A Phase I, Randomized, Placebo Controlled, Double-Blind, Dose Escalation Trial to Evaluate the Safety and Immunogenicity of an Andes Virus DNA Vaccine for the Prevention of Hantavirus Pulmonary Syndrome Using the PharmaJet Stratis(R) Needle-Free Injection System in Normal Healthy Adults
Phase: PHASE1
Conditions: Hantavirus Pulmonary Infection, Immunisation
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

DMID #16-0119 
Version 4.0 
ANDV DNA Vaccines in Healthy Adults 
1 August 2019 
 
 
 
 
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A Phase I, Randomized, Placebo Controlled, Double-blind, Dose 
Escalation trial to Evaluate the Safety and Immunogenicity of an 
Andes Virus DNA Vaccine for the Prevention of Hantavirus 
Pulmonary Syndrome Using the PharmaJet Stratis® Needle-Free 
Injection System in Normal Healthy Adults 
DMID Protocol Number:  16-0119 
DMID Funding Mechanism: HHSN272201300016I 
Pharmaceutical Support Provided by: Department of Molecular Virology, United States Army 
Medical Research Institute of Infectious Diseases (USAMRIID), Aldevron 
IND Sponsor: Division of Microbiology and Infectious Diseases (DMID), National Institute of 
Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH) 
Principal Investigator:  Grant Paulsen, MD 
DMID Protocol Champion:  Gabriele Feolo, RN, MSN 
DMID Medical Officer:  Kay M. Tomashek, MD, MPH 
DMID Medical Monitor:  Jorge Mejia-Galvis, MD 
Version Number: 4.0 
Day Month Year 
1 August 2019 
 

DMID #16-0119 
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ANDV DNA Vaccines in Healthy Adults 
1 August 2019 
 
 
 
 
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STATEMENT OF COMPLIANCE 
This trial will be carried out in accordance with Good Clinical Practice (GCP) as required by the 
following: 
• United States (US) Code of Federal Regulations (CFR) applicable to clinical studies (45 
CFR Part 46; 21 CFR Part 50, 21 CFR Part 54, 21 CFR Part 56, and 21 CFR Part 312); 
• International Conference on Harmonisation: Good Clinical Practice (ICH E6); 62 Federal 
Register 25691 (1997); and future revisions; 
• The Health Insurance Portability and Accountability Act (HIPAA) Privacy Rule-Final 
Modification (45 CFR Parts 160 and 164); 
• National Institutes of Health (NIH) Clinical Terms of Award, as applicable. 
Compliance with these standards provides public assurance that the rights, safety and well-being 
of study subjects are protected, consistent with the principles that have their origin in the 
Declaration of Helsinki. 
All key personnel (all individuals responsible for the design and conduct of this trial) have 
completed Human Subjects Protection Training. 
 

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SIGNATURE PAGE 
The signature below constitutes the approval of this protocol and the attachments, and provides 
the necessary assurances that this trial will be conducted according to all stipulations of the 
protocol, including all statements regarding confidentiality, and according to local legal and 
regulatory requirements and applicable US federal regulations and ICH guidelines. 
 
Site Investigator: 
 
Signed: 
 
Date: 
 
 
Name 
Title 
 
 
 

DMID #16-0119 
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TABLE OF CONTENTS 
Statement of Compliance .................................................................................................................2 
Signature Page .................................................................................................................................3 
Table of Contents .............................................................................................................................4 
List of Tables ...................................................................................................................................8 
List of Figures ..................................................................................................................................9 
List of Abbreviations .....................................................................................................................10 
Protocol Summary .........................................................................................................................12 
1 
Key Roles ..............................................................................................................................18 
2 
Background Information and Scientific Rationale ...............................................................20 
2.1 
Background Information ...........................................................................................20 
2.1.1 Safety/Toxicology of similar vaccines..........................................................21 
2.1.2 Pre-Clinical Evaluations of ANDV vaccines ................................................22 
2.1.3 Vaccine Delivery System ..............................................................................23 
2.2 
Rationale ...................................................................................................................23 
2.3 
Potential Risks and Benefits .....................................................................................24 
2.3.1 Potential Risks ..............................................................................................24 
2.3.2 Known Potential Benefits .............................................................................26 
3 
Objectives .............................................................................................................................27 
3.1 
Study Objectives .......................................................................................................27 
3.1.1 Primary Study Objective ...............................................................................27 
3.1.2 Secondary Study Objectives .........................................................................27 
3.1.3 Exploratory Study Objectives .......................................................................27 
3.2 
Study Outcome Measures .........................................................................................27 
3.2.1 Primary Outcome Measures ..........................................................................27 
3.2.2 Secondary Outcome Measures ......................................................................28 
3.2.3 Exploratory Outcomes ..................................................................................28 
4 
Study Design .........................................................................................................................29 
5 
study enrollment and withdrawal ..........................................................................................33 
5.1 
Subject Inclusion Criteria .........................................................................................33 
5.2 
Subject Exclusion Criteria ........................................................................................35 
5.3 
Eligibility Criteria for Doses 2-4 ..............................................................................37 
5.4 
Treatment Assignment Procedures ...........................................................................37 
5.4.1 Enrollment and Randomization Procedures..................................................37 
5.4.2 Masking Procedures ......................................................................................38 
5.4.3 Discontinuation of Study Product .................................................................39 

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5.4.4 Study Withdrawal .........................................................................................41 
5.4.5 Handling of Withdrawals ..............................................................................42 
5.4.6 Termination of Study ....................................................................................43 
6 
Study Intervention/Investigational Product ..........................................................................44 
6.1 
Study Product Description ........................................................................................44 
6.1.1 Acquisition ....................................................................................................44 
6.1.2 Formulation, Packaging, and Labeling .........................................................45 
6.1.3 Product Storage and Stability........................................................................45 
6.2 
Dosage, Preparation and Administration of Study Intervention/Investigational 
Product ......................................................................................................................46 
6.3 
Modification of Study Intervention/Investigational Product for a Participant .........47 
6.4 
Accountability Procedures for the Study Intervention/Investigational Product(s) ...48 
6.5 
Assessment of Subject Compliance with Study Intervention/Investigational 
Product ......................................................................................................................49 
6.6 
Concomitant Medications/Treatments ......................................................................49 
7 
Study Schedule......................................................................................................................50 
7.1 
Screening...................................................................................................................50 
7.2 
Enrollment/Baseline ..................................................................................................51 
7.3 
Follow-up, Vaccination Visits ..................................................................................53 
7.4 
Follow-up, Day 8 post each vaccination ...................................................................54 
7.5 
Follow-up, Day 29 post each vaccination .................................................................55 
7.6 
Follow-up Visits........................................................................................................56 
7.7 
Final Study Visit .......................................................................................................56 
7.8 
Early Termination Visit ............................................................................................57 
7.9 
Unscheduled Visit .....................................................................................................57 
8 
Study Procedures/Evaluations ..............................................................................................59 
8.1 
Clinical Evaluations ..................................................................................................59 
8.2 
Laboratory Evaluations .............................................................................................60 
8.2.1 Clinical Laboratory Evaluations ...................................................................60 
8.2.2 Acceptable Laboratory Values for Eligibility and Defining Normal 
Values ...........................................................................................................62 
8.2.3 Special Assays or Procedures .......................................................................63 
8.2.4 Specimen Preparation, Handling, and Shipping ...........................................63 
9 
Assessment of Safety ............................................................................................................65 
9.1 
Specification of Safety Parameters ...........................................................................65 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters .................................................................................................................65 

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9.2.1 Adverse Events .............................................................................................65 
9.2.2 Reactogenicity...............................................................................................67 
9.2.3 Serious Adverse Events ................................................................................71 
9.2.4 Procedures to be Followed in the Event of Abnormal Laboratory Test 
Values or Abnormal Clinical Findings .........................................................72 
9.3 
Reporting Procedures ................................................................................................72 
9.3.1 Serious Adverse Events ................................................................................73 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored 
IND ...............................................................................................................74 
9.3.3 Reporting of Pregnancy ................................................................................74 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events .........................74 
9.5 
Halting Rules ............................................................................................................75 
9.5.1 Sentinel Subject Halting Rules .....................................................................75 
9.5.2 Study Halting Rules ......................................................................................76 
9.6 
Safety Oversight (ISM plus SMC) ............................................................................77 
9.6.1 Independent Safety Monitor (ISM) ...............................................................77 
9.6.2 Safety Monitoring Committee (SMC) ..........................................................78 
10 
Clinical Monitoring ...............................................................................................................80 
10.1 
Site Monitoring Plan .................................................................................................80 
11 
Statistical Considerations ......................................................................................................81 
11.1 
Study Hypotheses......................................................................................................81 
11.2 
Sample Size Considerations ......................................................................................81 
11.3 
Planned Interim Analyses (if applicable) ..................................................................81 
11.3.1 Interim Safety Review ..................................................................................81 
11.3.2 Interim Analysis of Safety and Immunogenicity Data .................................82 
11.4 
Final Analysis Plan ...................................................................................................82 
11.4.1 Analysis Populations .....................................................................................82 
11.4.2 Safety Data ....................................................................................................83 
11.4.3 Immunogenicity Data....................................................................................84 
11.4.4 Missing Values and Outliers .........................................................................84 
12 
Source Documents and Access to Source Data/Documents .................................................85 
13 
Quality Control and Quality Assurance ................................................................................86 
14 
Ethics/Protection of Human Subjects ...................................................................................87 
14.1 
Ethical Standard ........................................................................................................87 
14.2 
Institutional Review Board .......................................................................................87 
14.3 
Informed Consent Process ........................................................................................87 
14.4 
Exclusion of Women, Minorities, and Children (Special Populations) ....................89 

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14.5 
Subject Confidentiality .............................................................................................89 
14.6 
Study Discontinuation ...............................................................................................90 
14.7 
Future Use of Stored Specimens and Data ...............................................................90 
15 
Data Handling and Record Keeping .....................................................................................92 
15.1 
Data Management Responsibilities...........................................................................92 
15.2 
Data Capture Methods ..............................................................................................92 
15.3 
Types of Data ............................................................................................................93 
15.4 
Timing/Reports .........................................................................................................93 
15.5 
Study Records Retention...........................................................................................93 
15.6 
Protocol Deviations ...................................................................................................94 
16 
Publication Policy .................................................................................................................95 
17 
Literature References ............................................................................................................97 
18 
Supplements/Appendices ......................................................................................................99 
Appendix A: Schedule of Events ........................................................................................100 
Appendix B: Acceptable Ranges of Screening Laboratory Measurements ........................103 
 
 
 
 

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LIST OF TABLES 
Table 1: Treatment Arms .............................................................................................................. 31 
Table 2: Blood Volume (mL) – All Treatment Arms ................................................................... 62 
Table 3:  Local (Injection Site) Reactogenicity Grading .............................................................. 67 
Table 4:  Local (Injection Site) Reactogenicity Measurements .................................................... 68 
Table 5: Subjective Systemic Reactogenicity Grading ................................................................. 68 
Table 6: Quantitative Systemic Reactogenicity Grading .............................................................. 69 
Table 7: Blood Pressure and Pulse Grading ................................................................................. 69 
Table 8:  Clinical Screening & Safety Laboratory Adverse Event Grading (Hematology) ......... 70 
Table 9: Clinical Screening & Safety Laboratory Adverse Event Grading (Chemistry) ............. 70 
Table 10: Probability of Observing an Adverse Event for Various Event Rates .......................... 81 
 
 

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LIST OF FIGURES 
Figure 1: Schematic of Study Design ........................................................................................... 17 
 

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LIST OF ABBREVIATIONS 
AE 
Adverse Event/Adverse Experience 
ALT 
Alanine Aminotransferase Test 
ANC 
Absolute Neutrophil Count 
ANDV 
Andes Virus 
BUN 
Blood Urea Nitrogen 
CMS 
Clinical Materials Services 
CFR 
Code of Federal Regulations 
CRF 
Case Report Form 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH,  
eCRF 
Electronic Case Report Form 
EDCSM 
Electronic Data Capture System 
FDA 
Food and Drug Administration 
FWA 
Federalwide Assurance 
GCP 
Good Clinical Practice 
HBsAg 
Hepatitis B Surface Antigen 
HCPS 
Hantavirus Cardio/Pulmonary Syndrome 
HCV 
Hepatitis C Virus 
HFRS 
Hemorrhagic Fever with Renal Syndrome 
Hgb 
Hemoglobin 
HgbA1C 
Hemoglobin A1C 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HLA 
Human Leukocyte Antigen 
HPS 
Hantavirus Cardio/Pulmonary Syndrome 
HTNV 
Hantaan Virus 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
ICS 
Intracellular Staining 
IND 
Investigational New Drug Application 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
LPS 
Lymphoproliferation Assay 

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MedDRA 
Medical Dictionary for Regulatory Activities 
MOP 
Manual of Procedures 
N 
Number (typically refers to subjects) 
NDA 
New Drug Application 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, 
DHHS 
NIH 
National Institutes of Health 
OHRP 
Office for Human Research Protections 
PHI 
Protected Health Information 
PI 
Principal Investigator 
PK 
Pharmacokinetics 
PMED 
Particle Mediated Epidermal Delivery 
PRNT 
Plaque Reduction Neutralization Test 
PsVNA 
Pseudovirion Neutralization Assay 
PUUV 
Puumala Virus 
QA 
Quality Assurance 
QC 
Quality Control 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SDCC 
Statistical and Data Coordinating Center 
SMC 
Safety Monitoring Committee 
SOP 
Standard Operating Procedure 
US 
United States 
USAMRIID 
Unites States Army Medical Research Institute of Infectious 
Diseases 
WBC 
White Blood Cell 

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PROTOCOL SUMMARY 
Title:   
A Phase I, Randomized, Placebo Controlled, Double-blind, Dose Escalation trial 
to Evaluate the Safety and Immunogenicity of an Andes Virus DNA Vaccine for 
the Prevention of Hantavirus Pulmonary Syndrome Using the PharmaJet Stratis® 
Needle-Free Injection System in Normal Healthy Adults 
Phase: 
I 
Population: 
48 males and non-pregnant females, 18-49 years old, inclusive, who are in good 
health and meet all eligibility criteria 
Number of 
Sites: 
One Vaccine and Treatment Evaluation Unit (VTEU) site 
Study 
Duration: 
Approximately 23 months 
Subject 
Participation 
Duration: 
Approximately 12 months 
Description 
of Agent or 
Intervention: 
Andes virus (ANDV) DNA vaccine, constructed from recombinant gene 
segments, plasmid pWRG/AND-M (opt2), was developed by the Department of 
Molecular Virology, US Army Medical Research Institute of Infectious Diseases 
and manufactured by Aldevron (Fargo, ND). ANDV DNA vaccine will be 
administered by the PharmaJet Stratis® Needle-Free Injection System in a 3 or 4 
dose regimen on Days 1, 29, 57 and 169 at two doses, 2 or 4 mg. The placebo will 
be normal saline (0.9% Sodium Chloride, USP). 
Objectives: 
 
Primary: 
• 
Assess the safety and reactogenicity of the ANDV DNA vaccine by dosage 
cohort and treatment arm when administered using the PharmaJet Stratis® 
Needle-Free Injection system in normal, healthy adults. 
Secondary: 

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• 
Assess the immunogenicity of the ANDV DNA vaccine by dosage cohort and 
treatment arm. 
Exploratory: 
• 
Assess cellular immune response to ANDV DNA vaccine by dosage cohort 
and treatment arm. 
• 
Assess immunogenicity of the ANDV DNA vaccine by dosage cohort and 
treatment arm at additional time points. 
Outcome 
Measures: 
Primary: 
• Occurrence of vaccine-related Serious Adverse Events (SAEs) through 
approximately 6 months post last vaccination. 
• Occurrence of vaccine-related unsolicited Adverse Events (AEs) through 28 
days post last vaccination. 
• Occurrence of Serious Adverse Events (SAEs) through approximately 6 
months post last vaccination. 
• Occurrence of unsolicited Adverse Events (AEs) through 28 days post last 
vaccination. 
• Occurrence of solicited local and systemic AEs through 7 days after each 
study vaccination.  
• Occurrence of clinical safety laboratory AEs through 7 days after each study 
vaccination.  
Secondary: 
• Incidence of an ANDV-specific titer of ≥ 20 on Day 57 (3 dose regimen), Day 
85 (4 dose regimen) and Day 197 (3 and 4 dose regimen) as measured by: 
o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
• Incidence of seroconversion (defined as a post-vaccination ANDV-specific 
titer ≥40 if baseline titer <20 or a minimum four-fold rise compared to 
baseline if baseline titer ≥20) on Day 57 (3 dose regimen), Day 85 (4 dose 
regimen) and Day 197 (3 and 4 dose regimen) as measured by: 

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o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
• Geometric mean titer (GMT) of neutralizing antibodies to ANDV on Day 57 
(3 dose regimen), Day 85 (4 dose regimen) and Day 197 (3 and 4 dose 
regimen) as compared to baseline (Day 1) titer and measured by:  
o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
Exploratory: 
• Incidence of a >3 standard deviation increase in ANDV-specific, cytokine-
secreting CD4+ and CD8+ cells on Day 29 (3 and 4 dose regimen), Day 57 (3 
dose regimen), Day 85 (4 dose regimen), Day 169 (3 and 4 dose regimen), and 
Day 197 (3 and 4 dose regimen) compared to baseline (Day 1) as measured by 
flow-based intracellular staining (ICS). 
• Incidence of a >3 standard deviation increase in ANDV-specific CD4+ and 
CD8+ cells on Day 29 (3 and 4 dose regimen), Day 57 (3 dose regimen), Day 
85 (4 dose regimen), Day 169 (3 and 4 dose regimen), and Day 197 (3 and 4 
dose regimen) compared to baseline (Day 1) as measured by a 
lymphoproliferation assay (LPA). 
• Incidence of an ANDV-specific titer of ≥ 20 at additional time points as 
measured by: 
o Pseudovirion neutralization titers 
o Plaque reduction neutralization titers 
Description 
of Study 
Design: 
This is a Phase 1, randomized, placebo controlled, double-blind, dose escalation 
trial of 48 males and non-pregnant females, 18-49 years old, inclusive, who are in 
good health and meet all eligibility criteria. This trial is designed to assess the 
safety, reactogenicity and immunogenicity of an ANDV DNA vaccine for the 
prevention of Hantavirus Pulmonary Syndrome (HPS). ANDV DNA vaccine or 
placebo will be administered using the PharmaJet Stratis® Needle-Free Injection 
System. Subjects assigned to the 3 dose regimen will receive ANDV DNA 
vaccine on Days 1, 29 and 169, and placebo on Day 57. Subjects assigned to the 4 
dose regimen will receive ANDV DNA on Days 1, 29, 57 and 169. Two doses 
(i.e., 2 or 4 mg) of ANDV DNA vaccine will be evaluated.  

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Potential subjects will be screened by history, physical exam, vital signs, and 
clinical laboratory tests. Height and weight will be collected. A urinalysis will be 
done for urine protein and drug screening for opiates. Potential female subjects of 
childbearing potential will have a serum pregnancy test. In addition, potential 
subjects will be screened for Human Immunodeficiency Virus (HIV) type 1 & 2 
antibody, Hepatitis C Virus (HCV) antibody, and Surface Antigen for Hepatitis B 
Virus (HBsAg) prior to enrollment.  
The first 24 participants in Cohorts 1 and 2 will include two sentinel subjects 
since this is a first-in-human Phase 1 study. Sentinel subjects will receive 2 mg of 
study vaccine in an open label manner in a 3 or 4 dose regimen. One sentinel 
subject will be vaccinated, followed for one day for safety and reactogenicity, and 
if no halting rules are met, then the second sentinel subject will receive study 
vaccine in an open-label manner. The two sentinels will be followed for safety 
through Day 8 (laboratory and solicited/unsolicited AEs) and if no pre-defined 
halting rules are met and no safety concerns are identified, enrollment of the 22 
remaining subjects in Cohorts 1 and 2 will proceed, otherwise a Safety 
Monitoring Committee (SMC) meeting will be held to conduct an electronic 
review (E-Review) of clinical and laboratory safety and reactogenicity data. 
While safety data is being evaluated for the sentinel subjects, no new subjects will 
be enrolled, but screening may continue.  
The 22 non-sentinel subjects in Cohorts 1/2 will be randomized in a 9:2:9:2 ratio 
(Treatment Arms 1b:1c:2b:2c) to receive either placebo or study vaccine (at a 
dose of 2 mg) in a 3 or 4 dose regimen, in double-blind fashion. The SMC will 
review all available safety data through 7 days post second vaccination for all 24 
subjects in Cohorts 1 and 2 and provide recommendations to proceed to Cohorts 3 
and 4 or, if necessary, modify the protocol before proceeding. While safety data is 
being evaluated by the SMC, no new subjects will be enrolled, but screening for 
Cohorts 3 and 4 and vaccination of the remaining doses in Cohorts 1 and 2 may 
continue.  
The 24 participants in Cohorts 3 and 4 will follow the same schedule as outlined 
for Cohorts 1 and 2 above. Sentinel subjects will receive 4 mg of study vaccine 
open label in a 3 or 4 dose regimen and be followed for safety through Day 8. The 
remaining 22 non-sentinel subjects in Cohorts 3 and 4 will be randomized in a 

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9:2:9:2 ratio (Treatment Arms 3b:3c:4b:4c) to receive placebo or study vaccine at 
a dose of 4 mg in a 3 or 4 dose regimen, in double blind fashion.   
Doses 2-4 for all treatment arms will be administered per the schedule outlined in 
Section 4,  
Table 1, assuming no halting rules are met. As the vaccine is expected to be well-
tolerated based on previously published studies in humans demonstrating the 
safety of Hantavirus DNA vaccines using the same plasmid backbone1, more than 
one dose will be studied to best investigate safety and immunogenicity. 
Reactogenicity will be measured by the occurrence of solicited injection site and 
systemic AEs. Subjects will maintain a memory aid through 7 days post each 
vaccination, recording temperature, solicited local and systemic symptoms. 
Unsolicited non-serious AEs will be collected from the time of first study 
vaccination through approximately 28 days after last vaccination. SAEs occurring 
from the time of study vaccination through approximately 6 months after the last 
study vaccination will be collected. Clinical safety labs will be collected on Day 1 
and 169, and 7 days after each vaccination; if labs are abnormal, they will be 
repeated at the next scheduled visit (sooner if medically indicated) and followed 
to normal or stabilization. 
Immunogenicity testing will include evaluation of the cellular immune response 
to ANDV peptide pools and assessment of humoral immune response as 
measured by ANDV Pseudovirion Neutralization Assay (PsVNA) and by ANDV 
Plaque Reduction Neutralization Test (PRNT). PsVNA will be measured prior to 
study vaccination (Day 1) and approximately on Days 29, 57, 85, 169, 197, 253, 
and 337. ANDV PRNT will be measured immediately prior to study vaccination 
(Day 1) and approximately on Days 57, 85, 169 and 197. PBMCs to be used in 
the cellular assays, flow-based ICS and LPA, will be obtained at day 1, 29, 57, 85, 
169 and 197. Blood will also be obtained at Days 85, 197 and 337 (PBMCs) for 
potential future exploratory assays. The duration of this trial for each subject will 
be approximately 12 months. 
Estimated 
Time to 
Complete 
Enrollment: 
Approximately 6 months 
 
 

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Figure 1: Schematic of Study Design 
Consent & Assess Eligibility 
Planned Enrollment: Total N=48 Subjects 
Males and non-pregnant females, 18 to 49 years old, inclusive 
Cohorts 1 and 2 sentinel subjects, dose 1 (n=2). 
Continue recruitment and screening 
Randomization and Vaccination of remaining subjects (Cohorts 
1and 2, 2mg Dose) 
Administer ANDV vaccine or placebo per Subject 
Assignment on Day 1 
If no halting rules are met through 7 days post second 
vaccination, proceed with Cohort 3 and 4 
Assessment of   Final 
Outcome Measures 
Randomized 9:2:9:2  
1b. Vaccine 3-dose regimen: 1c..Placebo: 2b. Vaccine 4-dose regimen: 2c placebo 
Randomization and Vaccination of remaining subjects (Cohorts 3 
and 4, 4mg Dose) 
Cohort 3 and4 sentinel subjects, dose 1 (n=2). 
Continue recruitment and screening 
Administer ANDV vaccine or placebo 
per Subject Assignment on Day 1 
Follow-up for Safety, Reactogenicity, and 
Immunogenicity 
Randomized 9:2:9:2 
3b. Vaccine 3-dose regimen: 3c. Placebo: 4b. Vaccine 4-dose regimen: 4c. placebo 

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1 
KEY ROLES 
Individuals: 
Protocol Champion:   
Gabriele Feolo, RN,MSN 
Virology Branch 
NIAID, NIH  
 
Principal Investigator:   
Grant Paulsen, MD 
Cincinnati Children’s Hospital Medical Center 
 
Medical Officer:  
Kay M. Tomashek, MD, MPH, DTM 
Division of Microbiology and Infectious Diseases 
NIAID, NIH 
 
Medical Monitor:  
Jorge Mejia-Galvis, MD,  
Office of Clinical Research Affairs 
Division of Microbiology and Infectious Diseases 
National Institute of Allergy and Infectious Diseases 
National Institutes of Health 
Institutions: 
Cincinnati Children’s Hospital Medical Center 
Local (Clinical) 
Laboratory: 
Cincinnati Children’s Hospital Medical Center 
CORE Laboratories  
Cellular Assays 
Laboratory: 
The Singh Laboratory 
Karnail Singh, PhD 
ANDV Neutralization 
Assays Laboratory: 
USAMRIID  
Jay Hooper, PhD 
Study Agent Repository: 
Fisher BioServices 
c/o DMID Clinical Materials Services Contract 

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Statistical and Data 
Coordinating Center 
The Emmes Corporation 
Safety and 
Pharmacovigilance 
Contractor: 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support 
(CROMS) 
6500 Rock Spring Dr., Suite 650 
Bethesda, MD 20814 
SAE Hot Line: 1-800-537-9979 (US) 
SAE Fax: 800-275-7619 (US) 
SAE Email: PVG@dmidcroms.com 
 

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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
Hantaviruses are members of the Bunyaviridae family and are found worldwide. Hantaviruses 
are negative-sense, single-stranded RNA viruses with three segments:  small (S), medium (M), 
and large (L). The M-segment encodes the envelope glycoproteins (Gn and Gc). Over 50 species 
of hantaviruses have been identified, causing a wide spectrum of disease. Some infections result 
in a mild disease, while others can be severe, ultimately leading to death. Pathogenic 
hantaviruses primarily infect vascular endothelial cells in humans causing dysfunction and 
increased vascular permeability2. The two primary disease presentations in humans are due to 
vascular leakage; hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardio/ 
pulmonary syndrome (HCPS or HPS). HPS was first described in 1993 in the southwestern 
United States, with subsequent identification of Sin Nombre virus (SNV) as the causative agent. 
HPS typically presents as a non-specific viral illness with cough, fever, chills, nausea/vomiting, 
diarrhea, myalgia and/or headache for 3-6 days, followed by abrupt onset of pulmonary edema 
and shock3. HFRS is more common (>20,000 cases/year), and is generally limited to Asia, 
Europe, Scandinavia, and Russia. HFRS presents with similar prodromal symptoms, and less 
commonly leads to hypotension, shock and acute renal failure4,5. 
 
In South America, the Andes virus (ANDV) is the predominant cause of HPS with the majority 
of cases occurring in Brazil, Chile and Argentina; there have been a few case reports from 
Bolivia, Paraguay and Uruguay.6-8 HPS due to ANDV is primarily a zoonotic disease in humans, 
spread via rodent excreta. While rodents, shrews, moles and bats can all be reservoir hosts for 
hantaviruses, the long-tailed rat (Oligoryzomys longicaudatus) is the host of ANDV and the deer 
mouse (Peromyscus maniculatus) remains the reservoir of the SNV2. Both are relatively 
common rodents and can be trapped in the wild. Unlike the other known hantaviruses, ANDV 
remains the only one with persuasive evidence of person-to-person transmission7,9. 
 
While HPS is relatively uncommon (approximately 600 US cases & 4,000 South American from 
1993-2012/13), it remains a serious infection, with mortality rates due to ANDV infection of 35-
40%, despite modern critical care support3,10,11. Additionally, there is no vaccine or effective 
therapy for HPS and treatment remains supportive and symptomatic. In the United States, due to 
the relative infrequency of hantavirus in general and HPS in particular, seroprevalence of IgG 
antibodies to hantaviruses are uncommon. Among individuals tested in a Baltimore, MD area 
study12,13 only 0.7 – 2.1% of the group had detectable serum antibodies to Hantaan, Seoul or 

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Puumala viruses, while no one had detectable antibodies against ANDV or SNV. Therefore, 
protective immunity against HPS is likely minimal in the North American population, and there 
is no evidence of cross protection between SNV and ANDV.14 
 
Due to issues associated with both production and formulation of an inactivated HRFS vaccine, 
DNA vaccines against the virus have been developed.4 A phase 1 clinical trial of Hantaan Virus 
(HTNV) and Puumala Virus (PUUV) DNA vaccines reported neutralizing antibodies of 30% and 
40% to HTNV and PUUV, respectively.4 In the combined vaccine group, 56% developed 
neutralizing antibodies to one or both viruses.4 When the same vaccine was administered using a 
different delivery system, and administering 2 doses of vaccine, seroconversion was increased to 
64% and 75% to HTNV and PUUV, respectively.1 Currently, there is an ongoing phase 2a trial 
comparing two different dosing schedules of a mixed HTNV/PUUV DNA vaccine administered 
via IM-EP (clinicaltrials.gov, NCT02116205) as well as a phase 1 trial evaluating HTNV and 
PUUV DNA vaccines delivered by the PharmaJet Stratis® Needle-Free Injection System 
(clinicaltrials.gov, NCT02776761).   
 
In order to develop a pan-hantavirus DNA vaccine, an ANDV vaccine has been developed using 
the same strategy and plasmid backbone as the HTNV and PUUV DNA vaccines. The 
development of the HTNV DNA, PUUV DNA, and the ANDV DNA vaccines from the same 
plasmid backbone, combined with the immunogenicity results of the previous hantavirus DNA 
vaccines, provides the rationale for evaluating the ANDV DNA vaccine administered by the 
PharmaJet Stratis® Needle-Free Injection System in humans. The plasmid, pWRG/AND-M 
(opt2), which is the active ingredient for the ANDV DNA vaccine, was constructed on a well-
characterized plasmid backbone, pWRG7077, and synthesized elements of the ANDV DNA 
genome. The optimized ANDV M gene encodes viral GnGc envelope glycoproteins that have 
been shown to be the targets of neutralizing antibodies.14 The DNA vaccine was developed by 
the Department of Molecular Virology, US Army Medical Research Institute of Infectious 
Diseases (USAMRIID; Fort Detrick, Maryland), and the final vaccine was manufactured by 
Aldevron (Fargo, ND). The proposed trial would be a first in man vaccine trial for the prevention 
of HPS. 
2.1.1 Safety/Toxicology of similar vaccines 
The plasmid vector backbone, pWRG7077, has been used in several DNA vaccines, including 
HTNV and PUUV DNA vaccines and a Venezuelan Equine Encephalitis (VEE) DNA vaccine.  
In the VEE study, 41 subjects received one or more immunizations and there were no serious or 
unanticipated safety concerns identified.15 There were 674 AEs recorded for all subjects over the 

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course of the study, the majority were local with resolution within 24 hours. Systemic AEs were 
also generally mild and transient; the only Grade 3 possibly vaccine related AE of severe fatigue 
resolved within 1 day. This study did report 3 SAEs, all of which were GI-related, occurred in 
the same patient with a recurrence of known ulcerative colitis and all concluded to be unrelated 
to the vaccine or study-related procedure.15 
 
Investigators have previously evaluated plasmid DNA vaccine strategies for hantaviruses in both 
pre-clinical studies and human trials since 1999.16 As referenced above, two prior hantavirus 
DNA vaccine trials have been published in humans. The first involved 3 cohorts of volunteers 
vaccinated with a DNA vaccine expressing M segments of HTNV and/or PUUV delivered via 
disposable gene gun devices (a.k.a. particle mediated epidermal delivery, PMED). Among the 28 
subjects to receive at least one dose of vaccine, there were no SAEs related to the vaccine or 
study-related procedures and no subject withdrew due to adverse events.4 Injection site pain was 
noted following 48-56% of doses and injection site tenderness following 79-89% of doses. All 
injection site pain was mild and moderate tenderness was documented only following the 3rd 
dose in 7% of subjects. The most common systemic AEs were fatigue, malaise, myalgia and, 
headache. Less common was oropharyngeal pain noted in 3 subjects. With the exception of one 
severe headache, all events were graded as mild or moderate in severity. 
   
A subsequent human trial in 31 participants, receiving at least one immunization with HTNV 
DNA, PUUV DNA or mixture of both via intramuscular electroporation (IM-EP) was recently 
published.1 No SAEs related to the vaccine or study-related procedures were observed. The most 
common solicited local AE was pain at the injection site which occurred in 28/31 participants. 
The next most common AEs were headache and myalgia, and all AEs were mild or moderate in 
severity. A single participant was found to have unrelated Grade 4 hypoglycemia, felt to be the 
result of an unrecognized pre-existing condition.1 
2.1.2 Pre-Clinical Evaluations of ANDV vaccines 
Several pre-clinical studies using variations of the ANDV vaccine and different delivery systems 
have been published. Studies of the same plasmid product as the current proposal (pWRG/AND-
M (opt2)), demonstrated that a combined ANDV DNA and SNV DNA vaccine elicited 
neutralizing antibodies in rabbits and nonhuman primates17. In this study, significantly higher 
neutralizing titers were seen using a disposable syringe jet injection (DSJI) system as compared 
to needle/syringe delivery; anti-ANDV PRNT50 geometric mean titers (GMTs) were 349 versus 
42 (p = 0.01). No safety issues or adverse events were reported.  
 

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Subsequent studies in geese and transchromosomal bovine also have demonstrated that the 
ANDV DNA vaccines administered with a DSJI system elicit neutralizing antibodies. 
Administration of these purified antibodies protected hamsters from lethal ANDV infection18,19. 
As in the nonhuman primate study above, there were no safety concerns or adverse events 
reported with respect to the vaccine product or delivery method. 
2.1.3 Vaccine Delivery System 
Historically, due to poor uptake of the DNA following injection, DNA vaccination has failed to 
raise a strong immune response in humans.20 This has led to a progression of delivery systems to 
improved immunogenicity. The gene gun uses gold-coated DNA delivered with helium pressure 
and has shown promise. Electroporation (EP) was subsequently developed and it applies a 
current over cell membranes to increase cellular uptake. EP has been shown to increase uptake 
and immune response in humans, but can require a somewhat more complex apparatus21. DSJI 
systems have also been used successfully to administer DNA vaccines with good tolerability and 
effect (Hooper et al., unpublished data).22   
 
Needle-free jet injection has a well-documented record of successful use for the delivery of 
conventional vaccines over the past 60 years, with estimates of hundreds of millions of doses 
delivered, including ANDV DNA vaccine delivery in nonhuman primates, as noted above17. The 
PharmaJet Stratis® Needle-Free Injection System is a US Food and Drug Administration (FDA) 
510k-cleared for administering FDA- approved medicines and is the proposed delivery system in 
this trial.17  This delivery system avoids both the use of gold-coated DNA and the complexity of 
administering a simultaneous electrical current with the immunization. 
2.2 
Rationale 
ANDV is a Category A Bioterrorism Agent with high mortality and no effective prevention or 
specific treatment. An effective vaccine could protect against naturally occurring HPS as well as 
protecting soldiers from pathogens endemic to deployment areas as well as protection from a 
potential biologic weapon. This has spurred investigation into vaccine development. 
 
Currently there are no FDA-licensed vaccines for HFRS or HPS, although there are inactivated 
cell culture-derived and rodent-brain derived vaccines available in other countries.4,5 Due to 
issues associated with both production and formulation of an inactivated combination HFRS 
vaccine, hantavirus DNA vaccines have been developed. These include HTNV and PUUV, two 
of the predominant causes of HFRS. DNA vaccines were described in the early 1990s and have 

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since shown that by injecting a plasmid, expressing a viral antigen under the control of a 
promoter, an immune response to the viral antigen could be detected.23 The current hantavirus 
DNA vaccine candidates utilize recombinant glycoprotein gene segments that are then subcloned 
into a plasmid backbone (pWRG7077). Given that the HTNV DNA, PUUV DNA, and ANDV 
DNA vaccines are all designed on the same plasmid backbone, the results of these ongoing 
clinical trials coupled with the immunogenicity of these DNA vaccines in humans and animals, 
provide the rationale for evaluating the ANDV DNA vaccine administered by the PharmaJet 
Stratis® Needle-Free Injection System in humans. 
2.3 
Potential Risks and Benefits 
2.3.1 Potential Risks 
The potential risks of this trial are those associated with having blood drawn, intramuscular 
injections (IM), and possible reactions to the experimental ANDV vaccine. There may be 
potential risk related to breach of confidentiality, as well as other unknown risks, discomforts, or 
side effects. 
 
The ANDV vaccine to be used in this clinical trial has never been administered to humans. It is 
administered intramuscularly and it is possible that vaccination may cause pain, tenderness, 
erythema (redness), induration (hardness/swelling), bruising and skin discoloration at the 
injection site. The vaccine may also cause systemic reactions such as fever, feverishness (chills, 
shivering, sweating), fatigue (tiredness), malaise (general unwell feeling), myalgia (body 
aches/muscular pain), nausea, dizziness, and headache. A less likely symptom is tachypnea 
(rapid breathing). In addition, vaccines can cause a severe allergic reaction. Such reactions are 
rare and would happen within a few minutes to a few hours after vaccination. Signs of a severe 
allergic reaction can include rash or hives, swelling of the face and throat, difficulty breathing, a 
fast heartbeat, dizziness, and weakness.   
 
Based on the results of preclinical and clinical studies performed using DNA vaccines with the 
same plasmid backbone (pWRG7077), the likelihood of DNA integration or production of anti- 
DNA antibodies for the ANDV DNA vaccine is minimal.24,25 These possibilities cannot be 
completely ruled out, however, and should be considered. In addition, long-term effects of 
repeated dosing with DNA vaccines have not been studied. The risk of a more virulent reaction 
or greater tendency toward a similar disease may be possible in immunized individuals. 
  
It is unknown if the ANDV vaccine poses any risks to an unborn child. Women of childbearing 
potential must agree to practice effective contraception for a minimum of 30 days prior to study 

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product administration and agree to practice effective contraception until 90 days after the last 
study vaccination. If a female subject becomes pregnant while participating in this trial, we will 
ask permission to follow-up about her health through pregnancy outcome and the health of her 
baby. Men who are sexually active with a woman of childbearing potential and have not had a 
vasectomy performed more than 1 year prior to screening must agree not to father a child for 90 
days after receipt of the last study vaccination. Female subjects must agree to not donate eggs (ova, 
oocytes) and male subject agrees not to donate sperm from the start of screening period until at 
least 90 days after receiving the last vaccination.  
Drawing blood may cause transient discomfort and fainting. Fainting is usually transient and 
managed by having the subject lie down and elevate his/her legs. Bruising at the blood draw site 
may occur, but can be prevented or lessened by applying pressure to the blood draw site for a 
few minutes after the blood is taken. Drawing blood may also cause infection. The use of aseptic 
(sterile) technique will make infection at the site where blood will be drawn extremely unlikely. 
 
An IM injection may cause transient discomfort and fainting. Giving an IM injection may also 
predispose a subject to infection. However, the use of sterile technique will make an infection at 
the injection site extremely unlikely.  The Pharmajet needle free delivery of vaccines has been 
associated with more local reactogenicity (e.g. pain, tenderness itching, redness or swelling) that 
was mild or moderate in nature than was observed with needle and syringe injections of 
vaccines.  There were no differences in systemic adverse events 
  
Subjects will be asked to provide personal health information (PHI). All attempts will be made to 
keep this PHI confidential within the limits of the law. However, there is a chance that 
unauthorized persons will see the subjects’ PHI. All study records will be kept in a locked file 
cabinet or maintained in a locked room at the VTEU site. Electronic files will be password-
protected. Only people who are involved in the conduct, oversight, monitoring, or auditing of 
this trial will be allowed access to the PHI that is collected. Any publications from this trial will 
not use information that will identify subjects by name. Organizations that may inspect and/or 
copy research records maintained at the site for quality assurance and data analysis include 
groups such as the local Institutional Review Board (IRB), NIAID and their designees, 
USAMRIID and FDA. 
 
There may be other risks, discomforts, or side effects that are unknown at this time.  

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2.3.2 Known Potential Benefits 
There are no known benefits attributable to the receipt of the ANDV DNA vaccine. It is possible 
that the study will result in the availability of a vaccine against HPS due to ANDV and/or that 
vaccination will result in protection from HPS caused by ANDV. The duration of any such 
protection is currently unknown. 

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3 
OBJECTIVES 
3.1 
Study Objectives 
3.1.1 Primary Study Objective 
• 
Assess the safety and reactogenicity of the ANDV DNA vaccine by dosage cohort and 
treatment arm when administered using the PharmaJet Stratis® Needle-Free Injection system 
in normal, healthy adults. 
3.1.2 Secondary Study Objectives 
• 
Assess the immunogenicity of the ANDV DNA vaccine by dosage cohort and treatment arm. 
3.1.3 Exploratory Study Objectives 
• 
Assess cellular immune response to ANDV DNA vaccine by dosage cohort and treatment 
arm. 
• 
Assess immunogenicity of the ANDV DNA vaccine by dosage cohort and treatment arm at 
additional time points. 
3.2 
Study Outcome Measures 
3.2.1 Primary Outcome Measures 
• 
Occurrence of vaccine-related Serious Adverse Events (SAEs) through approximately 6 
months post last vaccination. 
• Occurrence of vaccine-related unsolicited Adverse Events (AEs) through 28 days post last 
vaccination.  
• Occurrence of Serious Adverse Events (SAEs) through approximately 6 months post last 
vaccination. 
• Occurrence of unsolicited Adverse Events (AEs) through 28 days post last vaccination. 
• 
Occurrence of solicited local and systemic AEs through 7 days after each study vaccination.  
• 
Occurrence of clinical safety laboratory AEs through 7 days after each study vaccination.  

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3.2.2 Secondary Outcome Measures 
• 
Incidence of an ANDV-specific titer of ≥ 20 on Day 57 (3 dose regimen), Day 85 (4 dose 
regimen) and Day 197 (3 and 4 dose regimen) as measured by: 
o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
• 
Incidence of seroconversion (defined as a post-vaccination ANDV-specific titer ≥40 if 
baseline titer <20 or a minimum four-fold rise compared to baseline if baseline titer ≥20) on 
Day 57 (3 dose regimen), Day 85 (4 dose regimen) and Day 197 (3 and 4 dose regimen) as 
measured by: 
o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
• 
Geometric mean titer (GMT) of neutralizing antibodies to ANDV on Day 57 (3 dose 
regimen), Day 85 (4 dose regimen) and Day 197 (3 and 4 dose regimen) as compared to 
baseline (Day 1) titer and measured by:  
o Plaque reduction neutralization titers 
o Pseudovirion neutralization titers 
3.2.3 Exploratory Outcomes 
• 
Incidence of a >3 standard deviation increase in ANDV-specific, cytokine-secreting CD4+ 
and CD8+ cells on Day 29 (3 and 4 dose regimen), Day 57 (3 dose regimen), Day 85 (4 dose 
regimen), Day 169 (3 and 4 dose regimen) and Day 197 (3 and 4 dose regimen) compared to 
baseline (Day 1) as measured by flow-based ICS. 
• 
Incidence of a >3 standard deviation increase in ANDV-specific CD4+ and CD8+ cells on 
Day 29 (3 and 4 dose regimen), Day 57 (3 dose regimen), Day 85 (4 dose regimen), Day 169 
(3 and 4 dose regimen) and Day 197 (3 and 4 dose regimen) compared to baseline (Day 1) as 
measured by a LPA. 
• 
Incidence of an ANDV-specific titer of ≥20 at additional time points as measured by: 
o Pseudovirion neutralization titers 
o Plaque reduction neutralization titers 

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4 
STUDY DESIGN 
This is a Phase 1, randomized, placebo controlled, double-blind, dose escalation trial of 48 males 
and non-pregnant females, 18-49 years old, inclusive, who are in good health and meet all 
eligibility criteria. This trial is designed to assess the safety, reactogenicity and immunogenicity 
of an ANDV DNA vaccine for the prevention of Hantavirus Pulmonary Syndrome (HPS). 
ANDV DNA vaccine or placebo will be administered using the PharmaJet Stratis® Needle-Free 
Injection System. Subjects assigned to the 3 dose regimen will receive ANDV DNA vaccine on 
Days 1, 29 and 169, and placebo on Day 57. Subjects assigned to the 4 dose regimen will receive 
ANDV DNA on Days 1, 29, 57 and 169. Two doses (i.e., 2 or 4 mg) of ANDV DNA vaccine 
will be evaluated.  
Potential subjects will be screened by history, physical exam, vital signs, and clinical laboratory 
tests including white blood cells (WBC), hemoglobin (Hgb), platelet count, absolute neutrophil 
count (ANC), Hemoglobin A1C (HgbA1C), total bilirubin, alanine aminotransferase (ALT), 
sodium, potassium, and creatinine. Blood urea nitrogen (BUN) will be obtained only if creatinine 
is above the normal range. Height and weight will be obtained. A urinalysis will be done for 
urine protein and drug screening for opiates. Potential female subjects of childbearing potential 
will have a serum pregnancy test. In addition, potential subjects will be screened for HIV – 1/2 
antibody, HCV antibody, and HBsAg prior to enrollment.  
The first 24 participants in Cohorts 1 and 2 will include one sentinel subjects in each cohort since 
this is a first-in-human Phase 1 study. Sentinel subjects will receive 2 mg of study vaccine in an 
open label manner but will be randomized and blinded to the dose regimen. Subjects randomized 
to the 3 dose regimen (Cohort 1) will receive ANDV DNA vaccine on Days 1, 29 and 169, and 
placebo on Day 57. Subjects randomized  to the 4 dose regimen (Cohort 2) will receive ANDV 
DNA on Days 1, 29, 57 and 169. One sentinel subject will be vaccinated, followed for one day 
for safety and reactogenicity, and if no halting rules are met, then the second sentinel subject will 
receive study vaccine in an open-label manner. The two sentinels will be followed for safety 
through Day 8 (laboratory and solicited/unsolicited AEs) and if no pre-defined halting rules are 
met and no safety concerns are identified, enrollment of the 22 remaining subjects in Cohort 1 
and 2 will proceed, otherwise a Safety Monitoring Committee (SMC) meeting will be held to 
conduct an electronic review (E-Review) of clinical and laboratory safety and reactogenicity 
data. While safety data is being evaluated for the sentinel subjects, no new subjects will be 
enrolled, but screening may continue.  
The 22 non-sentinel subjects in Cohorts 1 and 2 will be randomized in a 9:2:9:2 ratio (Treatment 
Arms 1b:1c:2b:2c) to receive either study vaccine (at a dose of 2 mg) or placebo in a 3 or 4 dose 

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regimen, in double-blind fashion. The SMC will review all available safety data through 7 days 
post second vaccination for all 24 subjects in Cohort 1 and 2 and provide recommendations to 
proceed to Cohort 3 and 4 or, if necessary, modify the protocol before proceeding. While safety 
data is being evaluated by the SMC, no new subjects will be enrolled, but screening for Cohorts 
3 and 4 and vaccination of the remaining doses in Cohorts 1 and 2 may continue.  
The 24 participants in Cohorts 3 and 4 will follow the same schedule as outlined for Cohorts 
1and 2 above. Sentinel subjects will be randomized to a dose regimen and will receive 4 mg of 
study vaccine open label and be followed for safety through Day 8. Subjects assigned to the 3 
dose regimen will receive ANDV DNA vaccine on Days 1, 29 and 169, and placebo on Day 57. 
Subjects assigned to the 4 dose regimen will receive ANDV DNA on Days 1, 29, 57 and 169. 
The remaining 22 non-sentinel subjects in Cohorts 3 and 4 will be randomized in a 9:2:9:2 ratio 
(Treatment Arms 3b:3c:4b:4c) to receive study vaccine or placebo at a dose of 4 mg in a 3 or 4 
dose regimen, in double blind fashion.   
Doses 2-4 for all treatment arms will be administered per the schedule outlined in the table 
below, assuming no halting rules are met. 
 
 

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Table 1: Treatment Arms 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Cohort 
Subjects 
Enrolled 
Randomization/ 
Treatment 
Arm*# 
Day 1 
Day 29 
Day 57 
Day 169 
Cohort 1 
2mg^ 
12 
1a*  
n=1  
Sentinel 
ANDV 
DNA 
ANDV 
DNA 
Placebo 
ANDV 
DNA 
1b# 
N=9 
ANDV DNA 
ANDV 
DNA 
ANDV 
DNA 
Placebo 
ANDV 
DNA 
1c# 
n=2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Cohort 2 
2mg^ 
12 
2a*  
n=1  
Sentinel 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
2b# 
N=9 
ANDV DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
2c# 
n=2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Cohort 3 
4mg+ 
 
12 
3a*  
n=1  
Sentinel 
ANDV 
DNA 
ANDV 
DNA 
Placebo 
ANDV 
DNA 
3b# 
N=9 
ANDV DNA 
ANDV 
DNA 
ANDV 
DNA 
Placebo 
ANDV 
DNA 
3c# 
n=2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Cohort 4 
4mg+ 
 
12 
4a*  
n=1  
Sentinel 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
4b# 
N=9 
ANDV DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
ANDV 
DNA 
4c# 
n=2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
TOTAL 
48 subjects 
* All sentinel subjects and study personnel will be unblinded to dose and blinded to treatment 
schedule. 
# All non-sentinel subjects will be blinded to dose and treatment schedule. 
^  1mg ANDV DNA administered into the left and right deltoid 
+  2mg ANDV DNA administered into the right and left deltoid. 

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As the vaccine is expected to be well tolerated based on previously published studies in humans 
demonstrating the safety of hantavirus DNA vaccines using the same plasmid backbone, more 
than one dose will be studied to best investigate safety and immunogenicity. 
On vaccination days, all females of childbearing potential will have a urine pregnancy test done. 
All subjects will have vital signs and a targeted physical examination pre-vaccination. Blood will 
be collected pre-vaccination for immunogenicity testing. On the first and fourth vaccination 
days, clinical safety laboratory tests will be collected pre-vaccination. Vaccine and/or placebo 
will then be administered by intramuscular injection using the PharmaJet Stratis® Needle-Free 
Injection System, into both the left and right deltoid muscle one after another or simultaneously. 
In this way, subjects will be blinded to cohort (i.e., dosage group) and treatment arm within a 
cohort.   
Subjects will be observed for 30 minutes after the last injection is given. Reactogenicity will be 
measured by the occurrence of solicited injection site and systemic AEs. Subjects will maintain a 
memory aid through 7 days post each vaccination recording temperature, solicited local and 
systemic symptoms. Unsolicited non-serious AEs will be collected from the time of first study 
vaccination through approximately 28 days after the last vaccination. SAEs occurring from the 
time of study vaccination through approximately 6 months after the last study vaccination will be 
collected. Clinical safety labs will be collected on Days 1 and 169, and 7 days after each 
vaccination; if labs are outside the clinical laboratory reference range, they will be repeated at the 
next scheduled visit (sooner if medically indicated) and, followed to normal or stabilization. 
Immunogenicity testing will include evaluation of the cellular immune response to ANDV 
neutralizing antibodies peptide pool and assessment of humoral immune response as measured 
by ANDV Pseudovirion Neutralization Assay (PsVNA) and by ANDV Plaque Reduction 
Neutralization Test (PRNT). PsVNA will be measured prior to study vaccination (Day 1) and 
approximately on Days 29, 57, 85, 169, 197, 253, and 337. PRNT will be measured immediately 
prior to study vaccination (Day 1) and approximately on Days 57, 85, 169 and 197. PBMCs to be 
used in the cellular assays (ICS+ LPA) will be obtained at day 1, 29, 57, 85, 169 and 197. Blood 
will also be obtained at Days 85, 197 and 337 (PBMCs) for potential future exploratory assays. 
The duration of this trial for each subject will be approximately 12 months. 
For additional details on study procedures and evaluations by study visits/days, see Appendix A: 
Schedule of Events . 

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5 
STUDY ENROLLMENT AND WITHDRAWAL 
Forty-eight males and non-pregnant females, 18-49 years old, inclusive, who are in good health 
and meet all eligibility criteria, will be enrolled from a single VTEU site. The target population 
should reflect the community at large at the VTEU site. Estimated time to complete enrollment 
in this trial is approximately 6 months. Information regarding this trial may be provided to 
potential subjects who have previously participated in vaccine trials conducted at the VTEU site. 
Recruitment materials may be used however, the Institutional Review Board (IRB) will approve 
all materials prior to use. 
Subject Inclusion and Exclusion Criteria must be assessed by a study clinician licensed to make 
medical diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
No exemptions are granted on Subject Inclusion or Exclusion Criteria in DMID-sponsored 
studies. Questions about eligibility should be directed toward the DMID Medical Officer. 
5.1 
Subject Inclusion Criteria 
Subjects eligible to participate in this study must meet all of the following inclusion criteria: 
1. Provide written informed consent before initiation of any study procedures. 
2. Are able to understand and comply with planned study procedures and be available for all 
study visits/phone calls. 
3. Males or non-pregnant females ages 18-49, inclusive. 
4. Are in good health * 
* As determined by medical history and physical examination to evaluate acute or currently ongoing 
chronic medical diagnoses or conditions, defined as those that have been present for at least 90 
days which would affect the assessment of the safety of subjects or the immunogenicity of study 
vaccinations. Chronic medical diagnoses or conditions should be stable for the last 60 days (no 
hospitalizations, ER or urgent care for condition and no adverse symptoms that need medical 
intervention such as medication change/supplemental oxygen). This includes no change in chronic 
prescription medication, dose, or frequency as a result of deterioration of the chronic medical 
diagnosis or condition in the 60 days prior to enrollment. Any prescription change that is due to 
change of health care provider, insurance company, etc., or that is done for financial reasons, as 
long as in the same class of medication, will not be considered a deviation of this inclusion 
criterion. Any change in prescription medication due to improvement of a disease outcome, as 

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determined by the site principal investigator or appropriate sub-investigator, will not be considered 
a deviation of this inclusion criterion. Subjects may be on chronic or as needed (prn) medications 
if, in the opinion of the site principal investigator or appropriate sub-investigator, they pose no 
additional risk to subject safety or assessment of reactogenicity and immunogenicity and do not 
indicate a worsening of medical diagnosis or condition. Similarly, medication changes subsequent 
to enrollment and study vaccination are acceptable provided there was no deterioration in the 
subject’s chronic medical condition that necessitated a medication change, and there is no 
additional risk to the subject or interference with the evaluation of responses to study vaccination. 
Note: Topical, nasal, and inhaled medications (apart from steroids as outlined in the Subject 
Exclusion Criteria), herbals, vitamins, and supplements are permitted. 
5. Oral temperature is less than 100.0 °F (37.8oC). 
6. Pulse is 47 to 105 beats per minute (bpm), inclusive. 
7. Systolic blood pressure (BP) is 85 to 150mm Hg, inclusive. 
8. Diastolic blood pressure (BP) is 55 to 95 mm Hg, inclusive. 
9. Have acceptable screening laboratories*,** within 28 days prior to enrollment. 
* Refer to Appendix B for range of acceptable laboratory values. 
*  Screening laboratory values that are outside acceptable range but are thought to be due to an acute 
condition or due to laboratory error may be repeated once. [see Manual of Procedures (MOP)] 
10. Urine protein screen is negative or trace. 
11. Drug screen for opiates is negative. 
12. HgbA1C <6.3% at screening. 
13. HIV – 1/2 antibody negative. 
14. HCV antibody negative. 
15. HBsAg negative. 
16. Women of childbearing potential*, must be using an effective method of contraception** from 
30 days prior to the first study vaccination until 90 days after the last study vaccination. 
*  Women of childbearing potential are defined as those who have not been sterilized via tubal 
ligation, bilateral oophorectomy, hysterectomy, or successful Essure® placement (permanent, non-
surgical, non-hormonal sterilization) with history of documented radiological confirmation test at 
least 90 days after the procedure (or with use of another birth control method if history of 
confirmation test not confirmed), AND are still menstruating or < 1 year since the last menses if 
perimenoapausal.  
** For this study, we define an effective contraceptive method as one that results in a failure rate of 
less than 1% per year when it is used consistently and correctly. This includes, but is not limited to, 

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non-male sexual relationships, abstinence from sexual intercourse with a male partner, monogamous 
relationship with a vasectomized partner, male condoms with the use of applied spermicide, 
intrauterine devices, NuvaRing®, and licensed hormonal methods such as implants, injectables or 
oral contraceptives (“the pill”).   
17. Women of childbearing potential* must have a negative serum pregnancy test at screening 
and a negative urine pregnancy test within 24 hours prior to each study vaccination. 
* see definition of women of childbearing potential above. 
18. Sexually active male participants whose partner is a woman of childbearing potential* and 
has not had a vasectomy**  must agree not to father a child until 90 days after the last 
vaccination***.  
* see definition of women of childbearing potential above. 
** performed > 1 year prior to screening 
***must agree to use a barrier method of birth control e.g., either condom with spermicidal 
foam/gel/film/cream or partner reports usage of occlusive cap (diaphragm or cervical/vault caps) 
with spermicidal foam/gel/film/cream/suppository.   
19. Women agree to not donate eggs (ova, oocytes) and male subject agrees not to donate sperm 
from the start of screening onwards until at least 90 days after the last vaccination. 
20. Agree not to participate in another clinical trial during the study period. 
21. Agree not to donate blood to a blood bank for 3 months after receiving the last study vaccine. 
5.2 
Subject Exclusion Criteria 
Subjects eligible to participate in this study must not meet any of the following exclusion 
criteria: 
1. Women who are pregnant, planning to become pregnant or lactating*. 
* Includes breastfeeding or planning to breastfeed at any given time from the receipt of study 
vaccination through the 12-month trial period. 
2. Known allergy or history of anaphylaxis, severe local or other serious adverse reactions to 
vaccines or vaccine products*, or history of severe allergic reactions. 
* This includes a known allergy to an aminoglycoside (e.g., gentamicin, tobramycin, neomycin, 
streptomycin).  
3. Received an experimental agent* within 3 months prior to study vaccination, or expects to 
receive an experimental agent** during the 12-month trial-reporting period. 
* Including vaccine, drug, biologic, device, blood product, or medication. 
** Other than from participation in this study. 

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4. Received any licensed live vaccine within 28 days prior to or after each study vaccination. 
5. Received a licensed inactivated vaccine within 14 days prior to or after each study 
vaccination.*  
* Allowable exception for inactivated seasonal influenza vaccine received more than 7 days prior to 
or after a study vaccination. 
6. Individuals in whom the ability to observe possible local reactions at the eligible injection 
sites (deltoid region) is, unacceptably obscured due to a physical condition or permanent 
body art. 
7. Have an acute illness*, as determined by the site PI or appropriate sub-investigator, within 72 
hours prior to study vaccination. 
* An acute illness which is nearly resolved with only minor residual symptoms remaining is 
allowable if, in the opinion of the site PI or appropriate sub-investigator, the residual symptoms 
will not interfere with the ability to assess safety parameters as required by the protocol. Subjects 
may re-screen after an acute illness is resolved 
8. Any confirmed or suspected immunosuppressive or immunodeficient condition* or use of 
anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study 
vaccination. 
* including HIV infection 
9. Administration of chronic (defined as more than 14 days) immunosuppressants or other 
immune modifying drugs within 6 months of receipt of study vaccine* 
*~ For corticosteroids, this means prednisone, or equivalent, greater than or equal to 0.5 mg/kg/day.  
Intranasal and topical steroids ARE allowed; daily inhaled steroids for treatment of asthma NOT 
allowed. 
10. History of receipt of a Hantavirus vaccine, including vaccines for Hantaan virus, Puumala 
virus, or combination of both. 
11. Exposed to ANDV* or plans to travel to an endemic area^^ from enrollment through 6 
months post last vaccination. 
*$ Residence in an ANDV endemic area in the last 3 years or >2 consecutive weeks of travel to an 
ANDV endemic area^^ in the last 3 years. 
**ANDV endemic areas include Chile, Brazil and Argentina6,7  
12. Any chronic or active neurologic disorder, including seizures and epilepsy, excluding febrile 
seizures as a child. 
13. History of receiving immunoglobulin or other blood product within the 3 months before 
enrollment in this study. 

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14. Current or past history of alcohol or drug abuse in the last 5 years. 
15. Subjects with autoimmune disorders, chronic inflammatory disorders or neurological 
disorders with a potential autoimmune correlation. 
16. Have any diagnosis, current or past, of schizophrenia, bipolar disease, or other psychiatric 
diagnosis that may interfere with subject compliance or safety evaluations. 
17. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for 
danger to self or others within 10 years prior to study vaccination. 
18. Have received any antiviral within 3 days of study vaccination. 
19. A diagnosis of Type I or II diabetes. 
20. Current employee or staff paid entirely or partially by the contract for this trial, or staff who 
are supervised by the PI or Sub-Investigators. 
21. Any condition that would, in the opinion of the Site Investigator or appropriate sub-
investigator, is a contraindication to study participation. * 
* Including acute or chronic (persisting for at least 90 days) clinically significant medical disease or 
condition, that would place the subject at an unacceptable risk of injury, render the subject unable 
to meet the requirements of the protocol, or may interfere with the evaluation of responses or the 
subject’s successful completion of the study. 
5.3 
Eligibility Criteria for Doses 2-4 
Subjects may not receive subsequent vaccinations if any of the criteria in section 5.4.3 are met.  
Subsequent vaccinations may be deferred in subjects with transient acute illness as described in 
Section 5.4.3.  
5.4 
Treatment Assignment Procedures 
5.4.1 Enrollment and Randomization Procedures 
Per International Council for Harmonisation of Technical Requirements for Pharmaceuticals for 
Human Use (ICH) guideline E6: Good Clinical Practice (GCP), screening records will be kept at 
the VTEU site to document the reason why an individual was screened, but failed trial entry 
criteria. The reasons why individuals failed screening will be recorded in the Statistical and Data 
Coordinating Center’s (SDCC) Advantage eClinicalSM (Electronic Data Capture System). 
 

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Once consented and upon entry of demographic data and confirmation of eligibility for this trial, 
the subjects will be enrolled. The first 24 participants in Cohorts 1 and 2 will include two 
sentinel subjects since this is a first-in-human Phase 1 study  
 

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Table 1: Treatment Arms. Sentinel subjects will be randomized 1:1 to dose regimen and will 
receive 2 mg of study vaccine open label. The 22 non-sentinel subjects in Cohorts 1 and 2 will be 
randomized in a 9:2:9:2 ratio (Treatment Arms 1b:1c:2b:2c) to receive either study vaccine or 
placebo at a dose of 2 mg in a 3 or 4 dose regimen, in double-blind fashion.   
 
The 24 participants in Cohorts 3 and 4 will follow the same schedule as outlined for Cohorts 1 
and 2 above. Sentinel subjects will receive 4 mg of study vaccine open label and be followed for 
safety through Day 8. The remaining 22 non-sentinel subjects in Cohort 3 and 4 will be 
randomized in a 9:2:9:2 (Treatment Arms 3b:3c:4b:4c) ratio to receive study vaccine or placebo 
at a dose of 4 mg in a 3 or 4 dose regimen, in double blind fashion.   
 
Enrollment of subjects will be done online using the enrollment module of Advantage 
eClinicalSM. The randomization code will be prepared by statisticians at the SDCC and included 
in the enrollment module for this trial. Advantage eClinicalSM will assign each subject to a 
treatment arm after the demographic and eligibility data have been entered into the system. A 
designated individual at the VTEU site will be provided with a code list for emergency 
unblinding purposes, which will be kept in a secure place. 
 
Instructions for use of the enrollment module are included in the Advantage eClinicalSM User’s 
Guide. Manual back-up randomization procedures and instructions are provided for use in the 
event that the VTEU site temporarily loses access to the Internet or the online enrollment system 
is unavailable. 
 
Subjects who sign the informed consent form (ICF) and are randomized but do not receive study 
vaccine or placebo may be replaced. Subjects who sign the ICF, and are randomized and 
vaccinated, and subsequently withdraw, or are withdrawn or terminated from this trial, or are lost 
to follow-up will not be replaced. 
5.4.2 Masking Procedures 
Except for the sentinel subjects who will receive study vaccine open label, this is a double-blind 
clinical trial within each dose escalation group (i.e., double blinded within Cohorts 1 and 2 and 
double blinded within Cohorts 3 and 4). 
Investigators, study personnel performing any study-related assessments following study vaccine 
administration, and laboratory personnel performing antibody assays will be blinded to treatment 
arm assignment (i.e., placebo vs. study vaccine, and dose regimen). 

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Sentinel subjects will be blinded to cohort, but not dosage group. Non-sentinel subjects will be 
blinded to treatment arm assignment and cohort. 
The randomization scheme will be generated by the SDCC and provided to unblinded study 
personnel (i.e., research pharmacists performing study vaccination preparations and unblinded 
study vaccine administrators) at the VTEU site. 
The unblinded study vaccine administrator is a study personnel member credentialed to 
administer vaccines and may also participate in dose preparation, but will not be involved in 
study-related assessments or have subject contact for data collection following study vaccine 
administration.  
 
The SMC may receive data in aggregate and presented by treatment arm. The SMC may also be 
provided with expected and observed rates of the expected AEs in an unblinded fashion, and may 
request the treatment arm be unblinded for an individual subject if required for safety 
assessment. The SMC will review grouped and unblinded data in the closed session only. Refer 
to the protocol-specific Manual of Procedures (MOP) for unblinding procedures. 
5.4.3 Discontinuation of Study Product  
There are several reasons why a study subject may be prevented from receiving the second, third 
or fourth vaccination. If a subject’s study vaccine must be discontinued before the end of the 
vaccination schedule, this will not result in automatic withdrawal of the subject from the study.  
Subjects who discontinue study vaccine will be encouraged to remain in this trial for the 
following follow-up assessments:   
• 
Safety assessments: Venous blood sample at approximately 7 days after the last study 
vaccination. Subjects will be asked to complete the remaining study assessments per 
Appendix A, Schedule of Events which may be conducted by phone call/electronic 
communication (e.g., email, text message) rather than in person continuing through 
approximately 6 months after the last study vaccination.  
• 
Immunogenicity assessments: Venous blood sample at Day 29, 85 and approximately 6 
months after the last study vaccination See the protocol-specific Manual of Procedures 
(MOP) for alternate follow-up requirements. 
Follow-up study vaccinations (2nd, 3rd and/or 4th dose) will not be administered to a subject if 
any of the following criteria are met: 

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• 
Pregnancy 
• 
Receipt of disallowed licensed vaccine, experimental product or medication (see Section 5.2) 
• 
New onset of illness or condition that meets the Exclusion Criteria (see Section 5.2) 
• 
Medical condition or medication change for which continued participation, in the opinion of 
the site principal investigator or appropriate sub-investigator, would pose a risk to the subject 
or would likely confound interpretation of the results. Note: Medication changes subsequent 
to the first study vaccination are not exclusionary for receipt of the follow-up study 
vaccination provided there was no deterioration in the subject’s chronic medical condition 
that necessitated a medication change, and there is no additional risk to the subject or 
interference with the evaluation of responses to study vaccination. 
• 
Presence of signs or symptoms that could confound or confuse assessment of study vaccine 
reactogenicity.   
• 
Any laryngospasm, bronchospasm, or anaphylaxis within 1 day after administration of study 
product that is considered related to study product. 
• 
Any generalized urticaria after administration of study product that is considered related to 
study product. 
• 
Serious adverse event related to the study vaccination. 
• 
Grade 3 solicited or unsolicited (clinical sign or symptom) adverse event that occurs in the 7 
days following study vaccination, lasts for 24 hours or more without decreasing to a Grade 1 
or Grade 2, and does not have an alternative etiology. If the subject has a Grade 3 adverse 
event as per Reactogenicity Tables 3, 4, 5 or 6 in the seven days following study vaccination 
and it lasts < 24 hours before decreasing to a Grade 2 or Grade 1 adverse event, see 
discontinuation criteria for Grade 2 below.  
• 
Grade 3 solicited or unsolicited (clinical sign or symptom) adverse event that occurs >7 days 
following study vaccination and is ongoing at the time of the subsequent study vaccination 
(i.e., Day 29, Day 57 or Day 169).  Conversely, if this Grade 3 adverse event was resolved or 
reduced to a Grade 1 or Grade 2 adverse event at the time of the subsequent study 
vaccination, IP may only be given, following the documented determination by the site 
principal investigator or appropriate sub-investigator, that it would not render study 
vaccination unsafe or interfere with the evaluation of adverse events or immunologic 
response. 
• 
Grade 3 clinical safety laboratory value (according to the toxicity table, Section 9.2.2) that 
does not decrease to Grade 1 or less prior to the follow-up study vaccination (i.e., Day 29, 

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Day 57 or Day 169). Any clinical safety laboratory parameter may be re-evaluated only once 
at the local clinical laboratory to assess eligibility prior to the follow-up study vaccination. If 
the clinical safety laboratory value decreases to Grade 1 or less, the subject may receive the 
follow-up study vaccination.  The study vaccination should be scheduled to occur within the 
acceptable protocol-specified window for that visit. 
• 
Subjects who experienced any Grade 2 adverse event that is an unresolved Grade 2 or a 
Grade 1 at the time of the next vaccination, IP may only be given following the documented 
determination by the site principal investigator or appropriate sub-investigator, that it would 
not render study vaccination unsafe or interfere with the evaluation of adverse events or 
immunologic response. 
• 
As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
• 
Subject refusal of further study vaccination. 
• 
Termination of this trial. 
• 
New information becomes available that makes further administration of the study vaccine 
unsafe. 
Subjects with transient injection site or systemic signs or symptoms, or with an acute illness, 
including an oral temperature greater than or equal to 100.0°F, follow-up study vaccination/s 
should be postponed/deferred until signs, symptoms, or acute illness have resolved, or are 
improving as further specified below. Study vaccine given outside the protocol-specified window 
must be approved by the DMID Medical Officer.   
• 
Note for afebrile, acute illness only: If a subject is afebrile, his/her acute illness is nearly 
resolved with only minor residual symptoms remaining, this occurs within the acceptable 
protocol-specified window for that visit, and, in the opinion of the site principal investigator 
or appropriate sub-investigator, the residual symptoms will not interfere with the ability to 
assess safety parameters as required by the protocol, the subject may receive the follow-up 
study vaccination without DMID Medical Officer approval.   
5.4.4 Study Withdrawal 
Subjects may voluntarily withdraw their consent for trial participation at any time and for any 
reason, without penalty. The site principal investigator or appropriate sub-investigator may also 
choose to remove a subject from the study. A subject may withdraw or be withdrawn from this 
trial for any of the following reasons: 

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• 
Medical disease or condition, or any new clinical finding for which continued participation, 
in the opinion of the site principal investigator or appropriate sub-investigator, would 
compromise the safety of the subject, or would interfere with the subject's successful 
completion of this trial, or would interfere with the evaluation of adverse events or 
immunologic response. 
• 
As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
• 
Subject withdrawal of consent. 
• 
Subject lost to follow-up. 
• 
Subject dies. 
• 
Termination of this trial. 
• 
New information becomes available that makes further participation unsafe. 
5.4.5 Handling of Withdrawals 
The primary reason for withdrawal from this trial will be recorded on the Study Status data 
collection form. Subjects will be encouraged to complete the Early Termination Visit. The Early 
Termination Visit procedures are listed in Section 7.8. 
In the case of subjects who are lost to follow-up, extensive effort (i.e., three documented contact 
attempts via phone calls, emails, text messages, etc., made on separate occasions and followed 
by a certified letter) will be made to locate or recall them, or at least to determine their health 
status. These efforts will be documented in the subject’s study records. Every attempt will be 
made to follow all adverse events, including solicited local site and systemic AEs, unsolicited 
non-serious adverse events and SAEs, ongoing at the time of early withdrawal through resolution 
as per applicable collection times defined for the specific type of adverse event. 
Subjects who withdraw, or are withdrawn or terminated from this trial, or are lost to follow-up 
after signing the ICF, randomization, and receipt of study vaccine will not be replaced. Subjects 
who withdraw, or are withdrawn or terminated from this trial, or are lost to follow-up after 
signing the ICF and randomization but before receipt of study vaccine may be replaced. 
5.4.6 Termination of Study 
Although the sponsor has every intention of completing this trial, it reserves the right to 
terminate this trial at any time for clinical or administrative reasons. Reasons for termination 

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include, but are not limited to, study closure due to the recommendation after a SMC review and 
at the discretion of DMID. 

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6 
STUDY INTERVENTION/INVESTIGATIONAL PRODUCT 
6.1 
Study Product Description 
ANDV DNA vaccine is a deoxyribonucleic acid (DNA) vaccine. The ANDV DNA vaccine 
plasmid is constructed on a well-characterized plasmid backbone, pWRG7077, and elements of 
the ANDV genome. The plasmid used for the ANDV DNA vaccine, pWRG/AND-M (opt2), is 
optimized for homo sapien codon usage and mRNA stability by Genewiz. The vaccine was 
developed by the Department of Molecular Virology, US Army Medical Research Institute of 
Infectious Diseases (USAMRIID; Fort Detrick, Maryland), and the final vaccine was 
manufactured by Aldevron (Fargo, ND).  
The ANDV DNA vaccine drug substance, plasmid pWRG/AND- M(opt2), Lot # 87763, date of 
manufacture 01 Dec 2017, was diluted to a concentration of 4.0 +/- 0.2 mg/mL in phosphate-
buffered saline (PBS) at a pH of 7.2, sterile-filtered, and stored as bulk after aliquots were 
removed for stability, sterility, and quality control (QC) testing. 
The ANDV DNA plasmid that forms the active component for the vaccine is suspended in PBS. 
Each injection will contain 1 or 2 mg of DNA in 0.5 mL of PBS depending on randomization 
assignment; it is clear, colorless and free of visible particulate. 
6.1.1 Acquisition 
The ANDV DNA vaccine will be provided by Aldevron (Fargo, ND). Placebo and Diluent, 0.9% 
Sodium Chloride Injection, USP will be provided by the DMID Clinical Materials Services 
(CMS), Fisher BioServices, and will be shipped to the investigational site upon request and 
approval by DMID. 
Upon request by DMID, ANDV DNA Vaccine will be shipped to the following address: 
DMID Clinical Materials Services  
Fisher BioServices 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 
 
ANDV DNA vaccine, placebo and diluent will be provided through the DMID Clinical Materials 
Services (CMS) to the VTEU site prior to the start of this trial upon request and with prior 
approval from DMID. Should the site principal investigator require additional ANDV vaccine 
during this trial, further instructions are provided in the protocol-specific MOP. 

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6.1.2 Formulation, Packaging, and Labeling 
ANDV DNA Vaccine 
The ANDV DNA vaccine comes in single-use vials. Vials for the Phase 1 clinical trial will be 
fill-finished at 1.3 mL per vial. This will allow two 0.5 mL PharmaJet Stratis® Needle-Free 
Injection System syringes to be filled from each vial.  
 
The vaccine formulates will be administered using the PharmaJet Stratis® Needle-Free Injection 
System. This device delivers a 0.5 mL jet of liquid at high pressure that penetrates through the 
skin into the muscle. The device is FDA 510K cleared for use with approved medicines.   
 
The study product will be labeled according to manufacturer specifications and include the 
statement “Caution: New Drug – Limited by Federal Law to Investigational Use”. 
Placebo/Diluent 
 
Placebo and diluent will be supplied as 0.9% Sodium Chloride Injection, USP which is a sterile, 
nonpyrogenic, isotonic solution of sodium chloride and water for injection (WFI). Each mL 
contains sodium chloride 9 mg without any preservative. It contains no bacteriostatic, 
antimicrobial agent, or added buffer and is supplied only in single-dose containers. The solution 
may contain hydrochloric acid and/or sodium hydroxide for pH adjustment (pH 5.3 [4.5 to 7.0]). 
The placebo will be administered using the PharmaJet Stratis® Needle-Free Injection System. 
This device delivers a 0.5 mL jet of liquid at high pressure that penetrates through the skin into 
the muscle. The device is FDA 510K cleared for use with approved medicines.17   
6.1.3 Product Storage and Stability 
ANDV DNA Vaccine 
The vials containing study product must be stored at ≤ - 65º C. Vials are removed from the 
product freezer and thawed at room temperature for at least 30 minutes and administered within 
1 hour after thawing.  
 
 

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Placebo/Diluent 
Placebo and diluent must be stored at 20ºC to 25ºC (68°F to 77ºF) [See USP Controlled Room 
Temperature; excursions between 15°C and 30°C (59°F and 86°F) are permitted]. See protocol-
specific MOP for further instructions. 
The temperature of the storage unit must be recorded daily (excluding non-business days and 
holidays as applicable), continuously monitored and recorded during the duration of this trial per 
the VTEU site’s standard operating procedures (SOP), and documentation will be maintained. If 
the temperature fluctuates outside of the required range, the affected study product(s) must be 
quarantined at the correct storage temperature and labeled as ‘Do Not Use’ (until further notice). 
The research pharmacist must alert the site principal investigator and study coordinator, if the 
temperature fluctuates outside of the required range. In the event the temperature fluctuates 
outside of the required range, the affected study product(s) must not be administered. The site 
principal investigator or responsible person should immediately contact the DMID Product 
Support Team (see MOP for contact information) for further instructions before any additional 
study vaccinations are administered. Based on the information collected, DMID and/or the 
manufacturer will determine whether the affected study product(s) can be used. If it cannot be 
used, the site will receive specific instructions on how to return the affected study product(s) to 
the DMID CMS or destroy it on site. Additional instructions for quarantine are provided in the 
protocol-specific MOP. 
6.2 
Dosage, Preparation and Administration of Study 
Intervention/Investigational Product 
See the protocol-specific MOP for detailed information on the preparation, labeling, storage, and 
administration of study product for each cohort. Study product preparation will be performed by 
the VTEU site’s research pharmacist on the same day of study vaccine administration. 
 
Visually inspect the ANDV DNA vaccine upon receipt and prior to use. If the study product(s) 
appear(s) to have been damaged, contaminated or discolored, contain visible particulate matter or 
if there are any concerns regarding its integrity, do NOT use the affected study product(s). The 
affected study product(s) must be quarantined at ≤ - 65 ºC and labeled as ‘Do Not Use’ (until 
further notice). The site principal investigator or responsible person should immediately contact 
the DMID Product Support Team (see MOP for contact information) and DMID Clinical Project 
Manager for further instructions before any additional study vaccinations are administered. 
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affected study product(s) can be used. If it cannot be used, the site will receive specific 
instructions on how to return the affected study product(s) to the DMID CMS or destroy it on 
site. If the ANDV vaccine is unusable, study personnel will use another vial from the study 
supply. Replacement vials may be requested by contacting DMID. Additional instructions for 
quarantine and DMID contact information are provided in the protocol-specific MOP. 
 
The study product will be prepared for administration by thawing at room temperature for at 
least 30 minutes. Vials should be gently inverted at least 15 times once thawed and administered 
within 1 hour after thawing. The study product should have a clear appearance after thawing.  
 
Dilution is not required for subjects allocated to the 4 mg dosage cohorts, the dose will be 
divided into 2 mg per 0.5 mL administration into the left and right deltoid muscles. Dilution with 
0.9% Sodium Chloride Injection, USP is required for subjects allocated to the 2 mg dosage 
cohorts, the dosage will be divided into 1 mg per 0.5 mL administration into the left and right 
deltoid muscles. See protocol-specific MOP for further instructions on dilution.    
 
The volume administered to Cohort 1 and 2 subjects will be a total of 1.0 ml; 0.5 ml of ANDV 
DNA vaccine (diluted to 1.0 mg/0.5 mL) administered into both the left and right deltoid 
muscles.     
 
The volume administered to Cohort 3 and 4 subjects will be a total of 1.0 ml; 0.5 ml ANDV 
DNA vaccine (2.0 mg/0.5 ml) administered into both the left and right deltoid muscles.  
 
Study product administration will be performed by an unblinded study personnel member who is 
credentialed to administer vaccines and may also participate in dose preparation, but will not be 
involved in study-related assessments or have subject contact for data collection following study 
vaccine administration.  
6.3 
Modification of Study Intervention/Investigational Product for a 
Participant 
Subjects who do not receive follow-up vaccinations (2, 3 or 4) within the protocol defined 
window will be followed for safety and immunogenicity. 
Outside of protocol defined windows, there will be no dose modifications. 

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If a subject’s follow-up vaccinations are deferred, attempts will be made to reschedule the 
vaccination to occur within the acceptable protocol-specified window for that visit.   
Subjects who do not receive the follow-up study vaccination will continue with follow-up safety 
and immunogenicity assessments continuing through approximately 6 months after their last 
study vaccination. See Sections 5.4.3, 5.4.4 and 5.4.5 for reasons for and handling of 
withdrawals and discontinuation of treatment. See the protocol-specific MOP for alternate 
follow-up requirements.   
6.4 
Accountability Procedures for the Study Intervention/Investigational 
Product(s) 
After receipt of the ANDV vaccine, the site principal investigator is responsible for study 
product distribution and disposition and has ultimate responsibility for study product 
accountability.   As this is a blinded study, the site PI will delegate this responsibility to the 
unblinded site pharmacist. Study vaccine records must be maintained and document logs of 
receipt, accountability, and storage temperature conditions. The study product accountability 
records and dispensing logs will also capture vial numbers, including final vial number, date of 
study vaccine preparation/ administration, time of study vaccine preparation, time study vaccine 
is drawn into the syringe, and amount of study vaccine withdrawn for administration. Time of 
study vaccine administration to the subject will be captured on the appropriate data collection 
form. All study product(s), whether administered or not, must be documented on the appropriate 
study product accountability record or dispensing log. The sponsor's monitoring staff will verify 
the VTEU site’s study product accountability records and dispensing logs per the site monitoring 
plan. 
Used and unused vials of vaccine, will be retained until monitored and released for disposition as 
applicable. The injectors are re-usable, syringes are disposable. Used syringes post dosing may 
be disposed of upon completion of administration. Filled, un-used syringes will be retained until 
monitored and released for disposition as applicable. Final disposition of the unused vaccine will 
be determined by DMID and communicated to the enrolling site. 

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6.5 
Assessment of Subject Compliance with Study 
Intervention/Investigational Product  
Study products will be administered to subjects via intramuscular injection according to subject 
treatment assignment and as described in Section 6.2. Thus, subject compliance is not anticipated 
to be an issue. Deviations from the dose schedule may only occur as described in Section 6.3. 
6.6 
Concomitant Medications/Treatments 
Administration of any medications, therapies, or non-study vaccines will be recorded on the 
appropriate data collection form. Concomitant medications will include all current medications 
and medications taken within 60 days prior to signing the ICF through approximately 28 days 
after the last study vaccination or early termination (if prior to 28 days after the last study 
vaccination), whichever occurs first. Medications will include prescription and over-the-counter 
drugs as well as herbals, vitamins, and supplements. Medications and non-study vaccines 
reported in the electronic case report form (eCRF) are limited to those received within 30 days 
prior to the first study vaccination through approximately 28 days after the last study 
vaccination.  
Medications that might interfere with the evaluation of the investigational product(s) should not 
be used from time of study vaccination through 28 days post the last vaccination unless clinically 
indicated as part of the subject’s health care. Medications in this category include the prohibited 
medications per the Subject Exclusion Criteria (see Section 5.2). In addition, the site principal 
investigator or appropriate sub-investigator may identify other medications that should not be 
used due to a risk to subject safety or assessment of reactogenicity and immunogenicity. Use of 
medications (anti-inflammatories or analgesics) as prophylaxis prior to study vaccination is 
prohibited. There are no known drug-vaccine interactions with the study vaccine and subjects are 
not being asked to discontinue current medications not listed in the exclusion criteria.  
In the event medical conditions dictate use of medications, subjects are encouraged to obtain 
adequate care, comply with the course of therapy as prescribed by their physician and inform the 
site as soon as practicable. Details of all medications taken during the medication reporting 
period for this study (date, indication, brand or generic name) will be recorded. Use of new 
medication should prompt evaluation for the occurrence of an AE or worsening of a pre-existing 
medical condition. 
 

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7 
STUDY SCHEDULE 
Complete study schedule details listed by type of visit are described below. Refer also to 
Sections 4 and 8, and Appendix A: Schedule of Events . 
7.1 
Screening  
Day -28 to -1, Visit 00  
Potential subjects will be screened for eligibility within 28 days prior to the administration of 
study vaccination. The following activities will be performed at screening and may be done all at 
one visit or split into separate visits for all Treatment Arms: 
• 
Subjects will be provided with a description of this trial (purpose and study procedures) and 
asked to read and sign the ICF. The ICF will be signed prior to performing any study 
procedures, including any screening procedures. 
• 
Demographic information will be obtained by interview of subjects. 
• 
Eligibility criteria will be reviewed with subjects. The following screening procedures will be 
completed to ensure that the potential subject is eligible for the study: 
• 
Complete medical history will be obtained by interview of subjects. 
• 
History of all concomitant medications taken within 60 days prior to signing the ICF will be 
reviewed with subjects. Medications reported in the eCRF are limited to those taken within 
30 days prior to study vaccination. Assessment of study eligibility will include a review of all 
permitted and prohibited medications per the Subject Inclusion and Exclusion Criteria (see 
Section 5.1, 5.2, 5.3).  
• 
Vital signs, including oral temperature, pulse, and blood pressure, will be obtained. Subjects 
must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral 
temperature. 
• 
Height and weight will be collected. 
• 
A physical examination will be performed to include the following organs and organ 
systems: general appearance, skin, head and neck, lungs, heart, liver, spleen, extremities, 
musculoskeletal, lymph nodes, and nervous system by a study clinician licensed to make 
medical diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 

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• 
A serum pregnancy test will be performed on all females of childbearing potential and must 
be negative. 
• 
Venous blood will be collected for screening laboratory tests to be run at the local clinical 
laboratory. These include: WBC, Hgb, platelet count, ANC, sodium, potassium, and 
creatinine, HgbA1C, total bilirubin, and ALT. BUN will be obtained if creatinine is above 
the normal range. In addition, participants will be screened for HIV – 1/2 antibody, HCV 
antibody, and HBsAg. Testing results must be confirmed to meet the eligibility criteria as 
outlined in the Inclusion Criteria (see Section 5.1 and Appendix B) prior to randomization.   
• 
Urine will be collected for screening urinalysis (dipstick) for urine protein and urine 
toxicology screen for opiates. Values must meet the eligibility criteria (see Section 5) prior to 
randomization. 
7.2 
Enrollment/Baseline  
Day 1, Visit 1  
Forty-eight subjects who meet all inclusion and no exclusion criteria will be administered study 
product. The following procedures will occur for all Treatment Arms: 
• 
Subject’s willingness to participate will be reconfirmed prior to performing any further study 
procedures, including administration of the study vaccination. 
• 
Eligibility criteria, including results of all clinical screening laboratory evaluations, will be 
reviewed with subjects prior to study vaccination to ensure continued eligibility. 
• 
Complete medical history and any updates obtained by interview of subjects since the 
screening visit will be reviewed with subjects prior to study vaccination to ensure continued 
eligibility. 
• 
All concomitant medications will be reviewed with subjects prior to study vaccination for 
accuracy and completeness. Any new concomitant medications taken since the screening 
visit will be reviewed with subjects and assessed for continued eligibility prior to study 
vaccination. Medications reported in the eCRF are limited to those taken within 30 days prior 
to study vaccination. 
• 
Vital signs, including oral temperature, pulse, and blood pressure, will be obtained to ensure 
eligibility. Vital signs assessed on Day 1 prior to the first study vaccination will be 
considered as baseline. Subjects must not eat or drink anything hot or cold, or smoke within 
10 minutes prior to taking oral temperature. 

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• 
A targeted physical examination may be performed prior to study vaccination, if indicated 
based on review of complete medical history and any updates obtained by interview of 
subjects since the screening visit, by a study clinician listed on the Form FDA 1572. 
• 
A urine pregnancy test will be performed within 24 hours prior to study vaccination on all 
females of childbearing potential. Results must be negative and known prior to 
randomization and study vaccination. 
• 
Subjects will be enrolled in Advantage eClinicalSM and assigned randomly to a treatment 
arm prior to study vaccination. 
• 
Venous blood will be collected immediately prior to the study vaccination for baseline 
ANDV PRNT and PsVNA antibody assays. 
• 
Venous blood will be collected immediately prior to the study vaccination for baseline safety 
labs.  
• 
Venous blood will be collected immediately prior to the study vaccination for baseline ICS 
and LPA cellular assays. 
• 
Pre-administration systemic reactogenicity assessments will be performed prior to study 
vaccination to establish a baseline. Subjects will then receive study vaccination via 
intramuscular injection into the left and right deltoid one after another or simultaneously. The 
time of administration will be recorded on the appropriate data collection form. Subjects will 
be observed for at least 30 minutes after the last study vaccination was given. Post-
administration local and systemic reactogenicity assessments will be performed. Any 
AE/SAEs will be recorded on the appropriate data collection form prior to discharge from the 
clinic.   
• 
Subjects will be provided with a memory aid and other study-related materials to record daily 
oral temperature, solicited site and systemic AEs, unsolicited AEs, and concomitant 
medications. Subjects will be encouraged to take their oral temperature around the same time 
each day. Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes 
prior to taking oral temperature. Subjects will be instructed on how to use their memory aid 
and how to measure and record AEs prior to discharge from the clinic. Subjects will be 
instructed to notify the study center if they develop any severe reactions after study 
vaccination. If the site principal investigator or appropriate sub-investigator deems the 
reaction severe enough, further instructions will be given to the subject on the proper course 
of action, including a return to the clinic for immediate evaluation if appropriate. 

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7.3 
Follow-up, Vaccination Visits  
Day 29 (+2), 57 (+2), 169 (+5)  
Follow-up visits are scheduled in reference to study vaccination date as indicated for each visit 
window, see Appendix A, SOE. All Treatment Arms will return for vaccination on D29 (V3), 
D57 (V5) and D169 (V9).   
• 
Eligibility criteria will be reviewed with subjects prior to study vaccination to ensure 
continued eligibility. 
• 
Interim medical history will be obtained by interview of subjects to ensure continued 
eligibility prior to study vaccination. Any changes since the previous clinic visit or contact 
will be noted at each clinic visit. 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form at each clinic visit. Any new 
concomitant medications taken since the screening visit will be reviewed with subjects and 
assessed for continued eligibility prior to study vaccination. 
• 
Vital signs, including oral temperature, pulse, and blood pressure, will be obtained to ensure 
eligibility. Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes 
prior to taking oral temperature. 
• 
A targeted physical examination may be performed prior to study vaccination, if indicated 
based on review of complete medical history and any updates obtained by interview of 
subjects, by a study clinician listed on the Form FDA 1572. 
• 
A urine pregnancy test will be performed within 24 hours prior to study vaccination on all 
females of childbearing potential. Results must be negative and known prior to 
administration of study vaccination. 
• 
Venous blood will be collected immediately prior to the study vaccination (D29) for ANDV 
PsVNA antibody assays. 
• 
Venous blood will be collected at Days 57 and 169 immediately prior to the study 
vaccination for ANDV PsVNA and PRNT antibody assays. 
• 
Venous blood will be collected immediately prior to the study vaccination for safety labs on 
Day 169 only.  
• 
Venous blood will be collected immediately prior to the study vaccination for ICS and LPA 
cellular assays. 

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• 
Pre-administration systemic reactogenicity assessments will be performed prior to study 
vaccination to establish a baseline. Subjects will then receive the dose of study vaccine via 
intramuscular injection into the left and right deltoid one after another or simultaneously. The 
time of administration will be recorded on the appropriate data collection form. Subjects will 
be observed for at least 30 minutes after the last study vaccination was given. Post-
administration local and systemic reactogenicity assessments will be performed. Any 
AE/SAEs will be recorded on the appropriate data collection form prior to discharge from the 
clinic.   
• 
Subjects will be provided with a memory aid and other study-related materials to record daily 
oral temperature, solicited site and systemic AEs, unsolicited AEs, and concomitant 
medications. Subjects will be encouraged to take their oral temperature around the same time 
each day. Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes 
prior to taking oral temperature. Subjects will be instructed on how to use their memory aid 
and how to measure and record AEs prior to discharge from the clinic. Subjects will be 
instructed to notify the study center if they develop any severe reactions after study 
vaccination. If the site principal investigator or appropriate sub-investigator deems the 
reaction severe enough, further instructions will be given to the subject on the proper course 
of action, including a return to the clinic for immediate evaluation if appropriate. 
• 
All AEs and SAEs will be collected and recorded on the appropriate data collection form at 
each clinic visit. 
7.4 
Follow-up, Day 8 post each vaccination  
Day 8 (+2), 36 (+2), 64 (+2), 176 (+2)  
Follow-up visits are scheduled in reference to study vaccination date as indicated for each visit 
window, see Appendix A, SOE. All Treatment Arms will return on D8 (V2), D36 (V4), D64 
(V6) and D176 (V10). 
• 
Interim medical history will be obtained by interview of subjects to ensure continued 
eligibility. Any changes since the previous clinic visit or contact will be noted at each clinic 
visit. 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form at each clinic visit. Any new 
concomitant medications taken since the last clinic visit or phone contact will be reviewed 
with subjects and assessed for continued eligibility. 

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• 
A targeted physical examination may be performed, if indicated based on review of complete 
medical history and any updates obtained by interview of subjects, by a study clinician listed 
on the Form FDA. 
• 
Venous blood will be collected for safety labs.  
• 
All AEs and SAEs will be collected and recorded on the appropriate data collection form at 
each clinic visit. 
• 
Memory aid information will be reviewed with subjects at each clinic visit. 
7.5 
Follow-up, Day 29 post each vaccination 
Day 85, 197  
Follow-up visits are scheduled in reference to study vaccination date as indicated for each visit 
window, see Appendix A, SOE. All Treatment Arms will return 28 days post vaccination on D85 
(V7) and D197 (V11). 
• 
Interim medical history will be obtained by interview of subjects to ensure continued 
eligibility. Any changes since the previous clinic visit or contact will be noted at each clinic 
visit. 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form at Day 85 and Day 197 visit. Any 
new concomitant medications taken since the previous clinic visit or contact will be reviewed 
with subjects and assessed for continued eligibility. 
• 
A targeted physical examination may be performed, if indicated based on review of complete 
medical history and any updates obtained by interview of subjects, by a study clinician listed 
on the Form FDA 1572. 
• 
Venous blood will be collected for ANDV PRNT and PsVNA antibody assays. 
• 
Venous blood will be collected for future exploratory assays 
• 
Venous blood will be collected for ICS and LPA cellular assays. 
• 
All AEs and SAEs will be collected and recorded on the appropriate data collection form at 
each clinic visit. 
 

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7.6 
Follow-up Visits 
Day 141 & 253  
Follow-up visits are scheduled in reference to study vaccination date as indicated for each visit 
window, see Appendix A, SOE. All Treatment Arms will return on D141 (V8) and D253 (V12). 
• 
Interim medical history will be obtained by interview of subjects to ensure continued 
eligibility. Any changes since the previous clinic visit or contact will be noted at each clinic 
visit. 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form at the Day 141 clinic visit. Any new 
concomitant medications taken since the previous clinic visit or contact will be reviewed 
with subjects and assessed for continued eligibility. 
• 
A targeted physical examination may be performed, if indicated based on review of complete 
medical history and any updates obtained by interview of subjects, by a study clinician listed 
on the Form FDA 1572. 
• 
Venous blood will be collected for ANDV PsVNA antibody assays at Day 253. 
• 
All AEs will be collected and recorded on the appropriate data collection form at day 141.   
• 
SAEs will be collected and recorded on the appropriate data collection form at each clinic 
visit. 
7.7 
Final Study Visit  
Day 337  
Final visits are scheduled in reference to study vaccination date as indicated for each visit 
window, see Appendix A, SOE. All treatment Arms will return on D337 (V13). 
• 
Interim medical history will be obtained by interview of subjects. Any changes since the 
previous clinic visit or contact will be noted. 
• 
A targeted physical examination may be performed, if indicated based on review of complete 
medical history and any updates obtained by interview of subjects, by a study clinician listed 
on the Form FDA 1572. 
• 
Venous blood will be collected for future exploratory assays.  
• 
Venous blood will be collected for ANDV PsVNA antibody assays. 

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• 
All SAEs will be collected and recorded on the appropriate data collection form.  
7.8 
Early Termination Visit 
The following activities will be performed on all Treatment Arms at the early termination visit 
on subjects who withdraw, or are withdrawn or terminated from this trial: 
• 
Interim medical history will be obtained by interview of subjects and any changes since the 
previous clinic visit or contact will be noted (if indicated). 
• 
Memory aid information will be reviewed with subjects (if within 7 days post study 
vaccination). 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form (if prior to 28 days post last study 
vaccination). 
• 
All AE/SAEs will be recorded on the appropriate data collection form. AEs will be recorded 
if prior to 28 days post last study vaccination. AEs will be limited to SAEs, if after 28 days 
post last study vaccination. 
• 
Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated. Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes 
prior to taking oral temperature. 
• 
A targeted physical examination may be performed, if indicated based on review of interim 
medical history, by a study clinician licensed listed on the Form FDA 1572. 
• 
Post-administration reactogenicity assessments will be performed (if within 7 days post study 
vaccination). 
• 
Venous blood will be collected for safety laboratory evaluations and performed by the local 
(clinical) laboratory (if within 7 days post last study vaccination). 
• 
Venous blood will be collected for cellular (ICS, LPA) assays (if prior to 28 days post last 
study vaccination). 
• 
Venous blood will be collected for PsVNA antibody assays. 
7.9 
Unscheduled Visit  
Unscheduled visits may occur at any time during this trial. Any of the following activities may 
be performed: 

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• 
Interim medical history will be obtained by interview of subjects and any changes since the 
previous clinic visit or contact will be noted (if indicated). 
• 
Memory aid information will be reviewed with subjects (if within 7 days post study 
vaccination). 
• 
All concomitant medications (including solicitation for receipt of any non-study vaccines) 
will be recorded on the appropriate data collection form (if prior to 28 days post last study 
vaccination). 
• 
All AE/SAEs will be recorded on the appropriate data collection form. AEs will be recorded 
if prior to 28 days post last study vaccination. AEs will be limited to SAEs, if after 28 days 
post last study vaccination. 
• 
Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated. Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes 
prior to taking oral temperature. 
• 
A targeted physical examination may be performed, if indicated based on review of interim 
medical history, by a study clinician listed on the Form FDA 1572. 
• 
Post-administration reactogenicity assessments will be performed (if within 7 days post study 
vaccination). 
• 
Venous blood will be collected for safety laboratory evaluations and performed by the local 
(clinical) laboratory (if within 7 days post last study vaccination). 
• 
Venous blood will be collected for cellular (ICS, LPA) assays (if prior to 28 days post last 
study vaccination). 
• 
Venous blood will be collected for PsVNA antibody assays. 
 

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8 
STUDY PROCEDURES/EVALUATIONS 
8.1 
Clinical Evaluations 
Complete medical history will be obtained by interview of subjects at the screening visit and will 
be reviewed and/or updated on Day 1 prior to study vaccination. Subjects will be queried 
regarding a history of significant medical disorders of the head, eyes, ears, nose, throat, mouth, 
cardiovascular system, lungs, gastrointestinal tract, liver, pancreas, kidney, urologic system, 
nervous system, blood, lymph nodes, endocrine system, musculoskeletal system, skin, and 
genital/reproductive tract. A history of any allergies, cancer, immunodeficiency, psychiatric 
illness, substance abuse, and autoimmune disease will be solicited. At follow-up visits after study 
vaccination, an interim medical history will be obtained by interview of subjects noting any 
changes since the previous clinic visit or contact. The interim medical history should include an 
assessment for new medical conditions. 
 
Medication history (concomitant medications) will include a review of all current medications 
and medications taken within 60 days prior to signing the ICF through approximately 28 days 
after the last study vaccination or early termination (if prior to 28 days after the last study 
vaccination), whichever occurs first. Medications reported in the eCRF are limited to those taken 
within 30 days prior to the first study vaccination through approximately 28 days after the last 
study vaccination. Prescription and over-the-counter drugs will be included as well as herbals, 
vitamins, and supplements. Use of new medication should prompt evaluation for the occurrence 
of any AE. Assessment of eligibility will include a review of all permitted and prohibited 
medications per the Subject Inclusion and Exclusion Criteria (see Sections 5.1 and 5.2). In 
addition, the site principal investigator or appropriate sub-investigator may identify other 
medications that should not be used due to a risk to subject safety or assessment of reactogenicity 
and immunogenicity. History of non-study vaccines received from 30 days prior to the first study 
vaccination through approximately 28 days after the last study vaccination will be solicited and 
recorded on the eCRF. 
 
A full physical examination will be performed on all subjects at the screening visit. This exam 
will include the following organs and organ systems: general appearance, skin, head and neck, 
lungs, heart, liver, spleen, extremities, musculoskeletal, lymph nodes and nervous system, by a 
study clinician licensed to make medical diagnoses and listed on the Form FDA 1572 as the site 
principal investigator or sub-investigator.  
 

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Vital signs (oral temperature, pulse, and blood pressure) will be collected at the screening visit. 
Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking 
oral temperature. Height and weight will be collected at the screening visit prior to study 
vaccination. 
 
On Day 1 prior to the study vaccination and at follow-up visits after the study vaccination, a 
targeted physical examination may be performed, if indicated based on the subject’s interim 
medical history, by a study clinician licensed to make medical diagnoses and listed on the Form 
FDA 1572 as the site principal investigator or sub-investigator.  
 
On Day 1 and all vaccination visits, vital signs (oral temperature, pulse, and blood pressure) will 
be collected prior to study vaccination. A urine pregnancy test will be performed within 24 hours 
prior to study vaccination on all females of childbearing potential. Pre-administration 
reactogenicity assessments will be performed prior to each study vaccination to establish 
baseline, then the study vaccination will be given. Subjects will be observed in the clinic for at 
least 30 minutes after each study vaccination. The study vaccination site will be examined, post-
administration reactogenicity assessments will be performed and any AE/SAEs will be recorded 
on the appropriate data collection form prior to discharge from the clinic. All subjects will 
receive and complete a subject memory aid from the time of study vaccination through 7 days 
after study vaccination. Subject memory aids will be reviewed with the subjects for adverse 
events, (solicited local and systemic AEs and unsolicited AEs) approximately 7 days after each 
vaccination. 
 
Reactogenicity assessments will include an assessment of solicited adverse events occurring 
from the time of study vaccination through 7 days after study vaccination. These include local 
reactions such as pain, tenderness, erythema (redness), induration (hardness/swelling), bruising 
and skin discoloration as well as systemic AEs such as fever, feverishness (chills, shivering, 
sweating), fatigue (tiredness), malaise (general unwell feeling), myalgia (body aches/muscular 
pain), nausea, dizziness and headache. 
8.2 
Laboratory Evaluations 
8.2.1 Clinical Laboratory Evaluations 
Serum pregnancy tests will be done at the screening visit and processed locally by the site 
laboratory at the screening visit and a urine pregnancy test will be performed within 24 hours 
prior to study vaccination on all females of childbearing potential. Urine will be collected in a 

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sterile urine cup for processing by research staff using a CLIA waived urine pregnancy test kit. 
Results must be negative and known prior to randomization on Day 1 and prior to administration 
of study vaccination on Day 29, Day 57 and Day 169. 
 
Clinical screening laboratory tests will be performed at the screening visit and processed by the 
local (clinical) laboratory. Parameters to be evaluated to confirm trial eligibility as per Section 
5.1 and Appendix B, include the following: 
• 
Hematology: WBC, Hgb, platelet count, ANC, and HgbA1c.   
• 
Biochemistry: Total bilirubin, ALT, creatinine, sodium, and potassium. BUN will be done if 
creatinine is above the normal range.    
• 
Serum serology testing for HIV – 1/2 antibody, HCV antibody, and HBsAg. 
• 
Serum βHCG pregnancy test (all females of childbearing potential) at screening and urine 
pregnancy test as described above.   
• 
Urinalysis will be done to test for urine protein via dipstick in the clinic. Clean-catch, mid-
stream, urine specimen will be collected in a sterile urine cup. 
• 
Urine test for opiates: A clean-catch, mid-stream urine specimen will be collected in a sterile 
urine cup and transported to the clinical laboratory for processing and examination.  
 
Clinical safety labs will be performed at Day 1 and 169, and approximately 7 days after each 
study vaccination. If they are abnormal, tests will be repeated at the next scheduled visit (or 
sooner if medically indicated) and followed to normal or stabilization. Clinical safety laboratory 
parameters to be evaluated will include WBC, Hgb, platelet count, ANC, sodium, potassium, 
Creatinine, total bilirubin and ALT. These evaluations will be performed by the local (clinical) 
laboratory and will not be used for screening purposes prior to dosing. 
 
The volume of venous blood to be collected for clinical screening, immunologic assessments and 
safety laboratory evaluations is presented in the tables below: 
 
 
 

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Table 2: Blood Volume (mL) – All Treatment Arms 
Study Visit (V)  
00 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
U/S 
or ET 
Study Day and 
Window 
Screen 
D1 
D8 
D29 
D36 
D57 
D64 
D85 
D141 
D169 
D176 
D197 
D253 
D337 
 
Screening Labs 
20 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical Safety 
Evaluations 
 
6.5 
6.5 
 
6.5 
 
6.5 
 
 
6.5 
6.5 
 
 
 
6.5 
PBMCs (ICS, 
LPA) 
 
40 
 
40 
 
40 
 
40 
 
40 
 
40 
 
 
40 
Immunogenicity 
(ANDV PRNT) 
 
10 
 
 
 
10 
 
10 
 
10 
 
10 
 
 
 
Immunogenicity 
(ANDV PsVNA) 
 
10 
 
10 
 
10 
 
10 
 
10 
 
10 
10 
10 
10 
Future Exploratory 
Assays 
 
 
 
 
 
 
 
40 
 
 
 
40 
 
40 
 
Total volume of 
blood 
collected/visit 
20 
66.5 
6.5 
50 
6.5 
60 
6.5 
100 
0 
66.5 
6.5 
100 
10 
50 
56.5 
Cumulative 
volume of blood 
20 
86.5 
93 
143 
149.
5 
209.5 
216 
316 
316 
382.5 
389 
489 
499 
549 
N/A 
8.2.2 Acceptable Laboratory Values for Eligibility and Defining Normal Values 
If laboratory screening tests are out of acceptable range, repeat of screening tests is permitted once, 
provided there is an alternative explanation for the out of range value.  
Refer to Appendix B for acceptable values for study eligibility and clinical laboratory reference 
ranges. 
• 
Absolute neutrophil counts (ANC) below the laboratory normal reference range will be 
allowable if they are in keeping with the levels seen in a condition that is prevalent in our 
population known as “benign ethnic neutropenia.” For subjects who are of African American 
or Middle Eastern decent, ANC must be greater than or equal to 0.8 K/mcL for inclusion.  
• 
Creatinine, BUN, and ALT values that are below the lower limit of normal as presented in 
Appendix B are not exclusionary as these values are considered to be not clinically 
significant (NCS).  

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• 
If a creatinine level is abnormal (elevated above the clinical laboratory normal reference 
range) at screening, a BUN will be collected to accompany the repeat creatinine. If the BUN 
is within normal range (or less than the lower limits of normal), then the out of range 
screening and/or repeat creatinine value will be considered NCS and subject will be 
acceptable for study inclusion (see Appendix B for acceptable values for study eligibility). 
8.2.3 Special Assays or Procedures 
Cellular Immunology Assays 
Functional T cell immune responses in PBMCs collected from the study participants at different 
time points during the course of study will be studied by flow-based ICS and LPA. These assays 
will be performed at CCHMC. Venous blood samples will be collected from each subject at day 
1, 29, 57, 85, 169 and 197. PBMCs will be isolated and cryopreserved for future use. Batched 
PBMCs from each subject will be thawed and stimulated with ANDV GP peptide pools and the 
frequencies of CD4 and CD8 T cell secreting IFNg, IL-2 and TNF will be measured by ICS. In 
separate experiments thawed PBMCs will be stained with CFSE dye and stimulated with ANDV 
GP peptide pools. Five days later degree of proliferation of CD4 and CD8 T cells will be 
determined by measuring the dilution of CFSE on these cells by flow cytometry.  
ANDV, PsVNA and PRNT Assays 
Serum from ANDV DNA vaccinated individuals will be evaluated for immunogenicity by a 
neutralizing antibody response to ANDV measured by PsVNA and PRNT. These assays will be 
performed at USAMRIID. Venous blood samples will be collected from each subject prior to 
study vaccination (Day 1) and on Day 29, 57, 85, 169, 197, 253, and 337 (all Treatment Arms). 
Serum from all time points will be analyzed by ANDV PsVNA. Serum from Day 1, 57, 85, 169 
and 197 will be analyzed by ANDV PRNT. Those time points represent prior to the initial 
vaccination (Day 1), 1 month after the initial series of vaccinations (Day 57 and 85), prior to the 
six-month boost (Day 169) and 1 month after the six-month boost (Day 197). 
8.2.4 Specimen Preparation, Handling, and Shipping 
8.2.4.1 Instructions for Specimen Preparation, Handling, and Storage 
Instructions for specimen preparation, handling, and storage are included in the protocol-specific 
MOP as appropriate. 

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8.2.4.2 Specimen Shipment 
Specimen shipment will occur when all specimens are available following all applicable 
International Air Transport Association (IATA) requirements and according to the specifics for 
storage temperature and documentation as detailed in the local (clinical) laboratory manual and 
protocol-specific MOP as appropriate. 
 
All specimens for clinical screening and safety laboratory evaluations will be transported from 
the VTEU site to the local (clinical) laboratory. 
 
Specimens for cellular assays (ICS, LPA) will be transported from the VTEU site to the local 
(research) laboratory for processing.    
 
Specimens for antibody assays (ANDV, PsVNA and PRNT) will be shipped from the VTEU site 
to the Clinical Materials Services (CMS). 
 
Further instructions for specimen shipment are included in the local (clinical) laboratory manual 
and protocol-specific MOP, as appropriate. 

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9 
ASSESSMENT OF SAFETY 
9.1 
Specification of Safety Parameters 
Safety will be assessed by the occurrence of: 
1. Serious adverse events occurring from the time of study vaccination through approximately 6 
months after the last study vaccination. 
2. Solicited Adverse Events – reactogenicity events occurring from the time of study 
vaccination through 7 days after each study vaccination: 
a) Local AEs including pain, tenderness, erythema (redness), induration 
(hardness/swelling), bruising and skin discoloration.  
b) Systemic AEs including fever, feverishness (chills, shivering, sweating), fatigue 
(tiredness), malaise (general unwell feeling), myalgia (body aches/muscular pain), 
nausea, dizziness and headache. 
3. Clinical safety laboratory adverse events occurring from the time of study vaccination 
through approximately 7 days after each study vaccination. Parameters to be evaluated 
include WBC, Hgb, platelet count, ANC, sodium, potassium, Creatinine, total bilirubin and 
ALT.   
4. Unsolicited Adverse Events –non-serious adverse events occurring from the time of study 
vaccination through approximately 28 days after the last study vaccination. 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
9.2.1 Adverse Events 
Adverse Event (AE): ICH E6 defines an AE as any untoward medical occurrence in a patient or 
clinical investigation subject administered a pharmaceutical product regardless of its causal 
relationship to the study treatment. An AE can therefore be any unfavorable and unintended sign 
(including an abnormal laboratory finding), symptom, or disease temporally associated with the 
use of a medicinal (investigational) product. The occurrence of an AE may come to the attention 
of study personnel during study visits and interviews of a study recipient presenting for medical 
care, or upon review by a study monitor. 

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AEs, including solicited local (application site) and systemic (subjective and quantitative) 
reactions, not meeting the protocol-defined criteria for SAEs will be captured on the appropriate 
data collection form and eCRF.  
 
Information to be collected for unsolicited non-serious AEs includes event description, date of 
onset, licensed study clinician’s assessment of severity and relationship to study product and 
alternate etiology (if not related to study product) (assessed only by those with the training and 
authority to make a diagnosis and listed on the Form FDA 1572 as the site principal investigator 
or sub-investigator), date of resolution of the event, seriousness and outcome. AEs occurring 
during the trial collection and reporting period will be documented appropriately regardless of 
relationship. AEs will be followed through resolution. 
 
Any medical condition that is present at the time that the subject is screened will be considered 
as baseline and not reported as an AE. However, if the severity of any pre-existing medical 
condition increases from the time of study vaccination through 28 days post study vaccination, it 
will be recorded as an AE. 
 
AEs must be graded for severity and assessed for relationship to study product (see definitions 
below). Adverse events characterized as intermittent require documentation of onset and duration 
of each episode. The start and stop date of each reported AE will be recorded on the appropriate 
data collection form and eCRF. 
 
FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether or not considered drug related. 
 
Severity of Event: AEs will be assessed by a licensed study clinician listed on the Form FDA 
1572 as the site principal investigator or appropriate sub-investigator using a protocol-defined 
grading system (see Section 9.2.2). For events not included in the protocol-defined grading 
system, the following guidelines will be used to quantify severity: 
• 
Mild (Grade 1): Events require minimal or no treatment and do not interfere with the 
subject’s daily activities. 
• 
Moderate (Grade 2): Events result in a low level of inconvenience or concern with 
therapeutic measures. Moderate events may cause some interference with functioning and 
daily activities. 
• 
Severe (Grade 3): Events interrupt the subject’s daily activities and may require systemic 
drug therapy or other treatment. Severe events are usually incapacitating. 
 

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Relationship to Study Product: The licensed study clinician’s assessment of an AE's 
relationship to study product is part of the documentation process, but it is not a factor in 
determining what is or is not reported in this trial. If there is any doubt as to whether a clinical 
observation is an AE, the event should be reported. The relationship to study product must be 
assessed for AEs using the terms: related or not related. In a clinical trial, the study product must 
always be suspect. To help assess, the following guidelines are used: 
• Related – There is a reasonable possibility that the study product caused the adverse event. 
Reasonable possibility means that there is evidence to suggest a causal relationship between 
the study product and the adverse event. 
• Not Related – There is not a reasonable possibility that the administration of the study 
product caused the event. 
9.2.2 Reactogenicity  
Reactogenicity events are AEs that are common and known to occur following administration of 
this type of study vaccine. The following Toxicity Grading Scales will be used to grade solicited 
local (application site) and systemic (subjective and quantitative) AEs: 
Table 3:  Local (Injection Site) Reactogenicity Grading 
Local (Injection Site) 
Reaction 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Pain – experienced 
without touching the 
injection site 
(spontaneous discomfort) 
Subject is aware of pain, 
but it does not interfere 
with daily activity, and 
no pain medication is 
taken 
Subject is aware of pain; 
there is interference with 
daily activity or it 
requires repeated use of a 
non-narcotic pain reliever 
for >24 hours 
Subject is aware of pain, 
and it prevents daily 
activity or requires any use 
of a prescription medication 
Tenderness – hurts only 
when injection site is 
touched or the arm is 
moved 
The area immediately 
surrounding the injection 
site hurts only when 
touched or with arm 
motion, and it does not 
interfere with daily 
activity 
The area immediately 
surrounding the injection 
site hurts when touched or 
with arm motion, and it 
interferes with daily 
activity 
The area immediately 
surrounding the injection 
site hurts when touched or 
with arm motion, and it 
prevents daily activity 
Erythema (Redness)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Induration 
(Hardness/Swelling)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Skin Discoloration 
Barely perceptible 
Clearly discernable 
difference compared to 
Unsightly difference 

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Local (Injection Site) 
Reaction 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
difference compared to 
surrounding skin 
surrounding skin 
when compared to 
surrounding skin 
Ecchymosis (Bruising)* 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
Ecchymosis (bruising), erythema (redness) and induration (hardness)/swelling as analyzed by 
measurement will be graded as follows: 
Table 4:  Local (Injection Site) Reactogenicity Measurements 
Local (Injection Site) Reaction 
Small 
Medium 
Large 
Ecchymosis (Bruising)* 
<20 mm 
20 mm – 50 mm 
>50 mm 
Erythema (Redness)* 
<20 mm 
20 mm – 50 mm 
>50 mm 
Induration (Hardness)/Swelling* 
<20 mm 
20 mm – 50 mm 
>50 mm 
* In addition to grading the measured local reaction at the greatest single diameter, they should be evaluated and 
graded using the functional scale as well as the actual measurement.  Size will not be used as halting criteria by 
itself. 
Table 5: Subjective Systemic Reactogenicity Grading 
Systemic (Subjective) 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Feverishness 
(chills/shivering/sweating) 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference in 
that it prevents daily activity 
Malaise (General Unwell 
Feeling) 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference, 
prevents daily activity 
Fatigue (Tiredness) 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference, 
prevents daily activity 
Myalgia (Body 
Aches/Muscular Pain)* 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference, 
prevents daily activity 

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Systemic (Subjective) 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Headache 
noticeable but does not 
interfere with daily 
activity 
Any use of pain reliever or 
interferes with daily 
activity 
Prevents daily activity or 
requires use of a 
prescription medication  
Nausea 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference, 
prevents daily activity 
Dizziness 
noticeable but does not 
interfere with daily 
activity  
interferes with daily 
activity 
Significant interference, 
prevents daily activity 
* Not at injection site. 
Oral temperature# will be graded as follows: 
Table 6: Quantitative Systemic Reactogenicity Grading 
Systemic (Quantitative) 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Fever* - oral† 
37.8°C – 38.4°C 
100.00°F – 101.1°F 
38.5°C – 38.9°C 
101.2°F – 102.0°F 
>38.9°C 
>102.0°F 
# Oral temperature assessed on Day 1 prior to study vaccination will be considered as baseline. 
* A fever can be considered not related to the study product if an alternative etiology can be documented. 
† Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral temperature. 
Pulse and blood pressure# will be graded as follows: 
Table 7: Blood Pressure and Pulse Grading 
Physiologic Parameter 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Bradycardia - beats per minute 
45-46 
40 – 44 
<40 
Tachycardia - beats per minute 
106 – 130 
131 – 155 
>155 
Hypotension (systolic) mmHg 
80 – 84 
75 – 79 
<75 
Hypotension (diastolic) mmHg 
50 – 54 
45 – 49 
<45 
Hypertension (systolic) mmHg 
151 – 155 
156 – 160 
>160 

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Hypertension (diastolic) mmHg 
96 – 100 
101 – 105 
>105 
# Pulse and blood pressure assessed on Day 1 prior to study vaccination will be considered as baseline. 
Clinical safety laboratory results# will be graded as follows: 
Table 8:  Clinical Safety Laboratory Adverse Event Grading (Hematology) 
Hematology 
Clinical 
Laboratory 
Reference 
Range 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
WBC K/mcL (Decrease, 18 to < 21 years) 
4.5 – 13.0 
2.5 – 4.4 
1.5 – 2.4 
<1.5 
WBC K/mcL (Decrease, ≥ 21 years) 
4.5 – 11.0 
2.5 – 4.4 
1.5 – 2.4 
<1.5 
WBC K/mcL (Increase 18 to < 21 years) 
4.5 – 13.0 
13.1 – 15.0 
15.1 – 20.0 
>20.0 
WBC K/mcL (Increase > 21 years) 
4.5 – 11.0 
11.1 – 15.0 
15.1 – 20.0 
>20.0 
Hgb g/dL (Decrease) (Female) 
 11.7 – 15.7 
10.1 – 11.6 
8.5 – 10 
<8.5 
Hgb g/dL (Decrease) (Male) 
13.3 – 17.7 
11.0 – 13.2 
9.5 – 10.9 
<9.5 
Platelet count K/mcL (Decrease) 
135 - 466 
125 – 134 
100 – 124 
<100 
Platelet count K/mcL (Increase) 
135 - 466 
467 - 517 
518 – 750 
>750 
Absolute Neutrophil Count, K/mcL* (18 to 
< 21 years) 
1.80 – 8.00 
1.5-<1.8 
1.0-<1.5 
<1.0 
Absolute Neutrophil Count, K/mcL* (≥ 21 
years) 
1.80 – 7.70 
1.5-<1.8 
1.0-<1.5 
<1.0 
Absolute Neutrophil Count, K/mcL - 
Benign Ethnic Neutropenia* 
> 0.8 
0.6 – 0.7 
0.4 – 0.5 
< 0.4 
*  ANC for subjects that are of African American and Middle Eastern descent may have values as low as 0.8 K/mcL. 
Subjects of this descent must have an ANC ≥ 0.8 K/mcL to be eligible to participate in the study if all other study 
criteria are met.   
Table 9: Clinical Safety Laboratory Adverse Event Grading (Chemistry) 
Chemistry 
Clinical 
Laboratory 
Reference 
Range 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
ALT unit/L (Increase)  
≤49 
50-123 
124-245 
> 245 

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Total Bilirubin mg/dL (Increase) – when 
accompanied by any increase in ALT 
0.10 – 1.2 
1.3 – 1.5 
1.6 – 1.8 
>1.8 
Total Bilirubin mg/dL (Increase) – when 
ALT is normal 
0.10 – 1.2 
1.3 – 1.8 
1.9 – 2.4 
>2.4 
Creatinine mg/dL (Increase) (Female) 
0.50 – 0.80 
0.81 – 1.70 
1.71 – 2.00 
>2.00 
Creatinine mg/dL (Increase) (Male) 
0.60 – 1.10 
1.11 – 1.70 
1.71 – 2.00 
>2.00 
Sodium, low, mmol/L 
136 – 145 
130 - 135 
123-129 
<123 
Sodium, high, mmol/L 
136 - 145 
146 - 150 
151-157 
>157 
Potassium, high, mmol/L 
3.5 – 5.1 
5.2 - 6.0 
6.1-6.5 
>6.5 
Potassium, low, mmol/L 
3.5 – 5.1 
3.0 - 3.4 
2.5-2.9 
<2.5  
Blood Urea Nitrogen (BUN) mg/dL 
9.00 – 23.00 
24 – 26 
27 - 31 
>31 
# Clinical laboratory evaluations assessed at the Day 1 visit will be considered as baseline. 
9.2.3 Serious Adverse Events 
Serious Adverse Event (SAE): An adverse event or suspected adverse reaction is considered 
“serious” if, in the view of either the site principal investigator or sponsor, it results in any of the 
following outcomes: 
• death, 
• a life-threatening adverse event*, 
• inpatient hospitalization or prolongation of existing hospitalization, 
• a persistent or significant incapacity or substantial disruption of the ability to conduct normal 
life functions, or 
• a congenital anomaly/birth defect. 
• Important medical events that may not result in death, be life-threatening, or require 
hospitalizations may be considered serious when, based upon appropriate medical judgment 
they may jeopardize the patient or subject and may require medical or surgical intervention to 
prevent one of the outcomes listed in this definition. Examples of such medical events 
include allergic bronchospasm requiring intensive treatment in an emergency room or at 
home, blood dyscrasias or convulsions that do not result in inpatient hospitalization, or the 
development of drug dependency or drug abuse. 

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*  Life-threatening adverse event. An adverse event is considered “life-threatening” if, in the view of 
either the site principal investigator or sponsor, its occurrence places the subject at immediate risk of 
death. It does not include an adverse event that, had it occurred in a more severe form, might have 
caused death. 
SAEs will be: 
• 
Assessed for severity and relationship to study product and alternate etiology (if not 
related to study product) by a licensed study physician listed on the Form FDA 1572 as 
the site principal investigator or sub-investigator. 
• 
Recorded on the appropriate SAE form and eCRF. 
• 
Followed through resolution by a licensed study physician listed on the Form FDA 1572 
as the site principal investigator or sub-investigator. 
• 
Reviewed and evaluated by an Independent Safety Monitor (ISM) (as deemed necessary), 
the SMC (periodic review unless related), DMID, and the IRB. 
9.2.4 Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings 
The site principal investigator or appropriate sub-investigator is responsible for recording all 
AE/SAEs that are observed or reported during this trial, regardless of the relationship to study 
product. AE/SAEs, abnormal laboratory test values, or abnormal clinical findings will be 
collected, assessed, documented, reported, and followed appropriately. 
 
For baseline laboratory results that are abnormal according to the local laboratory reference 
range and fall within Grade 1 toxicity table range, these will not be considered laboratory 
adverse event (AE) and will thus not be graded.   However, if baseline clinical labs fall within 
Grade 1 range, then a laboratory AE is reported only if the value changes such that it falls into 
Grade 2 or higher when subsequent safety laboratory testing is done. 
9.3 
Reporting Procedures 
Solicited local and systemic AEs will be documented and reported from the time of study 
vaccination through 7 days after each study vaccination. 

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Clinical safety laboratory adverse events will be documented and reported at Day 1, 169 and 7 
days after each study vaccination. Additional safety labs will be reported if the subject returns to 
the clinic for re-evaluation of abnormal laboratory values through 7 days after the last study 
vaccination.  
Unsolicited non-serious AEs will be documented and reported from the time of study vaccination 
through approximately 28 days after the last study vaccination. 
SAEs (including lab values that meet SAE criteria) will be documented and reported from the 
time of study vaccination through approximately 6 months after the last study vaccination. 
At any time after completion of this trial, if the site principal investigator or appropriate sub-
investigator becomes aware of an SAE that is suspected to be related to study product, the site 
principal investigator or appropriate sub-investigator will report the event to the DMID 
Pharmacovigilance Group. 
9.3.1 Serious Adverse Events 
Any AE that meets a protocol-defined serious criteria must be submitted immediately (within 24 
hours of site awareness) on an SAE form to the DMID Pharmacovigilance Group at the 
following address: 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support (CROMS)  
6500 Rock Spring Dr. Suite 650 
Bethesda, MD 20814, USA 
SAE Hot Line: 1-800-537-9979 (US) or 1-301-897-1709 (outside US) 
SAE FAX: 1-800-275-7619 (US) or 1-301-897-1710 (outside US) 
SAE Email Address: PVG@dmidcroms.com 
In addition to the SAE form, selected SAE data fields must also be entered into Advantage 
eClinicalSM. Please see the protocol-specific MOP for details regarding this procedure. 
Other supporting documentation of the event may be requested by the DMID Pharmacovigilance 
Group and should be provided as soon as possible. 
The site will send a copy of the SAE report(s) to the ISM (as deemed necessary) when they are 
provided to the DMID Pharmacovigilance Group. The DMID Medical Monitor and DMID 
Clinical Project Manager will be notified of the SAE by the DMID Pharmacovigilance Group. 

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The DMID Medical Monitor will review and assess the SAE for regulatory reporting and 
potential impact on study subject safety and protocol conduct. 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND 
Following notification from the site principal investigator or appropriate sub-investigator, 
DMID, the Investigational New Drug (IND) sponsor, will report any suspected adverse reaction 
that is both serious and unexpected. DMID will report an adverse event as a suspected adverse 
reaction only if there is evidence to suggest a causal relationship between the drug and the 
adverse event. DMID will notify FDA and the site principal investigators (i.e., all principal 
investigators to whom the sponsor is providing drug under its IND(s) or under any principal 
investigator’s IND(s)) in an IND safety report of potential serious risks from clinical trials or any 
other source, as soon as possible, but in no case later than 15 calendar days after the sponsor 
determines that the information qualifies for reporting as specified in 21 CFR Part 312.32. DMID 
will also notify FDA of any unexpected fatal or life-threatening suspected adverse reaction as 
soon as possible, but in no case later than 7 calendar days after the sponsor’s initial receipt of the 
information. Relevant follow up information to an IND safety report will be submitted as soon as 
the information is available. Upon request from FDA, DMID will submit to FDA any additional 
data or information that the agency deems necessary, as soon as possible, but in no case later 
than 15 calendar days after receiving the request. 
All serious events designated as “not related” to study product(s), will be reported to the FDA at 
least annually in a summary format. 
9.3.3 Reporting of Pregnancy 
Pregnancies occurring in study subjects will be reported via Advantage eClinicalSM on the 
Pregnancy Report form. With the subject’s permission, all protocol-required venous blood 
samples will be obtained and the subject will continue to be followed for safety for the duration 
of this trial. Efforts will be made to follow all pregnancies reported during the course of this trial 
to pregnancy outcome pending the subject’s permission. 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events 
AEs will be collected, assessed, and followed through resolution from the time of study 
vaccination through approximately 28 days after the last study vaccination. 

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SAEs will be collected, assessed, and followed from the time of study vaccination through 
resolution even if this extends beyond the trial-reporting period (approximately 6 months after 
the last study vaccination). 
Resolution of an AE/SAE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic. 
If the site principal investigator or appropriate sub-investigator becomes aware of an acute 
febrile illness and the site principal investigator or appropriate sub-investigator decides to bring 
the subject in for an evaluation to determine etiology, then the site principal investigator or 
appropriate sub-investigator, at their own discretion, can determine if specific viral testing should 
be done by PCR to determine if the infectious agent was RSV. 
Follow-up procedures, evaluations, and outcomes will be recorded on the appropriate data 
collection form. 
9.5 
Halting Rules 
9.5.1 Sentinel Subject Halting Rules 
If any of these conditions are met, further vaccinations will be halted pending SMC electronic 
review. 
9.5.1.1 If none of the following conditions are met, may proceed to the second sentinel subject: 
• 
Any SAE regardless of the relationship to the study product 
• 
Any laryngospasm, bronchospasm, or anaphylaxis within 1 day after administration of study 
product that is considered related to study product. 
• 
One Grade 3 unsolicited adverse event that is considered related to study product. 
9.5.1.2 If none of the following conditions are met, may proceed to the remainder of the cohort: 
• 
Any SAE regardless of the relationship to the study product 
• 
Any laryngospasm, bronchospasm, or anaphylaxis within 1 day after administration of study 
product that is considered related to study product. 
• 
Any generalized urticaria after administration of study product that is considered related to 
study product. 

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• 
One Grade 3 unsolicited adverse event that is considered related to study product and not 
resolved or improved to lower grade within 2 days. 
• 
Same Grade 2 or higher solicited local adverse event in both sentinel subjects that is 
considered related to study product and not resolved or improved to lower grade within 2 
days. 
• 
Same Grade 2 or higher solicited systemic adverse event in both sentinel subjects that is 
considered related to study product and not resolved or improved to lower grade within 2 
days. 
• 
Same Grade 2 or higher laboratory adverse event in both sentinel subjects that is considered 
related to study product. 
9.5.2 Study Halting Rules 
Further enrollment and study vaccinations will be halted for SMC review/recommendation if any 
of the following are reported: 
• 
Any subject experiences a study product-related SAE from the time of the study product 
administration through the subject’s last study visit. 
• 
Any subject experiences laryngospasm, bronchospasm, or anaphylaxis within 1 day after 
administration of study product that is considered related to study product. 
• 
Two or more subjects experience generalized urticaria (defined as occurring at more than two 
body parts) after administration of study product that is considered related to study product. 
• 
Two or more subjects experience a grade 3 unsolicited AE (in the same MedDRA High 
Level Term) after administration of study product that is considered related to study product 
and not resolved or improved to lower grade within 2 days. 
This trial will also be halted for SMC review/recommendation if, within 7 days after 
administration of any study vaccination, any of the following occurs: 
• 
Two or more subjects experience the same grade 3 solicited local adverse event that is 
considered related to study product and not resolved or improved to lower grade within 2 
days. 
• 
Two or more subjects experience the same grade 3 solicited systemic adverse event that is 
considered related to study product and not resolved or improved to lower grade within 2 
days. 

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• 
Two or more subjects experience the same grade 3 laboratory adverse event that is 
considered related to study product. 
Grading scales for solicited local (application site) and systemic (subjective and quantitative) 
AEs are included in Section 9.2.2 
Grading scales for clinical safety laboratory adverse events are included in Section 9.2.2. 
If any of the halting rules are met following any subject receipt of any study vaccination, then 
this trial will not continue with the remaining enrollments or study vaccinations without a review 
by and recommendation from the SMC to proceed. 
DMID retains the authority to suspend additional enrollment and study interventions/ 
administration of study product during the entire trial, as applicable. 
9.6 
Safety Oversight (ISM plus SMC) 
9.6.1 Independent Safety Monitor (ISM) 
An ISM is a physician with relevant expertise whose primary responsibility is to provide to 
DMID an independent safety assessment in a timely fashion. Participation is for the duration of 
the DMID study and is a voluntary position that does not receive payment. The ISM must meet 
the requirements of the NIAID conflict of interest policy.   
The ISM: 
• 
Is in close proximity to the study site and has the authority and ability to readily access 
study participant records in real time. 
• 
May be a member of the participating institution’s staff but preferably be from a different 
organizational group within the institution. 
• 
Should not be in a direct supervisory relationship with the investigator. 
• 
Should have no direct involvement in the conduct of the study. 
The ISM will: 
• 
Sign a Conflict of Interest (COI) certification at the time they are asked to participate and 
provide updates to this information as needed. 
• 
Receive reports of Serious Adverse Events (SAEs) from the site investigator and will be 
notified by email when DMID is notified of the SAE. 

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• 
Evaluate the SAE and report their clinical assessment to DMID, through DMID-CROMS 
SOCS in a timely manner using the attached report form and email the report to DMID-
CROMS SOCS.   
• 
Communicate with the investigator at the site as needed. 
• 
Review additional safety related events at the request of DMID. 
• 
Provide additional information to DMID and/or the SMC by teleconference as requested. 
9.6.2 Safety Monitoring Committee (SMC) 
This clinical study will utilize an SMC, which is an independent group of experts that advises 
DMID. The primary responsibility of the SMC is to monitor subject safety. The SMC is external 
to the DMID and comprises at least 3 voting members. The SMC will consist of members with 
appropriate phase 1 study expertise to contribute to the interpretation of the data from this trial. 
Its activities will be delineated in a SMC charter that will describe the membership, 
responsibilities, and the scope and frequency of data reviews. The SMC will operate on a 
conflict-free basis independently of the study team. The DMID or the SMC may convene ad hoc 
meetings of the SMC according to protocol criteria or if there are concerns that arise during the 
study. 
 
The SMC will review the safety data at the following milestones: 
• 
Organizational meeting (prior to start of the study)   
• 
Data Review meeting (DRM) will be held if any sentinel halting rules are met for Cohort 1 
and 2 sentinel subjects. The SMC will conduct an electronic review of available safety data 
for the first 2 sentinel subjects in Cohorts 1 and 2 (2 mg dose group) that have completed 7 
days of the safety assessment after receiving study product. No new subjects may be enrolled 
during this follow-up and review period, but screening may continue.   
• 
DRM: The SMC will conduct a review of available safety data for the first 24 subjects in 
Cohort 1 and 2 (2 mg dose group) that have completed 7 days of the safety assessment after 
receiving the second dose of study product. No new subjects may be enrolled during this 
follow-up and review period, but screening for Cohort 3 and 4 and vaccination of the 
remaining doses in Cohorts 1 and 2 may continue.   
• 
DRM will be held if any sentinel halting rules are met for the Cohort 3 and 4 sentinel 
subjects.  The SMC will conduct an electronic review of available safety data for the first 2 
sentinel subjects in Cohort 3 and 4 (4 mg dose group) that have completed 7 days of the 
safety assessment after receiving study product. No new subjects may be enrolled during this 
follow-up and review period, but screening may continue.   

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• 
An Ad Hoc meeting will be convened when a halting rule is met, or at the request of the 
investigator and/or DMID if there are safety concerns during the course of the study. 
• 
Final review meeting: approximately 6 to 8 months after final clinical database lock to 
review the cumulative safety and immunogenicity data for this trial. The data will be 
provided in a standard summary format. The SMC may be asked to provide 
recommendations in response to questions posed by DMID. 
The SMC will operate under the rules of a DMID-approved charter that will be written at the 
organizational meeting of the SMC. At this time, each data element that the SMC needs to assess 
will be clearly defined. Procedures for SMC reviews/meetings will be defined in the charter. The 
SMC will review applicable data to include, but not limited to, study progress and participant 
clinical, safety, and reactogenicity data. Reports may include enrollment and demographic 
information, medical history, concomitant medications, physical assessments, clinical laboratory 
values, dosing compliance, and solicited and unsolicited AE/SAEs. Additional data may be 
requested by the SMC, and interim statistical reports may be generated as deemed necessary and 
appropriate by DMID. The SMC may receive data in aggregate and presented by cohort. The 
SMC may also be provided with expected and observed rates of the expected AEs in an 
unblinded fashion, and may request the treatment assignment be unblinded for an individual 
subject if required for safety assessment. The SMC will review grouped and unblinded data in 
the closed session only. The SMC will meet and review this data at scheduled time points or ad 
hoc as needed during this trial as defined in the SMC charter. As an outcome of each 
review/meeting, the SMC will make a recommendation as to the advisability of proceeding with 
study vaccinations (as applicable), and to continue, modify, or terminate this trial. 
DMID or the SMC chair may convene the SMC on an ad hoc basis according to protocol criteria 
or if there are immediate concerns regarding observations during the course of this trial. The 
DMID Medical Monitor is empowered to stop enrollment and study vaccinations if adverse 
events that meet the halting criteria are reported. The DMID Medical Monitor and the ISM (as 
deemed necessary) will be responsible for reviewing SAEs in real time. The SMC will review 
SAEs on a regular basis and ad hoc during this trial. 

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

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11 
STATISTICAL CONSIDERATIONS  
11.1 Study Hypotheses 
This is a phase I, randomized, placebo controlled, double-blind, dose escalation trial and is not 
designed to test a specific hypothesis. Rather, it is intended to obtain preliminary estimates in 
healthy adults of the safety, reactogenicity, and immunogenicity of the ANDV vaccine in healthy 
adults. 
11.2 Sample Size Considerations  
Rare adverse events are not demonstrable in a clinical study of this size; however, the 
probabilities of observing one or more AEs given various true event rates are presented in Table 
10. 
Table 10: Probability of Observing an Adverse Event for Various Event Rates 
N 
"True" Event 
Rate 
Probability of 
Observation (%) 
N 
"True" Event 
Rate 
Probability of 
Observation (%) 
10 
0.1% 
1.0 
20 
0.1% 
2.0 
0.5% 
4.9 
0.5% 
9.5 
1.0% 
9.6 
1.0% 
18.2 
2.0% 
18.3 
2.0% 
33.2 
3.0% 
26.3 
3.0% 
45.6 
4.0% 
33.5 
4.0% 
55.8 
5.0% 
40.1 
5.0% 
64.2 
10.0% 
65.1 
10.0% 
87.8 
20.0% 
89.3 
20.0% 
98.8 
11.3 Planned Interim Analyses (if applicable) 
11.3.1 Interim Safety Review  
After the completion of enrolling 24 subjects in Cohorts 1 and 2 (2 mg dose group), the SMC 
will review the data for Cohorts 1 and 2 through 7 days post the second vaccination.  
These reviews, however, may not involve any hypothesis testing and will not be considered in 
estimating the precision of any estimates made at the conclusion of the study. 

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11.3.2 Interim Analysis of Safety and Immunogenicity Data 
An interim analysis of safety, reactogenicity, and immunologic response data (excluding 
exploratory outcomes) is planned once all subjects (Cohort 1-4) have completed the Day 197 
visit and the data are entered in the database, validated and monitored according to the clinical 
monitoring plan. The study statistician will provide the analysis of aggregate data unblinded at 
the group level to the investigators and sponsor staff for the purpose of manuscript, abstract 
preparation or presentation. The information from the primary immunogenicity and safety review 
may be published or otherwise presented, pursuant upon executed agreements between NIAID 
and VTEU investigators. The laboratory staff that run the assays for the Day 253 and 337 visits 
will remain blinded, and the assessment of relationship to study product of any SAEs that may be 
reported at the Day 253 and 337 visits will be delegated to blinded sub-investigators at the 
clinical site. While the results will not be used to make any decisions concerning the conduct of 
this trial, they may be used to make decisions on activities external to this trial such as the design 
of future trials of this vaccine. Since this early analysis of the data is not intended to impact the 
conduct of the trial, it has no impact on Type I error and adjustments are not planned.  
11.4 Final Analysis Plan 
The final analysis will be performed and clinical study report completed when all primary safety 
endpoint data and all secondary immunogenicity endpoint data are available. The interim 
analysis based on the primary clinical data base will be included in the final clinical study report 
(CSR). The CSR will be completed after the final data lock (through approximately Day 337 
follow-up) and when all primary endpoint data are received by SDCC. Any available data from 
the exploratory immunogenicity endpoints may also be included or if agreed upon, in an 
addendum to the CSR. A formal statistical analysis plan will be developed by SDCC and 
finalized prior to the primary data lock. 
11.4.1 Analysis Populations 
The Safety Analysis population includes all subjects who received one dose of study vaccine. 
The intent-to-treat (ITT) population includes all subjects who received one dose of study vaccine 
and contributed both pre- and at least one post-study vaccination venous blood samples for 
immunogenicity testing for which valid results were reported. For analyses using the ITT 
population, subjects will be grouped based on randomized treatment arm.   

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In the final analysis, if there are major protocol deviations, a per-protocol (PP) analysis may be 
performed. The per protocol (PP) population includes all subjects in the ITT subset with the 
following exclusions: 
• 
Data from all available visits for subjects found to be ineligible at baseline. 
• 
Data from all visits subsequent to major protocol deviations, such as: 
• 
Receipt of non-study licensed live vaccine within 28 days before or after study 
vaccination, 
• 
Receipt of non-study licensed inactivated vaccine within 14 days before or after study 
vaccination,*  
*Allowable exception for inactivated seasonal influenza vaccine received more than 7 days prior 
to or after a study vaccination. 
• 
Receipt of immunosuppressive therapy (e.g., corticosteroids) within 30 days before or 
after study vaccination. 
• 
Data from any visit that occurs substantially out of window. 
For analyses using the PP population, subjects will be grouped based on study vaccinations 
received. 
11.4.2 Safety Data 
Summaries and analysis of safety data will be presented for the Safety Analysis Population. 
Solicited AEs will be summarized by severity for each day after vaccination (Days 1-7 post 
study vaccination) and as the maximum severity over all 7 days. Additionally, solicited AEs will 
be analyzed by taking the most severe response over the follow-up period, dichotomizing into a 
binary variable (none versus mild, moderate, or severe) and using standard techniques, such as 
exact confidence intervals (CI), to summarize the proportion of subjects reporting each symptom, 
any application site symptom, and any systemic symptom.  
Unsolicited non-serious AEs will be collected from the time of first study vaccination through 
approximately 28 days post last vaccination. Unsolicited AEs will be coded by Medical 
Dictionary for Regulatory Activities (MedDRA) for preferred term and system organ class 
(SOC). SAEs will be collected from the time of first study vaccination through approximately 6 
months post last vaccination. The numbers of SAEs are likely to be small in this trial and will be 
reported by detailed listings showing the event description, MedDRApreferred term and SOC, 
relevant dates (study vaccinations and AEs), severity, relatedness, and outcome for each event. 

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Non-serious unsolicited AEs will be summarized as number and percentage of subjects reporting 
at least one event in each MedDRApreferred term and SOC, cross tabulated by severity and 
relationship to study product. Additionally, the proportion of subjects and exact 95% confidence 
intervals of AEs in aggregate and by MedDRAcategories will be computed. 
Clinical laboratory data will be summarized by severity for each visit, and as the maximum over 
all post-study vaccination visits. Graphical presentations may include box plots and shift plots. 
11.4.3 Immunogenicity Data  
11.4.3.1  Cellular Assays (ICS, LPA) 
Summaries and analysis of cellular assay data will be presented for the ITT population. If there 
are major protocol deviations, a per-protocol (PP) analysis may also be performed. 
 
Immune responses in terms of incidence of a cellular immune response to ANDV peptide pool 
>3 standard deviations (SD) above that on Day 1 will be summarized. Cellular responses will be 
measured by mean intracellular cytokine-secreting CD4+ and CD8+ T Cell responses and mean 
CD4+ and CD8+ T Cell lymphoproliferation. Analyses will include response (>3 SD) for the 
different groups at the different time points. 
11.4.3.2  Immunogenicity (PsVNA, PRNT) 
Summaries and analysis of immunogenicity data will be presented for the ITT population. If 
there are major protocol deviations, a per-protocol (PP) analysis may also be performed.  
Immune responses in terms of a titer > 20 in antibody post vaccination, measured by PsVNA and 
PRNT will be summarized. Analyses will include incidence of seroconversion at different time 
points and geometric mean titers for the different groups at the different time points will be 
determined and compared. The correlation between PsVNA and PRNT titers will be evaluated. 
11.4.4 Missing Values and Outliers 
All attempts will be made to collect all data per protocol. As missing data are expected to be 
minimal, no imputation will be performed for missing values. Any data point that appears to be 
erroneous or inexplicable based on clinical judgment will be investigated as a possible outlier. If 
data points are identified as outliers, sensitivity analyses will be performed to examine the 
impact of including or excluding the outliers. Any substantive differences in these analyses will 
be reported. 

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

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

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14 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 Ethical Standard 
The site principal investigator will ensure that this trial is conducted in full conformity with 
principles of the Belmont Report: Ethical Principles and Guidelines for the Protection of Human 
Subjects of Research of the National Commission for the Protection of Human Subjects of 
Biomedical and Behavioral Research (April 18, 1979) and codified in 45 CFR 46, 21 CFR 50 
and 56, and ICH E6; 62 Federal Regulations 25691 (1997), if applicable. The site principal 
investigator’s institution will hold a current Federalwide Assurance (FWA) issued by the Office 
of Human Research Protection (OHRP) for federally funded research. 
14.2 Institutional Review Board 
Prior to enrollment of subjects into this trial, the protocol and ICF will be reviewed and approved 
by the appropriate IRB listed on its FWA. 
The responsible official for the IRB will sign the IRB letter of approval of the protocol prior to 
the start of this trial and a copy will be provided to DMID. The IRB FWA number will be 
provided to DMID. 
Should amendments to the protocol be required, the amendments will be written by the principal 
investigator and provided to the site IRB. Any amendments to the protocol or consent materials 
will be approved by the IRB before they are implemented. 
14.3 Informed Consent Process 
The site principal investigator will choose subjects in accordance with the eligibility criteria 
detailed in Section 5. Before any study procedures are performed, subjects must sign an ICF that 
complies with the requirements of ICH E6, 21 CFR Part 50 and 45 CFR 46 and the local IRB. 
Study personnel may employ recruitment efforts prior to obtaining study consent if a patient-
specific screening consent is on record or if the IRB has agreed that chart review is allowed 
without a fully executed screening consent. In cases where there is not a patient-specific 
screening consent on record, site clinical staff may pre-screen via chart review and refer potential 
subjects to the research staff. Research staff would obtain written consent per the standard 
informed consent process before conducting protocol-specific screening activities. 

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

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14.4 Exclusion of Women, Minorities, and Children (Special Populations) 
This trial will be inclusive of all adults who meet the Subject Inclusion Criteria (see Section 5.1) 
and do not meet the Subject Exclusion Criteria (see Section 5.2), regardless of religion, sex, or 
ethnic background. Should the outcome of this trial be deemed acceptable, additional trials may 
be initiated including those in other populations. Children will not be included in this trial as they 
are not the target population at this time. 
14.5 Subject Confidentiality 
Subject confidentiality is strictly held in trust by the site principal investigators, other study 
personnel, the sponsor, and their agents. This confidentiality is extended to cover testing of 
biological samples, in addition to the clinical information relating to participating subjects. 
Subjects will have code numbers and will not be identified by name. 
The study protocol, documentation, data, and all other information generated will be held in 
strict confidence. No information concerning this trial or the data will be released to any 
unauthorized third party without prior written approval of the sponsor. 
All information provided by the sponsor and all data and information generated by the VTEU 
site as part of this trial (other than a subject’s medical records) will be kept confidential by the 
site principal investigators and other study personnel to the extent permitted by law. This 
information and data will not be used by the site principal investigators or other study personnel 
for any purpose other than conducting this trial. These restrictions do not apply to: (1) 
information which becomes publicly available through no fault of the site principal investigators 
or other study personnel; (2) information which is necessary to disclose in confidence to an IRB 
solely for the evaluation of this trial (3) information which is necessary to disclose in order to 
provide appropriate medical care to a study subject; or (4) study results which may be published 
as described in Section 16.  
In addition, this research is covered by a Certificate of Confidentiality from the NIH. The 
investigators and their staff may not disclose or use information documents, or biospecimens that 
may identify the subjects in any federal, state, or local civil, criminal, administrative, legislative 
or other action, or be used as evidence unless the subject has consented. This does not apply to 
requests for information from the NIH or its representatives that are needed to monitor or audit 
the study, or for information that must be disclosed in order to meet FDA requirements. 

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The study monitor, applicable regulatory authorities, such as the FDA, or other authorized 
representatives of the sponsor may inspect all documents and records required to be maintained 
by the site principal investigators. This includes, but is not limited to, medical records (office, 
clinic, or hospital) and pharmacy records for the subjects in this trial. The VTEU site will permit 
access to such records. 
14.6 Study Discontinuation 
If this trial is discontinued, subjects, who have signed the ICF and are randomized and 
vaccinated, will continue to be followed for safety for the duration of this trial. No further study 
vaccinations will be administered. 
14.7 Future Use of Stored Specimens and Data 
Residual samples are those that are left over after the study has been completed and are stored 
for possible use in future research studies. Future use samples are extra tube/s of blood collected 
and stored for possible use in future research studies. Retention of residual samples for future use 
and the collection and storage of samples for use in future research studies is a condition of study 
participation.  Subjects who sign the informed consent form for the study are consenting to allow 
the collection, storage and use of both residual and future use samples.  
Residual samples (serum or cells) derived from venous blood samples will be maintained for 
possible use in future research studies, such as examining additional immunological assessments 
or testing for antibodies against other viruses or bacteria. These samples may be shared for 
purposes other than per protocol analysis with investigators at the participating VTEU site and 
with other investigators at other institutions once the clinical study report has been finalized.   
Future use samples (whole blood, serum or cells) will be collected at Day 85, 197 and 337. It is 
anticipated that approximately two to five aliquots of serum (4 ml/each) from venous blood 
samples will be available specifically for the purpose of future research at Day 85, 197 and 337. 
PBMCs collected at day 337 will also be processed for future use. Future use research studies 
may include but are not limited to non-traditional immune assay development, assessing innate 
immune factors, cytokines and other virologic evaluations and Human Leukocyte Antigen 
(HLA) testing to explore the impact of host genetics on immune responses to ANDV DNA 
vaccine. If HLA testing is performed, results will be part of research records and will not become 
part of a subject’s medical record and will not be shared with their doctor.  

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Residual and future use research samples will be stored indefinitely at a DMID designated 
central clinical storage facility. These samples will not be sold or used directly for production of 
any commercial product. Genetic tests may be performed on samples. Each sample will be 
encoded (labeled) only with a barcode and a unique tracking number to protect subject 
confidentiality. There are no benefits to subjects in the collection, storage and subsequent use of 
their specimens for future research. Reports about future research done with subjects’ samples 
will NOT be kept in their health records.  
Residual samples will be available upon the completion of the study; however, future use 
samples may be requested from DMID and shipped from the DMID CMS at any time. 
Subjects may change their decision to participate in the study at any time by notifying the study 
doctors or nurses in writing. However, any data from a previously collected sample prior to the 
withdrawn consent will not be removed. Any future use of residual samples or collection of 
samples specifically for future research not yet collected/stored will not be stored for 
future/residual use. Any data from a previously collected sample prior to the withdrawn consent 
will not be removed. 

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

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range checks, to identify data that appear inconsistent, incomplete, or inaccurate. Clinical and 
reactogenicity data will be entered directly from the data collection forms completed by the study 
personnel. 
15.3 Types of Data 
Data for this trial will include clinical, safety, and outcome measures (e.g., clinical laboratory 
values, reactogenicity, and immunogenicity data). 
15.4 Timing/Reports 
See Section 9.3 for further reporting details. 
Interim statistical reports may be generated as outlined in the protocol or as deemed necessary 
and appropriate by DMID. Upon request and DMID approval, the SDCC may provide the study 
team with interim summary results for purposes of designing future trials or for 
publication/presentation of the data. Safety and immunogenicity summary reports may be 
generated for the SMC. 
After full analysis and final reporting is complete, and upon request and DMID approval, the 
SDCC will provide the VTEU site with a summary of results by treatment arm and/or subject 
treatment assignments. In this regard, the VTEU site requesting such information to share with 
study subjects must do so in compliance with their respective IRB guidelines. 
15.5 Study Records Retention 
Study records and reports including, but not limited to, eCRFs, source documents, ICFs, and 
study drug disposition records shall be maintained for 2 years after a marketing application is 
approved for the drug for the indication for which it is being investigated; or, if no application is 
to be filed or if the application is not approved for the drug, until 2 years after the investigation is 
discontinued and the FDA has been notified. ICFs for future use will be maintained as long as 
the sample exists. 
The VTEU site must contact DMID for authorization prior to the destruction of any study 
records. 

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15.6 Protocol Deviations 
A protocol deviation is any noncompliance with the study protocol, GCP, or protocol-specific 
MOP requirements. The noncompliance may be either on the part of the subject, the site 
principal investigator, or other study personnel. As a result of deviations, corrective actions are 
to be developed by the site and implemented promptly. 
These practices are consistent with ICH E6: 
4.5 
Compliance with Protocol, Sections 4.5.1, 4.5.2, and 4.5.3 
5.1 
Quality Assurance and Quality Control, Section 5.1.1 
5.20 
Noncompliance, Sections 5.20.1, and 5.20.2 
It is the responsibility of the site principal investigator and other study personnel to use 
continuous vigilance to identify and report deviations within five working days of identification 
of the protocol deviation, or within five working days of the scheduled protocol-required 
activity. All deviations must be promptly reported to DMID, via the SDCC’s Advantage 
eClinicalSM. 
All protocol deviations, as defined above, must be addressed in study subject data collection 
forms. A completed copy of the DMID Protocol Deviation Form must be maintained in the 
Regulatory File as well as in the subject’s chart. Protocol deviations must be sent to the local 
IRB per its guidelines. The site principal investigator and other study personnel are responsible 
for knowing and adhering to their IRB requirements. 

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16 
PUBLICATION POLICY 
All investigators funded by the NIH must submit or have submitted for them to the National 
Library of Medicine’s PubMed Central (http://www.ncbi.nlm.nih.gov/pmc/) an electronic 
version of their final, peer-reviewed manuscripts upon acceptance for publication, to be made 
publicly available no later than 12 months after the official date of publication. The NIH Public 
Access Policy ensures the public has access to the published results of NIH funded research. It 
requires investigators to submit final peer-reviewed journal manuscripts that arise from NIH 
funds to the digital archive PubMed Central upon acceptance for publication. Further, the policy 
stipulates that these papers must be accessible to the public on PubMed Central no later than 12 
months after publication. 
Refer to: 
NIH Public Access Policy, http://publicaccess.nih.gov/ 
NIH OER Grants and Funding, http://grants.nih.gov/grants/oer.htm 
As of January 2018, all clinical trials supported by the NIH must be registered on 
ClinicalTrials.gov, no later than 21 days after the enrollment of the first subject. Results of all 
clinical trials supported by the NIH, generally, need to be submitted no later than 12 months 
following the primary completion date. A delay of up to 2 years is available for trials that meet 
certain criteria and have applied for certification of delayed posting. 
As part of the result posting a copy of this protocol (and its amendments) and a copy of the 
Statistical Analysis Plan will be posted on ClincialTrials.gov. 
For this trial the responsible party is DMID which will register the trial and post results.   
The responsible party does not plan to request certification of delayed posting. 
Refer to: 
Public Law 110-85, Section 801, Clinical Trial Databases 
42CFR11 
NIH NOT-OD-16-149 

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Results of any exploratory immunogenicity analysis will not be published prior to publication of 
the primary immunogenicity results for this trial. 
*Journal Citation: De Angelis C, Drazen JM, Frizelle FA, Haug C, Hoey J, Horton R, et al. 
Clinical trial registration: a statement from the International Committee of Medical Journal 
Editors. N Engl J Med. 2004; 351:1250-1. 

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17 
LITERATURE REFERENCES 
1. 
Hooper JW, Moon JE, Paolino KM, et al. A Phase 1 clinical trial of Hantaan virus and 
Puumala virus M-segment DNA vaccines for haemorrhagic fever with renal syndrome delivered 
by intramuscular electroporation. Clinical microbiology and infection : the official publication of 
the European Society of Clinical Microbiology and Infectious Diseases 2014;20 Suppl 5:110-7. 
2. 
Jiang H, Zheng X, Wang L, Du H, Wang P, Bai X. Hantavirus infection: a global 
zoonotic challenge. Virologica Sinica 2017;32:32-43. 
3. 
Simpson SQ, Spikes L, Patel S, Faruqi I. Hantavirus pulmonary syndrome. Infectious 
disease clinics of North America 2010;24:159-73. 
4. 
Boudreau EF, Josleyn M, Ullman D, et al. A Phase 1 clinical trial of Hantaan virus and 
Puumala virus M-segment DNA vaccines for hemorrhagic fever with renal syndrome. Vaccine 
2012;30:1951-8. 
5. 
Zhang YZ, Zou Y, Fu ZF, Plyusnin A. Hantavirus infections in humans and animals, 
China. Emerging infectious diseases 2010;16:1195-203. 
6. 
Limongi JE, Oliveira RC, Guterres A, et al. Hantavirus pulmonary syndrome and rodent 
reservoirs in the savanna-like biome of Brazil's southeastern region. Epidemiology and infection 
2016;144:1107-16. 
7. 
Martinez-Valdebenito C, Calvo M, Vial C, et al. Person-to-person household and 
nosocomial transmission of andes hantavirus, Southern Chile, 2011. Emerging infectious 
diseases 2014;20:1629-36. 
8. 
Padula PJ, Colavecchia SB, Martinez VP, et al. Genetic diversity, distribution, and 
serological features of hantavirus infection in five countries in South America. Journal of clinical 
microbiology 2000;38:3029-35. 
9. 
Wells RM, Sosa Estani S, Yadon ZE, et al. An unusual hantavirus outbreak in southern 
Argentina: person-to-person transmission? Hantavirus Pulmonary Syndrome Study Group for 
Patagonia. Emerging infectious diseases 1997;3:171-4. 
10. 
Nunez JJ, Fritz CL, Knust B, et al. Hantavirus infections among overnight visitors to 
Yosemite National Park, California, USA, 2012. Emerg Infect Dis 2014;20:386-93. 
11. 
Figueiredo LT, Souza WM, Ferres M, Enria DA. Hantaviruses and cardiopulmonary 
syndrome in South America. Virus research 2014;187:43-54. 
12. 
Auwaerter PG, Oldach D, Mundy LM, et al. Hantavirus serologies in patients 
hospitalized with community-acquired pneumonia. The Journal of infectious diseases 
1996;173:237-9. 
13. 
Diglisic G, Rossi CA, Doti A, Walshe DK. Seroprevalence study of Hantavirus infection 
in the community based population. Maryland medical journal (Baltimore, Md : 1985) 
1999;48:303-6. 
14. 
Hooper JW, Josleyn M, Ballantyne J, Brocato R. A novel Sin Nombre virus DNA 
vaccine and its inclusion in a candidate pan-hantavirus vaccine against hantavirus pulmonary 
syndrome (HPS) and hemorrhagic fever with renal syndrome (HFRS). Vaccine 2013;31:4314-
21. 

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15. 
Hannaman D, Dupuy LC, Ellefsen B, Schmaljohn CS. A Phase 1 clinical trial of a DNA 
vaccine for Venezuelan equine encephalitis delivered by intramuscular or intradermal 
electroporation. Vaccine 2016;34:3607-12. 
16. 
Hooper JW, Kamrud KI, Elgh F, Custer D, Schmaljohn CS. DNA vaccination with 
hantavirus M segment elicits neutralizing antibodies and protects against seoul virus infection. 
Virology 1999;255:269-78. 
17. 
Kwilas S, Kishimori JM, Josleyn M, et al. A hantavirus pulmonary syndrome (HPS) 
DNA vaccine delivered using a spring-powered jet injector elicits a potent neutralizing antibody 
response in rabbits and nonhuman primates. Current gene therapy 2014;14:200-10. 
18. 
Haese N, Brocato RL, Henderson T, et al. Antiviral Biologic Produced in DNA 
Vaccine/Goose Platform Protects Hamsters Against Hantavirus Pulmonary Syndrome When 
Administered Post-exposure. PLoS neglected tropical diseases 2015;9:e0003803. 
19. 
Hooper JW, Brocato RL, Kwilas SA, et al. DNA vaccine-derived human IgG produced in 
transchromosomal bovines protect in lethal models of hantavirus pulmonary syndrome. Science 
translational medicine 2014;6:264ra162. 
20. 
Li L, Petrovsky N. Molecular mechanisms for enhanced DNA vaccine immunogenicity. 
Expert review of vaccines 2016;15:313-29. 
21. 
Sallberg M, Frelin L, Ahlen G, Sallberg-Chen M. Electroporation for therapeutic DNA 
vaccination in patients. Medical microbiology and immunology 2015;204:131-5. 
22. 
Beckett CG, Tjaden J, Burgess T, et al. Evaluation of a prototype dengue-1 DNA vaccine 
in a Phase 1 clinical trial. Vaccine 2011;29:960-8. 
23. 
Ulmer JB, Donnelly JJ, Parker SE, et al. Heterologous protection against influenza by 
injection of DNA encoding a viral protein. Science (New York, NY) 1993;259:1745-9. 
24. 
Faurez F, Dory D, Le Moigne V, Gravier R, Jestin A. Biosafety of DNA vaccines: New 
generation of DNA vectors and current knowledge on the fate of plasmids after injection. 
Vaccine 2010;28:3888-95. 
25. 
Wang Z, Troilo PJ, Wang X, et al. Detection of integration of plasmid DNA into host 
genomic DNA following intramuscular injection and electroporation. Gene therapy 2004;11:711-
21. 
 
 

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18 
SUPPLEMENTS/APPENDICES 

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Appendix A: Schedule of Events  
All Treatment Arms 
Study Visit (V)  
00 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
U/S# 
ET# 
Study Day from Vaccine 1 
Day < -28 
Screen 
Day 1 
Vaccine 1 
Day 8 
+2 
Day 29 
+2 
Day 36 
Day 57 
Day 64 
 
Day 85 
 
Day 141 Day 169 Day 176 Day 197 Day 253 
Day 
337 
- 
- 
Weeks from Vaccine 1 
 
0 
1 
4 
5 
8 
9 
12 
20 
24 
25 
28 
36 
48 
- 
- 
Study Day from Vaccine 2 
 
 
 
Day 1 
Vaccine 2 
Day 8 
+2 
Day 29 
+2 
Day 36 
Day 57 
Day 113 Day 141 Day 148 Day 169 Day 225 
Day 
309 
- 
- 
Study Day from Vaccine 3 
 
 
 
 
 
Day 1 
Vaccine 3 
Day 8 
+2 
Day 29 
+2 
Day 85 
+/-5 
Day 
113+5 
Day 120 Day 141 Day 197 
Day 
281 
- 
- 
Study Day from Vaccine 4 
 
 
 
 
 
 
 
 
 
Day 1 
Vaccine 4 
Day 8 
+2 
Day 29 
+2 
Day 85 
+/-5 
Day 
169 
+/-7 
- 
- 
Visit Type 
Screen  
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Obtain Informed Consent∞ 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Collect Demographic Information 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Height & Weight 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Physical Examination - Full 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine Dipstick, Opioid testing 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Screening Labs~  
20= & 2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Enrollment/Randomization 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Review Eligibility Criteria 
X 
X†¬1 
 
X†¬ 
 
X†¬ 
 
 
 
X†¬ 
 
 
 
 
 
 
Medical History@ 
X 
X†¬1 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Concomitant Medications 4 
X√ 
X†¬ 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
X 
X 
Vital Signs (oral temp, pulse, BP)% 
X 
X†$ 
 
X† 
 
X† 
 
 
 
X† 
 
 
 
 
X 
X 
Physical Examination – Targeted0 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Pregnancy Test^ 
X 
X† 
 
X† 
 
X† 
 
 
 
X† 
 
 
 
 
 
 
Clinical Safety Evaluations* 
 
6.5 
6.5 
 
6.5 
 
6.5 
 
 
6.5 
6.5 
 
 
 
6.5 
6.5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Study Visit (V)  
00 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
U/S# 
ET# 
Study Day from Vaccine 1 
Day < -28 
Screen 
Day 1 
Vaccine 1 
Day 8 
+2 
Day 29 
+2 
Day 36 
Day 57 
Day 64 
 
Day 85 
 
Day 141 Day 169 Day 176 Day 197 Day 253 
Day 
337 
- 
- 
Weeks from Vaccine 1 
 
0 
1 
4 
5 
8 
9 
12 
20 
24 
25 
28 
36 
48 
- 
- 
Study Day from Vaccine 2 
 
 
 
Day 1 
Vaccine 2 
Day 8 
+2 
Day 29 
+2 
Day 36 
Day 57 
Day 113 Day 141 Day 148 Day 169 Day 225 
Day 
309 
- 
- 
Study Day from Vaccine 3 
 
 
 
 
 
Day 1 
Vaccine 3 
Day 8 
+2 
Day 29 
+2 
Day 85 
+/-5 
Day 
113+5 
Day 120 Day 141 Day 197 
Day 
281 
- 
- 
Study Day from Vaccine 4 
 
 
 
 
 
 
 
 
 
Day 1 
Vaccine 4 
Day 8 
+2 
Day 29 
+2 
Day 85 
+/-5 
Day 
169 
/
- 
- 
Visit Type 
Screen  
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
Clinic 
PBMCs (ICS, LPA) 
 
40 
 
40 
 
40 
 
40 
 
40 
 
40 
 
 
40 
40 
Immunogenicity ANDV PRNT 
 
10 
 
 
 
10 
 
10 
 
10 
 
10 
 
 
 
 
Immunogenicity ANDV PsVNA 
 
10 
 
10 
 
10 
 
10 
 
10 
 
10 
10 
10 
10 
10 
Future Exploratory Assays  
 
 
 
 
 
 
 
40 
 
 
 
40 
 
40 
 
 
Pre-administration reactogenicity 
assessment 
 
X 
 
X 
 
X 
 
 
 
X 
 
 
 
 
 
 
Study Vaccination3 
 
X 
 
X 
 
X 
 
 
 
X 
 
 
 
 
 
 
30-minute evaluation post vax> 
 
X 
 
X 
 
X 
 
 
 
X 
 
 
 
 
 
 
Distribute Memory Aid/Materials 
 
X 
 
X 
 
X 
 
 
 
X 
 
 
 
 
 
 
Review Memory Aid/Site assessment 
 
 
X 
 
X 
 
X 
 
 
 
X 
 
 
 
X 
X 
Assessment of Adverse Events 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
X 
X 
Assessment of SAEs 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
∞  Prior to study procedures. 
†  Prior to study vaccination. 
¬  Review/confirm information or activity in subjects previously consented and screened. 
1  Review results of clinical screening laboratory evaluations. 
@  Complete medical history will be obtained by interview of subjects at the screening visit and will be updated on Day 1 prior to the first study vaccination and interim medical history will be obtained 
by interview of subjects at follow-up visits after the first study vaccination. 
%  Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to taking oral temperature. 
$  Vital signs assessed on Day 1 prior to the first study vaccination will be considered as baseline. 
√  All current medications and medications taken within 60 days prior to signing the ICF. Concomitant medications taken within 30 days of enrollment through Day 28 post last vaccination are collected 
in Advantage eClinicalSM. 
> Observation period will begin after the study vaccination has been given.   
^  Serum pregnancy test will be performed on all females of childbearing potential at screening. A urine pregnancy test will be performed within 24 hours prior to study vaccination and results must be 
negative and known prior to each study vaccination. 

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~  Screening laboratories include: WBC, Hgb, platelet count, ANC, sodium, potassium, Creatinine (Blood Urea Nitrogen [BUN] will be obtained only if creatinine is above the normal range), HgbA1C, 
total bilirubin, Alanine aminotransferase (ALT), hepatitis B surface antigen, hepatitis C antibody, HIV types 1 and 2 
=  All clinical screening laboratory evaluations are to be performed at screening and the values are to be reviewed prior to the first study vaccination on Day 1; no additional blood draws for this are 
required on Day 1.   
&  Retesting of values that lead to exclusion is allowed once using an unscheduled visit during the screening period, provided there is an alternative explanation for the out of range value.  
2  If the initial laboratory screening occurred more than 28 days before baseline (Day 1) but the subject was unable to be vaccinated within the 28-day window (e.g., due to meeting Exclusion Criteria or 
for other reasons), the subject must have laboratories repeated. 
* Safety laboratories include: WBC, Hgb, platelet count, ANC, sodium, potassium, Creatinine, total bilirubin, Alanine aminotransferase (ALT). 
#  Refer to protocol for specific procedures and time points for ET or U/S visit. 
3  The 2mg dosage cohort will receive 1mg/0.5ml administered into the left and right deltoid muscles, one after another or simultaneously. The 4mg dosage cohort will receive 2mg/0.5ml administered into the left and right deltoid 
muscles, one after another or simultaneously.   
()  A targeted physical examination may be performed, if indicated based on review of medical history and any updates obtained by interview of subjects. 
4 Including solicitation for receipt of any non-study vaccines 
 
 

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Appendix B: Acceptable Ranges of Screening Laboratory Measurements 
Lab Test Name 
Clinical 
Laboratory 
Reference Range 
Study Eligibility 
Acceptable 
Lower Limit 
Study 
Eligibility 
Acceptable 
Upper Limit 
Lab 
Unit 
Hemoglobin, male 
13.30 - 17.70 
13.30 
17.70 
g/dL 
Hemoglobin, female 
11.70 - 15.70 
11.70 
15.70 
g/dL 
HgbA1C  
na 
na 
<6.3 
% 
White blood cell count (WBC) (18 to <21 years)^ 
4.5 – 13.0 
3.6  
13.0 
K/mcL 
White blood cell count(WBC) (≥21 years)^ 
4.5 – 11.0 
3.6 
11.0 
K/mcL 
Absolute Neutrophil count* 
1.80 – 7.70 
1.80 
7.70 
K/mcL 
Platelet count 
135 - 466 
135 
466 
K/mcL 
Sodium 
136.0 – 145.0 
136.0 
145.0 
mmol/L 
Potassium 
3.5 – 5.1 
3.5 
5.1 
mmol/L 
Blood urea nitrogen (BUN)† 
9.0 – 23.00 
<9.0 
23.00 
mg/dL 
Serum creatinine† ^, male  
0.60 – 1.10 
<0.60 
1.3 
mg/dL 
Serum creatinine† ^, female 
0.50 – 0.80 
<0.50 
1.2 
mg/dL 
Total bilirubin  
0.10 – 1.2  
0.10 
1.2 
mg/dL 
Alanine transferase† (ALT) 
na 
na 
≤49 
unit/L 
Hepatitis B surface antigen 
Negative 
Negative 
Negative 
 n/a 
Hepatitis C antibodies 
Negative 
Negative 
Negative 
 n/a 
HIV -  1/2 Ab 
Negative 
Negative 
Negative 
 n/a 
Serum hCG (females only) 
Negative 
Negative 
Negative 
n/a 
*  ANC for subjects that are of African American and Middle Eastern descent may have values as low as 0.8 K/mcL. 
Subjects of this descent must have an ANC ≥ 0.8 K/mcL to be eligible to participate in the study if all other study 
criteria are met.   
†  Values of serum creatinine and blood urea nitrogen (BUN) below the lower limit of normal (LLN) are acceptable 
for study enrollment.