VRC 316: A Phase I Open-Label Clinical Trial To Evaluate Dose, Safety, Tolerability, And Immunogenicity Of An Influenza HA Ferritin Vaccine, Alone Or In Prime-Boost Regimens With An Influenza DNA Vaccine In Healthy Adults
Phase: PHASE1
Conditions: Influenza
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

Official Title: A Phase I Open-Label Clinical Trial to Evaluate Dose, Safety, Tolerability, and 
Immunogenicity of an Influenza HA Ferritin Vaccine Alone or in Prime-Boost 
Regimen with an Influenza DNA Vaccine in Healthy Adults  
 
ClinicalTrials.gov Identifier: NCT03186781 
 
Document Type: 
• 
Redacted Final Version of the Study Protocol (Statistical Analysis Considerations located in 
Section 6 of the Protocol)  
• 
Redacted Final Version of IRB-approved Informed Consents  
 
Document/Date: 
• 
Protocol Version 6.0: March 11, 2019 
• 
Version 5.0 IRB-approved Informed Consents – Main and Supplemental: November 13, 2018 

 
 
Version 6.0 
March 11, 2019 
 
VACCINE RESEARCH CENTER 
 
Protocol VRC 316 
(NIH 17-I-0110) 
 
A PHASE I OPEN-LABEL CLINICAL TRIAL TO EVALUATE DOSE, SAFETY, TOLERABILITY, AND 
IMMUNOGENICITY OF AN INFLUENZA HA FERRITIN VACCINE, ALONE OR IN PRIME-BOOST 
REGIMENS WITH AN INFLUENZA DNA VACCINE IN HEALTHY ADULTS 
 
Vaccine Provided by 
National Institute of Allergy and Infectious Diseases (NIAID) 
Vaccine Research Center (VRC) 
Bethesda, Maryland  
 
Clinical Trial Sponsored by: 
National Institute of Allergy and Infectious Diseases (NIAID) 
Vaccine Research Center (VRC) 
Bethesda, Maryland 
BB-IND 17399 – held by VRC, NIAID, NIH 
 
Principal Investigator: 
Grace Chen, M.D., M.P.H 
Vaccine Research Center, National Institute of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
Bethesda, MD 20892 
 
IRB Initial Review Date:  March 13, 2016 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 2 of 62 
TABLE OF CONTENTS 
TABLE OF ABBREVIATIONS ................................................................................................................... 6
PRÉCIS ........................................................................................................................................................... 8

INTRODUCTION AND RATIONALE ........................................................................................ 10
 BACKGROUND ........................................................................................................................... 10
 Rationale For Development Of Influenza HA-F A/Sing Vaccine, VRC-FLUNPF081-00-VP ..... 11
 Rationale For Development of Influenza Plasmid DNA A/Sing Vaccine, VRC-FLUDNA082-00-
VP and its use with VRC-FLUNPF081-00-VP in Prime-Boost Regimens ................................... 11
1.3.1
DNA vaccines developed by VRC, NIAID, NIH ................................................................ 12
 Rationale for VRC-FLUDNA082-00-VP Vaccine Administration Method.................................. 12
 Rationale For Study Population Proposed In The Clinical Trial ................................................... 13
 Assessment of Vaccine Immunogenicity ....................................................................................... 13

STUDY AGENTS ........................................................................................................................... 14
 VRC-FLUNPF081-00-VP ............................................................................................................. 14
 VRC-FLUDNA082-00-VP ............................................................................................................ 14
 VRC-PBSPLA043-00-VP, PHOSPHATE BUFFERED SALINE (DILUENT) ........................... 14
 Preclinical Studies with VRC-FLUNPF081-00-VP ...................................................................... 14
 Preclinical Studies with VRC-FLUDNA082-00-VP ..................................................................... 15

STUDY OBJECTIVES ................................................................................................................... 15
 Primary Objectives......................................................................................................................... 15
 Secondary Objectives..................................................................................................................... 15
 Exploratory Objectives .................................................................................................................. 16

STUDY DESIGN AND METHODS .............................................................................................. 16
 Study Population ............................................................................................................................ 16
4.1.1
Inclusion Criteria .............................................................................................................. 16
4.1.2
Exclusion Criteria ............................................................................................................. 17
 Clinical Procedures and Evaluations.............................................................................................. 19
4.2.1
Screening .......................................................................................................................... 19
4.2.2
Study Schedule .................................................................................................................. 19
4.2.3
Enrollment and Randomization ........................................................................................ 20
4.2.4
Administration of Study Vaccines ..................................................................................... 20
4.2.5
7-Day Solicited Reactogenicity and Follow-up ................................................................ 21
4.2.6
Follow-Up through End of Study ...................................................................................... 21
4.2.7
Blood Sample Collection .................................................................................................. 21
4.2.8
Apheresis ........................................................................................................................... 21

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 3 of 62 
4.2.9
Concomitant Medications ................................................................................................. 22
 Criteria for Dose Escalation and Dose Continuation ..................................................................... 22
 Criteria for Discontinuing Study Injections or Protocol Participation ........................................... 23
4.4.1
Discontinuation of Study Injections .................................................................................. 23
4.4.2
Discontinuation of Protocol Participation ....................................................................... 24
 Criteria for Pausing and Resuming the Study ................................................................................ 24
4.5.1
Plan for Pausing the Study ............................................................................................... 24
4.5.2
Plan for Review of Pauses and Resuming the Study ......................................................... 24

SAFETY AND ADVERSE EVENT REPORTING ..................................................................... 24
 Adverse Events .............................................................................................................................. 24
 Serious Adverse Events ................................................................................................................. 25
 Adverse Event Reporting to the IND Sponsor ............................................................................... 25
5.3.1
IND Sponsor Reporting to the FDA .................................................................................. 26
 Reporting to the Institutional Review Board ................................................................................. 26
5.4.1
Unanticipated Problem Definition .................................................................................... 26
5.4.2
Protocol Deviation Definition........................................................................................... 27
5.4.3
Non-Compliance Definition .............................................................................................. 27
5.4.4
Expedited Reporting to the NIAID IRB ............................................................................. 27
5.4.5
Annual Reporting to the NIAID IRB ................................................................................. 28
 Serious Adverse Event Reporting to the Institutional Biosafety Committee (IBC) ...................... 28

STATISTICAL CONSIDERATIONS .......................................................................................... 28
 Overview ........................................................................................................................................ 28
 Objectives ...................................................................................................................................... 28
 Endpoints ....................................................................................................................................... 29
6.3.1
Primary Endpoints: Safety ................................................................................................ 29
6.3.2
Secondary Endpoints: Immunogenicity ............................................................................ 29
6.3.3
Exploratory Endpoints: Immunogenicity .......................................................................... 29
 Sample Size and Accrual ............................................................................................................... 29
6.4.1
Power Calculations for Safety .......................................................................................... 29
6.4.2
Power Calculations for Immunogenicity .......................................................................... 31
6.4.3
Power for Comparison ...................................................................................................... 31
 Statistical Analysis ......................................................................................................................... 32
6.5.1
Analysis Variables ............................................................................................................ 32
6.5.2
Baseline Demographics .................................................................................................... 32
6.5.3
Safety Analysis .................................................................................................................. 33

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 4 of 62 
6.5.4
Immunogenicity Analysis .................................................................................................. 33
6.5.5
Missing Data ..................................................................................................................... 33
6.5.6
Interim Analyses ............................................................................................................... 34
 Intervention Assignments .............................................................................................................. 34

PHARMACY AND VACCINE ADMINISTRATION PROCEDURES .................................... 34
 Study Product ................................................................................................................................. 34
 Study Product Presentation and Storage ........................................................................................ 35
7.2.1
Study Product Labels ........................................................................................................ 35
7.2.2
Study Product Storage ...................................................................................................... 35
7.2.3
Temperature excursions .................................................................................................... 35
 Preparation of Study Products for Injection ................................................................................... 35
7.3.1
Preparation of VRC-FLUNPF081-00-VP to be administered via Needle and Syringe 
Injection ............................................................................................................................ 36
7.3.2
Preparation of VRC-FLUDNA082-00-VP to be administered via PharmaJet Injection .. 36
 Study Product Accountability ........................................................................................................ 36
7.4.1
Documentation .................................................................................................................. 36
7.4.2
Disposition ........................................................................................................................ 36

HUMAN SUBJECT PROTECTIONS AND ETHICAL OBLIGATIONS ................................ 36
 Institutional Review Board ............................................................................................................ 37
 Subject Identification and Enrollment of Study Participants ......................................................... 37
8.2.1
Participation of Children .................................................................................................. 37
8.2.2
Participation of NIH Employees ....................................................................................... 37
 Informed Consent .......................................................................................................................... 37
 Subject Confidentiality .................................................................................................................. 38
 Risks and Benefits ......................................................................................................................... 38
8.5.1
Risks of the VRC-FLUNPF081-00-VP vaccine ................................................................ 38
8.5.2
Risks of the VRC-FLUDNA082-00-VP Vaccine ............................................................... 38
8.5.3
Other Risks ........................................................................................................................ 39
8.5.4
Study Benefits .................................................................................................................... 40
 Plan for Use and Storage of Biological Samples ........................................................................... 40
8.6.1
Use of Samples, Specimens and Data ............................................................................... 40
8.6.2
Storage and Tracking of Blood Samples and Other Specimens ........................................ 40
8.6.3
Disposition of Samples, Specimens and Data at Completion of the Protocol .................. 41
8.6.4
Loss or Destruction of Samples, Specimens or Data ........................................................ 41
 Compensation ................................................................................................................................ 41

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 5 of 62 
 Safety Monitoring .......................................................................................................................... 42
8.8.1
Protocol Safety Review Team ........................................................................................... 42

ADMINISTRATIVE AND LEGAL OBLIGATIONS ................................................................. 42
 Protocol Amendments and Study Termination .............................................................................. 42
 Study Documentation and Storage ................................................................................................. 42
 Data Collection, Data Sharing and Protocol Monitoring ............................................................... 43
9.3.1
Data Collection ................................................................................................................. 43
9.3.2
Data Sharing Plan ............................................................................................................ 43
9.3.3
Source Documents ............................................................................................................ 43
9.3.4
Protocol Monitoring Plan ................................................................................................. 43
 Language ........................................................................................................................................ 43
 Policy Regarding Research-Related Injuries ................................................................................. 44

REFERENCES ................................................................................................................................ 45
APPENDIX I: CONTACT INFORMATION ........................................................................................... 48
APPENDIX II: SCHEDULE OF EVALUATIONS .................................................................................. 51
APPENDIX III: ASSESSMENT OF RELATIONSHIP TO VACCINE AND GRADING SEVERITY 
OF ADVERSE EVENTS ............................................................................................................................. 55
 
 
 
 
 
 
 
 
 
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 6 of 62 
TABLE OF ABBREVIATIONS 
Abbreviation 
Term 
AE 
adverse event 
AdEDC 
Advantage Electronic Data Capture 
ALT 
alanine aminotransferase 
AoU 
assessment of understanding 
BMI 
body mass index 
CBC 
complete blood count 
cGMP 
current Good Manufacturing Practices 
dbGAP 
Database of Genotypes and Phenotypes 
DNA 
deoxyribonucleic acid 
DTM 
Department of Transfusion Medicine 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practices 
GLP 
Good Laboratory Practices 
HA 
influenza hemagglutinin protein  
HAI 
hemagglutination inhibition 
HIV 
human immunodeficiency virus 
HLA 
human leukocyte antigen 
HRPP 
Human Research Protection Program 
IB 
Investigator’s Brochure 
ICS 
intracellular cytokine staining 
IM 
Intramuscular 
IND 
investigational new drug application 
IRB 
Institutional Review Board 
LIMS 
Laboratory Information Management System 
MedDRA 
Medical Dictionary for Regulatory Activities 
MIV 
Monovalent Inactivated Vaccine 
NA 
neuraminidase  
NAb 
Neutralizing antibody 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
NIH CC 
NIH Clinical Center 
NSAID 
nonsteroidal anti-inflammatory drug 
PBMC 
peripheral blood mononuclear cells 
PI 
Principal Investigator 
PSRT 
Protocol Safety Review Team 
RNA 
ribonucleic acid 
SAE 
serious adverse event 
SARS 
Severe acute respiratory syndrome 
SUSAR 
serious and unexpected suspected adverse reaction 
TNF 
tumor necrosis factors 
ULN 
upper limit of normal 
UP 
Unanticipated Problem 
VCMP 
Vaccine Clinical Materials Program 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 7 of 62 
Abbreviation 
Term 
VITL 
Vaccine Immunology Testing Laboratory 
VRC 
Vaccine Research Center 
WBC 
white blood cell 
WHO 
World Health Organization 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 8 of 62 
PRÉCIS 
VRC 316: 
A Phase I Open-Label Clinical Trial to Evaluate Dose, Safety, Tolerability, and 
Immunogenicity of an Influenza HA Ferritin Vaccine Alone or in Prime-Boost 
Regimen with an Influenza DNA Vaccine in Healthy Adults. 
Study Design: This is a Phase I open-label, dose escalation study to evaluate the dose, safety, 
tolerability, and immunogenicity of VRC-FLUNPF081-00-VP (HA-F A/Sing) 
vaccine alone or in prime-boost regimens with VRC-FLUDNA082-00-VP 
(DNA A/Sing) vaccine. The hypotheses are that VRC-FLUNPF081-00-VP and 
VRC-FLUDNA082-00-VP vaccines are safe, well-tolerated, and induce an 
immune response to the H2 antigen. The primary objectives are to evaluate the 
safety and tolerability of two different doses of the HA-F A/Sing vaccine alone 
and in prime-boost regimens in healthy adults. Secondary objectives are related 
to the evaluation of the immunogenicity of the HA-F A/Sing and DNA A/Sing 
vaccines in prime-boost regimens. 
Study Products: The investigational HA-F A/Sing vaccine, developed by the Vaccine Research 
Center (VRC), National Institute of Allergy and Infectious Diseases (NIAID), is 
composed of Helicobacter pylori non-haem ferritin with an influenza virus H2 
hemagglutinin (HA) insert to form a nanoparticle displaying eight HA trimers 
from A/Singapore/1/57 (H2N2) influenza. 
                           The investigational DNA A/Sing vaccine, developed by the VRC, NIAID, is 
composed of a single closed-circular DNA plasmid that encodes the H2 protein 
of A/Singapore/1/57 influenza.  
Subjects: 
Up to 80 healthy adults ages 18-70 will be enrolled; adults born between 1966 
and 1969 will be excluded from the trial. 
Study Plan: 
Vaccines will be administered intramuscularly (IM) in the deltoid muscle.  This 
study has two parts: 
Part I will evaluate the safety, tolerability, and immunogenicity of 1 or 2 doses 
of the HA-F A/Sing vaccine in a dose-escalation design. In Group 1, five 
subjects will receive a low dose of the HA-F A/Sing vaccine via needle and 
syringe on Day 0. If this low dose is assessed as safe and well tolerated, 
enrollment will begin for Group 2. In Group 2, five subjects will receive the 
higher dose of the HA-F A/Sing vaccine via needle and syringe on Day 0 and 
Week 16. If this higher dose is assessed as safe, enrollment will begin for Part 
II. 
 
Part II will evaluate the safety, tolerability, and immunogenicity of HA-F 
A/Sing vaccine in prime-boost regimens. Subjects will be stratified by age and 
randomized equally into Groups 3 and Group 4 as shown in the vaccination 
schema. In Group 3, subjects will receive DNA A/Sing vaccine via PharmaJet 
Needle-Free Injector on Day 0 and HA-F A/Sing vaccine via needle and syringe 
on Week 16. In Group 4, subjects will receive HA-F A/Sing vaccine via needle 
and syringe on Day 0 and Week 16. 
  
For Group 1, the protocol requires about 8 clinic visits and 1 telephone follow-

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 9 of 62 
up contact after the injection.  
 
For Group 2, Group 3 and Group 4, the protocol requires about 10 clinic visits 
and 2 telephone follow-up contacts after each injection. 
 
For all Groups, solicited reactogenicity will be evaluated using a 7-day diary 
card. Assessment of vaccine safety will include clinical observation and 
monitoring of hematological and chemical parameters at clinical visits 
throughout the study.  
 
VRC 316 Vaccination Schema 
PART I: Dose Escalation of HA-F A/Sing 
Group  
Dose Level  
Age Cohorts 
Subjects 
Day 0 
Prime 
Week 16  
(±4 weeks) 
Boost 
1 
Low dose 
Ages 18-47 
5 
HA-F A/Sing, 20 mcg 
 
2 
High dose 
Ages 18-47 
5 
HA-F A/Sing, 60 mcg 
HA-F A/Sing, 60 mcg 
Total 
10* 
*Enrollment of up to 15 subjects is permitted if additional 
subjects are needed for safety or immunogenicity 
evaluations.  
 
 
HA-F A/Sing injections are administered by needle and 
syringe. 
PART II: Evaluation of HA-F A/Sing and DNA A/Sing in Prime-Boost Regimens 
Group  
Regimen 
Age Cohorts 
Subjects 
Day 0 
Prime 
Week 16  
(±4 weeks) 
Boost 
3 
DNA A/Sing 
HA-F A/Sing 
Group 3A; 
18-47 
10 
DNA A/Sing, 4 mg 
HA-F A/Sing, 60 mcg 
Group 3B; 
52-70 
10 
4 
HA-F A/Sing 
HA-F A/Sing 
Group 4A; 
18-47 
10 
HA-F A/Sing, 60 mcg 
HA-F A/Sing, 60 mcg 
Group 4B; 
52-70 
10 
Total 
40* 
*Enrollment of up to 65 subjects is permitted if additional 
subjects are needed for safety or immunogenicity 
evaluations.  
 
 
DNA A/Sing injections are administered by PharmaJet. 
HA-F A/Sing injections are administered by needle and 
syringe. 
 
Study  
Duration:  
Subjects will be evaluated for 40 weeks following the first vaccine 
administration.  
 
 
 
 
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 10 of 62 
 INTRODUCTION AND RATIONALE 
 BACKGROUND 
Influenza is an enveloped, negative single-stranded (-ss) ribonucleic acid (RNA) virus that 
belongs to the family Orthomyxoviridae. Of the five genera of influenza circulating in nature, 
only influenza A and B are known to cause epidemics in humans [1]. Influenza A viruses consist 
of 8 RNA gene segments and are classified on the basis of the antigenicity of their surface 
glycoproteins: hemagglutinin (HA) and neuraminidase (NA). There are 18 different HA subtypes 
(H1 through H18) and 11 NA subtypes known to exist, but only three HA subtypes (H1, H2, and 
H3) and two NA subtypes (N1 and N2) have caused significant human epidemics [2]. The HA is 
the predominant viral antigen target for antibody neutralization [3].  
Influenza virus causes seasonal epidemics and pandemics at irregular intervals that result in 
significant morbidity and mortality. According to the World Health Organization, the annual 
global attack rate of influenza is estimated to be 5%–10% in adults and 20%–30% in children; 
worldwide these annual epidemics result in about 3 to 5 million cases of severe illness and about 
250,000 to 500,000 deaths [4]. The US Centers for Disease Control and Prevention estimates that 
a pandemic Influenza outbreak costs the United States between $71 billion and $167 billion 
without counting the financial impact on commerce and society [5]. 
Over a million human deaths worldwide were attributed to the H2N2 influenza that occurred 
from 1957 until 1968. H2 influenza remains a potential public health threat because H2 influenza 
viruses continue to circulate in avian and swine reservoirs worldwide. Many humans would 
likely be susceptible to infection as H2 influenza has not circulated in humans since 1968 and 
herd immunity has therefore diminished [6]. 
The emergence of new influenza strains, continuous mutations and reassortment of circulating 
virus diminish the effectiveness of annual influenza vaccines [7, 8]. Furthermore, in the United 
States, the current manufacturing process in egg-based systems can lead to lower yields and 
significant lag times due to virus strain identification and vaccine production, availability, and 
distribution [9, 10]. These limitations have raised the need for developing a universal influenza 
vaccine that can provide durable, cross-strain protection against different influenza viruses, with 
a rapid manufacturing process in which large vaccine quantities could be produced under well-
controlled conditions. A universal influenza vaccine would eliminate the need for annual 
reformulation and revaccination, and improve pandemic preparedness [11]. 
With the goal of developing a universal influenza vaccine, the Vaccine Research Center (VRC), 
National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health 
(NIH) has engineered a ferritin vaccine (VRC-FLUNPF081-00-VP) that enhances the 
presentation of the H2 glycoprotein to the immune system, improving neutralization potency and  
breadth against influenza virus subtype H2. VRC-FLUNPF081-00-VP (HA-F A/Sing) is an 
investigational vaccine that has not been administered to humans before this study. The HA-F 
A/Sing vaccine could play an important role in the planning and preparation for future influenza 
pandemics. 
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 11 of 62 
 RATIONALE FOR DEVELOPMENT OF INFLUENZA HA-F A/SING VACCINE, VRC-
FLUNPF081-00-VP 
New vaccine platforms and production technologies directed toward the goal of a universal 
influenza vaccine include cell-culture-based manufacturing processes, novel live attenuated 
vaccines, recombinant proteins, recombinant DNA-based vaccines, and nanoparticles [9, 10]. 
Ferritin is a ubiquitous iron storage protein, highly conserved, and found in various species from 
Archaea, Eukarya, and Bacteria domains [12], that can be used in vaccines for antigen 
presentation of influenza HA [13]. Ferritin self-assembles into a nearly-spherical nanoparticle, 
composed of  24 subunits organized in an octahedral symmetry with a hollow interior, that 
mimics the structure of the viral antigen and mediates the interaction with the immune system 
[13, 14]. The advantages of using ferritin as a vaccine platform to improve antigen presentation 
and immune stimulation against different strains of influenza viruses rely on the ability to obtain 
higher levels of protein quaternary structures and the capacity to display heterologous antigens 
on their surface [15] [13]. Furthermore, as the self-assembly process requires no energy and can 
be manufactured from simple expression vectors (without relying on egg-based systems), 
vaccine manufacturing timelines would be potentially shortened which would improve the 
response to an influenza pandemic [13]. 
Investigators at the VRC, NIAID, NIH, Bethesda, MD have identified a non-haem ferritin from 
Helicobacter pylori (H. pylori) as a protein able to display eight trimeric influenza HA spikes 
that mimics the physiological HA structure of A/Singapore/1/1957 influenza. [13].  Kanekiyo, et 
al. genetically fused the ectodomain of A/New Caledonia/20/1999 (H1N1) HA to H. pylori 
ferritin, creating a synthetic ferritin that antigenically resembles the native HA viral spikes on the 
surface of the ferritin spherical core [13].  In preclinical immunogenicity studies, this HA ferritin 
vaccine has shown to elicit two types of broadly neutralizing antibodies against the highly 
conserved HA stem and to the conserved receptor binding site (RBS) of the viral HA, which are 
structures of major interest for the development of a universal vaccine against influenza [16]. 
Also, this HA ferritin vaccine possesses the desired structural properties, and has been shown to 
have the capacity to enhance the potency and breadth of neutralizing antibodies in pre-clinical 
studies when it is compared to the current commercial trivalent inactivated vaccine (TIV) 
containing the same 1999 H1N1 HA [13]. 
Based on data reported with the HA ferritin vaccine in animal studies (Section 2.3), it is expected 
that the HA -F A/Sing vaccine will be safe and immunogenic in this Phase I study in humans. H2 
was selected for evaluation of this vaccine concept both because of the low seroprevalence in 
large segments of the human population and the potential for future outbreaks in humans [17]. 
This is the first study to test the VRC-FLUNPF081-00-VP vaccine, and there is no previous 
human experience with this product.  
 RATIONALE FOR DEVELOPMENT OF INFLUENZA PLASMID DNA A/SING VACCINE, VRC-
FLUDNA082-00-VP AND ITS USE WITH VRC-FLUNPF081-00-VP IN PRIME-BOOST 
REGIMENS 
VRC-FLUDNA082-00-VP was developed by the VRC and is a plasmid DNA vaccine that 
encodes the H2 protein of Influenza A/Singapore/1/1957.  This is the first study to test the VRC-
FLUDNA082-00-VP vaccine in a clinical trial, and there is no previous human experience with 
this product. 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 12 of 62 
In this clinical trial, we will evaluate a prime-boost regimen based on data generated by the VRC 
in assessing candidate H5 DNA and H7 DNA vaccines [18-23].  DNA A/Sing-prime and HA-F 
A/Sing-boost will be evaluated and compared to HA-F A/Sing-prime and HA-F A/Sing-boost in 
a 16-week interval.  In previous studies using a boost interval of 12-24 weeks for a DNA 
vaccine-prime and Monovalent Inactivated Vaccine (MIV) boost regimen, the VRC has found 
that antibody responses were higher in the DNA-MIV regimen compared to the MIV-MIV 
vaccination regimen[24] [23].  Based on these studies, priming with a DNA A/Sing or HA-F 
A/Sing vaccine followed by a boost with HA-F A/Sing vaccine in a 16 week interval is being 
evaluated in this trial. 
1.3.1 DNA vaccines developed by VRC, NIAID, NIH 
DNA vaccines have been shown to be an efficient platform technology to respond to emerging 
infectious diseases [25]- they have provided a new alternative to induce humoral and cellular 
immune responses, they are cost-effective to produce and store, and they are more stable than 
traditional vaccines and have an excellent safety profile [26].  VRC plasmid DNA vaccines have 
demonstrated protective immune responses against influenza in preclinical studies [27, 28]. VRC 
DNA vaccines have been evaluated for safety and immunogenicity in human clinical trials, and 
shown to be safe, well tolerated, and capable of eliciting immune responses to targeted influenza 
antigens [24]. 
Cumulatively, since the first clinical trial using the VRC DNA vaccine platform in October 2001, 
more than 3,000 study subjects, 18-70 years of age, have received one or more injections of VRC 
DNA vaccines, which have all been evaluated as safe and immunogenic [18, 29-38]. 
Although a human efficacy study has not yet been completed with any DNA vaccine given 
alone, both VRC clinical trials of West Nile Virus (WNV) vaccines [29, 30], a severe acute 
respiratory syndrome (SARS) virus DNA vaccine (VRC-SRSDNA015-00-VP) [32], an H5 
influenza DNA vaccine (VRC-AVIDNA036-00-VP) [18, 19], and an H7 influenza DNA vaccine 
(VRC-FLUDNA071-00-VP) [23], provided evidence that DNA vaccines can induce neutralizing 
antibodies (NAbs) against targeted pathogens. 
 RATIONALE FOR VRC-FLUDNA082-00-VP VACCINE ADMINISTRATION METHOD 
VRC DNA vaccines have been administered at dosages of up to 8 mg, however the majority of 
injections have been given at 4 mg. Data from two dose-escalation studies (VRC 004 and VRC 
204) indicate that a 4 mg dosage offers the combination of a good safety profile, greater ease of 
administration than an 8 mg dosage, and reliable immunogenicity as indicated by laboratory 
measures of immune response. Preclinical and clinical evaluations to date of plasmid DNA 
vaccines support the safety and immunogenicity of DNA vaccines at 4 mg. 
In previous studies, DNA vaccines have been administered by intramuscular (IM), intradermal or 
subcutaneous routes using either standard needle and syringe or Biojector (Biojector 2000® 
needle-free injection system, Bioject Medical Technologies, Tigard, OR). It was demonstrated 
that the use of a needle-free injection system is superior to needle and syringe by offering a 
significant contribution to the elicited immunogenicity [39]. 
In March 2016, Biojector became unavailable. Consequently, another needle-free injection 
system from PharmaJet Stratis® (Golden, CO) is available. The PharmaJet Stratis® 0.5 mL 
Needle-Free Jet Injector (PharmaJet) is FDA approved for use in children and adults. PharmaJet 
holds a General Use 510k marketing clearance from FDA and received the Performance, Quality 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 13 of 62 
and Safety (PQS) pre-qualified certification from the WHO (#E008/050). PharmaJet provides an 
opportunity to deliver the vaccine by a needle-free device, to simplify clinical operations and 
potentially increase elicited immunogenicity. Immunogenicity studies have been conducted in 
rhesus macaques in which animals received doses of the Zika virus (ZIKVwt) DNA vaccine via 
PharmaJet showed an increase in NAb production following immunization in both groups [40]. 
In this Phase I study in healthy adults, study subjects enrolled only in Group 3 will receive 1 IM 
vaccination with the VRC-FLUDNA082-00-VP vaccine via PharmaJet on Day 0 as per the 
Schedule of Evaluations (Appendix II). 
 RATIONALE FOR STUDY POPULATION PROPOSED IN THE CLINICAL TRIAL  
To assess safety, tolerability, and immunogenicity of the novel  HA-F A/Sing vaccine platform, 
two age cohorts will be evaluated in part II of this study. These age ranges are based on the 
likelihood of prior H2 exposure; human H2 influenza last circulated as a seasonal strain between 
1957 and 1968.  Therefore, the younger age cohort, age 18 through adults born after 1969, 
should have no pre-existing immunity to H2.  The older age cohort, adults born in or before 1965 
through age 70, may have some immunity. The age gap in which subjects are ineligible, adults 
born between January 1, 1966 and December 31, 1969, excludes subjects with unknown 
exposure. H2 immunity will not be evaluated as an eligibility criterion. 
 ASSESSMENT OF VACCINE IMMUNOGENICITY  
In this study, specimens to evaluate immunogenicity will be collected at baseline and at specified 
time points. The primary immunogenicity time point is two weeks after the boost vaccination. 
HA-specific antibody as measured by HAI assay is the traditional benchmark measure of 
immune response to influenza vaccines and will be conducted on stored samples obtained 
throughout the study. Additional measurements of human ferritin antibody, H. pylori ferritin 
antibody, and T cell responses may also be assessed from stored samples for time points 
throughout the study as exploratory evaluations. 
The detection of antibody by HAI or neutralization assay is based on validated laboratory 
methods. Exploratory T cell assays may include the intracellular cytokine staining (ICS) assay 
which is based on previously published methods [41] and quantitates of the frequency of CD4+ 
and CD8+ cells that produce interleukin-2, interferon-gamma or tumor necrosis factor (TNF)-
alpha in response to pools of overlapping peptides representing HA antigens. 
Research samples for immunogenicity assays will be processed by the Vaccine Immunology 
Testing Laboratory (VITL) in Gaithersburg, MD, where many of the immunogenicity assays will 
also be performed. Some immunogenicity assays may be performed by VRC laboratories in 
Bethesda, MD, approved contract laboratories, or approved research collaborators. 
Results from this study are expected to provide a foundation for building a universal influenza 
vaccine candidate as well as show proof-of-concept for engineered HA-ferritin as a vaccine 
platform.  
 
  

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 14 of 62 
 STUDY AGENTS 
The two study agents (VRC-FLUNPF081-00-VP and VRC-FLUDNA082-00-VP) are 
manufactured under current Good Manufacturing Practice (cGMP) by the Vaccine Clinical 
Materials Program (VCMP) operated under contract by Leidos Biomedical Research, Inc., 
Frederick, MD.  
 VRC-FLUNPF081-00-VP 
VRC-FLUNPF081-00-VP (HA-F A/Sing) is a vaccine composed of HA protein from Influenza 
A/Singapore/1/1957 genetically fused to ferritin protein from H. pylori. 
The purified HA-F A/Sing displays eight well-formed HA trimers that antigenically resembles 
the native H2 viral spikes.  
VRC-FLUNPF081-00-VP vaccine is a sterile, aqueous buffered solution.  Vials are aseptically 
filled to a volume of 0.7 mL into a single dose vials at 180 mcg/mL.  The buffer consists of 
20mM NaPhosphate, 100mM NaCl, and 5% Sucrose, at pH 7.2. 
More details related to vaccine production, preparation, quality control and preclinical studies 
performed with HA-F A/Sing vaccine can be found in the Investigator’s Brochure (IB). 
 VRC-FLUDNA082-00-VP 
VRC-FLUDNA082-00-VP (DNA A/Sing) is a DNA vaccine that encodes the hemagglutinin 
(H2) protein of Influenza A/Singapore/1/1957. 
VRC-FLUDNA082-00-VP is a single closed circular plasmid DNA (plasmid VRC 9265) that 
contains a human cytomegalovirus regulatory R region (CMV/R) as a promoter.  The CMV/R 
promoter consists of a translational enhancer region of the CMV immediate-early region one 
enhancer (CMV-IE) substituted with the 5 '-untranslated human T-cell leukemia virus type 1 
(HTLV-1) R-U5 region (HTLV-1 R-U5) long terminal repeat (LTR).  This promoter has been 
shown to optimize gene expression [42] and it has been evaluated in preclinical safety studies 
[43], as well as several clinical trials of DNA plasmid vaccines for Human Immunodeficiency 
Virus (HIV) (BB-IND 11750), WNV (BB-IND 12933), Ebola virus (BB-IND 11294), SARS 
(BB-IND 11995), avian influenza virus (BB-IND 13197), and Zika virus (BB-IND 17080). 
VRC-FLUDNA082-00-VP vaccine is formulated in phosphate buffered saline (PBS) solution. 
Vials are aseptically filled to a volume of 1.2 mL with 4 mg/mL of the plasmid. 
More details related to vaccine production, preparation and quality control of VRC-
FLUDNA082-00-VP can be found in the IB. 
 VRC-PBSPLA043-00-VP, PHOSPHATE BUFFERED SALINE (DILUENT) 
Diluent consists of sterile phosphate buffered saline (PBS) at pH 7.2, aseptically filled at 1.2 mL 
in a 3 mL glass vial for single use.  Diluent vials will be stored until use at -45°C to -10°C. 
 PRECLINICAL STUDIES WITH VRC-FLUNPF081-00-VP 
Non-clinical immunogenicity studies in mice confirmed that the VRC-FLUNPF081-00-VP Drug 
Product administered IM is immunogenic as detected by hemagglutinin inhibition (HAI) and 
neutralization assay. 

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PROTOCOL: VRC 316, VERSION 6.0 
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A repeat dose toxicity study was performed in New Zealand white rabbits in accordance with the 
Good Laboratory Practices (GLP) Regulations for nonclinical laboratory studies. IM 
administration of either the monovalent influenza vaccine (60 µg of VRC-FLUNPF081-00-VP) 
or the trivalent influenza vaccine (60 µg VRC-FLUNPF081-00-VP; 60 µg VRC-FLUNPF088-
00-VP; 60 µg VRC-FLUNPF089-00-VP) once every three weeks for three injections (Study 
Days 1, 22, and 43) was well tolerated as there were no adverse or limiting toxicity seen 
throughout the study and any findings noted were considered to be of minimal toxicological 
significance (e.g., noted in only one sex, reversible, transient, no alteration in organ function, 
etc.). 
More details on pre-clinical studies with VRC-FLUNPF081-00-VP can be found in the IB.  
 PRECLINICAL STUDIES WITH VRC-FLUDNA082-00-VP 
No preclinical pharmacology, toxicology, pharmacokinetic, or metabolism studies were 
conducted for VRC-FLUDNA082-00-VP.  VRC DNA products have been extensively tested in 
animals and humans, alone and as a prime for various vaccine boosts, and have shown a robust 
safety profile.  Early biodistribution studies showed complete DNA clearance by three months 
[43].  
More details on pre-clinical studies with DNA vaccines can be found in the IB.  
 STUDY OBJECTIVES 
 PRIMARY OBJECTIVES 
• To evaluate the safety and tolerability of the HA-F A/Sing vaccine, administered as a 
single dose at 20 mcg IM via needle and syringe at Day 0 to healthy adults. 
 
• To evaluate the safety and tolerability of the HA-F A/Sing vaccine, administered at 60 
mcg IM via needle and syringe to healthy adults by repeat dosing at Day 0 and Week 16 
for a total of 2 injections. 
 
• To evaluate the safety and tolerability of the DNA A/Sing vaccine, administered as a 
single dose at 4 mg IM via needle-free injection device at Day 0 to healthy adults in the 
prime-boost regimen. 
 SECONDARY OBJECTIVES 
• To evaluate the antibody response to two prime-boost vaccine regimens in healthy adults:  
 
1) DNA A/Sing vaccine administered at 4 mg IM via needle-free injection device at Day 
0 followed by HA-F A/Sing vaccine administered at 60 mcg IM via needle and 
syringe at Week 16. 
2) HA-F A/Sing vaccine administered at 60 mcg IM via needle and syringe by repeat 
dosing at Day 0 and Week 16. 
• To compare the magnitude and the frequency of H2-specific antibody response as 
measured by HAI at four weeks after the prime vaccine and two weeks after the boost 
vaccine for each treatment and age group. 

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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
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 EXPLORATORY OBJECTIVES 
• To evaluate the magnitude, specificity and frequency of the H2-specific T-cell, B-cell 
responses and antibody responses at indicated time points throughout the study. 
 STUDY DESIGN AND METHODS 
This is a Phase I, open-label, dose escalation study to evaluate the dose, safety, tolerability, and 
immunogenicity of the VRC-FLUNPF081-00-VP vaccine alone or in prime-boost regimens with 
the VRC-FLUDNA082-00-VP vaccine in healthy adults. The study schema is shown in Table 2. 
The hypotheses are that the vaccines are safe, well-tolerated, and will induce an antibody 
response to influenza virus subtype H2. The study will be conducted by the VRC Clinical Trials 
Program at a single site at the NIH Clinical Center (NIH CC). 
Table 2: VRC 316 Vaccination Schema 
PART I: Dose Escalation of HA-F A/Sing 
Group  
Dose Level  
Age Cohorts 
Subjects 
Day 0 
Prime 
Week 16  
(±4 weeks) 
Boost 
1 
Low dose 
Ages 18-47 
5 
HA-F A/Sing, 20 mcg 
 
2  
High dose 
Ages 18-47 
5 
HA-F A/Sing, 60 mcg 
HA-F A/Sing, 60 mcg 
Total 
10* 
*Enrollment of up to 15 subjects is permitted if 
additional subjects are needed for safety or 
immunogenicity evaluations. 
 
 
HA-F A/Sing injections are administered by needle and 
syringe. 
PART II: Evaluation of HA-F A/Sing and DNA A/Sing in Prime-Boost Regimens 
Group  
Regimen 
Age Cohorts 
Subjects 
Day 0 
Prime 
Week 16  
(±4 weeks) 
Boost 
3  
DNA A/Sing 
HA-F A/Sing 
Group 3A; 
18-47 
10 
DNA A/Sing, 4 mg 
HA-F A/Sing, 60 mcg 
Group 3B; 
52-70 
10 
4 
HA-F A/Sing 
HA-F A/Sing 
Group 4A; 
18-47 
10 
HA-F A/Sing, 60 mcg 
HA-F A/Sing, 60 mcg 
Group 4B; 
52-70 
10 
Total 
40* 
*Enrollment of up to 65 subjects is permitted if 
additional subjects are needed for safety or 
immunogenicity evaluations.  
 
 
 DNA A/Sing injections are administered by PharmaJet. 
 HA-F A/Sing injections are administered by needle and 
syringe. 
 STUDY POPULATION 
All inclusion and exclusion criteria must be evaluated for eligibility. 
4.1.1 Inclusion Criteria 
A subject must meet all of the following criteria: 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
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1. Healthy subjects aged 18-70 (excluding subjects born between 1966-1969) 
2. Based on history and examination, must be in good general health and without history of 
any of the conditions listed in the exclusion criteria 
3. Received at least one licensed influenza vaccine from 2014 to the present 
4. Able and willing to complete the informed consent process 
5. Available for clinic visits for 40 weeks after enrollment 
6. Willing to have blood samples collected, stored indefinitely, and used for research 
purposes 
7. Able to provide proof of identity to the satisfaction of the study clinician completing the 
enrollment process  
8. Physical examination and laboratory results without clinically significant findings and a 
BodyMassIndex(BMI)≤40withinthe84 days before enrollment 
Laboratory Criteria within 84 days before enrollment 
9. White blood cells (WBC) and differential either within institutional normal range or 
accompanied by the site Principal Investigator (PI) or designee approval 
10. Totallymphocytecount≥800cells/mm3 
11. Platelets = 125,000 – 500,000/mm3 
12. Hemoglobin within institutional normal range 
13. Serum iron either within institutional normal range or accompanied by the site PI or 
designee approval 
14. Alanineaminotransferase(ALT)≤1.25xinstitutional upper limit of normal (ULN) 
15. Aspartateaminotransferase(AST)≤1.25xinstitutionalULN 
16. Alkalinephosphatase(ALP)≤1.1xinstitutional ULN 
17. Total bilirubin within institutional upper limit of normal (ULN) 
18. Serum creatinine≤1.1xinstitutional ULN 
19. Negative for HIV infection by an FDA approved method of detection 
Criteria applicable to women of childbearing potential: 
20. Negative beta-humanchorionicgonadotropin(β-HCG) pregnancy test (urine or serum) 
on the day of enrollment  
21. Agrees to use an effective means of birth control from at least 21 days prior to enrollment 
through the end of the study 
4.1.2 Exclusion Criteria 
A subject will be excluded if one or more of the following conditions apply: 
1. Breast-feeding or planning to become pregnant during the study.  
Subject has received any of the following substances: 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 18 of 62 
2. More than 10 days of systemic immunosuppressive medications or cytotoxic medications 
within the 4 weeks prior to enrollment or any within the 14 days prior to enrollment  
3. Blood products within 16 weeks prior to enrollment 
4. Live attenuated vaccines within 4 weeks prior to enrollment 
5. Inactivated vaccines within 2 weeks prior to enrollment.  
6. Investigational research agents within 4 weeks prior to enrollment or planning to receive 
investigational products while on the study 
7. Current allergy treatment with allergen immunotherapy with antigen injections, unless on 
maintenance schedule 
8. Current anti-TB prophylaxis or therapy 
9. Previous H2 influenza investigational vaccine 
10. Receipt of a licensed influenza vaccine within 6 weeks before trial enrollment 
Subject has a history of any of the following clinically significant conditions: 
11. Serious reactions to vaccines that preclude receipt of study vaccinations as determined by 
the investigator  
12. Hereditary angioedema, acquired angioedema, or idiopathic forms of angioedema 
13. Asthma that is not well controlled  
14. Diabetes mellitus (type I or II), with the exception of gestational diabetes 
15. Thyroid disease that is not well controlled 
16. Idiopathic urticaria within the past year 
17. Evidence of autoimmune disease or immunodeficiency 
18. Hypertension that is not well controlled 
19. Bleeding disorder diagnosed by a doctor (e.g. factor deficiency, coagulopathy, or platelet 
disorder requiring special precautions) or significant bruising or bleeding difficulties with 
IM injections or blood draws 
20. Malignancy that is active or history of malignancy that is likely to recur during the period 
of the study. 
21. Seizure disorder other than 1) febrile seizures, 2) seizures secondary to alcohol 
withdrawal more than 3 years ago, or 3) seizures that have not required treatment within 
the last 3 years 
22. Asplenia, functional asplenia or any condition resulting in the absence or removal of the 
spleen 
23. Guillain-Barré Syndrome 
24. Psychiatric condition that precludes compliance with the protocol; past or present 
psychoses; or within 5 years prior to enrollment, a history of suicide plan or attempt 

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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 19 of 62 
25. Any medical, psychiatric, social condition, occupational reason or other responsibility 
that, in the judgment of the investigator, is a contraindication to protocol participation or 
impairs a subject’s ability to give informed consent. 
 CLINICAL PROCEDURES AND EVALUATIONS 
Evaluation of this vaccine will include laboratory tests, medical history, physical assessment by 
clinicians, and subject self-assessment recorded on a diary card for 7 days after each injection. 
Potential adverse reactions will be further evaluated prior to continuing the vaccination schedule. 
The schedule of study evaluations is described in this section and shown in table format in 
Appendix II. Total blood volume drawn from each subject will not exceed the NIH CC 
Guidelines. 
4.2.1 Screening 
Screening for this study will be completed through the VRC’s screening protocol, VRC 500 
(NIH 11-I-0164). Subjects will be recruited through Institutional Review Board (IRB)-approved 
advertising. The evaluations and sample collection that will be included in screening are a 
medical history, physical exam, any laboratory tests needed to confirm eligibility, and pregnancy 
test for females of reproductive potential.  
If screening evaluations suggest a current concerning health condition or infection, such as 
hepatitis, then appropriate laboratory tests may be conducted to evaluate for these conditions. 
Additional assessments of health may be conducted at screening based on clinical judgment. 
Screening evaluations for specific eligibility criteria (Section 4.1) must be completed within the 
time interval specified prior to enrollment for the given parameter and may be repeated, as 
needed, to confirm eligibility. Research blood samples will be collected during screening; 
although generally collected in the 84 days prior to enrollment; a particular interval of time prior 
to enrollment for collection of these samples is not specified. 
The informed consent form (ICF) will be reviewed and counseling related to pregnancy 
prevention will be provided. 
Subjects who are not up to date on standard vaccinations may receive these, if available, during 
their participation in the screening protocol or at a later date during study participation. As part 
of the informed consent process, an Assessment of Understanding (AoU) is completed on the 
day the subject is scheduled to enroll prior to signing the VRC 316 protocol informed consent 
document. Incorrect answers will be explained to the volunteer, who will sign the ICF only after 
the study clinician is satisfied with the volunteer’s understanding of the study. 
4.2.2 Study Schedule  
The Schedule of Evaluations in Appendix II provides details on the study schedule and the 
permitted visit windows. A clinician will discuss the target dates and timing of the study 
vaccination(s) and sample collections before completing an enrollment to help ensure that the 
subject can comply with the projected schedule. 
The schedule of study visits, permitted windows for completing the visits, and evaluations 
performed at each visit are shown in Appendix II. After enrollment, deviations from the visit 
windows in completing study visits and study injections are discouraged and will be recorded as 
protocol deviations but are permitted at the discretion of the PI (or designee) in the interest of 
completing the vaccination schedule and obtaining subject safety and immunogenicity 
evaluations. 

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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
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4.2.3 Enrollment and Randomization 
Day 0 is defined as the day of protocol enrollment and first injection for all groups. Protocol-
specific eligibility is reviewed on Day 0 as part of the enrollment process, but eligibility 
evaluations conducted during a screening visit are routinely used for eligibility if evaluations are 
within the specified window prior to Day 0.  However, if clinical assessment on Day 0 suggests 
significant changes may have occurred since the screening visit, then evaluations done on Day 0 
are used for eligibility. Day 0 evaluations and medical history prior to the first injection are the 
baseline for subsequent safety assessments.  
Part I includes staged enrollment with required interim safety reviews before proceeding with the 
next dose group. In Group 1, five subjects will be enrolled with no more than one subject per 
day. Following dose escalation review, five subjects will be enrolled in Group 2 with no more 
than one subject per day. An internal safety review will determine whether enrollment can begin 
in Part II.   
In Part II subjects will be stratified by age and randomized equally into Group 3 and Group 4.  
Neither the clinic staff nor the subjects will know in advance which group will be assigned to the 
enrollees in the study. The Protocol Statistician will prepare the randomization plan in advance 
and provide it to the site Pharmacy and Data Management Center. 
The pharmacy database and study randomization will be set up prior to opening the study to 
accrual. The group assignment will become known to the staff and subject immediately after 
completing the electronic enrollment into the study on Day 0. The visit schedule is based on 
intervals of time after each study injection (Appendix II). 
4.2.4 Administration of Study Vaccines 
All study injections will be completed according to the assigned group and vaccines will be 
administered IM in the deltoid muscle.  Blood draws must be completed before vaccinations. 
On the injection day (prior to injection), study subjects will be clinically evaluated and samples 
will be collected as per Schedule of Evaluations (Appendix II). A subject who arrives at the 
clinic with fever or evidence of an acute illness that precludes administration of the vaccine may 
be rescheduled within the allowed study visit window. 
Pregnancy test results for women of reproductive potential must be obtained on each injection 
day prior to the study injection and the results must be negative to proceed. 
When choosing an arm for injection, clinicians should consider whether there is an arm injury, 
local skin problem or significant tattoo that precludes administering the injection or will interfere 
with evaluating the arm after injection.  
Subjects enrolled in Group 1, Group 2, and Group 3 will be observed for a minimum of 1 hour. 
Vital signs (temperature, blood pressure, pulse and respiratory rate) will be collected at least 1 
hour after the injection, prior to subject departure from the clinic. Subjects enrolled in Group 4 
will be observed for a minimum of 30 minutes; vital signs will be collected after at least 30 
minutes. The injection site will be inspected for evidence of local reaction.  
Subjects randomized in Group 3 will receive the DNA A/Sing prime vaccine with the needle-free 
injection device, PharmaJet. The PharmaJet device requires that the DNA A/Sing product be 
administered as two 0.5 mL injections. When the PharmaJet device is used, subjects and 
clinicians will be asked to wear safety glasses during product administration. (See Section 8.5.2) 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 21 of 62 
In keeping with the NIH CC policy and good medical practice, acute medical care will be 
provided to subjects for any immediate allergic reactions or other injury resulting from 
participation in this research study.  
4.2.5 7-Day Solicited Reactogenicity and Follow-up 
Subjects will be given a “Diary Card” to use as a memory aid, on which to record temperature 
and symptoms daily for 7 days after each injection. Subjects will be trained and encouraged to 
use the secure electronic database but will have the option to complete a paper diary card. When 
the diary card parameters are recorded directly by the subject in the electronic database, the 
subject’s electronic record will be the source for these data. When collected on paper, the paper 
diary card will be used as a source document. When neither paper nor electronic diary is 
available from the subject, the study clinician will document the source of reactogenicity 
information recorded in the study database. 
The solicited signs and symptoms on the diary card will include the following parameters: 
unusually tired/feeling unwell, muscles aches (other than at injection site), headache, chills, 
nausea, joint pain, and pain/tenderness at injection site. Subjects will also record the day’s 
highest measured temperature and measurement of largest diameter for redness and swelling at 
injection site.  
Follow-up on subject well-being will be performed by telephone on the first day following all 
injections and by clinic visits as shown in the Schedule of Evaluations (Appendix II) 
Events following any study injection that may require clinical evaluation include rash, urticaria, 
fever of38.5˚C(Grade2)orhigherlastinggreaterthan24hours,orsignificantimpairmentinthe
activities of daily living. Additionally, other clinical concerns may prompt a study visit based on 
the judgment of a study clinician. 
4.2.6 Follow-Up through End of Study 
Study follow-up will continue via clinic visits through 40 weeks following the first vaccine 
administration. Refer to Section 4.4 which describes the criteria for discontinuing product 
administration and/or study participation. 
4.2.7 Blood Sample Collection 
At intervals throughout the study, blood will be drawn for safety and immunologic assays. Blood 
will be drawn from the arm veins of subjects by standard phlebotomy procedures. Total blood 
volume drawn from each subject will not exceed the NIH CC Guidelines. 
4.2.8 Apheresis 
Apheresis will be offered as an optional procedure at Visit 08 (except for Group 1). Each 
apheresis procedure will be carried out by trained Department of Transfusion Medicine (DTM) 
medical staff using automated cell separator devices.  
All apheresis procedures performed under this protocol will be performed solely for research 
purposes. Prior to the scheduling apheresis, the subject must have a venous assessment 
performed by the DTM staff.  
In order to undergo apheresis procedures, a subject must meet the apheresis eligibility criteria 
and have no medical contraindications, as determined by the DTM staff.  A VRC study clinician 
will complete a checklist for apheresis eligibility before referring a subject for the procedure.   

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4.2.8.1 Apheresis Procedures 
All study subjects will be treated according to standard whole blood and apheresis donation 
policies and procedures operative in the DTM. 
For women of reproductive potential, a pregnancy test by blood or urine will be performed by a 
VRC study clinician within 72 hours prior to the apheresis procedure. Results must be negative 
to proceed with apheresis.   
Prior to beginning the apheresis procedure, a study clinician may request in advance that other 
laboratory tests be collected as needed to monitor the well-being of the subject or samples as 
needed by a research laboratory.  
The Dowling Apheresis Clinic staff at the NIH Clinical Center routinely performs a hemoglobin 
test prior to initiating apheresis, per DTM Apheresis Clinic standard policies. If a subject is 
found to have a hemoglobin value less than permitted by the Apheresis Clinic, then the apheresis 
will not be initiated and the ordering provider will be notified.  The VRC Study Coordinator will 
provide the Apheresis center with a request for numbers and types of tubes of blood to be 
collected prior to beginning the apheresis.  
In this study, the procedure will require two antecubital venous access sites and will involve 
processing 1 to 4 liters of whole blood. The expected mononuclear cell yield is approximately 
0.5 to 1.0 x 109 cells per liter processed, and the apheresis device can process about 2-3 liters per 
hour. Thus, 1 to 2 hours are required to process 1 to 4 liters of blood and obtain about 1-4 x 109 
leukocytes. The packed red cell loss during the procedure is the equivalent of a 6 mL blood 
draw; this is the volume that will be used for the purposes of calculating cumulative blood draw 
when apheresis is performed. 
During or following an apheresis visit, if there is any concern about the well-being of the subject, 
the DTM clinic may conduct appropriate medical evaluations by history-taking, physical 
examination, laboratory tests, and/or other testing. 
Research blood samples will be processed and stored at VITL or a collaborating research 
laboratory. Stored samples may be used later to further evaluate immune responses and to 
elucidate genetic factors associated with immune response. 
4.2.9 Concomitant Medications 
Only routine prescription medications will be entered in the database at the time of enrollment.  
Subsequently, concomitant medications associated with an AE requiring expedited reporting or 
the development of a new chronic condition requiring ongoing medical management will be 
recorded or updated in the database. Antiviral medications taken for influenza or influenza-like 
illnesses will be recorded in the database. Receipt of a FDA-approved vaccine at any time during 
the study will be recorded in the database (clinicians should work with subjects regarding the 
timing of licensed vaccines relative to study injection). Otherwise, concomitant medications 
taken throughout the study will be recorded in the subject’s chart as needed for general medical 
documentation but will not be recorded in the database.   
 CRITERIA FOR DOSE ESCALATION AND DOSE CONTINUATION  
There will be one dose escalation in this study. The Protocol Safety Review Team (PSRT, 
Section 8.8) will conduct an interim safety data review before dose escalation and repeat dosing 

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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 23 of 62 
may occur. The PSRT must assess the data as showing no significant safety concerns before 
enrollment of the next dose level and repeat dosing at the same level may proceed. 
Enrollment will begin in Group 1 (20 mcg of VRC-FLUNPF081-00-VP) with no more than one 
subject enrolled per day. Two weeks after vaccination of the fifth volunteer, there will be an 
internal safety review on all five subjects to determine whether to proceed to the next dose level. 
If the 20 mcg dose of VRC-FLUNPF081-00-VP vaccine is assessed as safe, enrollment will 
begin for Group 2. 
In Group 2 (60 mcg of VRC-FLUNPF081-00-VP), enrollment continues with no more than one 
subject enrolled per day. Two weeks after vaccination of the fifth volunteer, there will be an 
internal safety review on all five subjects to determine whether to proceed to Part II. If the 60 
mcg dose of VRC-FLUNPF081-00-VP vaccine is assessed as safe, subjects in Group 2 will 
proceed to receive a second vaccination at week 16 and enrollment can begin for all groups in 
Part II. 
If a first vaccination is not completed or there are discontinuations from the study before there 
are sufficient data to conduct the dose escalation review for a group, then extra subjects may be 
enrolled into that group in order to have the requisite data on at least 5 subjects. 
The IRB will be provided with documentation of the safety review process and notification of 
the dose escalation. Consultation with the IRB and FDA, if needed, as per study pause criteria 
(Section 4.5) will occur if indicated by the review. One outcome of a dose escalation review may 
be to recommend evaluation of additional subjects at the current dose level and reassess for 
safety before proceeding to a higher dose level and repeat dosing at the same dose level.  
 CRITERIA FOR DISCONTINUING STUDY INJECTIONS OR PROTOCOL PARTICIPATION 
Decisions on discontinuation of study injections or protocol participation for a subject will be 
made by the site PI or designee. 
4.4.1 Discontinuation of Study Injections 
Subjects who receive at least one injection will continue follow-up according to the Schedule of 
Evaluations., except that the research sample collections will be discontinued for pregnant 
women or others in which it is contraindicated A subject may be discontinued from receiving 
study product for the following reasons: 
1. 
Pregnancy. 
2. 
Grade 3 AE assessed as related to study product (with the exception that self-limited 
Grade 3 solicited reactogenicity does not require discontinuation). 
3. 
Grade 4 AE assessed as related to study product.  
4. 
Immediate hypersensitivity reaction associated with study product.  
5. 
Intercurrent illness that is not expected to resolve prior to the next scheduled study 
injection. 
6. 
Treatment with systemic glucocorticoids (e.g., prednisone or other glucocorticoid) or 
other immunomodulators (other than nonsteroidal anti-inflammatory drugs [NSAIDs]), 
with the exception that study injections may continue per investigator discretion if the 
next dose occurs at least 2 weeks following completion of glucocorticoid treatment. 

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7. 
The study PI assesses that it is not in the best interest of the subject to continue on the 
vaccination schedule. 
4.4.2 Discontinuation of Protocol Participation 
A subject may be discontinued from protocol participation for the following reasons: 
1. Subject decides to discontinue participation; 
2. Subject develops a medical condition that is a contraindication to continuing study 
participation; 
3. The IND sponsor or regulatory authority stops the protocol; 
4. The PI assesses that is it not in the best interest of the subject to continue participation in 
the study or that the subject’s compliance with the study is not sufficient. 
 CRITERIA FOR PAUSING AND RESUMING THE STUDY 
4.5.1 Plan for Pausing the Study 
The PI and Protocol Safety Review Team (PSRT) will closely monitor and analyze study data as 
they become available and will make determinations regarding the presence and severity of AEs. 
The administration of study injections and new enrollments will be paused and the IND Sponsor 
will be promptly notified according to the following criteria: 
• One (or more) subject experiences a SAE assessed as related to study product. 
• Two (or more) subjects experience the same Grade 3 or higher unsolicited AE assessed 
as related to study product. 
• Three (or more) subjects experience the same Grade 2 or higher laboratory 
abnormality assessed as related to study product.   
 
4.5.2 Plan for Review of Pauses and Resuming the Study 
The IND Sponsor, with participation by the PI and PSRT, will conduct the review and make the 
decision to resume, amend or close the study.  As part of the pause review, the reviewers will 
also advise on whether the study needs to be paused again for any subsequent AEs of the same 
type.  The pause criterion for the SAE will continue to apply. 
The administration of study injections and new enrollments would resume only if review of the 
AEs that caused the pause resulted in a recommendation to permit further study injections and 
enrollments. Safety data reports and changes in study status will be submitted to relevant 
regulatory authorities in accordance with Section 5 and institutional policy. 
 SAFETY AND ADVERSE EVENT REPORTING 
 ADVERSE EVENTS 
An adverse event (AE) is any untoward or unfavorable medical occurrence in a human subject, 
including any abnormal sign (e.g., abnormal physical exam or laboratory finding), symptom, or 
disease temporally associated with the use of study treatment, whether or not considered related 
to the study treatment.   

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Each AE will be graded according to the Table for Grading Severity of Adverse Events 
(Appendix III). The following guidelines will be used to determine whether or not an AE is 
recorded in the study database: 
• Solicited AEs (i.e., reactogenicity parameters) will be recorded by the subject in paper or 
electronic diary cards for 7 days after each injection. If paper diary card is completed by 
subject, data will be transcribed by clinician into the study database without attribution 
assessments. Clinicians will follow and collect resolution information for any 
reactogenicity symptoms that are not resolved within 7 days. 
• All unsolicited AEs will be recorded in the study database from receipt of first study 
injection through the “Week 4” visit after each study injection with attribution 
assessments. At other time periods between injections and when greater than 4 weeks 
after the booster injection, only SAEs (Section 5.2), new chronic medical conditions, and 
influenza or influenza-like illness (ILI) will be recorded through the last study visit.  
• Cases of influenza or influenza-like illness will be evaluated using clinical judgment and 
current CDC definition of influenza symptoms 
(https://www.cdc.gov/flu/about/disease/complications.htm) and recorded on a case report 
form rather than on an AE form. ILI is defined as fever (temperature of 100F [37.8C] or 
greater) and a cough and/or sore throat in the absence of a known cause other than 
influenza. 
 
 SERIOUS ADVERSE EVENTS 
The term “Serious Adverse Event” (SAE) is defined in 21 CFR 312.32 as follows: “An adverse 
event or suspected adverse reaction is considered serious if, in the view of either the investigator 
or the 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 hospitalization 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.” 
“Life-threatening” refers to an AE that at occurrence represents an immediate risk of death to the 
subject. An event that may cause death if it occurs in a more severe form is not considered life-
threatening. Similarly, a hospital admission for an elective procedure is not considered a SAE. 
 ADVERSE EVENT REPORTING TO THE IND SPONSOR 
AEs that meet SAE Reporting Requirements must be reported and submitted by the clinical site 
on an expedited basis to the IND Sponsor, VRC/NIAID/NIH, according to sponsor guidelines as 
follows:  
• results in death 
• is life threatening 

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• results in persistent or significant disability/incapacity 
• requires unplanned inpatient hospitalization or prolongation of existing hospitalization 
• is a congenital anomaly/birth defect in the offspring of a study subject 
• is an important medical event that may jeopardize the subject or may require intervention 
to prevent one of the other outcomes listed above.  
In addition, any event, regardless of severity, which in the judgment of the PI represents a SAE, 
may be reported on an expedited basis. 
An investigator will communicate an initial SAE report within 24 hours of site awareness of 
occurrence to the IND Sponsor by email to the VRC Protocol Operations Office (Appendix I).  
A written report by PI should be submitted to the IND Sponsor within 3 working days. In order 
for the IND Sponsor to comply with regulations mandating sponsor notification of specified 
SAEs to the FDA within 7 or 15 calendar days, the investigator must submit additional 
information as soon as it is available.   
5.3.1 IND Sponsor Reporting to the FDA 
It is the responsibility of the IND Sponsor to make the determination of which SAEs are “serious 
and unexpected suspected adverse reactions” (SUSARs) as defined in 21 CFR 312.32. 
• Suspected adverse reaction means any AE for which there is a reasonable possibility that 
the drug caused the AE. 
• Unexpected Adverse Event means an AE that is not listed in the Investigator’s Brochure 
or is not listed at the specificity or severity that has been observed.   
All SUSARs, as determined by the IND Sponsor, will be reported to the FDA as IND Safety 
Reports and IND Safety Reports will be provided to the IRB. 
The IND Sponsor will also submit an IND Annual Report of the progress of the investigation to 
the FDA as defined in 21 CFR 312.33. 
 REPORTING TO THE INSTITUTIONAL REVIEW BOARD 
5.4.1 Unanticipated Problem Definition 
An  unanticipated problem (UP) is defined as any incident, experience, or outcome that meets all 
three of the following criteria: 
• unexpected in nature, severity, or frequency in relation to the research risks that are 
described in the protocol, informed consent, Investigator’s Brochure, other study 
documents or in consideration of the characteristics of the subject population being 
studied; and 
• related to participation in the research; and 
• suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized. 
Serious UP: An UP that meets the definition of a SAE or compromises the safety, welfare or 
rights of subjects or others. 

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An UP that is not an Adverse Event (UPnonAE) is an unanticipated problem that does not fit the 
definition of an AE, but which may, in the opinion of the investigator, involve risk to the subject, 
affect others in the research study, or significantly impact the integrity of research data. Such 
events would be considered a non-serious UP. For example, we will report occurrences of 
breaches of confidentiality, accidental destruction of study records or samples, or unaccounted-
for study drug. 
5.4.2 Protocol Deviation Definition 
A protocol deviation is defined as any change, divergence, or departure from the IRB-approved 
study procedures in a research protocol. Protocol deviations are designated as serious or non-
serious and further characterized as follows: 
• Those that occur because a member of the research team deviates from the protocol. 
• Those that are identified before they occur, but cannot be prevented. 
• Those that are discovered after they occur. 
Serious Protocol Deviation: A deviation that meets the definition of a SAE or compromises the 
safety, integrity of the data, welfare or rights of subjects or others. 
5.4.3  Non-Compliance Definition 
Non-compliance is the failure to comply with applicable NIH Human Research Protections 
Program (HRPP) policies, IRB requirements, or regulatory requirements for the protection of 
human subjects. Non-compliance is further characterized as serious, continuing or minor. 
“Serious non-compliance” is defined as non-compliance that: 
• Increases risks, or causes harm, to participants 
• Decreases potential benefits to participants 
• Compromises the integrity of the NIH-HRPP 
• Invalidates the study data 
“Continuing non-compliance” is non-compliance that is recurring. 
“Minor non-compliance” is non-compliance that is neither serious nor continuing. 
5.4.4 Expedited Reporting to the NIAID IRB 
The following will be reported within 7 calendar days of investigator awareness: 
• Serious and non-serious UP 
• Deaths 
• Serious protocol deviations 
• Serious or continuing non-compliance  
• SAEs that are possibly, probably, or definitely related to the research regardless of 
expectedness 
The following waiver applies to reporting anticipated protocol deviations and expected 
UPnonAEs: Anticipated deviations in the conduct of the protocol will not be reported to the IRB 
unless they occur at a rate greater than anticipated by the study team. Expected AEs will not be 

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Page 28 of 62 
reported to the IRB unless they occur at a rate greater than that known to occur in healthy adults. 
If the rate of these events exceeds the rate expected by the study team, the events will be 
classified and reported as though they are unanticipated problems.  
5.4.5 Annual Reporting to the NIAID IRB 
The following will be reported to the NIAID IRB in summary at the time of Continuing Review: 
• Serious and non-serious UPs  
• Expected SAEs that are possibly, probably, or definitely related to the research 
• SAEs that are not related to the research 
• All AEs, except expected AEs granted a waiver of reporting  
• Serious and non-serious Protocol Deviations  
• Serious, continuing, and minor non-compliance  
• Any trends or events which in the opinion of the investigator should be reported 
 SERIOUS ADVERSE EVENT REPORTING TO THE INSTITUTIONAL BIOSAFETY COMMITTEE 
(IBC) 
The NIH Institutional Biosafety Committee (IBC) (Bethesda, MD) has the responsibility to 
review research using recombinant DNA for compliance with NIH Guidelines. In keeping with 
IBC requirements, any SAE reports sent to the IRB will be subsequently provided to the IBC by 
the investigators. 
 STATISTICAL CONSIDERATIONS 
 OVERVIEW 
This is a Phase I, open-label, dose escalation study to evaluate the dose, safety, tolerability, and 
immunogenicity of VRC-FLUNPF081-00-VP vaccine alone or in prime-boost regimens with 
VRC-FLUDNA082-00-VP vaccine. 
 OBJECTIVES 
The primary objectives are to evaluate the safety and tolerability of VRC-FLUNPF081-00-VP 
vaccine when administered as a single dose at 20 mcg, and to evaluate the safety and tolerability 
of two prime-boost vaccine regimens: 
• VRC-FLUNPF081-00-VP administered at 60 mcg by repeat dosing at Day 0 and Week 16. 
• VRC-FLUDNA082-00-VP vaccine administered at 4 mg at Day 0 and VRC-FLUNPF081-
00-VP administered at 60 mcg at Week 16.  
Secondary objectives include the evaluation of antibody responses to two prime-boost vaccine 
regimens in healthy adults at two weeks after the boost vaccine for each treatment and age group.  
Exploratory objectives include the evaluation of the magnitude, the specificity, and the frequency 
of H2-specific T-cell, B-cell responses and antibody responses at indicated time points 
throughout the 40 weeks of the study. 

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 ENDPOINTS 
6.3.1 Primary Endpoints: Safety 
Assessment of product safety will include clinical observation and monitoring of hematological 
and chemical parameters. Reactogenicity will be closely monitored for 7 days after each 
injection and safety evaluated by clinical visits throughout 40 weeks. See Section 4.2 and 
Appendix II for details and specified time points.  
The following parameters will be assessed for all study groups: 
• Occurrence of solicited local reactogenicity symptoms for 7 days following each injection 
• Occurrence of solicited systemic reactogenicity symptoms for 7 days following each 
injection 
• Change from baseline in safety laboratory measures 
• Occurrence of AEs of all severities through the “Week 4” visit after each injection 
• Occurrence at any time throughout the study of SAEs or new chronic medical conditions 
that require ongoing medical management. 
6.3.2 Secondary Endpoints: Immunogenicity 
The principal immunogenicity endpoints are magnitude and frequency of antibody responses 
measured by neutralization assay and/or HAI assay at Week 0, at 4 weeks (W4) after the prime 
vaccination, and at 2 weeks (W18) after the boost vaccination. 
6.3.3 Exploratory Endpoints: Immunogenicity  
The magnitude, the specificity, and the frequency of H2-specific neutralizing antibodies that are 
four-fold greater than baseline at 2 weeks after the boost for each group will be evaluated. 
Evaluation of antibody and T cell responses at time points throughout the study will be 
completed as an exploratory evaluation. 
For the H2 neutralization assay, a positive response for a subject is defined as the subject 
achieving a four-fold rise in the H2-specific neutralization antibody titer from baseline. 
 SAMPLE SIZE AND ACCRUAL 
Recruitment will target 50 healthy adult participants 18 to 70 years of age. Adults born between 
1966 and 1969 will be excluded from the trial. 
6.4.1 Power Calculations for Safety 
The goal of the safety evaluation for this study is to identify safety concerns associated with 
injections of the investigational vaccines. Primary sample size calculations for safety are 
expressed in terms of the ability to detect serious adverse experiences. Other sample size 
calculations for comparing the two vaccination groups on adverse experiences are similar to the 
calculations for immunogenicity (Section 6.4.3). 
The ability of the study to identify SAEs will be expressed in terms of the probability of 
observing a certain number of SAEs. Useful values are the minimum true rate such that the 
probability of observing at least one event is at least 90%, and the maximum true rate such that 
the probability of not observing any event is at least 90%.  

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For Part I (dose-escalation) within each group (n=5), there is a 90% chance to observe at least 1 
SAE if the true rate is at least 0.369 and over 90% chance to observe no SAE if the true rate is 
less than 0.021. 
For Part II (prime-boost) within each group (n=10), there is over a 90% chance to observe at 
least 1 SAE if the true rate is at least 0.206 and over a 90% chance to observe no SAE if the true 
rate is no more than 0.010. 
Probabilities of observing 0 or more than 1 SAE within each group are presented in Table 6-1 
for a range of possible true event rates. These calculations provide a complete picture of the 
sensitivity of this study design to identify potential safety problems with the vaccine. 
Table 6-2 gives the upper and lower bounds for 95% exact binomial confidence intervals of the 
true SAE rate at all possible numbers of events within each group (n=5 and n=10).  
For Part I (n=5): If none of the 5 participants receiving the vaccines experience SAEs, the 95% 
exact 2-sided upper confidence bound for the SAE would range from 0 to 0.52. 
For Part II (n=10): If none of the 10 participants receiving the vaccines experience SAEs, the 
95% exact 2-sided upper confidence bound for the SAE rate is 0.308. 
 
Table 6-1: Probability of Events for Different Safety and Immunogenicity Scenarios 
 
Part I (dose-escalation): N=5 
Part II (prime-boost): N=10 
True 
Event 
Rate 
Pr  
(observing 0 
event) 
Pr  
(observing more 
than 1 event) 
Pr  
(observing 0 
event) 
Pr  
(observing more 
than 1 event) 
0.01 
0.951 
0.001 
0.904 
0.004 
0.02 
0.904 
0.004 
0.817 
0.016 
0.05 
0.774 
0.023 
0.599 
0.086 
0.1 
0.590 
0.081 
0.349 
0.264 
0.15 
0.444 
0.165 
0.197 
0.456 
0.2 
0.328 
0.263 
0.107 
0.624 
0.3 
0.168 
0.472 
0.028 
0.851 
0.4 
0.078 
0.663 
0.006 
0.954 
0.5 
0.031 
0.813 
0.001 
0.989 
 

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Table 6-2: 95% Confidence Intervals for the True Rate at All Possible Observed Rates 
within a Group (n=5 and n=10) 
 
Part I: N=5 
95% Confidence Interval 
 
 
Part II: N=10 
95% Confidence Interval 
Observed 
Rate 
Lower 
Bound 
Upper 
Bound 
 
Observed 
Rate 
Lower 
Bound 
Upper 
Bound 
0/5 
0 
0.522 
 
0/10 
0 
0.308 
1/5 
0.005 
0.716 
 
1/10 
0.003 
0.445 
2/5 
0.053 
0.853 
 
2/10 
0.025 
0.556 
3/5 
0.147 
0.947 
 
3/10 
0.067 
0.652 
4/5 
0.284 
0.995 
 
4/10 
0.122 
0.738 
5/5 
0.478 
1 
 
5/10 
0.187 
0.813 
 
 
 
 
6/10 
0.262 
0.878 
 
 
 
 
7/10 
0.348 
0.933 
 
 
 
 
8/10 
0.444 
0.975 
 
 
 
 
9/10 
0.555 
0.997 
 
 
 
 
10/10 
0.692 
1 
6.4.2 Power Calculations for Immunogenicity 
Table 6-2 is applicable to the immunogenic response rates, and gives the exact 95% confidence 
interval of the true response rate over possible number of responses out of the 5 and 10 subjects.  
Table 6-1 gives the probabilities of observing 0 or more than 1 response over a range of 
underlying response rates.  
6.4.3 Power for Comparison 
As exploratory analyses, we will compare the four groups in Part II of the study for possible 
differences in immunogenicity; a simple comparison on the positive response rate will be used. 
Table 6-3 gives the power of Fisher’s exact test to compare any two schedules over a range of 
possible response rates. It indicates that a comparison between two groups is not powered by the 
study design with 10 samples per group. Table 6-3 also gives the power of Fisher’s exact test 
when the age cohorts within each group are combined, resulting in a comparison between two 
groups with 20 samples per group. It indicates that with age cohorts combined, the comparison 
would be powered for larger differences between groups. 
 

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Table 6-3: Power to Detect Difference in Response Rates between the groups in Part II by 
Fisher’s Exact Test, Where Both Groups are of Size n=10 or n=20 
Group 4A (N=10) 
Group 3A (N=10) 
 
0.1 
0.2 
0.3 
0.4 
0.5 
0.6 
0.1 
 
1 
6 
15 
30 
47 
0.2 
1 
 
2 
6 
13 
25 
0.3 
6 
2 
 
2 
5 
12 
0.4 
15 
6 
2 
 
2 
5 
0.5 
30 
13 
5 
2 
 
2 
0.6 
47 
25 
12 
5 
2 
 
 
Group 4B (N=20) 
Group 3B (N=20) 
 
0.1 
0.2 
0.3 
0.4 
0.5 
0.6 
0.1 
 
7 
24 
49 
74 
91 
0.2 
7 
 
6 
19 
41 
65 
0.3 
24 
6 
 
6 
16 
35 
0.4 
49 
19 
6 
 
5 
14 
0.5 
74 
41 
16 
5 
 
5 
0.6 
91 
65 
35 
14 
5 
 
 STATISTICAL ANALYSIS  
All statistical analyses will be performed using Statistical Analysis System (SAS), R, or S-Plus 
statistical software. No formal multiple comparison adjustments will be employed for safety 
endpoints or secondary endpoints. 
6.5.1 Analysis Variables 
The analysis variables consist of baseline variables and safety variables for primary and 
secondary objective analyses. 
6.5.2 Baseline Demographics 
Baseline characteristics including demographics and laboratory measurements will be 
summarized using descriptive statistics. 

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6.5.3 Safety Analysis 
6.5.3.1 Reactogenicity 
The number and percentage of participants experiencing each type of reactogenicity sign or 
symptom will be tabulated by severity. For a given sign or symptom, each participant’s 
reactogenicity will be counted once under the maximum severity for all assessments. 
6.5.3.2 Adverse Experiences 
Adverse experiences are coded into Medical Dictionary for Regulatory Activities (MedDRA) 
preferred terms. The number and percentages of participants experiencing each specific AE will 
be tabulated by severity and relationship to treatment. For the calculations in these tables, each 
participant’s adverse experience will be counted once under the maximum severity or strongest 
recorded causal relationship to treatment. 
A complete listing of adverse experiences for each participant will provide details including 
severity, relationship to treatment, onset, duration and outcome. 
6.5.3.3 Local Laboratory Values 
Boxplots of local laboratory values will be generated for baseline values and for values measured 
during the course of the study. Each boxplot will show the 1st quartile, the median, and the 3rd 
quartile. Outliers, or values outside the boxplot, will also be plotted. If appropriate, horizontal 
lines representing boundaries for abnormal values will be plotted. 
6.5.4 Immunogenicity Analysis 
The statistical analysis for immunogenicity will employ the intent-to-treat principle whereby all 
data from enrolled subjects will be analyzed according to the group assignment. However, if 
during immune assessment on stored samples, a subject is found to have a positive antibody 
response at baseline, the vaccine immune responses assessment for these subjects will not be 
included in the final immunogenicity analysis. If needed, a per-protocol analysis will be 
performed as secondary analysis where subjects will be analyzed according to their actual 
vaccination scheme if it is different from the assigned or up to the last visit in the study if there 
are early dropouts. The study is not designed to power the comparison in immune responses 
between vaccine dosages. 
If assay data are qualitative (i.e., positive or negative) then analyses will be performed by 
tabulating the frequency of positive response for each assay at each time point that an assessment 
is performed. Binomial response rates will be presented with their corresponding exact 95% 
confidence interval estimates. 
Some immunologic assays have underlying continuous or count-type readout that is often 
dichotomized into responder/non-responder categories. For these assays, graphical and tabular 
summaries of the underlying distributions will be made. These summaries may be performed on 
transformed data (e.g., log transformation) to better satisfy assumptions of symmetry and 
homoscedasticity. 
6.5.5 Missing Data 
Missing responses will be assumed to be missing completely at random. Analyses will include 
all samples available at each study time point. 

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6.5.6 Interim Analyses 
Safety Reviews: The PSRT will review safety data routinely throughout the study. The study 
will utilize both electronic database features and reviews by designated safety review personnel 
to identify in a timely manner if any of the safety pause rules of the study are met.  
Immunogenicity Review: The analyses of immunogenicity may be performed when the HAI 
assays of samples are collected at 4 weeks after the prime vaccination and at 2 weeks after the 
boost vaccine. This may occur prior to completion of safety follow-up visits or collection of data 
for secondary and exploratory immunogenicity endpoints. Such an analysis would constitute the 
final analysis for the primary immunogenicity endpoint, so sample size adjustments are not 
required. Reports providing results by study group may be provided to VRC solely for the 
purpose of informing decisions related to future trials in a timely manner. The results should in 
no way influence the conduct of the VRC 316 trial in terms of early termination or later safety or 
immunogenicity endpoint assessments. Analyses of secondary and exploratory immunogenicity 
assays may also be performed as data become available. 
 INTERVENTION ASSIGNMENTS 
For Part I (dose-escalation): Subjects between 18-47 years will be assigned to the dose group 
that is open to accrual at the time of enrollment. When a subject is enrolled into a group but does 
not begin study injections or if a subject withdraws, a new eligible subject may be enrolled into 
the same group in order to complete the safety data set as planned. 
For Part II (prime-boost): Subjects will be stratified by age and randomized equally into Group 3 
or Group 4.  
The subject and the study clinicians will be informed on the subject’s randomization assignment 
upon completing of enrollment in Advantage Electronic Data Collection (AdEDC) system. 
To decrease the potential for participant dropouts during the period between randomization and 
initial vaccination, randomization will occur on Day 0 after the study consent is signed and 
eligibility is confirmed. If subjects accrued to a study group do not complete the number of 
vaccinations and follow-up duration specified, then additional subjects may be accrued in that 
group. 
 
PHARMACY AND VACCINE ADMINISTRATION PROCEDURES  
The study groups and study agent dosing schedule are shown in Table 2 in Section 4. Refer to 
the IB for further information about the investigational study products.   
 STUDY PRODUCT 
The study includes two investigational vaccines described as follows: 
• VRC-FLUNPF081-00-VP at 180 mcg/mL. Vials contain 0.7 mL of a clear, colorless, 
isotonic buffered (pH=7.2) sterile solution. The formulation buffer consists of 20mM 
Sodium Phosphate, 100mM Sodium Chloride and 5% Sucrose. Vials are intended for 
single use only and do not contain a preservative.  
• VRC-FLUDNA082-00-VP at 4 mg/mL. DNA A/Sing vaccine is supplied as a 3 mL glass 
vial containing a clear, colorless, isotonic, PBS-buffered (pH=7.2), sterile solution. Vials 
are intended for single use only and do not contain a preservative. Each vial (4 mg/mL) 
contains a volume of 1.2 mL (4.8 mg). 

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• VRC-PBSPLA043-00-VP (PBS), diluent, is a clear, colorless, sterile solution at pH 7.2, 
aseptically filled to a volume of 1.2 mL in a 3 mL glass vial. Vials are intended for single 
use only and do not contain preservative. Vials must not be refrozen or reused after 
thawing. 
 STUDY PRODUCT PRESENTATION AND STORAGE 
7.2.1 Study Product Labels 
VRC-FLUNPF081-00-VP and VRC-FLUDNA082-00-VP will have specific product information 
(e.g., part number, lot number, fill volume, storage temperature) included on the product vial 
labels. The label will contain an Investigational Use Statement (“Limited by Federal Law to 
Investigational Use”), and manufacturer information. 
7.2.2 Study Product Storage 
VRC-FLUNPF081-00-VP and VRC-FLUDNA082-00-VP vials will be shipped within the 
recommended temperature range using appropriate shipping configurations, to the study 
pharmacist or designee. 
VRC-FLUNPF081-00-VP (HA-F A/Sing): Vials of vaccine are stored until use at -35°C to          
-15°C in a qualified, continuously monitored, temperature-controlled freezer. As freezer 
temperatures may fluctuate, a temperature range of -45°C to -15°C is acceptable. 
VRC-FLUDNA082-00-VP (DNA A/Sing): Vials of vaccine are stored until use at -35°C to         
-15°C in a qualified, continuously monitored, temperature-controlled freezer. As freezer 
temperatures may fluctuate, a temperature range of -45°C to -10°C is acceptable. Storage below  
-45ºC is not permitted because of the stopper limitation. 
VRC-PBSPLA043-00-VP (PBS): Vials of PBS are stored until use at the target temperature of -
35°C to -15°C in a qualified, continuously monitored, temperature-controlled freezer. As freezer 
temperatures may fluctuate, a temperature range of -45°C to -10°C is acceptable based upon 
historic stability data from studies of similar products. 
7.2.3 Temperature excursions  
If deviations in storage temperature occur from the normal allowance for the pharmacy freezer, 
the site pharmacist or designee must report the storage temperature excursion promptly to the PI 
and IND Sponsor. The excursion must be evaluated and investigated and action must be taken to 
restore and maintain the desired temperature limits. Pending the outcome of the investigation, the 
IND Sponsor will notify the pharmacist if continued clinical use of the product is acceptable. 
 PREPARATION OF STUDY PRODUCTS FOR INJECTION  
This section describes how the site pharmacist will prepare study injections. Clinician 
instructions on how to select an arm and administer an injection are in Section 4.2.4. Refer to the 
group assignment for the study subject to select the proper vial type.  
For all study groups, the prepared syringe will be labeled with the subject identifier and the date 
and time after which the preparation may not be used. Allow the vial to equilibrate to room 
temperature (15 to 30º C). Swirl the contents gently. Filled syringes should be kept at room 
temperature and out of direct sunlight until the product is administered. All injections must be 
administered within 8 hours after removing the vaccine vial from the freezer. 

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7.3.1 Preparation of VRC-FLUNPF081-00-VP to be administered via Needle and 
Syringe Injection 
Subjects will receive the VRC-FLUNPF081-00-VP vaccine via needle and syringe injection. The 
following instructions apply for VRC-FLUNPF081-00-VP vaccine preparation: 
Group 1 (20 mcg): 
1. Thaw 1 vial of HA-F A/Sing at room temperature until all ice crystals have melted.  Swirl 
gently to mix. 
2. Thaw 1 vial of PBS diluent at room temperature until all ice crystals have melted.  Swirl 
gently to mix. 
3. Withdraw 0.11 mL of HA-F A/Sing into the syringe. 
4. Withdraw 0.2 mL of PBS diluent directly into the syringe. 
5. Invert syringe 5x to mix. 
Group 2 and Groups 4A/B (60 mcg, prime and boost); Groups 3A/B (60 mcg, ONLY boost): 
1. Thaw 1 vial of HA-F A/Sing at room temperature until all ice crystals have melted.  Swirl 
gently to mix. 
2. Withdraw 0.33 mL of HA-F A/Sing into the syringe. 
Please refer to the IB for more information on vaccine preparation for injection. 
7.3.2 Preparation of VRC-FLUDNA082-00-VP to be administered via PharmaJet 
Injection 
For subjects randomized to Group 3A/B (PharmaJet), remove a single dose vial of the DNA 
vaccine 4 mg/mL from the freezer. Using aseptic technique, withdraw 0.5 mL of the DNA 
vaccine from the vial into the PharmaJet 0.5 mL needle-free syringe and remove all air bubbles. 
Two 0.5 mL injections of the 4 mg/mL preparation will be administered for each 4mg dose with 
the PharmaJet. 
 STUDY PRODUCT ACCOUNTABILITY 
7.4.1 Documentation 
The study pharmacist or designee will be responsible for maintaining an accurate record of the 
codes, inventory, and an accountability record of vaccine supplies for this study. Electronic 
documentation as well as paper copies will be used. 
7.4.2 Disposition 
The empty vials and the unused portion of a vial will be discarded in a biohazard containment 
bag and incinerated or autoclaved following the injection. Any unopened vials that remain at the 
end of the study will be discarded at the discretion of the VRC in accordance with policies that 
apply to investigational products. Partially used vials or expired prepared doses cannot be 
administered to other subjects nor used for in vitro experimental studies and will be discarded as 
indicated above.  
 HUMAN SUBJECT PROTECTIONS AND ETHICAL OBLIGATIONS 
This research study will be conducted in compliance with the protocol, International Council for 
Harmonisation Good Clinical Practices (ICH-GCP) guidance, and all applicable regulatory 
requirements. 

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 INSTITUTIONAL REVIEW BOARD 
A copy of the protocol, ICF, other written subject information, and any advertising material will 
be submitted to the NIAID IRB for written approval prior to use. 
The PI must submit and, where necessary, obtain approval from the IRB for all subsequent 
protocol amendments and changes to the ICF. The PI will notify the IRB of unanticipated 
problems, non-compliance, deviations from the protocol, and serious SAEs as described in 
Section 5.4. 
The investigator will be responsible for obtaining IRB approval of the annual Continuing Review 
throughout the duration of the study. 
 SUBJECT IDENTIFICATION AND ENROLLMENT OF STUDY PARTICIPANTS  
Study subjects will be recruited through on-site and off-site advertising done for the screening 
protocol, VRC 500 (11-I-0164). All study activities will be carried out at the NIH CC. Effort will 
be made to include women and minorities in proportions similar to that of the community from 
which they are recruited. 
8.2.1 Participation of Children 
Children are not eligible to participate in this clinical trial because the investigational vaccine 
regimens have not been previously evaluated in adults. If the product is assessed as safe and 
immunogenic, other protocols designed for children may be conducted in the future. 
8.2.2 Participation of NIH Employees 
NIH employees and members of their immediate families may participate in this protocol. NIH 
staff may be a vulnerable class of subjects. The VRC will follow the Guidelines for the Inclusion 
of Employees in NIH Research Studies and will give each employee a copy of the “NIH 
Information Sheet on Employee Research Participation” and a copy of the “Leave Policy for 
NIH Employees Participating in NIH Medical Research studies.” 
Neither participation nor refusal to participate will have an effect, either beneficial or adverse, on 
the participant’s employment or work situation. The NIH information sheet regarding NIH 
employee research participation will be distributed to all potential subjects who are NIH 
employees. The employee subject’s privacy and confidentiality will be preserved in accordance 
with NIH CC and NIAID policies. For NIH employee subjects, consent will be obtained by an 
individual who is independent of the employee’s team. If the individual obtaining consent is a 
co-worker to the subject, independent monitoring of the consent process will be included through 
the Bioethics Consultation Service. Protocol study staff will be trained on obtaining potentially 
sensitive and private information from co-workers or subordinates. 
 INFORMED CONSENT 
The study ICF, , describes the investigational products to be used and all aspects involved in 
protocol participation. 
Before a subject’s participation in the study, it is the investigator’s responsibility to obtain 
written informed consent from the subject, after adequate explanation of the aims, methods, 
anticipated benefits, and potential hazards of the study and before any protocol-specific 
procedures or study products are administered. 

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An IRB-approved AoU quiz, intended to assist in the evaluation of the subject’s understanding 
of this study, is administered as part of the consent process. 
The acquisition of informed consent will be documented in the subject’s medical records, as 
required by 21 CFR 312.62, and the ICF will be signed and personally dated by the subject and 
by the person who conducted the informed consent discussion. The original signed ICF will be 
placed in the medical record and a copy of the signed ICF will be provided to the subject. 
 SUBJECT CONFIDENTIALITY 
The investigator must ensure that the subject’s anonymity is maintained and will ensure that no 
information identifying the subject will be released to any unauthorized party. Subjects will not 
be identified in any reports on this study. All records will be kept confidential to the extent 
provided by federal, state and local law. Medical records are made available for review when 
required by the FDA or other authorized users, such as the vaccine manufacturer, only under the 
guidelines set by the Federal Privacy Act. Direct access includes examining, analyzing, 
verifying, and reproducing any records and reports that are important to the evaluation of the 
study. The investigator is obligated to inform the subjects that the above-named representatives 
will review their study-related records without violating the confidentiality of the subjects. 
Stored study research samples are labeled by a code (such as a number) that only the study team 
can link to the subject. The requirement to maintain subject confidentiality is included in the 
study ICF. 
 RISKS AND BENEFITS  
8.5.1 Risks of the VRC-FLUNPF081-00-VP vaccine 
This is the first study in humans of this investigational VRC-FLUNPF081-00-VP vaccine. This 
product is derived from H. pylori ferritin and not human ferritin, for this reason it is not expected 
to interfere with iron storage, transport or iron serum levels [13]. 
Potential side effects resulting from IM injection include stinging, arm discomfort, redness of the 
skin or mild bruising at vaccine injection sites.   
Subjects may exhibit general signs and symptoms associated with administration of a vaccine, 
including fever, chills, rash, aches and pains, nausea, headache, dizziness and fatigue.  These 
side effects will be monitored, but are generally short term, mild to moderate severity and 
usually do not require treatment. 
There may be other unknown side effects. 
8.5.2 Risks of the VRC-FLUDNA082-00-VP Vaccine 
This is the first study in humans of the investigational DNA vaccine, VRC-FLUDNA082-00-VP. 
The risks noted are based on risks of vaccines in general, and results of previous studies with 
other investigational DNA vaccines. 
Potential side effects resulting from IM injection with PharmaJet include stinging, arm 
discomfort, redness of the skin, mild bruising or a small laceration at vaccine injection sites.  
There is also a small chance that the needle-free device could not work properly.  As of June 26, 
2017, there have been 8 out of 426 (1.9%) injections that involved device malfunction in VRC 
705 Part A. This includes the syringe breaking and study product spraying during 
administration. No harm to subjects has occurred when this happened. Investigations are ongoing 

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to determine the factors leading to malfunction. As a result, Subjects in Group 3A/B and 
clinicians administering the vaccine will be asked to wear safety glasses when using PharmaJet. 
Subjects may exhibit general signs and symptoms associated with administration of a vaccine 
injection, including fever, chills, rash, aches and pains, nausea, headache, dizziness and fatigue.  
These side effects will be monitored, but are generally short-term, mild to moderate severity and 
usually do not require treatment. 
In previous VRC DNA vaccine studies, placebo and vaccine recipients were noted to have 
occasional asymptomatic and self-limited changes in laboratory tests such as temporary drop in 
white blood cell count. Urticaria has been reported as possibly related to DNA vaccines in rare 
cases. 
Investigational DNA vaccines administered via needleless injection device have been associated 
with mild skin lesions (0.5-1.0 cm diameter) at the vaccination site. In these cases a small scab 
formed within 1-2 weeks after immunization and came off after a few days. The skin healed 
without treatment within a few weeks. We do not know if the side effects from PharmaJet will be 
the same as Biojector. There may be other unknown side effects. 
8.5.3 Other Risks 
Risks of Specimen Collections: 
• Blood drawing: The risks of blood sample collection are minimal and consist of mild 
discomfort at the sample collection site. The procedure may cause pain, bruising, 
fainting, and, rarely, infection at the site where the blood is taken. 
 
• Apheresis: donations may cause pain, bruising, and discomfort in the arms where the 
needles are placed. It may also cause chills, nausea, heartburn, mild muscle cramps and 
tingling sensation around the mouth or in the fingers, however this can usually be 
relieved by slowing or temporarily interrupting the apheresis or taking a calcium 
containing antacid, such as Tums®. Other possible side effects are anxiety, vomiting and 
lightheadedness. Temporary lowering of the blood pressure may develop. There is the 
rare possibility of infection, fainting or seizure. Very rarely a nerve problem at the needle 
placement site may occur. Also, very rarely, a machine malfunction may occur, resulting 
in the loss of about one unit of blood. There may be additional risks of apheresis that are 
unknown at this time. 
Risks of study vaccines on the fetus or nursing infant: 
We do not know the possible effects of the study vaccines on the fetus or nursing infant. Women 
of reproductive potential will be required to agree to use birth control beginning 21 days prior to 
enrollment and continuing through end of study.  
Because this is a research study, women of reproductive potential will be tested for pregnancy 
prior to administration of each study injection and asked to notify the site immediately upon 
learning of a pregnancy during this study. In the case of pregnancy, subjects will no longer 
receive additional vaccine product and continue to be followed for safety. Research sample 
collections will be discontinued for pregnant women. The subject will be contacted to ask about 
the outcome of a pregnancy that begins during the study. 
 

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Risks of new diagnoses: 
It is possible that the standard medical tests performed as part of this research protocol will result 
in new diagnoses. Depending upon the medical findings and consequences of being provided 
with the new medical information about health status, the study subject may view this aspect of 
study participation as either a risk or a benefit. Any such information will be shared and 
discussed with the subject and, if requested by the subject, will be forwarded to the subject’s 
primary health care provider for further workup and management. 
8.5.4 Study Benefits  
Study subjects will not receive direct health benefit from study participation. This protocol is not 
designed to provide treatment for any condition. Others may benefit from knowledge gained in 
this study that may aid in the development of an H2 (or universal) influenza virus vaccine. 
Neither of the two investigational vaccines are expected to provide protection from influenza. 
 PLAN FOR USE AND STORAGE OF BIOLOGICAL SAMPLES 
The plan for use and storage of biological samples from this protocol is as outlined in the 
following sections. 
8.6.1 Use of Samples, Specimens and Data 
Samples, specimens and data collected under this protocol may be used to conduct protocol 
related safety and immune response evaluations, exploratory laboratory evaluations related to the 
type of infection the study product was designed to prevent, exploratory laboratory evaluations 
related to vaccine or infectious disease research in general and for research assay validation. 
Genetic testing may be performed in accordance with the genetic testing information that was 
included in the study ICF. HLA testing may be done in association with identifying factors 
linked with the immune response development or progression of infections.  
Additional optional genetic testing, including transcriptome sequencing may be done on 
collected specimens in an effort to assess the expression of genes involved in the immune 
response to vaccination.  
Results of genetic testing may have psychological implications for patients such as revelations 
regarding future health risks, incurable conditions, and/or information contradictory to stated 
biological relationships. Genetics counseling and advice will be available from the NIH to help 
subjects at all study sites with the implications of findings, where appropriate.  
Following genetic testing, the data will be shared in a controlled-access public database for other 
investigators to benefit from it (e.g. the Database of Genotypes and Phenotypes dbGAP). 
However no personal, identifiable information will be shared in this process as the results will 
only be shared with a code. 
Other optional analysis, including proteome, lipidome, metabolome, and exosome may be done 
on collected specimens to evaluate some proteins, lipids, metabolites, and low molecular weight 
molecules involved in the immune response to vaccination.  
8.6.2 Storage and Tracking of Blood Samples and Other Specimens 
All of the stored study research samples are labeled by a code that only the site can link to the 
subject. Samples are stored at the VITL, Gaithersburg, MD or VRC Laboratories in Building 40, 
Bethesda, MD, which are both secure facilities with limited access. Data will be kept in 

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password-protected computers. Only investigators or their designees will have access to the 
samples and data. Samples will be tracked in the Laboratory Information Management System 
(LIMS) database or using another software designed for this purpose (e.g., Freezerworks). 
8.6.3 Disposition of Samples, Specimens and Data at Completion of the Protocol  
In the future, other investigators (both at NIH and outside) may wish to study these samples 
and/or data. IRB approval must be sought prior to any sharing of samples. Any clinical 
information shared about those samples would similarly require prior IRB approval. The 
research use of stored, unlinked or unidentified samples may be exempt from the need for 
prospective IRB review and approval. Exemption requests will be submitted in writing to the 
NIH Office of Human Subjects Research, which is authorized to determine whether a research 
activity is exempt. 
At the time of protocol termination, samples will remain in the VITL facility or VRC 
laboratories or, after IRB approval, transferred to another repository. Regulatory oversight of the 
stored samples and data may be transferred to a stored samples protocol as part of the IRB-
approved termination plan. Data will be archived by the VRC in compliance with requirements 
for retention of research records, or after IRB and study sponsor approval, it may be either 
destroyed or transferred to another repository. 
8.6.4 Loss or Destruction of Samples, Specimens or Data 
The NIH Intramural Protocol Deviation definition related to loss of or destruction of samples or 
data will be followed. Any loss or unanticipated destruction of samples (for example, due to 
freezer malfunction) or data (for example, misplacing a printout of data with identifiers) that 
compromises the scientific integrity of the study will be reported to the IRB in accordance with 
institutional policies. The PI will also notify the IRB if the decision is made to destroy the 
remaining samples. 
 COMPENSATION 
Subjects will be compensated for time and inconvenience in accordance with the standards for 
compensation of the Clinical Research Volunteer Program. The compensation per visit will be 
$275 for visits that include injections and $175 for clinic visits which include a blood draw. The 
compensation for any clinic visit that does not include a blood draw or procedure will be $75 and 
compensation for timely completion of electronic diary card will be $25. Compensation will be 
$250 for apheresis if performed. The total compensation for the subject is based on the number 
of study clinic visits, injections completed and if optional research blood collections are 
performed.  
Subjects will receive compensation by direct deposit within approximately 1 or 2 weeks after 
each completed visit. 
Compensation may need to be reported to the Internal Revenue Service (IRS) as taxable income. 
The approximate total compensation is as follows: 
 
 
 
 

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Group 
Without  
Apheresis 
With  
Apheresis 
Group 1 
$1,525 
NA 
Group 2 
Group 3A-B 
Group 4A-B 
$2,000 
$2,075 
 SAFETY MONITORING  
8.8.1 Protocol Safety Review Team 
Close cooperation between the designated members of the Protocol Team will occur to evaluate 
and respond to individual AEs in a timely manner. The VRC designated Safety Officer for the 
day conducts a daily safety review of clinical data per VRC Standard Operating Procedures. The 
PSRT, comprised of the PI, Associate Investigators, Study Coordinator, Protocol Specialists, and 
other Study Clinicians, will review the summary study safety data reports on a weekly basis 
through 4 weeks after the last subject receives the final study injection. After this time, the PSRT 
will monitor the safety data reports on a monthly basis through completion of the last study visit.   
 ADMINISTRATIVE AND LEGAL OBLIGATIONS 
 PROTOCOL AMENDMENTS AND STUDY TERMINATION 
Protocol amendments may be made only with the prior approval of the IND sponsor 
representative, VRC, NIAID.  Agreement from the PI must be obtained for all amendments to 
the protocol and the informed consent document. All study amendments will be submitted to the 
IRB for approval. 
The VRC, NIAID IRB, Office of Human Research Protections, study PI, and FDA reserves the 
right to terminate the study. The PI will notify the IRB in writing of the study’s completion or 
early termination. 
 STUDY DOCUMENTATION AND STORAGE 
The PI will maintain a list of appropriately qualified persons to whom trial duties have been 
delegated. 
Source documents are original documents, data, and records from which the subject’s data are 
obtained. These include but are not limited to hospital records, clinical and office charts, 
laboratory and pharmacy records, microfiches, radiographs, and correspondence.  
The PI and staff are responsible for maintaining a comprehensive and centralized filing system of 
all study-related (essential) documentation, suitable for inspection at any time by representatives 
from the VRC, IRB, FDA, and/or applicable regulatory authorities. Elements include: 
• Subject files containing completed informed consent forms, and supporting copies of 
source documentation (if kept) 
• Study files containing the protocol with all amendments, Investigator Brochures, copies 
of all correspondence with the IRB and VRC 
In addition, all original source documentation must be maintained and be readily available. 

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All essential documentation should be retained by the institution for the same period of time 
required for medical records retention. The FDA requires study records to be retained for up to 
two years after marketing approval or refusal (21 CFR 312.62). No study document should be 
destroyed without prior written agreement between the VRC and the PI. Should the PI wish to 
assign the study records to another party or move them to another location, VRC must be 
notified in writing of the new responsible person and/or the new location. 
 DATA COLLECTION, DATA SHARING AND PROTOCOL MONITORING 
9.3.1 Data Collection 
Clinical research data will be collected in a secure electronic web-based clinical data 
management system (CDMS) through a contract research organization, EMMES (Rockville, 
MD). Extracted data without patient identifiers will be sent to the Protocol Statistician for 
statistical analysis. 
9.3.2 Data Sharing Plan 
Data generated in this study will be shared as de-identified data in the government-funded public 
repository, www.ClinicalTrials.gov.  Data may be shared prior to publication at approved public 
presentations or for collaborative development and will be shared at the time of publication or 
within 1 year of the primary completion date.  
9.3.3 Source Documents  
The site will maintain appropriate medical and research records for this trial, in compliance with 
ICH-GCP, regulatory and institutional requirements for the protection of confidentiality of 
subjects. 
Source data are all information, original records of clinical findings, observations, or other 
activities in a clinical trial necessary for the reconstruction and evaluation of the trial. Examples 
of these original documents and data records include, but are not limited to, medical records, 
laboratory reports, pharmacy records and other research records maintained for the clinical trial. 
9.3.4 Protocol Monitoring Plan 
The NIAID and VRC or their authorized representatives are responsible for contacting and 
visiting the PI for the purpose of inspecting the facilities and, upon request, inspecting the 
various records of the trial, provided that subject confidentiality is respected. 
Site investigators will allow the study monitors, the NIAID IRB, and the FDA to inspect study 
documents (e.g., consent forms, drug distribution forms, and case report forms) and pertinent 
hospital or clinic records for confirmation of the study data. 
Site visits by study monitors will be made in accordance with the study monitoring plan to 
monitor the following: study operations, the quality of data collected in the research records, the 
accuracy and timeliness of data entered in the database, and to determine that all process and 
regulatory requirements are met. Study monitoring visits will occur as defined by the IND 
Sponsor approved monitoring plan. 
 LANGUAGE 
All written information and other material to be used by subjects and investigative staff must use 
vocabulary and language that are clearly understood.  

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 POLICY REGARDING RESEARCH-RELATED INJURIES 
The NIH CC will provide short-term medical care for any injury resulting from participation in 
this research. In general, the NIH, the NIH CC, or the U.S. Federal Government will provide no 
long-term medical care or financial compensation for research-related injuries. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Identify the Minimum Interval Necessary to Observe the H5 DNA Influenza Vaccine 
Priming Effect. J Infect Dis, 2013. 208(3): p. 418-22. 
20. 
Ledgerwood, J.E., et al., DNA priming for seasonal influenza vaccine: a phase 1b 
double-blind randomized clinical trial. PLoS One, 2015. 10(5): p. e0125914. 
21. 
Khurana, S., et al., DNA Priming Prior to Inactivated Influenza A(H5N1) Vaccination 
Expands the Antibody Epitope Repertoire and Increases Affinity Maturation in a Boost-
Interval-Dependent Manner in Adults. J Infect Dis, 2013. 208(3): p. 413-7. 
22. 
Ledgerwood, J.E., et al., Phase I Clinical Evaluation of Seasonal Influenza 
Hemagglutinin (HA) DNA Vaccine Prime Followed by Trivalent Influenza Inactivated 
Vaccine (IIV3) Boost. Contemp Clin Trials, 2015. 
23. 
Adam D. DeZure, et al., Safety and Immunogenicity of H7 Influenza Prime-Boost 
Regimens in Healthy Adults, in National Foundation for Infectious Diseases. 2016: 
Baltimore, MD. 
24. 
Ledgerwood, J.E., et al., Influenza virus H5 DNA vaccination is immunogenic by 
intramuscular and intradermal routes in humans. Clinical and Vaccine Immunology, 
2012. 19(11): p. 1792-7. 
25. 
Ledgerwood, J.E. and B.S. Graham, DNA vaccines: a safe and efficient platform 
technology for responding to emerging infectious diseases. Hum Vaccin, 2009. 5(9): p. 
623-6. 
26. 
Cui, Z., DNA vaccine. Adv Genet, 2005. 54: p. 257-89. 
27. 
Wei, C.J., et al., Induction of broadly neutralizing H1N1 influenza antibodies by 
vaccination. Science, 2010. 329(5995): p. 1060-4. 
28. 
Wei, C.J., et al., Elicitation of broadly neutralizing influenza antibodies in animals with 
previous influenza exposure. Science Transl Med, 2012. 4(147): p. 147ra114. 
29. 
Ledgerwood, J.E., et al., A West Nile virus DNA vaccine utilizing a modified promoter 
induces neutralizing antibody in younger and older healthy adults in a phase I clinical 
trial. J Infect Dis, 2011. 203(10): p. 1396-404. 
30. 
Martin, J.E., et al., A West Nile virus DNA vaccine induces neutralizing antibody in 
healthy adults during a phase 1 clinical trial. J Infect Dis, 2007. 196(12): p. 1732-40. 
31. 
Graham, B.S., et al., Phase 1 safety and immunogenicity evaluation of a multiclade HIV-
1 DNA candidate vaccine. J Infect Dis, 2006. 194(12): p. 1650-1660. 
32. 
Martin, J.E., et al., A SARS DNA vaccine induces neutralizing antibody and cellular 
immune responses in healthy adults in a Phase I clinical trial. Vaccine, 2008. 26(50): p. 
6338-43. 
33. 
Martin, J.E., et al., A DNA vaccine for Ebola virus is safe and immunogenic in a phase I 
clinical trial. Clin Vaccine Immunol, 2006. 13(11): p. 1267-77. 
34. 
Tavel, J.A., et al., Safety and immunogenicity of a Gag-Pol candidate HIV-1 DNA 
vaccine administered by a needle-free device in HIV-1-seronegative subjects. J Acquir 
Immune Defic Syndr, 2007. 44(5): p. 601-5. 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 47 of 62 
35. 
Kibuuka, H., et al., A phase 1/2 study of a multiclade HIV-1 DNA plasmid prime and 
recombinant adenovirus serotype 5 boost vaccine in HIV-Uninfected East Africans (RV 
172). J Infect Dis, 2010. 201(4): p. 600-7. 
36. 
Jaoko, W., et al., Safety and immunogenicity study of Multiclade HIV-1 adenoviral vector 
vaccine alone or as boost following a multiclade HIV-1 DNA vaccine in Africa. PLoS 
One, 2010. 5(9): p. e12873. 
37. 
Koup, R.A., et al., Priming Immunization with DNA Augments Immunogenicity of 
Recombinant Adenoviral Vectors for Both HIV-1 Specific Antibody and T-Cell 
Responses. PLoS One, 2010. 5(2): p. e9015. 
38. 
Churchyard, G.J., et al., A Phase IIA Randomized Clinical Trial of a Multiclade HIV-1 
DNA Prime Followed by a Multiclade rAd5 HIV-1 Vaccine Boost in Healthy Adults 
(HVTN204). PLoS One, 2011. 6(8): p. e21225. 
39. 
Graham, B.S., et al., DNA vaccine delivered by a needle-free injection device improves 
potency of priming for antibody and CD8+ T-cell responses after rAd5 boost in a 
randomized clinical trial. PLoS One, 2013. 8(4): p. e59340. 
40. 
Dowd, K.A., et al., Rapid development of a DNA vaccine for Zika virus. Science, 2016. 
41. 
Betts, M.R., J.P. Casazza, and R.A. Koup, Monitoring HIV-specific CD8+ T cell 
responses by intracellular cytokine production. Immunol Lett, 2001. 79(1-2): p. 117-25. 
42. 
Barouch, D.H., et al., A human T-cell leukemia virus type 1 regulatory element enhances 
the immunogenicity of human immunodeficiency virus type 1 DNA vaccines in mice and 
nonhuman primates. J Virol, 2005. 79(14): p. 8828-34. 
43. 
Sheets, R.L., et al., Biodistribution of DNA Plasmid Vaccines against HIV-1, Ebola, 
Severe Acute Respiratory Syndrome, or West Nile Virus Is Similar, without Integration, 
despite Differing Plasmid Backbones or Gene Inserts. Toxicol Sci, 2006. 91(2): p. 610-
619. 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Page 48 of 62 
APPENDIX I: CONTACT INFORMATION 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
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PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 50 of 62 
 
 
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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 51 of 62 
 
APPENDIX II: SCHEDULE OF EVALUATIONS 

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PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 52 of 62 
 
VRC 500 
VRC  316 Schedule of Evaluations 
PART I: Group 1 
 
Visit Number 
*01 
02 
02A 
03 
04 
05 
06 
08 
10 
11 
Week of Study -12 to 0 
W0 
W1 
W1 
W2 
W4 
W16 
W18 
W28 
W40 
Day of Study -84 to 0 
1D0 
D1 
D6 
D14 
D28 
D112 
D126 
D196 
D280 
Clinical 
Tube  
 
 
 
 
*VRC 500 Screening Consent 
X 
 
 
 
 
VRC 316 AoU; Consent 
 
X 
 
 
 
 
2Physical exam for eligibility, height /weight/ vitals at screening; vital signs and 
targeted exam (as needed) other visits. 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Medical history targeted to eligibility at screening; then interim medical history 
X 
X 
X 
X 
X 
X 
X 
X 
X 
3Study Product Administration: 
Part I, Group 1 
 
HA-F 
A/Sing 
 
 
 
 
Phone evaluation (clinic visit as needed) 
 
X 
 
 
 
 
Begin diary card 
 
X 
 
 
 
 
4Pregnancy test: urine or serum 
X 
X 
 
 
X 
 
 
X 
4Pregnancy prevention counseling/ Reproductive Information Form 
X 
X 
 
 
X 
 
 
X 
CBC with differential 
EDTA 
3 
3 
 
3 
3 
 
 
3 
Iron 
 
X 
 
 
 
X 
 
 
 
Total bilirubin, AST, ALT, and ALP  
GLT 
4 
4 
 
4 
4 
 
 
4 
Creatinine 
X 
X 
4 
5HLA type 
EDTA  
 
 
20 
 
 
HIV (other tests, if needed) 
SST 
4 
 
 
 
 
Research Samples 
 
 
 
 
 
H. pylori ferritin antibody and Human ferritin antibody 
SST 
X 
 
 
 
X 
 
 
 
Antibody assays. Serum storage 
SST 
32 
32 
32 
 
32 
32 
16 
32 
32 
Immunological assays (T cell and antibody responses).  
PBMCs and plasma storage 
EDTA 
80 
80 
80 
 
80 
80 
40 
80 
80 
Daily Volume (mL) 
123 
119 
0 
116 
7 
139 
112 
56 
112 
119 
Max. Cumulative Volume (mL) 
123 
242 
242 
358 
365 
504 
616 
672 
784 
903 
* VRC 500: Most screening evaluations must be no more than 84 days prior to Day 0 to be used for eligibility (pregnancy test from Day 0 must be used for eligibility). If clinical assessment on Day 0 suggests 
significant changes may have occurred since screening, then physical examination & laboratory studies done on Day 0 are used for eligibility. 
1 Day 0=day of enrollment and first vaccine injection. Day 0 evaluations prior to first injection are the baseline for assessing adverse events subsequently. 
2 Screening visit includes physical exam with vital signs. At other visits, physical examination is done if indicated by interim history or laboratory test results. Otherwise only blood pressure (BP), pulse, and 
temperature are required. 
3 Study Product Administration: Part I: Group 1 will receive HA-F A/Sing 20 mcg IM at Day 0. Complete post vaccination evaluations (BP, pulse, temperature, respiration and injection site assessment) at 1 hour 
or longer after study injection. 
4 Negative pregnancy test results must be confirmed for women of reproductive potential prior to each study injection. 
5 HLA type blood sample is collected once at any time-point in the study and is shown as a Visit 05 evaluation for convenience; however, if HLA type is already available in the medical record, it does not need to 
be repeated. HLA type may also be obtained from a frozen sample. 
Visit windows: Schedule Visits 02A through 11 with respect to Day 0; the following visit windows apply: Visits 02A (+1 day). Visit 03 (±1 day). Visits 04 and 05 (±2 days). Visits 06 (± 7 days). Visit 08 (±14 
days). Visit 10 and 11 (± 14 days).  
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
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Page 53 of 62 
VRC 500 
VRC  316 Schedule of Evaluations 
PART I: Group 2 
 
Visit Number 
*01 
02 
02A 
03 
04 
05 
06 
06A 
07 
08 
09 
10 
11 
Week of Study -12 to 0 
W0 
W1 
W1 
W2 
W4 
W16 
W17 
W17 
W18 
W20 
W28 
W40 
Day of Study -84 to 0 
1D0 
D1 
D6 
D14 D28 
D112 D113 
6D118 
D126 
D140 
D196 
D280 
Clinical 
Tube 
 
 
*VRC 500 Screening Consent 
 
X 
 
 
VRC 316 AoU; Consent 
 
X 
 
 
2Physical exam for eligibility, height /weight/ vitals at screening; vital signs and 
targeted exam (as needed) other visits. 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Medical history targeted to eligibility at screening; then interim medical history 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
3Study Product Administration: 
Part I, Group 2 
 
HA-F 
A/Sing 
HA-F 
A/Sing 
 
 
Phone evaluation (clinic visit as needed) 
 
X 
X 
 
 
Begin diary card 
 
X 
 
 
4Pregnancy test: urine or serum 
 
X 
X 
X 
X 
X 
X 
X 
4Pregnancy prevention counseling/ Reproductive Information Form 
 
X 
X 
X 
X 
X 
X 
X 
CBC with differential 
EDTA 
3 
3 
3 
3 
3 
3 
3 
Iron 
 
X 
 
 
X 
 
 
X 
 
Total bilirubin, AST, ALT, and ALP 
GLT 
4 
4 
4 
4 
4 
 
4 
4 
Creatinine 
 
X 
X 
4 
 
 
5HLA type 
EDTA 
20 
 
 
HIV (other tests, if needed) 
SST 
4 
 
 
Research Samples 
H. pylori ferritin antibody and Human ferritin antibody 
SST 
X 
 
X 
 
 
X 
 
Antibody assays. Serum storage 
SST 
32 
32 
32 
32 
32 
32 
32 
32 
32 
32 
Immunological assays (T cell and antibody responses). Multi-OMICS analysis. 
PBMCs and plasma storage 
EDTA 
80 
80 
80 
80 
80 
680 
7120 or  
Apheresis (6) 
80 
80 
80 
Daily Volume (mL) 
 
123 
119 
0 
116 
7 
139 
119 
0 
112 
152 
119 
112 
119 
Max. Cumulative Volume (mL) 
 
123 
242 
242 
358 
365 504 
623 
623 
735 
887 
1006 
1118 
1237 
* VRC 500: Most screening evaluations must be no more than 84 days prior to Day 0 to be used for eligibility (pregnancy test from Day 0 must be used for eligibility). If clinical assessment on Day 0 suggests 
significant changes may have occurred since screening, then physical examination & laboratory studies done on Day 0 are used for eligibility. 
1 Day 0=day of enrollment and first vaccine injection. Day 0 evaluations prior to first injection are the baseline for assessing adverse events subsequently. 
2 Screening visit includes physical exam with vital signs. At other visits, physical examination is done if indicated by interim history or laboratory test results. Otherwise only blood pressure (BP), pulse, and 
temperature are required. 
3 Study Product Administration: Part I: Group 2 will receive HA-F A/Sing 60 mcg IM at Day 0 and at Week 16. Complete post vaccination evaluations (BP, pulse, temperature, respiration and injection site 
assessment) at 1 hour or longer after each study injection. 
4 Negative pregnancy test results must be confirmed for women of reproductive potential prior to each study injection. 
5 HLA type blood sample is collected once at any time-point in the study and is shown as a Visit 05 evaluation for convenience; however, if HLA type is already available in the medical record, it does not need to 
be repeated. HLA type may also be obtained from a frozen sample. 
6 Visit 07: Two tubes of PBMCs collected in EDTA tubes will be sent to Building 40, while the remainder of the blood collected will be sent to VITL. 
7 If optional apheresis occurs, ONLY draw 32mL in SST (DO NOT draw 120 ml in EDTA tubes). For women of reproductive potential, pregnancy test must be negative within 72 hours prior to apheresis procedure.  
Visit windows: Schedule Visits 02A through 06 with respect to Day 0; Schedule Visits 06A through 11 with respect to Visit 06. The following visit windows apply: Visits 02A and 06A (+1 day). Visit 03 and 07 (±1 
day). Visits 04, 05 and 09 (±2 days). Visit 06 (±28 days). Visits 08 (±3 days). Visits 10 and 11 (±14 days). 
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 54 of 62 
VRC 500 
VRC  316 Schedule of Evaluations 
PART II: Group 3A/B and Group 4A/B 
 
Visit Number 
*01 
02 
02A 
03 
04 
05 
06 
06A 
07 
08 
09 
10 
11 
Week of Study -12 to 0 
W0 
W1 
W1 
W2 
W4 
W16 
W17 
W17 
W18 
W20 
W28 
W40 
Day of Study -84 to 0 
1D0 
D1 
D6 
D14 D28 
D112 
D113 6D118 
D126 
D140 
D196 D280 
Clinical 
Tube 
 
 
 
 
*VRC 500 Screening Consent 
 
X 
 
 
 
 
VRC 316 AoU; Consent 
 
X 
 
 
 
 
2Physical exam for eligibility, height /weight/ vitals at screening; vital signs and 
targeted exam (as needed) other visits. 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
X 
X 
Medical history targeted to eligibility at screening; then interim medical history 
 
X 
X 
X 
X 
X 
X 
 
X 
X 
X 
X 
X 
3Study Product Administration: 
Part II, Group 3A/B 
 
DNA 
A/Sing 
HA-F 
A/Sing  
 
 
 
3Study Product Administration: 
Part II, Group 4A/B 
 
HA-F 
A/Sing 
HA-F 
A/Sing  
 
 
 
Phone evaluation (clinic visit as needed) 
 
X 
X  
 
 
Begin diary card 
 
X 
 
 
 
 
4Pregnancy test: urine or serum 
 
X 
X 
X 
X 
 
 
X 
X 
 
X 
4Pregnancy prevention counseling/ Reproductive Information Form 
 
X 
X 
X 
X 
 
 
X 
X 
 
X 
CBC with differential 
EDTA 
3 
3 
3 
3 
3 
 
 
3 
 
3 
Iron 
 
X 
 
 
X 
 
 
 
X 
 
 
Total bilirubin, AST, ALT, and ALP 
GLT 
4 
4 
4 
4 
4 
 
 
4 
 
4 
Creatinine 
 
X 
X 
4 
 
 
 
 
5HLA type 
EDTA 
20 
 
 
 
 
HIV (other tests, if needed) 
SST 
4 
Research Samples 
 
 
 
 
 
H. pylori ferritin antibody and Human ferritin antibody 
SST 
X 
 
X 
 
 
 
X 
 
 
Antibody assays. Serum storage 
SST 
32 
32 
32 
32 
32 
 
32 
32 
32 
32 
32 
Immunological assays (T cell and antibody responses). Multi-OMICS analysis. 
PBMCs and plasma storage 
EDTA 
80 
80 
80 
80 
80 
 
680 
7120 or 
Apheresis 
80 
80 
80 
Daily Volume (mL) 
123 
119 
0 
116 
7 
139 
119 
0 
112 
152 
119 
112 
119 
Max. Cumulative Volume (mL) 
 
123 
242 
242 
358 
365 504 
623 
623 
735 
887 
1006 
1118 
1237 
* VRC 500: Most screening evaluations must be no more than 84 days prior to Day 0 to be used for eligibility (pregnancy test from Day 0 must be used for eligibility). If clinical assessment on Day 0 suggests 
significant changes may have occurred since screening, then physical examination & laboratory studies done on Day 0 are used for eligibility. 
1 Day 0=day of enrollment and first vaccine injection. Day 0 evaluations prior to first injection are the baseline for assessing adverse events subsequently. 
2 Screening visit includes physical exam with vital signs. At other visits, physical examination is done if indicated by interim history or laboratory test results. Otherwise only blood pressure (BP), pulse, and 
temperature are required. 
3 Study Product Administration: Part II: Group 3A/B will receive DNA A/Sing HA-F A/Sing 4 mg IM at Day 0 and HA-F A/Sing 60 mcg IM at Week 16. Complete post vaccination evaluations (BP, pulse, 
temperature, respiration and injection site assessment) at 1 hour or longer after each study injection. Group 4A/B will receive HA-F A/Sing 60 mcg IM at Day 0 and at Week 16. Complete post vaccination 
evaluations (BP, pulse, temperature, respiration and injection site assessment) at 30 minutes or longer after each study injection. 
4 Negative pregnancy test results must be confirmed for women of reproductive potential prior to each study injection. 
5 HLA type blood sample is collected once at any time-point in the study and is shown as a Visit 05 evaluation for convenience; however, if HLA type is already available in the medical record, it does not need to 
be repeated. HLA type may also be obtained from a frozen sample. 
6 Visit 07: Two tubes of PBMCs collected in EDTA tubes will be sent to Building 40, while the remainder of the blood collected will be sent to VITL. 
7 If optional apheresis occurs, ONLY draw 32mL in SST (DO NOT draw 120 ml in EDTA tubes). For women of reproductive potential, pregnancy test must be negative within 72 hours prior to apheresis procedure.  
Visit windows: Schedule Visits 02A through 06 with respect to Day 0; Schedule Visits 06A through 11 with respect to Visit 06. The following visit windows apply: Visits 02A and 06A (+1 day). Visit 03 and 07 (±1 
day). Visits 04, 05 and 09 (±2 days). Visit 06 (±28 days). Visits 08 (±3 days). Visits 10 and 11 (±14 days). 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 55 of 62 
APPENDIX III: ASSESSMENT OF RELATIONSHIP TO VACCINE AND GRADING 
SEVERITY OF ADVERSE EVENTS

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 56 of 62 
Assessment of Relationship of an Adverse Event to Study Vaccine:  
 
The relationship between an AE and the vaccine will be assessed by the investigator on the basis 
of his or her clinical judgment and the definitions below. 
• Definitely Related. The AE and administration of study agent are related in time, and 
a direct association can be demonstrated. 
• Probably Related. The AE and administration of study agent are reasonably related 
in time, and the AE is more likely explained by study agent than other causes.  
• Possibly Related. The AE and administration of study agent are reasonably related in 
time, but the AE can be explained equally well by causes other than study agent. 
• Not Related. The AE is clearly explained by another cause not related to the study 
product.  
For purposes of preparing summary data reports in which AE attributions are simplified to 
“Related” or “Not Related”, in this protocol, the “Definitely, Probably and Possibly” attributions 
above will be mapped to the “Related” category, while the “Unlikely/Probably Not Related” and 
“Not Related” attributions above will be mapped to the “Not Related” category. The definitions 
that apply when these two attribution categories alone are used are as follows: 
• Related – There is a reasonable possibility that the AE may be related to the study 
product(s). 
• Not Related – There is not a reasonable possibility that the AE is related to the study 
product(s). 
 
Grading the Severity of Adverse Events: 
The FDA Guidance for Industry (September 2007): “Toxicity Grading Scale for Healthy Adult 
and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical Trials” is the basis for the 
severity grading of AEs in this protocol. Several modifications were made to the table as follows: 
• “Emergency room visit” is not automatically considered a life-threatening event; these 
words have been removed from any “Grade 4” definition where they appear in the table 
copied from the guidance document. 
• Laboratory value shown as a “graded” value in the table that is within the institutional 
normal range will not be severity graded or recorded as an AE. 
• Severity grading for hemoglobin decrease on the basis of the magnitude of decrease from 
baseline is not applicable at the Grade 1 level; only absolute hemoglobin will be used to 
define Grade 1. 
• Severity grading for Grade 4 local reaction to injectable product (Erythema/Redness and 
Induration/Swelling) refer to necrosis or exfoliative dermatitis “requiring medical 
attention.” 
 
When not otherwise specified, the following guidance will be used to assign a severity grade: 
• Grade 1 (Mild): No effect on activities of daily living 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
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Page 57 of 62 
• Grade 2 (Moderate): Some interference with activity not requiring medical intervention 
• Grade 3 (Severe): Prevents daily activity and requires medical intervention  
• Grade 4 (Life-threatening): Hospitalization; immediate medical intervention or therapy 
required to prevent death. 
• Grade 5 (Death): Death is assigned a Grade 5 severity. Only the single AE that is 
assessed as the primary cause of death should be assigned “Grade 5” severity. 
 
Toxicity Grading Scale for Healthy Adult and Adolescent Volunteers Enrolled in 
Preventive Vaccine Clinical Trials 
Modified from FDA Guidance - September 2007 
 
A. Tables for Clinical Abnormalities  
 
Local Reaction to  
Injectable Product  
Mild 
 (Grade 1)  
Moderate 
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening  
(Grade 4)  
Pain  
Does not 
interfere  
with activity  
Repeated use of 
non-narcotic 
pain reliever > 
24 hours or 
interferes with 
activity  
Any use of 
narcotic pain 
reliever or 
prevents daily  
activity  
Hospitalization  
Tenderness  
Mild discomfort 
to touch  
Discomfort with 
movement  
Significant  
discomfort 
at 
rest  
Hospitalization  
1Erythema/Redness   
2.5 – 5 cm  
5.1 – 10 cm  
> 10 cm  
Necrosis or  
exfoliative 
dermatitis 
requiring 
medical 
attention  
2 Induration/Swelling   2.5 – 5 cm and 
does 
not 
interfere  
with activity  
5.1 – 10 cm or  
interferes with  
activity  
> 10 cm or  
prevents daily  
activity  
Necrosis 
requiring 
medical 
attention  
 
3 Vital Signs 
Mild  
(Grade 1)  
Moderate 
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening  
(Grade 4)  
4 Fever (°C)  
            (°F)  
38.0 – 38.4  
100.4 – 101.1  
38.5 – 38.9  
101.2 – 102.0  
39.0 – 40  
102.1 – 104  
> 40  
> 104  
Tachycardia - beats 
per  
minute  
101 – 115  
116 – 130  
> 130  
Hospitalization 
for arrhythmia  

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
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Page 58 of 62 
5 Bradycardia - beats 
per Minute 
50 – 54  
45 – 49  
< 45  
Hospitalization 
for arrhythmia  
Hypertension 
(systolic) - mm Hg  
141 – 150  
151 – 155  
> 155  
Hospitalization 
for malignant  
hypertension  
Hypertension 
(diastolic) - mm Hg  
91 – 95  
96 – 100  
> 100  
Hospitalization 
for malignant  
hypertension  
Hypotension 
(systolic) – mm Hg  
85 – 89  
80 – 84  
< 80  
Hospitalization 
for hypotensive 
shock  
Respiratory Rate – 
breaths per minute  
17 – 20  
21 – 25  
> 25  
Intubation  
 
1. In addition to grading the measured local reaction at the greatest single diameter, the measurement 
should be recorded as a continuous variable.  
2. Induration/Swelling should be evaluated and graded using the functional scale as well as the actual 
measurement. 
3. Subject should be at rest for all vital sign measurements.  
4. Oral temperature; no recent hot or cold beverages or smoking.  
5. When resting heart rate is between 60 – 100 beats per minute. Use clinical judgment when 
characterizing  
Bradycardia among some healthy subject populations, for example, conditioned athletes. 
 
Systemic (General)  
Mild  
(Grade 1)  
Moderate 
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening  
(Grade 4)  
Nausea/vomiting  
No interference  
with activity or 
1 – 2  
episodes/24 
hours  
Some 
interference  
with activity or 
> 2  
episodes/24 
hours  
Prevents daily  
activity, requires  
outpatient IV  
hydration  
Hospitalization 
for hypotensive 
shock  
Diarrhea  
2 – 3 loose 
stools or 
< 400 gms/24 
hours 
4 – 5 stools or  
400 – 800 
gms/24  
hours  
6 or more 
watery  
stools or  
> 800gms/24 
hours or 
requires  
outpatient IV  
hydration  
Hospitalization  
Headache  
No interference 
with activity  
Repeated use of 
non-narcotic 
pain  
Significant; any  
use of narcotic  
pain reliever or  
Hospitalization  

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 59 of 62 
reliever > 24 
hours or some 
interference  
with activity  
prevents daily  
activity  
Fatigue  
No interference 
with activity  
Some 
interference  
with activity  
Significant;  
prevents daily  
activity  
Hospitalization  
Myalgia  
No interference 
with activity  
Some 
interference  
with activity  
Significant;  
prevents daily  
activity  
Hospitalization  
 
Systemic Illness  
Mild  
(Grade 1)  
Moderate 
(Grade 2) 
Severe 
 (Grade 3)  
Potentially Life  
Threatening  
(Grade 4)  
Illness or clinical 
adverse  
event (as defined  
according to 
applicable  
regulations)  
No interference 
with activity  
Some 
interference  
with activity not  
requiring 
medical  
intervention  
Prevents daily  
activity and  
requires medical  
intervention  
Hospitalization  
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 60 of 62 
B. Tables for Laboratory Abnormalities  
 
The laboratory values provided in the tables below serve as guidelines and are dependent upon 
the institutional normal parameters. Institutional normal reference ranges should be provided to 
demonstrate that they are appropriate. 
 
Serum *  
Mild  
(Grade 1)  
Moderate  
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening  
(Grade 4) ** 
Sodium – Hyponatremia 
mEq/L  
132 – 134  
130 – 131  
125 – 129  
< 125  
Sodium – Hypernatremia 
mEq/L  
144 – 145  
146 – 147  
148 – 150  
> 150  
Potassium – Hyperkalemia 
mEq/L  
5.1 – 5.2  
5.3 – 5.4  
5.5 – 5.6  
> 5.6  
Potassium – Hypokalemia 
mEq/L  
3.5 – 3.6  
3.3 – 3.4  
3.1 – 3.2  
< 3.1  
Glucose – Hypoglycemia 
mg/dL  
65 – 69  
55 – 64  
45 – 54  
< 45  
Glucose – Hyperglycemia  
Fasting – mg/dL  
Random – mg/dL  
 
100 – 110  
110 – 125  
 
111 – 125  
126 – 200  
 
>125  
>200  
Insulin 
requirements or 
hyperosmolar 
coma  
Blood Urea Nitrogen  
BUN mg/dL  
23 – 26  
27 – 31  
> 31  
Requires dialysis  
Creatinine – mg/dL  
1.5 – 1.7  
1.8 – 2.0  
2.1 – 2.5  
> 2.5 or requires 
dialysis  
Calcium – hypocalcemia 
mg/dL  
8.0 – 8.4  
7.5 – 7.9  
7.0 – 7.4  
< 7.0  
Calcium – hypercalcemia 
mg/dL  
10.5 – 11.0  
11.1 – 11.5  
11.6 – 12.0  
> 12.0  
Magnesium – 
hypomagnesemia mg/dL  
1.3 – 1.5  
1.1 – 1.2  
0.9 – 1.0  
< 0.9  
Phosphorous – 
hypophosphatemia mg/dL  
2.3 – 2.5  
2.0 – 2.2  
1.6 – 1.9  
< 1.6  
CPK – mg/dL  
1.25 – 1.5 x 
ULN***  
>1.5 – 3.0 x 
ULN  
>3.0 –10 x 
ULN  
> 10 x ULN  
Albumin – 
Hypoalbuminemia g/dL  
2.8 – 3.1  
2.5 – 2.7  
< 2.5  
--  
Total Protein – 
Hypoproteinemia g/dL  
5.5 – 6.0  
5.0 – 5.4  
< 5.0  
--  
Alkaline phosphate –  
increase by factor  
1.1 – 2.0 x 
ULN  
2.1 – 3.0 x 
ULN  
> 3.0 – 10 x 
ULN  
> 10 x ULN  

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 61 of 62 
Serum *  
Mild  
(Grade 1)  
Moderate  
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening  
(Grade 4) ** 
Liver Function Tests –
ALT, AST increase by 
factor  
1.1 – 2.5 x 
ULN  
> 2.6 – 5.0 x 
ULN  
> 5.1 – 10 x 
ULN  
> 10 x ULN  
Bilirubin – when 
accompanied by any 
increase in Liver Function 
Test  
increase by factor  
1.1 – 1.25 x 
ULN  
> 1.26 – 1.5 x 
ULN  
> 1.51 – 
1.75 x ULN  
> 1.75 x ULN  
Bilirubin – when Liver 
Function Test is normal; 
increase by factor  
1.1 – 1.5 x 
ULN  
1.6 – 2.0 x 
ULN  
2.0 – 3.0 x 
ULN  
> 3.0 x ULN  
Cholesterol  
201 – 210  
211 – 225  
> 226  
---  
Pancreatic enzymes – 
amylase, lipase  
1.1 – 1.5 x 
ULN  
1.6 – 2.0 x 
ULN  
2.1 – 5.0 x 
ULN  
> 5.0 x ULN  
* The laboratory values provided in the tables serve as guidelines and are dependent upon 
institutional normal parameters. Institutional normal reference ranges should be provided to 
demonstrate that they are appropriate. 
** The clinical signs or symptoms associated with laboratory abnormalities might result in 
characterization of the laboratory abnormalities as Potentially Life Threatening (Grade 4). For 
example, a low sodium value that falls within a Grade 3 parameter (125-129 mE/L) should be 
recorded as a Grade 4 hyponatremia event if the subject had a new seizure associated with the 
low sodium value. 
***ULN” is the upper limit of the normal range. 
 
 
 

PRODUCTS: VRC-FLUNPF081-00-VP AND VRC-FLUDNA082-00-VP 
 
PROTOCOL: VRC 316, VERSION 6.0 
PROTOCOL DATE: MARCH 11, 2019 
Page 62 of 62 
Hematology *  
Mild  
(Grade 1)  
Moderate  
(Grade 2)  
Severe  
(Grade 3)  
Potentially Life  
Threatening 
(Grade 4)  
Hemoglobin (Female) - 
gm/dL  
11.0 – 12.0  
9.5 – 10.9  
8.0 – 9.4  
< 8.0  
Hemoglobin (Female)  
decrease from baseline 
value - gm/dL  
not 
applicable 
1.6 – 2.0  
2.1 – 5.0  
> 5.0  
Hemoglobin (Male) - 
gm/dL  
12.5 – 13.5  
10.5 – 12.4  
8.5 – 10.4  
< 8.5  
Hemoglobin (Male)  
decrease from baseline 
value – gm/dL  
not 
applicable  
1.6 – 2.0  
2.1 – 5.0  
> 5.0  
WBC Increase - cell/mm
3 
10,800 – 
15,000  
15,001 – 
20,000  
20,001 – 25, 
000  
> 25,000  
WBC Decrease - cell/mm
3 
2,500 – 
3,500  
1,500 – 2,499  
1,000 – 
1,499  
< 1,000  
Lymphocytes Decrease - 
cell/mm
3 
750 – 1,000  500 – 749  
250 – 499  
< 250  
Neutrophils Decrease - 
cell/mm
3 
1,500 – 
2,000  
1,000 – 1,499  
500 – 999  
< 500  
Eosinophils - cell/mm
3 
650 – 1500  
1501 - 5000  
> 5000  
Hypereosinophilic  
Platelets Decreased - 
cell/mm
3 
125,000 – 
140,000  
100,000 – 
124,000  
25,000 – 
99,000  
< 25,000  
PT – increase by factor  
(prothrombin time)  
1.10 x 
ULN**  
> 1.11 – 1.20 x 
ULN  
1.21 – 1.25 
x ULN  
> 1.25 ULN  
PTT – increase by factor 
(partial thromboplastin 
time)  
1.10 – 1.20 
x ULN  
1.21 – 1.4 x 
ULN  
1.4 – 1.5 x 
ULN  
> 1.5 x ULN  
Fibrinogen increase - 
mg/dL  
400 – 500  
501 – 600  
> 600  
--  
Fibrinogen decrease - 
mg/dL  
150 – 200  
125 – 149  
100 – 124  
< 100 or 
associated with 
gross bleeding  
or disseminated 
intravascular 
coagulation (DIC)  
* The laboratory values provided in the tables serve as guidelines and are dependent upon 
institutional normal parameters. Institutional normal reference ranges should be provided to 
demonstrate that they are appropriate. 
**ULN” is the upper limit of the normal range. 

 
 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
               •  Adult Patient or     •  Parent, for Minor Patient 
 
 
INSTITUTE: 
National Institute of Allergy and Infectious Diseases 
 
 
STUDY NUMBER: 
17-I-0110 
 
PRINCIPAL INVESTIGATOR: Grace Chen, M.D., M.P.H. 
 
 
STUDY TITLE: 
VRC 316: A Phase I Open-Label Clinical Trial to Evaluate Dose, Safety, Tolerability, and 
Immunogenicity of an Influenza HA Ferritin Vaccine, Alone or in Prime-Boost Regimens with an 
Influenza DNA Vaccine in Healthy Adults 
 
 
Continuing Review Approved by the IRB on 04/23/18 
 
 
 
 
Amendment Approved by the IRB on 11/13/18 (J)                                         Date Posted to Web: 11/24/18 
Standard Study Consent, Version 5.0 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY 
 
•   Adult Patient or    •   Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent (1) 
 
 
INTRODUCTION 
 
We invite you to take part in a research study at the National Institutes of Health (NIH). 
 
First, we want you to know that: 
· 
Taking part in NIH research is entirely voluntary. 
· 
You may choose not to take part, or you may withdraw from the study at any time. In either case, you will not 
lose any benefits to which you are otherwise entitled. However, to receive care at the NIH, you must be taking 
part in a study or be under evaluation for study participation. 
· 
You may receive no benefit from taking part. The research may give us knowledge that may help people in the 
future. 
 
Second, some people have personal, religious or ethical beliefs that may limit the kinds of medical or research treatments 
they would want to receive (such as blood transfusions). If you have such beliefs, please discuss them with your NIH 
doctors or research team before you agree to the study. 
 
Now we will describe this research study. Before you decide to take part, please take as much time as you need to ask 
any questions and discuss this study with anyone at NIH, or with family, friends or your personal physician or other health 
professional. 
PURPOSE OF THE STUDY 
This is the first study in people of two experimental vaccines for prevention of H2 influenza (flu). “Experimental” means 
that the study vaccines have not been approved by the Food and Drug Administration (FDA). The FDA allows these 
vaccines to be used for research purposes only. These vaccines have never been given to humans before this study. We 
do not know if these vaccines work. The main purpose of this study is to see if the experimental vaccines are safe and if 
there are any side effects. We also want to study immune responses to these vaccines. 
STUDY VACCINES 
Vaccines are given to teach the body to prevent or fight an infection. In this study, we are testing two experimental 
vaccines that were developed by the Vaccine Research Center (VRC) at the NIH: VRC-FLUNPF081-00-VP, also called “HA-F 
A/Sing Vaccine” and VRC-FLUDNA082-00-VP, also called “DNA A/Sing Vaccine.” These vaccines are intended to help the 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  2  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
body to make an immune response to H2 flu. 
HA-F A/Sing Vaccine: Most vaccines are made of proteins that are injected into a muscle. Proteins are natural substances 
that the body uses as building blocks. This vaccine is made in the laboratory with two proteins: one protein from the H2 
flu virus and one protein from a type of bacteria called Helicobacter pylori (H. pylori). These two proteins have been 
modified in the laboratory. When combined, they make a particle that looks like the outside of the H2 flu virus. The body’s 
immune system may respond to this particle. There is no virus or bacteria in the vaccine.  
DNA A/Sing Vaccine: DNA (deoxyribonucleic acid) is a molecule that serves as nature’s code (instructions) for protein 
production in the body. The vaccine tested in this study is made from the DNA that is the code for an H2 flu virus protein. 
When you get the DNA vaccine, it tells your body to make a small amount of H2 protein. Your body may use this protein 
to build an immune response. There is no virus in the vaccine. 
Similar DNA vaccines for other types of influenza were tested in many people. There were no serious side effects related 
to these DNA vaccines. The NIH has tested several DNA vaccines made by the VRC. These vaccines were found to be safe 
and well-tolerated in people. 
This is the first study to give these two vaccines to humans. We do not know if the HA-F A/Sing or DNA A/Sing vaccine 
will protect you from the flu.  
There is no virus or bacteria in the vaccines, so you cannot get an H2 flu infection or bacterial infection from these 
vaccines. 
ELIGIBILITY 
You are eligible to take part in this study because you have completed the screening process and are: 
· 
18-47 or 52-70 years old, unless you were born in 1966-1969 
· 
In general good health without significant medical problems as determined at screening 
· 
Willing to get the experimental vaccines 
· 
Willing to donate blood samples for future research 
· 
Willing to use birth control from at least 21 days prior to enrollment through the end of the study, if female and 
able to become pregnant 
STUDY PLAN 
About 50 people will take part in this study at the NIH Clinical Center in Bethesda, MD. You will be in the study for about 
40 weeks (10 months) and get either 1 or 2 vaccinations. You will get the experimental vaccines by injections (shots) in 
the upper arm muscle. This is called an intramuscular “IM” injection. We will check you for any side effects from the 
vaccine.  

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  3  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
We will tell you if we learn anything new during this study that might cause you to change your mind about staying in the 
study. At the end of the study, we will tell you when study results may be available and how to learn about them. 
The study has two Parts:  
Part I will test a lower dose (20 mcg) and higher dose (60 mcg) of the HA-F A/Sing vaccine. The study will start by 
enrolling people to get the lower dose in Group 1. If there are no safety concerns at this dose, we will start enrolling 
people to get the higher dose in Group 2. If there are no safety concerns at this dose, we will start enrolling people in 
Part II. We will use a needle and syringe to give you the HA-F A/Sing vaccine. 
The table below shows the plan for Part I: 
 
Part I 
Group 
Age 
Subjects 
Day 0 
Week 16 
1 
18-47 years 
5 
HA-F A/Sing, 20 mcg 
2 
18-47 years 
5 
HA-F A/Sing, 60 mcg  
HA-F A/Sing, 60 mcg 
Total 
 
10 
Part II will test the DNA A/Sing vaccine and the higher dose of the HA-F A/Sing vaccine. People will be grouped by age 
and randomly assigned (like pulling a number from a hat) to 1 of 2 groups (in Group 3 or Group 4). This is done so that 
researchers may compare the immune response when people were either exposed or not exposed to the H2 flu virus 
during their lifetime. Once enrolled, you will know which group you are in. 
We will use a needle and syringe to give the HA-F A/Sing vaccine.  
We will use a needle-free injection device called the PharmaJet Stratis® Needle Free Jet Injector (PharmaJet) to give the 
DNA A/Sing vaccine, it will be given as two injections at the visit, one in each upper arm. The needle-free injection device 
delivers the vaccine through the skin without the use of a needle. Instead, it uses high pressure to push the vaccine 
through your skin into the muscle. The needle-free injection device is “needleless,” but may cause pain, some people have 
said that the sensation is similar of being snapped with a rubber band or as a mild punch in the arm. Needleless systems 
have been used to give vaccines and other medications since 1947. This system has been cleared by the FDA for giving 
vaccine injections into the muscle. Previous studies have shown that some people who get injections with PharmaJet have 
more local reactions at the injection site than people who get injections with a needle and syringe. These local reactions 
were mild. 
The table below shows the plan for Part II: 
Part II 
Group 
Age 
Subjects 
Day 0 
Week 16 
3 
A 
18-47 years 
10 
DNA A/Sing, 4 mg;  
needle-free injector 
HA-F A/Sing, 60 mcg; 
needle and syringe 
B 
52-70 years 
10 
4 
A 
18-47 years 
10 
HA-F A/Sing, 60 mcg;  
needle and syringe 
HA-F A/Sing, 60 mcg; 
needle and syringe 
B 
52-70 years 
10 
 
Total 
 
40 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  4  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
STUDY PROCEDURES 
If you agree to take part in the study and are enrolled in: 
· 
Group 1: you will have 1 vaccination visit. The HA-F A/Sing vaccine will be given as one injection at Day 0 
· 
Group 2: you will have 2 vaccination visits. The HA-F A/Sing vaccine will be given as one injection at Day 0 and at 
Week 16. 
· 
Group 3A/B: you will have 2 vaccination visits. The DNA A/Sing will be given as two injections at Day 0 and the 
HA-F A/Sing vaccine will be given as one injection Week 16. 
· 
Group 4A/B: you will have 2 vaccination visits. The HA-F A/Sing vaccine will be given as one injection at Day 0 
and at Week 16. 
The study duration will last for 40 weeks (10 months) with about 8-10 clinic visits and 1 or 2 telephone contacts, 
depending on the group. 
If you are a woman who could get pregnant, we will do a pregnancy test before each injection. The test must show that 
you are not pregnant to get the injection. 
Each vaccination visit will take about 4-6 hours. Most follow-up clinic visits will take about 1 to 2 hours; the optional 
apheresis visit will take 2-4 hours. 
If you are enrolled in Group 1, Group 2, or Group 3A/B, you will need to stay in the clinic for at least 1 hour after each 
vaccination. If you are enrolled in Group 4A/B, you will need to stay in the clinic for at least 30 minutes after each 
vaccination.  
After each vaccination, the clinic staff will call you to check on you. Also, after each vaccination you will need to complete 
a diary card for 7 days after each injection. The purpose of the diary card is to record any symptoms that you may have. 
We will give you a thermometer to record your temperature even if you feel well. We will also give you a ruler to measure 
any skin changes at the injection site(s). You will get a password to a secure website to enter this data online. If you do 
not have access to a computer, you may use a paper diary card instead. 
If you have any symptoms or feel unwell, you should tell a clinic nurse or doctor as soon as possible. You can reach the 
staff by phone 24 hours a day. If you have symptoms, you may be asked to come to the clinic for a checkup. It is very 
important that you follow the instructions from the clinic staff. 
At each visit, we will check you for any health changes or problems. We will ask you how you are feeling and if you have 
taken any new medications. We will draw your blood at each visit, taking about 2 to 16 tubes of blood. We will tell you 
right away if any of your test results show a health problem. Some blood samples will be used for research only, to study 
your immune response to the vaccine. Results of these research tests are not used to check your health and will not be 
given to you. 
Apheresis: After your final study injection we would like to collect your blood by a method called “Apheresis.” This 
procedure is optional, and it is only being offered to subjects enrolled in Group 2, Group 3A/B, and Group 4A/B. 

 
  
CONTINUATION SHEET for either: 
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NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  5  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
To be eligible for apheresis: 
· 
You must not have an unstable heart as indicated by your medical history and test results 
· 
You must not have blood pressure greater than 180/100 
· 
You must not have a known blood clotting disorder 
· 
You must not be pregnant or breast feeding 
· 
You must not have a condition that the attending physician or the apheresis clinic staff considers a reason to not 
do an apheresis procedure. 
Before apheresis, we will check your weight, pulse and blood pressure. We will ask questions about your general health 
and medical history. If you are a woman who could get pregnant, we will do a pregnancy test before the apheresis 
procedure. The test must show that you are not pregnant. 
During procedure, you will lie on a recliner, couch, or hospital bed. The procedure requires placing a needle into a vein in 
one or both arms using a sterile method. The kits used to collect apheresis samples are sterile, single-use, disposable sets 
that are not in contact with any person’s body fluids other than yours. No blood products are given to you during these 
procedures. The apheresis is done at the NIH Clinical Center, and a physician from the Department of Transfusion 
Medicine will be available in or near the apheresis donor area at all times. 
In the apheresis procedure, blood is removed through a needle in the vein of one arm, spun in a machine that permits 
separation of the desired blood component (white blood cells or plasma), and then the remainder is returned through a 
needle in the other arm. Citrate, a medication to prevent blood from clotting, is added to the blood while in the machine 
to prevent it from clotting. 
The purpose of these procedures is to allow the investigator to obtain and study a larger number of white blood cells or 
plasma than would otherwise be possible by simple blood drawing. The number of white blood cells or plasma collected is 
a small fraction of the total amount in your body. The body quickly replaces removed cells and plasma. Similar procedures 
are used daily in the Blood Bank of the Clinical Center and by other blood banks to obtain blood products from normal 
donors and as a form of therapy for certain diseases. However, we will not use your samples for transfusion or therapy. 
The procedure will take approximately 1-3 hours. 
MONITORING OF THE STUDY 
This study will be monitored by a group of physicians and scientists at NIH. This group will review the study information 
and will pay close attention to any reactions. If there are serious side effects, study injections may be delayed or 
canceled. 
GENETIC TESTING 
Some of the blood drawn from you during this study will be used for genetic tests. Some genetic tests are done in 
research studies to see if genetic differences in people cause different types of immune responses. Your blood sample 

 
  
CONTINUATION SHEET for either: 
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STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  6  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
used in these genetic tests will not have your name on it and the results will not be in your medical record except for 
Human Leukocyte Antigen (HLA) typing results. These tests are not used to check your health and we will not tell you the 
results.  
A special genetic test, called HLA typing, is done by the NIH Clinical Center medical laboratory. These results will be in 
your medical record but they will not be used to check your health. Any genetic testing, including HLA testing, is for 
research purposes only. Any genetic information collected or learned about you will be kept confidential. Medical records, 
including HLA test results are kept securely. We will not give any genetic information that is in your medical record to 
anyone without your permission. 
STORED SAMPLES 
We will collect blood samples from you during the study. We will keep these samples for future research indefinitely to 
learn more about flu virus, vaccines, the immune system, and other research questions. Results from research with your 
samples will not be in your medical record or reported to you. 
Labeling of Stored Samples: Your stored samples will be labeled by a special code or number and not your personal 
information. Only the study team can link this code to you. Any identifying information about you (like name or date of 
birth) will be kept as confidential as allowable by law. 
Risks of Stored Samples: There is a risk of unplanned release of information from your medical record. The chance 
that this information will be given to an unauthorized person without your permission is very small. Possible problems with 
the unplanned release of information include discrimination when applying for insurance or employment. Similar problems 
may occur if you give information about yourself or agree to have your medical record released. 
Future studies: In the future, other investigators at NIH or outside of NIH may wish to study your stored samples. When 
your stored samples are shared, they will be marked with a code.  
Your samples will not have any identifying information on them. Some information about you, like your gender, age, 
health history, or ethnicity may be shared with other researchers. Any future research studies using your samples will be 
conducted in a way that protects the rights and privacy of study participants. 
Your stored samples will only be used for research and will not be sold. The research done with your materials may be 
used to develop new products in the future but you will not receive payment for such products. 
Making your Choice: You cannot take part in this study if you do not want us to collect or store your blood samples. If 
you agree to take part in this study, you must also agree to let us keep any of your samples for future research. If you 
decide not to take part in this study, you may still take part in other studies at NIH. 
HUMAN DATA SHARING 
To advance science, it is helpful for researchers to share information they get from studying humans by putting it into 
shared scientific databases. Researchers can then study the information combined from many studies to learn even more 
about health and diseases. 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  7  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
If you agree to take part in this study, some of your data will be placed into one or more scientific databases. We will 
remove identifying information like your name, address, and birth date. The data may then be used for future research 
and shared broadly for research purposes. 
Only researchers who are approved to access the database may be able to see and use your information. You will not get 
any direct benefits from future research that uses your data and information. 
You may stop participating in this study at any time and withdraw permission for your individual data, specimens, and 
health information to be used for additional or future research. You may ask to have your research data destroyed. 
However, it may not be possible to withdraw or delete data once they have been shared with other researchers. 
POSSIBLE STUDY RISKS 
Possible risks of injections: Temporary stinging, pain, redness, soreness, itchiness, swelling, or bruising.  
A needleless injector system called the PharmaJet Stratis® Needle Free Jet Injector (PharmaJet) will be used to give the 
DNA vaccine given in Group 3A/B. It uses pressure to push the vaccine into the arm.  
There is a small chance the vaccine may spray on the arm and a small chance that the needle-free device could not work 
properly. As of June 26, 2017, in another research study called VRC 705 Part A, there have been 8 out of 426 (1.9%) 
injections that involved device malfunction. This includes the syringe breaking and study product spraying during 
administration. No harm to subjects has occurred when this happened. As a result, if you are enrolled in Groups 3A/B, we 
ask that you and the nurse wear safety glasses when PharmaJet is used for injection. 
There is a very small chance of infection.  
Vaccines given by needleless injector systems have been associated with mild skin lesions, and a small scab about 1-2 
weeks after being given the vaccine. 
Possible risks of any vaccine: Fever, chills, rash, aches and pains, nausea, headache, dizziness, and feeling tired 
and/or unwell. These types of reactions are usually greatest within the first 24 hours after vaccination and typically last 1 
to 3 days. Over-the-counter medicine, like acetaminophen (Tylenol) or ibuprofen, may be used to help these symptoms.  
Very rarely, a serious allergic reaction with symptoms like hives, trouble breathing, or sudden weakness may occur shortly 
after any vaccination. This is called “anaphylaxis” and may be life-threatening. While you are waiting in the clinic after the 
vaccination, we will monitor you for anaphylaxis. Treatment for anaphylaxis will be given right away if it occurs. 
Possible risks of the HA-F A/Sing vaccine: The HA-F A/Sing vaccine has not been given to people before. It may 
have unknown risks. It has been tested in mice and rabbits. The vaccine did not cause any unusual side effects and met 
the safety criteria to be tested in humans. 
Possible risks of DNA A/Sing vaccine: Temporary drop in white blood cell count, sore arm, skin rash or hives. Some 
people get a small red bump and then a scab for a few days where the shot is given. Possible side effects from 
intramuscular injections are stinging, arm discomfort, redness of the skin, or mild bruising. 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  8  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
Possible risks of blood drawing: Pain, bleeding, bruising, feeling lightheaded, and fainting. Rarely, infection may occur 
at the site where the blood is taken. 
Possible risks of apheresis: Apheresis is generally safe and side effects are rare. Pain, bruising or discomfort at the 
needle placement site may occur. Sometimes apheresis causes a tingling sensation around the lips, nose and mouth, 
coolness all over and slight nausea. This can usually be relieved by slowing or temporarily stopping the apheresis or taking 
an antacid with calcium pill, like Tums®. Other possible side effects are anxiety, vomiting and lightheadedness. 
Temporary lowering of the blood pressure may develop. There is the rare possibility of infection, fainting or seizure. Very 
rarely a nerve problem at the needle placement site may occur. Also, very rarely, a machine malfunction may occur and 
result in the loss of about one unit (one pint) of blood. 
There are theoretical risks from re-infusion of the blood after processing by the machine like infection or an adverse 
reaction to the blood components. However, this has not been seen in many thousands of volunteers who have 
undergone this or similar procedures to date. There may be other risks of apheresis that are unknown at this time. 
Unknown safety risks: There may be side effects from the study vaccines - even serious or life threatening ones- that 
we do not yet know about. Please tell the study staff about any side effect you think you are having. This is important for 
your safety. 
Possible risks of genetic testing: Unplanned release of information that could be used by insurers or employers to 
discriminate against you or your family; discovering a gene that suggests risk of disease for you or your family; 
discovering unknown family relationships. 
Possible risks of data sharing: Information in the shared databases could be linked back to you and used to 
discriminate against you or your family. State and federal laws provide some protections against genetic and pre-existing 
conditions discrimination. 
Possible risks during pregnancy: We do not know how the experimental vaccines may affect a fetus or nursing infant. 
Therefore, women who are able to become pregnant must have a negative pregnancy test before each injection and 
agree to use effective birth control beginning at least 21 days before the first injection until the end of the study. We will 
discuss effective methods of birth control with you. If you are pregnant or want to become pregnant in the next 40 
weeks, you cannot participate. 
You must tell the clinic staff right away if you become pregnant or think that you might be pregnant during the study. If 
you are pregnant, you will not get any more vaccinations. You will be asked to continue with follow-up visits so that we 
can check your health. We will ask you the outcome of the pregnancy. 
Possible other risks: We do not know if the study vaccine(s) will change how your body responds to flu virus infections 
in the future. 
You may not donate blood at a blood bank while taking part in this study. You may not donate blood for one year after 
the last experimental vaccine injection. 
 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  9  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
POSSIBLE BENEFITS 
This study is not designed to benefit you. We do not know if the vaccines will work. The study is not designed to protect 
you from flu. You and others may benefit in the future from the information that will be learned from the study. The study 
visits are used to check your health for research purposes, not to provide health care. However, we will tell you right 
away if any of your test results show a possible health problem. 
COSTS TO YOU FOR PARTICIPATION 
There are no costs to you for taking part in this study. We will not charge you or your insurance carrier for any health 
evaluations or services. You or your health insurance will have to pay for all costs for medical care that you get outside 
this study. It is possible that you may have some costs that are not covered by the study compensation we give you. 
COMPENSATION TO YOU FOR PARTICIPATION 
You will be compensated for your time and inconvenience by the NIH Clinical Research Volunteer Program. It is possible 
that you may have some expenses that are not covered by the compensation provided. 
The compensation for is $275 for vaccination visit(s). You will get $25 total for the timely completion of all 7 days of an 
electronic diary. You will get $175 for each scheduled follow-up visit that includes a research blood draw. You will get $75 
for all other visits that do not include research blood draws. You will get $250 for apheresis. 
If you are enrolled in Group 1, the total compensation for completion of all study visits is about $1,525. 
If you are enrolled in Groups 2-4 the total compensation for completion of all study visits is about $2,000 without 
apheresis and about $2,075 with apheresis. 
Actual compensation is based on the number and type of study visits you complete. Your compensation may need to be 
reported to the Internal Revenue Service (IRS) as taxable income. 
REASONS FOR STOPPING STUDY INJECTIONS 
You may not get all of your planned study vaccinations. Reasons for this may include: 
· 
You don't keep appointments or follow procedures. 
· 
You get a serious illness that needs ongoing medical care. 
· 
You have a serious side effect thought to be due to the study vaccine. 
· 
You become pregnant. 
· 
You need to get treatment with a medication that affects your immune system (such as a steroid like prednisone). 
· 
The study is stopped by regulatory agencies, the study sponsor or study investigators. If this happens, we will tell 
you why. 
 
If you agree to take part in this study, it is important for you to keep all of your appointments. Your participation in this 

 
  
CONTINUATION SHEET for either: 
  MEDICAL RECORD 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
 
STUDY NUMBER:  
17-I-0110 
 
CONTINUATION:  page  10  of  11  pages 
 
 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
 
NIH-2514-1 (10-84) 
 
NIH-2514-2 (10-84) 
 
P.A.: 09-25-0099 
 
 
study is completely voluntary. You can choose to stop taking part in the study at any time. There is no penalty or loss of 
benefits choosing to leave the study.  
If you get the first but not the second injection (for Groups 2, 3A/B, or 4A/B), we will ask you to continue with your 
planned follow-up visits until the end of the study. It is important that we continue to check your health even if you do not 
get a second injection. 
ALTERNATIVES 
This study is not designed to treat or prevent any disease. You may choose to not take part in this study. You may be 
eligible for other studies.  
CONFLICT OF INTEREST  
The NIH research staff is checked yearly for conflicts of interest. You may ask the research team for more information. 
This study may have investigators who are not NIH employees. Non-NIH investigators are expected to follow the 
principles of the Protocol Review Guide but are not required to report their personal financial holdings to the NIH. 
The NIH, including some members of the VRC scientific staff, developed the investigational vaccines being used in this 
research study. The results of this study could play a role in whether the FDA will approve the vaccine for sale at some 
time in the future. If approved, the future sale of the vaccine could lead to payments to NIH and some NIH scientists. By 
U.S. law, government scientists are required to receive such payments for their inventions. You will not get any money 
from the development or sale of the product.  
CLINICALTRIALS.GOV 
A description of this clinical trial will be available on www.ClinicalTrials.gov. This website will not include information that 
can identify you. At most, the website will include a summary of the results. You can search this website at any time. 

 
  
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
•  Adult Patient or    •  Parent, for Minor Patient 
 
STUDY NUMBER: 
17-I-0110 
 
CONTINUATION:  page  11  of  11  pages 
 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY (Continuation Sheet) 
 
•  Adult Patient or   •  Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent 
 
OTHER PERTINENT INFORMATION 
 
1. Confidentiality. When results of an NIH research study are reported in medical journals or at scientific meetings, the 
people who take part are not named and identified. In most cases, the NIH will not release any information about your 
research involvement without your written permission. However, if you sign a release of information form, for example, 
for an insurance company, the NIH will give the insurance company information from your medical record. This 
information might affect (either favorably or unfavorably) the willingness of the insurance company to sell you insurance. 
 
The Federal Privacy Act protects the confidentiality of your NIH medical records. However, you should know that the Act 
allows release of some information from your medical record without your permission, for example, if it is required by the 
Food and Drug Administration (FDA), members of Congress, law enforcement officials, or authorized hospital accreditation 
organizations. 
 
2. Policy Regarding Research-Related Injuries. The Clinical Center will provide short-term medical care for any 
injury resulting from your participation in research here. In general, no long-term medical care or financial compensation 
for research-related injuries will be provided by the National Institutes of Health, the Clinical Center, or the Federal 
Government. However, you have the right to pursue legal remedy if you believe that your injury justifies such action. 
 
3. Payments. The amount of payment to research volunteers is guided by the National Institutes of Health policies. In 
general, patients are not paid for taking part in research studies at the National Institutes of Health. Reimbursement of 
travel and subsistence will be offered consistent with NIH guidelines. 
 
4. Problems or Questions. If you have any problems or questions about this study, or about your rights as a research 
participant, or about any research-related injury, contact the Principal Investigator, 
 
 or the Study Coordinator, 
 
 
. 
You may also call the Clinical Center Patient Representative at 
. 
 
5. Consent Document. Please keep a copy of this document in case you want to read it again. 
 
COMPLETE APPROPRIATE ITEM(S) BELOW: 
 
A. Adult Patient’s Consent 
B. Parent’s Permission for Minor Patient. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby consent to take 
part in this study. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby give permission 
for my child to take part in this study.   
(Attach NIH 2514-2, Minor’s Assent, if applicable.) 
 
  
 
 
  
 
Signature of Adult Patient/Legal Representative 
Date 
 
 
 
Print Name 
Signature of Parent(s)/Guardian 
Date 
 
 
 
Print Name 
C. Child’s Verbal Assent (If Applicable) 
The information in the above consent was described to my child and my child agrees to participate in the study. 
 
 
 
  
 
 
 
 
Signature of Parent(s)/Guardian 
Date 
 
Print Name 
THIS CONSENT DOCUMENT HAS BEEN APPROVED FOR USE  
FROM APRIL 23, 2018 THROUGH APRIL 22, 2019. 
 
 
 
  
 
 
  
 
Signature of Investigator 
Date 
 
 
 
Print Name 
Signature of Witness 
Date 
 
 
 
Print Name 
 

 
 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
               •  Adult Patient or     •  Parent, for Minor Patient 
 
 
INSTITUTE: 
National Institute of Allergy and Infectious Diseases 
 
 
STUDY NUMBER: 
17-I-0110 
 
PRINCIPAL INVESTIGATOR:  Grace Chen, M.D., M.P.H. 
 
 
STUDY TITLE: 
VRC 316: A Phase I Open-Label Clinical Trial to Evaluate Dose, Safety, Tolerability, and 
Immunogenicity of an Influenza HA Ferritin Vaccine, Alone or in Prime-Boost Regimens with an 
Influenza DNA Vaccine in Healthy Adults 
 
 
Continuing Review Approved by the IRB on 04/23/18 
 
 
 
 
Amendment Approved by the IRB on 11/13/18 (J)                                         Date Posted to Web: 11/24/18 
Supplementary Study Consent, Version 5.0 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY 
 
•   Adult Patient or    •   Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent (2) 
 
 
Supplementary consent for an extended genetic testing (optional) 
You agreed to participate in VRC 316 study for testing of two investigational vaccines to prevent H2 influenza (flu) virus 
infection. This additional consent covers optional extended genetic testing that you may agree to. If you do not agree to 
this additional testing, you still can participate in the VRC 316 study. 
There is a new type of genetic test that lets us look at the expressions of genes, called transcriptome sequencing. This 
test lets us look at the genes that are actively expressed at any given moment. However, it does not measure the amount 
of protein produced. Also, this new genetic test is still in development and researchers are working on understanding the 
data and how that data can be utilized in various clinical applications, such as in medicine to help prevent disease.  
This test may take a long time to understand, and we may not have any news to give you about it. Since we are looking 
for genes that control infection or immune response, we will not report to you or your doctors things that we find that are 
not related to infection or immunity. However, if we find something in your DNA that we think is urgent to deal with 
because of your health, we will confirm the result and then tell you about it. We think this sort of problem will be rare. 
Genetic Data Sharing 
Following genetic testing for transcriptome sequencing, your sequence data will be shared in a controlled access public 
database, for other investigators to benefit from it. However, no personal, identifiable information will be shared in this 
process, as the shared results will be coded with no link back to you.  
 

 
  
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
   MEDICAL RECORD 
•  Adult Patient or    •  Parent, for Minor Patient 
 
STUDY NUMBER:   17-I-0110 
 
 
 
CONTINUATION: Page 2 of 2   
 
 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL 
 
RESEARCH STUDY (Continuation Sheet) 
 
•  Adult Patient or   •  Parent, for Minor Patient 
 
NIH-2514-1 (07-09) 
 
P.A.: 09-25-0099 
 
File in Section 4:  Protocol Consent 
 
 
OTHER PERTINENT INFORMATION 
 
1. Confidentiality. When results of an NIH research study are reported in medical journals or at scientific meetings, the 
people who take part are not named and identified. In most cases, the NIH will not release any information about your 
research involvement without your written permission. However, if you sign a release of information form, for example, 
for an insurance company, the NIH will give the insurance company information from your medical record. This 
information might affect (either favorably or unfavorably) the willingness of the insurance company to sell you insurance. 
 
The Federal Privacy Act protects the confidentiality of your NIH medical records. However, you should know that the Act 
allows release of some information from your medical record without your permission, for example, if it is required by the 
Food and Drug Administration (FDA), members of Congress, law enforcement officials, or authorized hospital accreditation 
organizations. 
 
2. Policy Regarding Research-Related Injuries. The Clinical Center will provide short-term medical care for any 
injury resulting from your participation in research here. In general, no long-term medical care or financial compensation 
for research-related injuries will be provided by the National Institutes of Health, the Clinical Center, or the Federal 
Government. However, you have the right to pursue legal remedy if you believe that your injury justifies such action. 
 
3. Payments. The amount of payment to research volunteers is guided by the National Institutes of Health policies. In 
general, patients are not paid for taking part in research studies at the National Institutes of Health. Reimbursement of 
travel and subsistence will be offered consistent with NIH guidelines. 
 
4. Problems or Questions. If you have any problems or questions about this study, or about your rights as a research 
participant, or about any research-related injury, contact the Principal Investigator 
 or the Study Coordinator, 
 
 
.  
You may also call the Clinical Center Patient Representative at 
. 
 
5. Consent Document. Please keep a copy of this document in case you want to read it again. 
 
COMPLETE APPROPRIATE ITEM(S) BELOW: 
 
A. Adult Patient’s Consent 
B. Parent’s Permission for Minor Patient. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby consent to take 
part in this study. 
I have read the explanation about this study and have been given the 
opportunity to discuss it and to ask questions.  I hereby give permission 
for my child to take part in this study.   
(Attach NIH 2514-2, Minor’s Assent, if applicable.) 
 
  
 
 
  
 
Signature of Adult Patient/Legal Representative 
Date 
 
 
 
Print Name 
Signature of Parent(s)/Guardian 
Date 
 
 
 
Print Name 
C. Child’s Verbal Assent (If Applicable) 
The information in the above consent was described to my child and my child agrees to participate in the study. 
 
 
 
  
 
 
 
 
Signature of Parent(s)/Guardian 
Date 
 
Print Name 
THIS CONSENT DOCUMENT HAS BEEN APPROVED FOR USE  
FROM APRIL 23, 2018 THROUGH APRIL 22, 2019. 
 
 
 
  
 
 
  
 
Signature of Investigator 
Date 
 
 
 
Print Name 
Signature of Witness 
Date 
 
 
 
Print Name