon Gamma-Induced Protein 10 Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 118: 
Interferon-Alpha Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 119: 
Interferon-Alpha Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
Table 120: 
Interferon-Beta Fold Change by Study Visit Day and Treatment Group in the Intent-To-Treat Population 
Table 121: 
Interferon-Beta Fold Change by Study Visit Day and Treatment Group in the Per-Protocol Population 
 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 122: 
Wilcoxon Signed-Rank Test Results for Comparing Cytokine/Chemokine Concentrations between Pre- and Post-Vaccination 
Time Points in the Intent-To-Treat Population 
  
  
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Cytokine 
Comparison 
Wilcoxon Statistic  
(P-value) 
Wilcoxon Statistic  
(P-value) 
Wilcoxon Statistic  
(P-value) 
Fractalkine 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 8 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
n/a 
n/a 
Fractalkine 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 8 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
n/a 
n/a 
GM-CSF 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 8 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x.xxx) 
n/a 
n/a 
Interferon-gamma 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
n/a 
x xxx (x xxx) 
x.xxx (x xxx) 
Colored in blue= p-value  < 0.05 


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Continue for the remaining cytokine/chemokines IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-23, ITAC, MIP 1a, MIP1b, MIP3a, TNF-alpha, IP-10, IFNa, IFNb 
Table with similar format: 
Table 123: 
Wilcoxon Signed-Rank Test Results for Comparing Cytokine/Chemokine Concentrations between Pre- and Post-Vaccination 
Time Points in the Per-Protocol Population 
 


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Table 124: 
Wilcoxon Rank Sum Test Results for Comparing Fold Changes between Study Groups in the Intent-To-Treat Population 
  
  
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Cytokine 
Comparison 
Wilcoxon Statistic  
(p-value) 
Wilcoxon Statistic  
(p-value) 
Wilcoxon Statistic  
(p-value) 
Fractalkine 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Fractalkine 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
n/a 
n/a 
Fractalkine 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Fractalkine 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Fractalkine 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
GM-CSF 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
GM-CSF 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
n/a 
n/a 
GM-CSF 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
GM-CSF 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
GM-CSF 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Interferon-gamma 
Day 2 (Post -Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 4 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
x xxx (x xxx) 
x.xxx (x xxx) 
Interferon-gamma 
Day 29 (Post-Vaccination 1) vs. Pre-Vaccination 1 
x xxx (x xxx) 
n/a 
n/a 
Interferon-gamma 
Day 2 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Interferon-gamma 
Day 4 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Interferon-gamma 
Day 8 (Post-Vaccination 2) vs. Pre-Vaccination 2 
x xxx (x xxx) 
n/a 
n/a 
Colored in blue= p-value < 0.05 
Continue for the remaining cytokine/chemokines IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-23, ITAC, MIP 1a, MIP1b, MIP3a, TNF-alpha, IP-10, IFNa, IFNb 


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Table with similar format: 
Table 125: 
Wilcoxon Rank Sum Test Results for Comparing Fold Changes between Study Groups in the Per-Protocol Population 
 


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14.3 
Transcriptomics Data 
Table 126: 
Genes Differentially Expressed Compared to Pre-Vaccination by Treatment Group, Cell Type, and Post-Vaccination Day 
 
 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Time Point 
Cell Type 
DE Genes [#] 
DE Genes [#] 
DE Genes [#] 
Day 2 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
xxx 
xxx 
 
Dendritic cells 
xxx 
xxx 
xxx 
 
Neutrophils 
xxx 
xxx 
xxx 
 
NK-cells 
xxx 
xxx 
xxx 
 
T-cells 
xxx 
xxx 
xxx 
 
B-cells 
xxx 
xxx 
xxx 
Day 4 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
xxx 
xxx 
 
Dendritic cells 
xxx 
xxx 
xxx 
 
Neutrophils 
xxx 
xxx 
xxx 
 
NK-cells 
xxx 
xxx 
xxx 
 
T-cells 
xxx 
xxx 
xxx 
 
B-cells 
xxx 
xxx 
xxx 
Day 8 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
n/a 
n/a 
 
Dendritic cells 
xxx 
n/a 
n/a 
 
Neutrophils 
xxx 
n/a 
n/a 
 
NK-cells 
xxx 
n/a 
n/a 
 
T-cells 
xxx 
n/a 
n/a 
 
B-cells 
xxx 
n/a 
n/a 
Day 29 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
n/a 
xxx 
xxx 
 
Dendritic cells 
n/a 
xxx 
xxx 
 
Neutrophils 
n/a 
xxx 
xxx 
 
NK-cells 
n/a 
xxx 
xxx 


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Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
 
T-cells 
n/a 
xxx 
xxx 
 
B-cells 
n/a 
xxx 
xxx 
Day 2 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
n/a 
xxx 
xxx 
 
Dendritic cells 
n/a 
xxx 
xxx 
 
Neutrophils 
n/a 
xxx 
xxx 
 
NK-cells 
n/a 
xxx 
xxx 
 
T-cells 
n/a 
xxx 
xxx 
 
B-cells 
n/a 
xxx 
xxx 
Day 4 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
n/a 
xxx 
xxx 
 
Dendritic cells 
n/a 
xxx 
xxx 
 
Neutrophils 
n/a 
xxx 
xxx 
 
NK-cells 
n/a 
xxx 
xxx 
 
T-cells 
n/a 
xxx 
xxx 
 
B-cells 
n/a 
xxx 
xxx 
Day 8 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
n/a 
xxx 
xxx 
 
Dendritic cells 
n/a 
xxx 
xxx 
 
Neutrophils 
n/a 
xxx 
xxx 
 
NK-cells 
n/a 
xxx 
xxx 
 
T-cells 
n/a 
xxx 
xxx 
 
B-cells 
n/a 
xxx 
xxx 
N=Number of subjects in the Omics analysis population 
DE Gene = differentially expressed gene 
Table 127: 
Genes Differentially Expressed Between Treatment Groups by Immune Cell Type, and Post-Vaccination Day 


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Table 127: 
Genes Differentially Expressed Between Treatment Groups by Immune Cell Type, and Post-Vaccination Day 
 
 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Time Point 
Cell Type 
DE Genes [#] 
DE Genes [#] 
DE Genes [#] 
Day 2 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
xxx 
xxx 
 
Dendritic cells 
xxx 
xxx 
xxx 
 
Neutrophils 
xxx 
xxx 
xxx 
 
NK-cells 
xxx 
xxx 
xxx 
 
T-cells 
xxx 
xxx 
xxx 
 
B-cells 
xxx 
xxx 
xxx 
Day 4 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
xxx 
xxx 
 
Dendritic cells 
xxx 
xxx 
xxx 
 
Neutrophils 
xxx 
xxx 
xxx 
 
NK-cells 
xxx 
xxx 
xxx 
 
T-cells 
xxx 
xxx 
xxx 
 
B-cells 
xxx 
xxx 
xxx 
Day 29 (Post Vaccination 1 vs. Pre-Vaccination 1) 
Monocytes 
xxx 
n/a 
n/a 
 
Dendritic cells 
xxx 
n/a 
n/a 
 
Neutrophils 
xxx 
n/a 
n/a 
 
NK-cells 
xxx 
n/a 
n/a 
 
T-cells 
xxx 
n/a 
n/a 
 
B-cells 
xxx 
n/a 
n/a 
Day 2 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
xxx 
n/a 
n/a 
 
Dendritic cells 
xxx 
n/a 
n/a 


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Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) vs. 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
 
Neutrophils 
xxx 
n/a 
n/a 
 
NK-cells 
xxx 
n/a 
n/a 
 
T-cells 
xxx 
n/a 
n/a 
 
B-cells 
xxx 
n/a 
n/a 
Day 4 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
xxx 
n/a 
n/a 
 
Dendritic cells 
xxx 
n/a 
n/a 
 
Neutrophils 
xxx 
n/a 
n/a 
 
NK-cells 
xxx 
n/a 
n/a 
 
T-cells 
xxx 
n/a 
n/a 
 
B-cells 
xxx 
n/a 
n/a 
Day 8 (Post Vaccination 2 vs. Pre-Vaccination 2) 
Monocytes 
xxx 
n/a 
n/a 
 
Dendritic cells 
xxx 
n/a 
n/a 
 
Neutrophils 
xxx 
n/a 
n/a 
 
NK-cells 
xxx 
n/a 
n/a 
 
T-cells 
xxx 
n/a 
n/a 
 
B-cells 
xxx 
n/a 
n/a 
N=Number of subjects in the Omics analysis population 
DE Gene = differentially expressed gene 
 


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Table 128: 
Genes Differentially Expressed at Day 2 Compared to Pre-First Vaccination (RNA-Seq, Monocytes, Study Arm 1 A/H3N2v, D1) 
Generate tables for each applicable post-vaccination day, treatment group, pairwise treatment group comparisons (Group 1 vs. 2, Group 1 vs. 3, Group 1 
vs. 3, and Group 2 vs. 3), and cell type (monocytes, neutrophils, dendritic cells, NK cells B cell, and T cells). For Group 1 include tables for days 2, 4, 
and 8 post-first vaccination. For Groups 2 and 3 provide tables for post-vaccination days 2, 4, 29, 30, 32, and 36 post-first vaccination. For pairwise 
group comparisons that include Group 1, generate tables for days 2 and 4 post-first vaccination. Otherwise, prepare comparison results for days 2,4, 29, 
30, 32, and 36 post-first vaccination.  
Ensembl 
Gene ID 
Ensembl 
Gene Name 
Ensembl 
Gene Description 
Gene 
Type 
Log2 Fold Change 
(Day 2 vs. Pre-First 
Vaccination) 
Average 
Log2 
CPM* 
Likelihood Ratio 
Test Statistic 
P-Value 
FDR 
Adjusted P-
value 
ENSG00000
115415 
STAT1 
signal transducer and activator of transcription 1, 91kDa 
[Source:HGNC Symbol;Acc:11362] 
protein 
coding 
1.94 
2.034 
5.324 
0.000034 
0.000613 
ENSG00000
125347 
IRF1 
interferon regulatory factor 1 [Source:HGNC 
Symbol;Acc:6116] 
protein 
coding 
0.672 
2.506 
4.324 
0.0343 
0.00234 
ENSG00000
247275 
AL160008.1 
lincRNA 
0.80 
0.96 
3.42 
0.043 
0.0483 
*: average gene expression levels across Day 2 and Pre-First Vaccination samples measured in average log2  CPM 
 
 


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Table 129: 
Clusters of Co-Expressed Differential Genes (Monocytes, Days 2-29) 
Generate tables for each fold change set (Day 2-29 post-first vaccination and Day 2-8 post-second vaccination) and cell type (monocytes, neutrophils, 
dendritic cells, NK cells B cell, and T cells) 
Cluster ID 
Cluster Size 
Ensembl 
Gene ID 
Ensembl Gene Name 
Ensembl 
Gene Description 
Gene Type 
KEGG Pathway 
CCD229-001 
3 
ENSG00000115415 
STAT1 
signal transducer and activator of 
transcription 1, 91kDa [Source:HGNC 
Symbol;Acc:11362] 
protein coding 
Jak-STAT signaling pathway 
ENSG00000125347 
IRF1 
interferon regulatory factor 1 [Source:HGNC 
Symbol;Acc:6116] 
protein coding 
Pertussis 
Prolactin signaling pathway 
Hepatitis C  
Human papillomavirus infection 
ENSG00000247275 
AL160008.1 
lincRNA 
 
CCD229-002 
2 
ENSG00000108984 
MAP2K6 
protein coding 
protein coding 
MAPK signaling pathway 
ENSG00000152689 
RASGRP3 
RAS guanyl releasing protein 3 (calcium and 
DAGregulated) 
protein coding 
MAPK signaling pathway 
Ras signaling pathway 
Rap1 signaling pathway 
B cell receptor signaling pathway 
Pathways in cancer 
 
 
 


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Table 130: 
KEGG Pathways Enriched in Differentially Expressed Genes at Day 2 Compared to Pre-First Vaccination (RNA-Seq, Monocytes, 
Study Arm 1 A/H3N2v, D1) 
Generate tables for each gene set category (KEGG Pathways, MSigDB Reactome Pathways, and MSigDB Immunologic Signatures), post-first 
vaccination day (Day 2, 4, 8, 29), Study Arm (Study Arm 1, 2, 3), all pairwise Study Arm Comparison (1 vs. 2, 1 vs. 3, 2 vs. 3), and cell type 
(monocytes, neutrophils, dendritic cells, NK cells B cell, and T cells). Repeat for post-second vaccination time points relative to pre-second vaccination 
for Study Arms 2 and 3. 
Gene Set ID 
Gene Set 
Genes # 
Differentially Expressed Genes N 
(%)  
Up-regulated Differentially 
Expressed Genes N (%) 
Down-regulated 
Differentially Expressed 
Genes N (%) 
Jaccard Index 
P-Value 
FDR Adjusted P-
value 
Influenza A 
62 
9 (15) 
9 (15) 
0 
0.234 
<0.0001 
0.0006 
Antigen processing and 
presentation 
59 
4 (7) 
3 (5) 
1 (2) 
0.231 
<0.0001 
0.0004 
 
 


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14.4 
Safety Data 
14.4.1 Adverse Events 
14.4.1.1 Adverse Events of Special Interest 
Table 131: 
Number and Percentage of Subjects Experiencing Adverse Events of Special Interest 
 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Category 
n 
% 
n 
% 
n 
% 
Serious Adverse Event (SAE) 
x 
xx 
x 
xx 
x 
xx 
Medically-Attended Adverse Event (MAAE) 
x 
xx 
x 
xx 
x 
xx 
New-Onset Chronic Medical Condition (NOCMC) 
x 
xx 
x 
xx 
x 
xx 
Potentially Immune-Mediated Medical Condition (PIMMC) 
x 
xx 
x 
xx 
x 
xx 
N = Number of subjects in the Safety Analysis Population 
 
 
 


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14.4.1.2 
Solicited Adverse Events 
Table 132: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom, and Treatment 
Group (Post-First Vaccination) 
  
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
n 
% 
95% CI 
Any Symptom 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
Any Systemic Symptom 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
[Systemic Symptom 1] 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
[Systemic Symptom 2] 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
Any Local Symptom 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
[Local Symptom 1] 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
[Local Symptom 2] 
x 
xx 
x x, x.x 
x 
xx 
x x, x x 
x 
xx 
x x, x x 
N= Number of subjects in the per safety analysis population. 
 
Tables with similar format: 
Table 133: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom, and Treatment 
Group (Post-Second Vaccination) 
Table 134: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom, and Treatment 
Group (Post-Any Vaccination) 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Table 135: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Maximum Severity, and 
Treatment Group (Post-First Vaccination) 
  
  
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Symptom 
Severity 
n 
% 
95% CI 
n 
% 
95% CI 
n 
% 
95% CI 
Any Symptom 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
Systemic Symptoms 
 
 
 
 
 
 
 
 
 
 
Any Systemic Symptom 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
[Systemic Symptom 1] 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
[Systemic Symptom 2] 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
Local Symptoms 
 
 
 
 
 
 
 
 
 
 
Any Local Symptom 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 


Statistical Analysis Plan - DMID Protocol:14-0015  
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Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
[Local Symptom 1] 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
[Local Symptom 2] 
None 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Mild 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Moderate 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
  
Severe 
x 
xx 
x x, x.x 
x 
xx 
x.x, x x 
x 
xx 
x x, x.x 
N = Number of subjects in the Safety Analysis Population who received the specified dose. 
Severity is the maximum severity reported over all solicited symptoms post dosing for each subject. 
 
Tables with similar format: 
Table 136: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Maximum Severity, and Treatment Group (Post-
Second Vaccination) 
Table 137: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Maximum Severity, and Treatment Group (Post-
Any Vaccination) 
 
 


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Table 138: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Severity, and Post-Vaccination Day (Post-First 
Vaccination, Study Arm 1 A/H3N2v, D1) 
Study Arm 1 A/H3N2v, D1 (N=X) 
Symptom 
Severity 
Pre-
Vaccination 1 
 
Post-
Vaccination 1 
 
Day 1 
 
Day 2 
 
Day 3 
 
Day 4 
 
Day 5 
 
Day 6 
 
Day 7+ 
 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any 
Symptom 
None 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Systemic Symptoms 
Any  
Systemic 
Symptom 
None 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
[Systemic 
Symptom 1] 
None 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
[Systemic 
Symptom 2] 
None 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Study Arm 1 A/H3N2v, D1 (N=X) 
Symptom 
Severity 
Pre-
Vaccination 1 
 
Post-
Vaccination 1 
 
Day 1 
 
Day 2 
 
Day 3 
 
Day 4 
 
Day 5 
 
Day 6 
 
Day 7+ 
 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Local Symptoms 
Any Local 
Symptom 
None 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
[Local 
Symptom 1] 
None 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
[Local 
Symptom 2] 
None 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Mild 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
N = Number of subjects in the Safety Analysis Population who received the specified dose. 
Severity is the maximum severity reported post dosing for each subject for each day. 
 
 


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Tables with similar format: 
Table 139: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Severity, and Post-Vaccination Day (Post-First 
Vaccination, Study Arm 2 A/H7N9 + AS03, D1, 29) 
Table 140: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Severity, and Post-Vaccination Day (Post-First 
Vaccination, Study Arm 3 A/H7N9 + PBS, D1, 29) 
Table 141: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Severity, and Post-Vaccination Day (Post-Second 
Vaccination, Study Arm 2 A/H7N9 + AS03, D1, D29) 
Table 142: 
Number and Percentage of Subjects Experiencing Solicited Events by Symptom, Severity, and Post-Vaccination Day (Post-Second 
Vaccination, Study Arm 3 A/H7N9 + PBS, D1, 29) 
 


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14.4.1.3 
Unsolicited Adverse Events 
Table 143: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events with 95% Confidence Intervals by MedDRA® 
System Organ Class and Preferred Term, and Treatment Group (Post-First Vaccination) 
MedDRA® System Organ Class 
MedDRA® 
Preferred 
Term 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
n 
% 
95% CI 
Any SOC 
Any PT 
x 
xx 
xx, xx 
x 
xx 
xx, xx 
x 
xx 
xx, xx 
[SOC 1] 
Any PT 
  
  
  
  
  
  
  
  
  
  
[PT 1] 
  
  
  
  
  
  
  
  
  
  
[PT 2] 
  
  
  
  
  
  
  
  
  
[SOC 2] 
Any PT 
  
  
  
  
  
  
  
  
  
  
[PT 1] 
  
  
  
  
  
  
  
  
  
  
[PT 2] 
  
  
  
  
  
  
  
  
  
Note: N = number of subjects in the Safety Analysis Population who received the specified dose.  
This table presents number and percentage of subjects.  A subject is only counted once per PT. 
 
Tables with similar format: 
Table 144: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events with 95% Confidence Intervals by MedDRA® 
System Organ Class and Preferred Term, and Treatment Group (Post-Second Vaccination) 
Table 145: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events with 95% Confidence Intervals by MedDRA® 
System Organ Class and Preferred Term, and Treatment Group (Post-Any Vaccination) 


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Table 146: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA® System Organ Class and Preferred 
Term, Maximum Severity, and Relationship (Study Arm 1 A/H3N2v, D1) 
Study Arm 1 A/H3N2v, D1 (N=X) 
MedDRA® System Organ Class 
MedDRA® Preferred Term 
Any Incidence 
Severity [1] 
Relationship to Treatment [2] 
Mild 
Moderate 
Severe 
Not Related 
Related 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
Any PT 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
x 
xx 
[SOC 1] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of subjects in the Safety Analysis Population.  
[1] For severity, a subject is counted once per preferred term and is summarized according to their highest severity. 
[2] For relationship, a subject is only counted once per preferred term and is summarized according to their closest relationship. 
 
Tables with similar format: 
Table 147: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA® System Organ Class and Preferred 
Term, Maximum Severity, Relationship (Study Arm 2 A/H7N9 + AS03, D1, 29) 
Table 148: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA® System Organ Class and Preferred 
Term, Maximum Severity, Relationship (Study Arm 3 A/H7N9 + PBS, D1, 29) 
 


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14.4.2 Listing of Deaths, Other Serious and Significant Adverse Events 
Table 149: 
Listing of Serious Adverse Events 
Adverse 
Event 
Associated 
with Dose # 
# of Days 
Post 
Associated 
Dose 
(Duration) 
# of Days 
Post Dose 
the Event 
Became 
Serious 
Reason 
Reported as 
an SAE 
Severity 
Relationship 
to Study 
Treatment 
In Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Subject 
Discontinue
d Due to 
AE 
Outcome 
MedDRA® 
System 
Organ 
Class 
MedDRA® 
Preferred 
Term 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
Table 150: 
Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events  
Adverse Event 
Associated 
with Dose # 
# of Days Post 
Associated 
Dose 
(Duration) 
Severity 
Relationship 
to Study 
Treatment 
In Not 
Related, 
Alternative 
Etiology 
Action Taken 
with Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA® 
System Organ 
Class 
MedDRA® 
Preferred 
Term 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
 


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Table 151: 
Listing of Adverse Events of Special Interest 
Adverse Event 
Vaccination 
Received at 
Time of Event? 
No. of Days 
Post 
Vaccination  
Duration of 
Event 
Severity 
MedDRA 
System Organ 
Class 
NOCMC 
 
? 
PIMMC 
Relationship 
Outcome 
 
Subject ID: , Study Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Study Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 


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14.4.3 Narratives of Deaths, Other Serious and Significant Adverse Events 
(not included in SAP, but this is a placeholder for the CSR) 
14.4.4 Displays of Vital Signs 
Table 152: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group – Any Assessment 
  
  
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Study Visit Day 
Severity 
n 
% 
n 
% 
n 
% 
Day -7 (Screening) 
None 
x 
xx 
x 
xx 
x 
xx 
  
Mild 
  
  
  
  
  
  
  
Moderate 
  
  
  
  
  
  
  
Severe 
  
  
  
  
  
  
  
Missing 
  
  
  
  
  
  
Day 1 (Pre-Vaccination 1) 
None 
x 
xx 
x 
xx 
x 
xx 
  
Mild 
  
  
  
  
  
  
  
Moderate 
  
  
  
  
  
  
  
Severe 
  
  
  
  
  
  
  
Missing 
  
  
  
  
  
  
Day 29 (Post-Vaccination 1) 
None 
n/a 
n/a 
x 
xx 
x 
xx 
  
Mild 
n/a 
n/a 
  
  
  
  
  
Moderate 
n/a 
n/a 
  
  
  
  
  
Severe 
n/a 
n/a 
  
  
  
  
  
Missing 
n/a 
n/a 
  
  
  
  
Maximum Post-Vaccination 1 
None 
x 
xx 
x 
xx 
x 
xx 
  
Mild 
  
  
  
  
  
  
  
Moderate 
  
  
  
  
  
  
  
Severe 
  
  
  
  
  
  
  
Missing 
  
  
  
  
  
  


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Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
Maximum Post-Vaccination 1 indicates the maximum severity experienced by each subject at any time point post baseline, including unscheduled assessments.  
N = Number of subjects in the Safety Analysis Population. 
 


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14.5 
Concomitant Medications 
Table 153: 
Number and Percentage of Subjects with Prior and Concurrent Medications by WHO Drug Classification and Treatment Group 
WHO Drug Code 
Level 1, Anatomic Group 
WHO Drug Code 
Level 2, Therapeutic Subgroup 
Study Arm 1  
A/H3N2v, D1  
(N=X) 
Study Arm 2 
A/H7N9 + AS03, D1, 29  
(N=X) 
Study Arm 3 
A/H7N9 + PBS, D1, 29 
(N=X) 
n 
% 
n 
% 
n 
% 
Any Level 1 Codes 
Any Level 2 Codes 
x 
xx 
x 
xx 
x 
xx 
[ATC Level 1 - 1] 
Any [ATC 1 – 1] 
 
 
 
 
 
 
 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
[ATC Level 1 – 2] 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
N = Number of subjects in the Safety Analysis Population 
n = Number of subjects reporting taking at least one medication in the specific WHO Drug Class. 
 


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APPENDIX 2. FIGURE MOCK-UPS 
 
 


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LIST OF FIGURES 
Figure 1: 
CONSORT Flow Diagram .........................................................................................94 
Figure 2: 
Hemagglutination Inhibition GMTs and 95% CIs by Study Arm and Post-
Vaccination Day .........................................................................................................94 
Figure 3: 
Barplot of Subject-Level Hemagglutination Inhibition Results at Day 29 
Post-Last Vaccination .................................................................................................95 
Figure 4: 
Median Fold Change in Fractalkine Concentration and Associated 95% CI 
by Study Arm and Post-Vaccination Day ...................................................................96 
Figure 5: 
Radar Plot of Median Fold Change by Study Arm and Post-Vaccination Day ..........96 
Figure 6: 
Multivariate starplots of reference alignment statistics ..............................................97 
Figure 7: 
Multivariate PCA, MDS, and hierarchical clustering plots ........................................98 
Figure 8: 
Reverse empirical cumulative distribution function plots to summarize the 
percentage of genes that pass a certain average LCPM expression cut off ................99 
Figure 9: 
MA and Volcano plots ..............................................................................................100 
Figure 10: DE gene overlap between post-first vaccination days (Study Arm 1, Day 2-
29 post-first vaccination) ..........................................................................................101 
Figure 11: Heatmap, Co-Expressed Gene Cluster Dendrograms, and Cluster Time 
Trend Visualizations .................................................................................................102 
Figure 12: Pathway Enrichment Visualizations .........................................................................103 
Figure 13: Maximum Severity of Solicited Systemic Symptoms per Subject by Day 
(Post-First Vaccination) ............................................................................................105 
Figure 14: Incidence of Unsolicited Adverse Events by MedDRA® System Organ 
Class and Severity .....................................................................................................106 
 


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10.1 
Disposition of Subjects 
Figure 1: 
CONSORT Flow Diagram 
14.2 
Immunogenicity Data 
Immunogenicity Response Figures by Measure, Treatment/Vaccination, and Time Point 
Figure 2: 
Hemagglutination Inhibition GMTs and 95% CIs by Study Arm and Post-Vaccination Day 
 
Repeat for neutralizing antibodies. Include all study arms. Label study arms as: Study Arm 1 A/H3N2v, D1; Study Arm 2 A/H7N9 + AS03, D1, 29; Study 
Arm 3 A/H7N9 + PBS, D1, 29. 


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Figure 3: 
Barplot of Subject-Level Hemagglutination Inhibition Results at Day 29 Post-Last 
Vaccination 
 
 
 
Repeat for neutralizing antibodies. Include all study arms. Label study arms as: Study Arm 1 A/H3N2v, D1; 
Study Arm 2 A/H7N9 + AS03, D1, 29; Study Arm 3 A/H7N9 + PBS, D1, 29. 
 
 
 


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Figure 4: 
Median Fold Change in Fractalkine Concentration and Associated 95% CI by Study 
Arm and Post-Vaccination Day  
 
Plot individual fold change observations using treatment group-specific symbols. Include Study Arm 1 A/H3N2v. Label study arms 
as: Study Arm 1 A/H3N2v, D1; Study Arm 2 A/H7N9 + AS03, D1, 29; Study Arm 3 A/H7N9 + PBS, D1, 29. Repeat for GM-CSF, 
IFN-γ, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-23, ITAC, MIP 1a, MIP1b, MIP3a, TNF-
alpha, IP-10, IFNa, IFNb. 
 
Figure 5: 
Radar Plot of Median Fold Change by Study Arm and Post-Vaccination Day 
 
Include Study Arm 1 A/H3N2v. Label study arms as: Study Arm 1 A/H3N2v, D1; Study Arm 2 A/H7N9 + AS03, D1, 29; Study Arm 3 
A/H7N9 + PBS, D1, 29. Include Fractalkine GM-CSF, IFN-γ, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-
17A, IL-21, IL-23, ITAC, MIP 1a, MIP1b, MIP3a, TNF-alpha, IP-10, IFNa, IFNb.  


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14.3 
Transcriptomics Data 
Figure 6: 
Multivariate starplots of reference alignment statistics 
Each sample is depicted by a star. Each ray represents one of the reference alignments statistics. Ray length is 
globally scaled so that the respective maximum alignment statistic across all samples is 1 and the minimum is 0. 
Stars will be color-coded by Study Arm. 
 


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Figure 7: 
Multivariate PCA, MDS, and hierarchical clustering plots 
Standardized LCPM distributions will be used as input for PCA (A), MDS (B, C), and hierarchical clustering (D, E). 
For MDS and hierarchical clustering, results will be provided for Euclidean (B, D) as well as 1-Spearman 
correlation distance (C, E). 
 
 
 
 
 


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Figure 8: 
Reverse empirical cumulative distribution function plots to summarize the 
percentage of genes that pass a certain average LCPM expression cut off 
The x-axis represents the log2 count per million cut off for identifying lowly expressed genes. The y-axis shows the 
percentage of all genes with a minimum gene expression level across all study samples that exceeds the respective 
cut off. The grey box indicates the target range of genes to be selected (between 10,000 and 14,000 genes). The 
black vertical lines represent the actual cut off that would be selected. 
 
 
 


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Figure 9: 
MA and Volcano plots 
The y-axis on the MA plot (left panel) represents the log2 fold change for the respective treatment effect. The x-axis 
shows the average log2 expression level. The y-axis on the Volcano plot (right panel) represents the –log10 adjusted 
p-value and the x-axis the log2 fold change for the respective treatment effect. Each black dot represents a gene. In 
red: DE genes. Blue dashed line: fold change cut off. MA and Volcano plots will be generated for each post-
vaccination day, study arm, and study arm comparison. 
 
Generate MA and Volcano plots for each post-first vaccination day (Day 2, 4, 8 or 29), Study 
Arm (Study Arm 1, 2, or 3), and pairwise Study Arm Comparison (1 vs. 2, 1 vs. 3, or 2 vs. 3). 
Repeat for post-second vaccination time points (Day 30, 32, and 36) relative to pre-second 
vaccination for Study Arms 2 and 3. 
 


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Figure 10: 
DE gene overlap between post-first vaccination days (Study Arm 1, Day 2-29 
post-first vaccination)  
This plot summarizes the DE gene overlap between post-vaccination days for all DE (shown to 
the right), for up-regulated DE genes (shown in the middle), and down-regulated DE genes 
(shown to the right). For example, in the left panel, the bottom left horizontal barplot labeled 
‘SDEG Set Size’ shows the total number of DE genes per post-vaccination time point. The black 
circles in the matrix represent what would be the different Venn diagram spaces (unique and 
overlapping DE genes). Connected black circles indicate a certain intersection of post-
vaccination days. In the left panel, the first vertical bar/column shows those DE genes that are 
unique to Day 6. The second shows those DE genes that are shared only between Day 6 and 7. 
The third are those DE genes that are shared between Days 6, 7, and 8, and so forth. 
 
Generate this plot for each study arm post-first vaccination. Repeat for post-second vaccination 
time points (Days 30, 32, and 36) relative to pre-second vaccination for Study Arms 2 and 3. 
 


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Figure 11: 
Heatmap, Co-Expressed Gene Cluster Dendrograms, and Cluster Time Trend 
Visualizations 
Left: Heatmap of log2 fold changes of DE genes color-coded by up/down–regulation. Center: Gene cluster 
dendrogram with bootstrap probabilities and identified co-expressed gene clusters. Right: Time trends for 
mean log2 fold changes and associated 95% bootstrap CI of co-expressed gene clusters. 
 
Left: Generate heatmaps for each post-first vaccination day (Day 2, 4, 8 or 29), Study Arm (Study Arm 1, 2, 
and 3) and pairwise Study Arm Comparison (1 vs. 2, 1 vs. 3, 2 vs. 3). Repeat for post-second vaccination time 
points (Days 30, 32, and 36) relative to pre-second vaccination for Study Arms 2 and 3.  
Center: Generate gene cluster dendrograms for each fold change set (Day 2-29 post-first vaccination and 
Day 2-8 post-second vaccination).  
Right: Generate cluster time trends for each cluster and fold change set (Day 2-29 post-first vaccination and 
Day 2-8 post-second vaccination). 
 
 


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Figure 12: 
Pathway Enrichment Visualizations 
Left: Pathway map of a significantly enriched pathway color-coded by up/down–regulation of genes. 
Center: Heatmap summarizing pathway enrichment across study arms and post-vaccination days. Cells are 
color-coded by enrichment score: -1 x log10(enrichment p-value). Numbers in each cell represent the number 
of DE genes. Two numbers will be printed in each cell: 1) the number of up-regulated DE genes and the 2) the 
number of down-regulated DE genes per pathway.  
Right: Protein-protein interaction network centered around proteins encoded by DE genes. Nodes will be 
labeled by corresponding gene name. Edges will represent experimentally determined (reported in ≥2 
publications) binary protein-protein interactions or protein complex membership. Color and size of individual 
nodes will indicate the mean log2 fold change between conditions. Further details are provided as part of the 
legend within the figure. 
 
Left: Generate KEGG pathway maps for each enriched KEGG pathway for each post-first vaccination day 
(Day 2, 4, 8 or 29), Study Arm (Study Arm 1, 2, and 3) and pairwise Study Arm Comparison (Study Arm 1 vs. 


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2, 1 vs. 3, 2 vs. 3). Repeat for post-second vaccination time points (Days 30, 32, and 36) relative to pre-
second vaccination for Study Arms 2 and 3. 
Center: Summarize for each gene set category (KEGG Pathways, MSigDB Reactome Pathways, MSigDB GO 
Biological Process, MSigDB Immunologic Signatures, and Blood Transcription Modules), for each post-first 
vaccination day (Day 2, 4, 8 or 29), Study Arm (Study Arm 1, 2, and 3) and pairwise Study Arm Comparison 
(1 vs. 2, 1 vs. 3, 2 vs. 3). Repeat for post-second vaccination time points (Days 30, 32, and 36) relative to pre-
second vaccination for Study Arms 2 and 3.  
Right: Generate protein-interaction networks for H7N9 Study Arm Comparisons (Study Arm 1 vs. 2). 
 
 


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14.4 
Safety Data 
Figure 13: 
Maximum Severity of Solicited Systemic Symptoms per Subject by Day (Post-First 
Vaccination) 
 
Repeat for post-second vaccination and for solicited local symptoms. Include all study arms. Label study arms 
as: Study Arm 1 A/H3N2v, D1; Study Arm 2 A/H7N9 + AS03, D1, 29; Study Arm 3 A/H7N9 + PBS, D1, 29. 


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Figure 14: 
Incidence of Unsolicited Adverse Events by MedDRA® System Organ Class and Severity 
 
Include all study arms. Label study arms as: Study Arm 1 A/H3N2v, D1; Study Arm 2 A/H7N9 + AS03, D1, 
29; Study Arm 3 A/H7N9 + PBS, D1, 29. 
 


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APPENDIX 3. LISTINGS MOCK-UPS 
 
 


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LISTINGS 
Listing 1: 16.1.6 Listing of Subjects Receiving Investigational Product ..................................109 
Listing 2: 16.2.1 Early Terminations or Discontinued Subjects ...............................................110 
Listing 3: 16.2.2.1 Subject-Specific Protocol Deviations .........................................................111 
Listing 4: 16.2.2.2 Non-Subject-Specific Protocol Deviations .................................................112 
Listing 5: 16.2.3 Analysis Populations Exclusions ...................................................................113 
Listing 6: 16.2.4.1 Demographic Data ......................................................................................114 
Listing 7: 16.2.4.2 Pre-Existing and Concurrent Medical Conditions ......................................115 
Listing 8: 16.2.6 Individual Immunogenicity Response Data – Antibody Assays ...................116 
Listing 9: 16.2.6 Individual Immunogenicity Response Data – Cytokine/Chemokine 
Assay .........................................................................................................................116 
Listing 10: 16.2.7.1 Solicited Events – Systemic Symptoms .....................................................117 
Listing 11: 16.2.7.2 Solicited Events – Local Symptoms ...........................................................118 
Listing 12: 16.2.7.3 Unsolicited Adverse Events ........................................................................119 
Listing 13: 16.2.9.1 Vital Signs ..................................................................................................120 
Listing 14: 16.2.9.2 Physical Exam Findings .............................................................................120 
Listing 15: 16.2.10 Concomitant Medications ............................................................................121 
 


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Listing 1: 
16.1.6 
Listing of Subjects Receiving Investigational Product 
Subject ID 
Randomized  
Study Group 
Product Received Study Vaccination 1 
Product Received Study Vaccination 2 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2 
Database Listings by Subject 
16.2.1 Discontinued Subjects 
Listing 2: 
16.2.1 
Early Terminations or Discontinued Subjects 
Treatment Group 
 Subject ID 
Category 
Reason for Early Termination or Treatment Discontinuation 
Study Day 
 
 
 
 
 
 
 
 


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16.2.2 Protocol Deviations 
Listing 3: 
16.2.2.1 
Subject-Specific Protocol Deviations 
Treatment 
Group 
Subject ID 
DV Number 
Deviation 
Deviation 
Category 
Study Day 
Reason for 
Deviation 
Deviation 
Resulted 
in AE? 
Deviation Resulted 
in Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 4: 
16.2.2.2 
Non-Subject-Specific Protocol Deviations 
Site 
Start Date  
Deviation 
End Date 
Reason for 
Deviation 
Deviation 
Resulted in 
Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Category 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.3 Subjects Excluded from the Efficacy Analysis 
Listing 5: 
16.2.3 
Analysis Populations Exclusions 
Treatment Group 
Subject ID 
Study Visit 
Analyses in which 
Subject is Included 
Analyses from which  
Subject is Excluded 
Results Available? 
Reason for Exclusion 
 
 
 
[e.g., Safety, ITT, PP] 
[e.g., Safety, ITT, PP, Day x] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: “Yes” in the “Results available” column indicates that available data were removed from the analysis. “No” indicates that no data were available for inclusion in the analysis. 
 
 


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16.2.4 Demographic Data 
Listing 6: 
16.2.4.1 
Demographic Data 
Treatment Group 
 Subject ID 
Sex 
Age at Enrollment (years) 
Ethnicity 
Race 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 7: 
16.2.4.2 
Pre-Existing and Concurrent Medical Conditions 
Treatment 
Group 
 Subject ID 
MH Number 
Medical History Term 
Condition Start Day  
Condition End Day 
MedDRA System Organ Class 
MedDRA Preferred Term 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.6 Individual Immunogenicity Response Data 
Listing 8: 
16.2.6 
Individual Immunogenicity Response Data – Antibody Assays 
Treatment Group  
Subject ID  
Planned Time Point 
Actual Study Day 
HAI Titer 
Neut Titer 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 9: 
16.2.6 
Individual Immunogenicity Response Data – Cytokine/Chemokine Assay 
Treatment Group  
Subject ID  
Planned Time Point 
Actual Study Day 
Cytokine/Chemokine 
Concentration 
 
 
 
 
[Cytokine/Chemokine]* 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*: Fractalkine, GM-CSF, IFN-γ, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IL-21, IL-23, ITAC, MIP 1a, MIP1b, MIP3a, TNF-alpha, IP-10, IFNa, IFNb 


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16.2.7 Adverse Events 
Listing 10: 
16.2.7.1 
Solicited Events – Systemic Symptoms 
Treatment 
Group  
Subject ID  
Dose Number 
Post Dose Day 
Assessmenta 
Symptom 
Severity 
Attributed to 
Alternate 
Etiology?b 
Alternate Etiology 
 
 
 
 
MA 
 
 
 
 
 
 
 
 
Clinic 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
a MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
b Grade 3 events only.  
Note: Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 
 
 


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Listing 11: 
16.2.7.2 
Solicited Events – Local Symptoms 
Treatment Group  
Subject ID  
Dose Number 
Post Dose Day 
Assessmenta 
Symptom 
Severity 
 
 
 
 
MA 
 
 
 
 
 
 
Clinic 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
a MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
Note: Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 


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Listing 12: 
16.2.7.3 
Unsolicited Adverse Events 
Adverse 
Event 
Associated 
with Dose 
No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
Severity 
SAE? 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System 
Organ Class 
MedDRA 
Preferred 
Term 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
Note: For additional details about SAEs, see Table 149.  
 
 
 


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16.2.9 Vital Signs and Physical Exam Findings 
Listing 13: 
16.2.9.1 
Vital Signs 
Treatment 
Group 
Subject ID 
Planned Time 
Point 
Actual Study 
Day 
Temperature 
(°C) 
Systolic Blood 
Pressure 
(mmHg) 
Diastolic Blood 
Pressure 
(mmHg) 
Heart Rate 
(beats/min) 
Respiratory Rate 
(breaths/min) 
Weight 
(kg) 
Height 
(cm) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 14: 
16.2.9.2 
Physical Exam Findings 
Treatment 
Group 
Subject ID 
Planned Time 
Point 
Actual Study 
Day 
Body System 
Abnormal Finding 
Reported as an AE? 
(AE Description; Number) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.10 Concomitant Medications 
Listing 15: 
16.2.10 
Concomitant Medications 
Treatment 
Group 
Subject ID 
CM 
Number 
Medication 
Medication Start 
Day 
Medication End 
Day 
Indication 
Taken for an AE? 
(AE Description; 
Number) 
Taken for a condition 
on Medical History? 
(MH Description; 
Number) 
ATC Level 1  
(ATC Level 2)