A Phase 1 Randomized Study to Evaluate the Safety, Tolerability, and Immunogenicity of Ranging Doses of ALFQ Adjuvant in a Candidate HIV Vaccine Containing A244 and B.65321
Phase: PHASE1
Conditions: Human Immunodeficiency Virus (HIV), AIDS Virus
Sponsor: U.S. Army Medical Research and Development Command

NCT05423418 
 
 
 
A Phase 1 Randomized Study to Evaluate the 
Safety, Tolerability, and Immunogenicity of 
Ranging Doses of ALFQ Adjuvant in a 
Candidate HIV Vaccine Containing A244 and 
B.65321 
 
 
January 7, 2025 

RV-575/WRAIR 2924 Protocol 
 
The Surgeon General 
IND 28352, S-21-03 
 
Department of the Army 
 
Version 2.2 (07JAN2025) 
1 
 
TITLE PAGE 
A Phase 1 Randomized Study to Evaluate the Safety, Tolerability, and 
Immunogenicity of Ranging Doses of ALFQ Adjuvant in a Candidate HIV 
Vaccine Containing A244 and B.63521 
 
Protocol Number: 
RV 575/WRAIR 2924 
 
Compound/Product Number/Name: 
A244, B.63521, and ALFQ Adjuvant 
 
Sponsor: 
The Surgeon General, Department of the Army 
 
Sponsor’s Representative: Senior Regulatory 
Advisor & IND Sponsor’s Representative: 
Emily D. Badraslioglu, MS, PMP, RAC 
US Army Medical Research and Development Command 
(USAMRDC) 
1430 Veterans Drive 
Fort Detrick, MD 21702-5009 
Tel: +1 301-619-0317 
DSN: 343-0317 
Fax Number: +1 301-619-0197 
E-mail: usarmy.detrick.medcom-
usamrmc.mbx.usamrmcregulatory- 
affairs@health.mil 
 
Funding Source:  
Division of AIDS, National Institute of Allergy and 
Infectious Disease, US National Institutes of Health and 
Cooperative Agreement (#W81XWH-18-2-0040) 
 
Regulatory Agency Identifying Number: 
IND 28352 
 
Clinical Trial Site: 
Clinical Trials Center, Room 2w78 
Center for Enabling Capabilities (CEC) 
Walter Reed Army Institute of Research 
503 Robert Grant Avenue 
Silver Spring, MD 20910 
Phone Number: 301-319-9660 
 
KEY ROLES 
 
 
Independent Safety Monitor 
 
 
 
 
 
 
 
 
Melanie McCauley, MD 
Research Physician | HJF 
in support of the Emerging Infectious Diseases Branch 
(EIDB) 
Walter Reed Army Institute of Research (WRAIR) 
6720A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
Phone Number:+1 301- 500-3636 
Email: mmccauley@eidresearch.org 

RV-575/WRAIR 2924 Protocol 
 
The Surgeon General 
IND 28352, S-21-03 
 
Department of the Army 
 
Version 2.2 (07JAN2025) 
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Alternate Independent Safety Monitor 
 
 
 
 
 
 
 
DAIDS Medical Officer: 
 
 
 
 
 
 
 
 
 
 
Roger Ying, MD  
Research Physician | HJF 
U.S. Military HIV Research Program (MHRP) 
6720A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
Email:RYing@global-id.org 
 
 
Laura Polakowski, MD, MSPH 
Medical Officer 
Vaccine Clinical Research Branch 
Vaccine Research Program 
Division of AIDS, NIAID, NIH 
5601 Fishers Lane, 9D11B 
Rockville, MD 20852 
Phone Number: 301-627-3040 
Principal Investigator: 
Brennan R. Cebula, MD, MTM&H  
MAJ, MC, USA 
U.S. Military HIV Research Program (MHRP)  
Walter Reed Army Institute of Research 
6720-A Rockledge Drive, Suite 400 
Bethesda, MD 20817  
Phone Number: 814-233-0841 
E-mail: BCebula@hivresearch.org 
 
Statistical Support Team: 
EMMES 
401 North Washington Street  
Rockville, MD 20852 
Phone Number: 301-251-1161 
 
Data Manager/Statistician: 
Data Coordinating and Analysis Center  
Henry M Jackson Foundation 
6720–A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
 
Research Laboratory: 
US Military HIV Research Program Walter Reed Army 
Institute of Research (WRAIR) 
503 Robert Grant Avenue  
Silver Spring, MD 20910 
 
 
Walter Reed Army Institute of Research 
Specimen Processing Laboratory and Biorepository 
(SPL/BioR) 
Diagnostics and Countermeasures Branch (DCB) 
Center for Infectious Disease Research (CIDR) 
13 Taft Ct., Suite 100 
Rockville, MD 20850 
 
External Research Collaborators are listed in Attachment 2. 

RV-575/WRAIR 2924 Protocol 
 
The Surgeon General 
IND 28352, S-21-03 
 
Department of the Army 
 
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Clinical Laboratories: 
Walter Reed Army Institute of Research 
HIV Diagnostics and Reference Laboratory (HDRL) 
Diagnostics and Countermeasures Branch (DCB) 
Center for Infectious Disease Research (CIDR) 
9100 Brookville Road, Building 508 
Silver Spring, MD 20910 
 
Quest Diagnostics Inc.  
1715 Twin Springs Road 
Baltimore, MD 21227-2997 
 
Institutional Review Boards: 
Walter Reed Army Institute of Research IRB 
US Army Garrison-Forest Glen 
503 Robert Grant Avenue 
Silver Spring, MD 20910  
FWA #00000015 
 

RV-575/WRAIR 2924 Protocol 
 
The Surgeon General 
IND 28352, S-21-03 
 
Department of the Army 
 
Version 2.2 (07JAN2025) 
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PROTOCOL AMENDMENT HISTORY 
 
Version No. Date 
Description 
0.5 
21JUN21 
Team review 
1.0 
04AUG21 
Medical writing review, original version submitted to the Protocol 
Review Board 
1.1 
27SEP21 
Included response to the FDA 
1.2 
14JAN22 
Included DAIDS comments and response to the IRB 
1.3 
04APR22 
WRAIR HSPB comments addressed 
1.4 
21JUN22 
Change in PI and IRB comments addressed 
1.5 
26JUL22 
Response to IRB stipulations 
1.6 
11OCT22 
Response to FDA non-hold comments and includes administrative 
updates (office name changes and contact information) and clinical 
updates 
1.7 
26OCT22 
Updated to clarify compensation language 
1.8 
09NOV22 
Response to WRAIR review: updated blood donation 
inclusion criterion 
1.9 
14MAR23 
Added Planned Interim Analyses - Section 9.6.6 
2.0 
15AUG24 
Updated study personnel to include change in PI from MAJ 
Adjei to MAJ Cebula. 
2.1 
24SEP24 
Response to WRAIR review: updated funding CA number 
2.2 
07JAN25 
Added exploratory objective and endpoint 

RV-575/WRAIR 2924 Protocol 
 
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Department of the Army 
 
Version 2.2 (07JAN2025) 
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INVESTIGATOR’S AGREEMENT 
1. I agree to follow this protocol version as approved by the IRB. 
2. I will conduct the study in accordance with applicable IRB requirements, Federal regulations, 
and state and local laws to maintain the protection of the rights and welfare of study 
participants. 
3. I certify that I, and the study staff, have received the requisite training to conduct this research 
protocol. 
4. I will not modify the protocol without first obtaining an IRB approved amendment and new 
protocol version unless it is necessary to protect the health and welfare of study participants. 
PRB approval of amended study documents prior to IRB submission is required. 
5. I have read and understand the information in the Investigators’ Brochure (or Manufacturer’s 
Brochure) regarding the risks and potential benefits. I agree to conduct the protocol in 
accordance with Good Clinical Practices (ICH-GCP), the applicable ethical principles, the 
Statement of Investigator (Form FDA 1572), and with local regulatory requirements. In 
accordance with the U.S. FDA Modernization Act and NIH policy, I will ensure the 
registration and reporting results of the trial on the www.clinicaltrials.gov website. 
6. I will ensure that the data (and/or specimens) are maintained in accordance with the data 
(and/or specimen) disposition outlined in the protocol. Any modifications to this plan should 
first be reviewed and approved by the WRAIR IRB. 
7. I will promptly report changes to the research or unanticipated problems to the WRAIR IRB 
immediately via the WRAIR Human Subjects Protection Branch at 301-319-9940 (during 
duty hours) or to usarmy.detrick.medcom-wrair.mbx.hspb@health.mil and submit a written 
report within 10 working days of knowledge of the event. 
8. I will prepare continuing review reports at an interval established by the IRB, and a study 
closure report when all research activities are completed. 
9. I will immediately report to the WRAIR Human Subjects Protection Branch knowledge of any 
pending compliance inspection by any outside governmental agency. 
10. I agree to maintain adequate and accurate records in accordance with IRB policies, Federal, 
state, and local laws and regulations. 
 
 
Brennan R. Cebula, MD, MTM&H Principal Investigator 
Date 
 
 
 

RV-575/WRAIR 2924 Protocol 
 
The Surgeon General 
IND 28352, S-21-03 
 
Department of the Army 
 
Version 2.2 (07JAN2025) 
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TBLE OF CONTENTS, LIST OF TABLES, AND LIST OF FIGURES 
TABLE OF CONTENTS 
TITLE PAGE ................................................................................................................................... 1 
PROTOCOL AMENDMENT HISTORY ....................................................................................... 4 
INVESTIGATOR’S AGREEMENT ............................................................................................... 5 
TABLE OF CONTENTS, LIST OF TABLES, AND LIST OF FIGURES .................................... 6 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ............................................... 11 
1. 
PROTOCOL SUMMARY ....................................................................................... 14 
1.1. 
Synopsis ....................................................................................................... 14 
1.2. 
Schema ......................................................................................................... 21 
1.3. 
Schedule of Activities .................................................................................. 22 
2. 
INTRODUCTION ................................................................................................... 27 
2.1. 
Study Rationale ............................................................................................ 27 
2.2. 
Background .................................................................................................. 28 
2.2.1. A244d11 gp120 ................................................................................ 28 
2.2.2. B.63521 ............................................................................................ 29 
2.2.2.1. 
Non-Human Primate Study ............................................ 29 
2.2.3. ALFQ ............................................................................................... 30 
2.3. 
Military Relevance ....................................................................................... 32 
2.4. 
Benefit/Risk Assessment ............................................................................. 32 
2.4.1. Local Reactions ................................................................................ 32 
2.4.2. Systemic Reactions .......................................................................... 33 
2.4.3. Risks to Study Personnel and the Environment ............................... 33 
2.4.4. Other Risks ....................................................................................... 33 
2.4.5. Unknown Risks ................................................................................ 34 
2.4.6. Alternative to this Product or Study ................................................. 35 
2.4.7. Benefits ............................................................................................ 35 
3. 
OBJECTIVES AND ENDPOINTS ......................................................................... 35 
4. 
STUDY DESIGN ..................................................................................................... 36 
4.1. 
Overall Study Design ................................................................................... 36 
4.2. 
Number of Participants ................................................................................ 37 
4.3. 
Sentinel Group Allocation ........................................................................... 37 
4.4. 
Scientific Rational for Study Design ........................................................... 37 
4.5. 
Justification for Dose ................................................................................... 38 
4.6. 
Method of Treatment Assignment ............................................................... 38 
4.7. 
End of Study Definition ............................................................................... 38 
5. 
STUDY POPULATION .......................................................................................... 38 
5.1. 
Participant Recruitment ............................................................................... 38 
5.2. 
Participant Identification Numbers .............................................................. 39 
5.3. 
Participant Eligibility ................................................................................... 40 
5.3.1. Inclusion Criteria ............................................................................. 40 
5.3.2. Exclusion Criteria ............................................................................ 41 

RV-575/WRAIR 2924 Protocol 
 
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5.4. 
Lifestyle and Reproduction Restrictions ...................................................... 43 
5.4.1. Contraception and Pregnancy .......................................................... 43 
5.4.1.1. 
Individuals of Childbearing Potential ............................ 43 
5.4.1.2. 
Participants of Childbearing Potential ........................... 44 
5.4.1.3. 
Effective Methods of Contraception .............................. 44 
5.4.1.4. 
Pregnancy Testing .......................................................... 45 
6. 
STUDY INTERVENTION ...................................................................................... 45 
6.1. 
Treatments Administered ............................................................................. 45 
6.2. 
Preparation, Handling, Storage, and Accountability .................................... 46 
6.2.1. Acquisition Accountability .............................................................. 46 
6.2.2. Product Storage and Stability ........................................................... 46 
6.2.3. Product Preparation .......................................................................... 47 
6.3. 
Study Intervention Compliance ................................................................... 47 
6.4. 
Measures to Minimize Bias: Randomization and Blinding ......................... 47 
6.5. 
Concomitant Therapy ................................................................................... 48 
6.6. 
Dose Modification ....................................................................................... 48 
6.7. 
Treatment After the End of the Study .......................................................... 48 
7. 
DISCONTINUATION CRITERIA ......................................................................... 48 
7.1. 
Screening Failures ........................................................................................ 48 
7.2. 
Discontinuation of Study Intervention ......................................................... 49 
7.2.1. PSRT ................................................................................................ 49 
7.2.2. Pregnancy ......................................................................................... 50 
7.3. 
Participant Discontinuation/Withdraw from the Study ................................ 50 
7.3.1. Replacement of Withdrawn Participants ......................................... 51 
7.3.2. Follow-up for Participants Who Withdraw Consent ....................... 51 
7.4. 
Lost to Follow-up ......................................................................................... 51 
8. 
STUDY ASSESSMENTS AND PROCEDURES ................................................... 51 
8.1. 
Safety Assessments ...................................................................................... 52 
8.1.1. Demographic/Medical History ......................................................... 52 
8.1.2. Physical Examination ....................................................................... 52 
8.1.3. Vital Signs ........................................................................................ 52 
8.1.4. Clinical Laboratory Assessments ..................................................... 52 
8.1.4.1. 
Hematology .................................................................... 53 
8.1.4.2. 
Blood Chemistry ............................................................ 53 
8.1.4.3. 
Laboratory assays ........................................................... 53 
8.1.4.4. 
Pregnancy Screen ........................................................... 54 
8.2. 
Adverse Events ............................................................................................ 54 
8.2.1. Definitions ........................................................................................ 54 
8.2.1.1. 
Adverse Event and Suspected Adverse Reaction .......... 54 
8.2.1.2. 
Solicited Adverse Event ................................................. 54 
8.2.1.3. 
Serious Adverse Event ................................................... 55 
8.2.1.4. 
HIV Diagnostic Algorithm ............................................ 56 
8.2.1.5. 
Medically Attended Adverse Events (MAAE) .............. 56 
8.2.1.6. 
Unexpected Adverse Event or Unexpected Suspected 
Adverse Reaction ........................................................... 57 
8.2.2. Timeframe for Collecting Adverse Event Information .................... 57 

RV-575/WRAIR 2924 Protocol 
 
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8.2.3. Methods for Obtaining AE and SAE Data ....................................... 57 
8.2.4. Recording Adverse Event and Serious Adverse Event Data ........... 58 
8.2.5. Severity Assessment ........................................................................ 58 
8.2.6. Estimating Severity Grade for Parameters Not Identified in the 
Grading Table .................................................................................. 58 
8.2.7. Exceptions to the Grading Table ...................................................... 59 
8.2.7.1. 
Creatinine ....................................................................... 59 
8.2.7.2. 
Injection Site Erythema and Induration ......................... 59 
8.2.7.3. 
Unintentional Weight Loss ............................................ 59 
8.2.8. Causality Assessment ....................................................................... 60 
8.2.8.1. 
Serious Adverse Events ................................................. 60 
8.2.9. Sponsor Independent Safety Monitor .............................................. 60 
8.2.10. Reporting Serious and Unexpected Adverse Events ....................... 61 
8.2.10.1. Reporting to WRAIR IRB ............................................. 62 
8.2.11. Additional Immediately Reportable Events ..................................... 62 
8.2.11.1. Potentially Immune-Mediated Medical Conditions 
(PIMMCs) ...................................................................... 62 
8.2.11.2. Unanticipated Problems Involving Risks to Subjects or 
Others ............................................................................. 63 
8.2.11.3. Study Pause or Termination ........................................... 63 
8.2.11.4. Participant Status Change: Incarceration ....................... 63 
8.2.11.5. AE-Related Withdrawal of Consent .............................. 64 
8.2.12. Pending Inspections/Issuance of Reports ......................................... 64 
8.2.13. IND Annual Report to the FDA ....................................................... 64 
8.2.14. Final Report...................................................................................... 64 
8.2.15. Pregnancy Reporting ........................................................................ 65 
8.2.16. Hepatitis B and C and HIV Reporting to the Local Health 
Authorities ........................................................................................ 66 
8.2.17. Serious Adverse Event Follow-up ................................................... 66 
8.2.18. Safety Monitoring Committee ......................................................... 66 
8.3. 
Immunogenicity Assessments ...................................................................... 67 
8.3.1. Specification of Immunogenicity Endpoints ................................... 67 
8.3.2. Immunogenicity Endpoints .............................................................. 67 
8.3.3. Humoral Immune Responses ........................................................... 68 
8.3.4. Cellular Immune Responses ............................................................. 68 
8.3.5. Innate Immunity ............................................................................... 69 
8.4. 
Specimen Archiving and Transfer ............................................................... 69 
8.5. 
Future Use and Storage of Data and Biological Samples ............................ 69 
9. 
STATISTICS ........................................................................................................... 70 
9.1. 
Statistical Criteria for the Termination of the Trial ..................................... 70 
9.2. 
Procedures for Reporting Deviations from the Statistical Plan ................... 70 
9.3. 
Accounting for Missing, Unused, and Spurious Data .................................. 70 
9.4. 
Sample size and Power calculation .............................................................. 71 
9.5. 
Populations for Analyses ............................................................................. 72 
9.6. 
Statistical Analyses ...................................................................................... 73 
9.6.1. Demographics and Baseline Characteristics .................................... 73 
9.6.2. Safety Analysis ................................................................................ 73 

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9.6.3. Immunogenicity Analysis ................................................................ 73 
9.6.4. Missing Values and Outliers ............................................................ 74 
9.6.5. Clinical Laboratory Data Analyses .................................................. 74 
10. 
ETHICS .................................................................................................................... 75 
10.1. Ethics Review .............................................................................................. 75 
10.1.1. Protocol Modifications ..................................................................... 75 
10.1.2. Protocol Deviations .......................................................................... 76 
10.2. Ethical Conduct of the Study ....................................................................... 76 
10.3. Confidentiality ............................................................................................. 77 
10.4. Compensation for Participation ................................................................... 77 
10.5. Medical Care for Research-Related Injury .................................................. 78 
10.6. COVID-19 Information ............................................................................... 79 
10.7. Source Documents ....................................................................................... 79 
10.8. Study and Site Closure ................................................................................. 80 
10.9. Written Informed Consent ........................................................................... 80 
11. 
ADMINISTRATIVE ............................................................................................... 81 
11.1. Study Monitoring ......................................................................................... 81 
11.2. Data Quality Assurance ............................................................................... 81 
11.3. Record Retention ......................................................................................... 82 
11.4. Pending Inspections/Issuance of Reports ..................................................... 82 
11.5. Reports and Publications .............................................................................. 82 
11.5.1. Annual Report to the FDA ............................................................... 82 
11.5.2. Final Clinical Study Report .............................................................. 82 
11.5.3. Continuing Review and Closeout Reports ....................................... 83 
11.5.4. Publication Policy ............................................................................ 83 
11.6. Roles and Responsibilities ........................................................................... 83 
12. 
LIST OF REFERENCES ......................................................................................... 84 
13. 
APPENDICES ......................................................................................................... 86 
APPENDIX A. 
DAIDS TABLE FOR GRADING THE SEVERITY OF ADVERSE 
EVENTS .................................................................................................................. 87 
APPENDIX B. 
ROLES AND RESPONSIBILITIES ........................................................... 88 
APPENDIX C. 
PARTICIPANT VISIT SCHEDULE ........................................................... 92 
APPENDIX D. 
INFORMED CONSENT FORM ................................................................. 93 
APPENDIX E. 
TEST OF UNDERSTANDING ................................................................... 94 
APPENDIX F. 
RECRUITMENT SCRIPT ........................................................................... 95 
APPENDIX G. 
INFORMATION SHEET REGARDING COMPENSATION TO 
FEDERAL PERSONNEL WHEN THEY PARTICIPATE IN RESEARCH ......... 96 
APPENDIX H. 
VISP LETTER ............................................................................................. 97 
APPENDIX I. 
UNBLINDING LETTER ............................................................................. 98 
APPENDIX J. 
EMERGENCY CONTACT CARD ............................................................. 99 
APPENDIX K. 
BRIEFING SLIDES ................................................................................... 100 

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Department of the Army 
 
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APPENDIX L. 
LIST OF POTENTIALLY IMMUNE-MEDIATED MEDICAL 
CONDITIONS ....................................................................................................... 101 
APPENDIX M. RISK ASSESSMENT TOOL............................................................................ 104 
ATTACHMENT 1: PROTOCOL TEAM ROSTER ................................................................... 105 
ATTACHMENT 2: TABLE OF EXTERNAL COLLABORATORS ........................................ 109 
 
LIST OF TABLES 
Table 1: 
Schedule of Evaluations (SOE) .................................................................................. 22 
Table 2: 
Protocol-Required Safety Laboratory Assessments ................................................... 26 
Table 3: 
Objectives and Endpoints ........................................................................................... 35 
Table 4: 
Products Administered ............................................................................................... 45 
Table 5: 
DAIDSAdverse Event Grading Scale ........................................................................ 59 
Table 6: 
Causality Relationships .............................................................................................. 60 
Table 7: 
Study Contacts for Reporting Serious and Unexpected Adverse Events ................... 61 
Table 8: 
Sponsor’s Adverse Event Reporting Requirements ................................................... 65 
Table 9: 
Summary of Information Required When Reporting Adverse Events to the 
Sponsor ....................................................................................................................... 65 
Table 10: Immunology Assays ................................................................................................... 67 
Table 11: Sample size per arm and minimum effect size with 80% power, assuming 
residual standard deviation of 0.2 ............................................................................... 71 
Table 12: Exact 95% Clopper-Pearson CI for the SAE Rate in Each Study Group ................... 72 
Table 13: Analysis Populations .................................................................................................. 72 
 
LIST OF FIGURES 
Figure 1: 
Study Design .............................................................................................................. 21 
Figure 2: 
Reactogenicity Results for FMP013 and FMP014 ..................................................... 31 
 
 

RV-575/WRAIR 2924 Protocol 
 
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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
Abbreviation 
or Specialist 
Term 
Explanation 
3D-PHAD® 
Synthetic monophosphoryl lipid A 
AAE 
Acquired angioedema 
Ab 
Antibody 
ADCC 
Antibody-Dependent Cell-Mediated Cytotoxicity 
ADCP 
Antibody-Dependent Cell-Mediated Phagocytosis 
AE 
Adverse event 
AESIs 
Adverse events of special interest 
AIDS 
Acquired immunodeficiency disease syndrome 
AHFG 
Aluminum Hydroxide Fluid Gel 
ALF 
Army Liposome Formulation 
ALFQ 
ALF mixed with the saponin QS21 
AlOH 
Aluminum hydroxide 
ALT 
Alanine aminotransferase 
ATO 
Army Technology Objective 
AUC 
Area under the curve 
CD4 
Cluster of Differentiation 
CFSE 
Carboxyfluorescein succinimidyl ester, a fluorescent cell staining dye 
CRFs 
Case Report Forms 
CRR 
Continuing review report 
CTC 
WRAIR Clinical Trials Center 
DAIDS 
Division of AIDS 
DODI 
Department of Defense Instruction 
DSMB 
Data Safety Monitoring Board 
eCRF 
Electronic Case Report Forms 
EDC 
Electronic Data Capture 
EDTA 
Ethylenediaminetetraacetic acid 
ELISPOT 
Enzyme-Linked Immunospot 
FDA 
U.S. Food and Drug Administration 
FSH 
Follicle stimulating hormone 
GBS 
Guillain-Barré syndrome 
GCLP 
Good Clinical Laboratory Practices 
GCP 
Good Clinical Practices 
HAE 
Hereditary angioedema 
HBsAg 
Hepatitis B surface antigen 
HIV-1 
Human Immunodeficiency Virus 1 
HRT 
Hormonal replacement therapy 
HSPB 
Human Subjects Protection Branch, WRAIR 
IBs 
Investigational brochures 
ICH 
International Conference on Harmonization 
ICS 
Intra-cellular cytokine staining 
ID 
Infectious disease 

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Abbreviation 
or Specialist 
Term 
Explanation 
IEC 
Independent Ethics Committee 
IFNγ 
Interferon gamma 
IgA 
Immunoglobulin A 
IgG 
Immunoglobulin G 
IL2 
Interleukin 2 
IM 
Intramuscular 
IND 
Investigational New Drug 
IP 
Investigational product 
IQR 
Interquartile range 
IRB 
Institutional Review Board 
ISM 
Sponsor Independent Safety Monitor 
IUD 
Intrauterine device 
IUS 
Intrauterine hormone-releasing system 
LPLV 
Last Participant Last Visit 
MAAE 
Medically Attended Adverse Events 
MHRP 
U.S. Military HIV Research Program 
MOP 
Manual of Procedures 
MPLA 
Monophosphoryl lipid A 
NAAT 
Nucleic Acid Testing 
NCO 
Non-commissioned officer 
NHP 
Non-human primate 
NK 
Natural Killer Cell 
NKT 
Natural Killer T cell 
NSAIDs 
Nonsteroidal anti-inflammatory drugs 
OAE 
Other significant adverse events 
OHARO 
Office of Human and Animal Research Oversight 
OHRO 
Office of Human Research Oversight 
ORA 
Office of Regulated Activities 
PBMC 
Peripheral blood mononuclear cell 
PI 
Principal Investigator 
PIMMCs 
Potentially Immune-Mediated Medical Conditions 
PIN 
Participant identification number 
PRB 
Protocol Review Board 
PSRT 
Protocol Safety Review Team 
PSSO 
Product Safety Surveillance Office 
PVG 
Pharmacovigilence 
RFADCC 
Rapid and fluorometric ADCC 
SAE 
 
Serious adverse event 
SD 
Standard deviation 
SGTP 
Serum glutamic pyruvic transaminase 
SOE 
Schedule of Evaluations 
SOP 
Standard operating procedure 
STI 
Sexually transmitted infection 

RV-575/WRAIR 2924 Protocol 
 
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Abbreviation 
or Specialist 
Term 
Explanation 
TOU 
Test of understanding 
UPIRTSO 
Unanticipated problems involving risks to subjects or others 
USAMRDC 
United States Army Medical Research and Development Command 
WRAIR 
Walter Reed Army Institute of Research 

RV-575/WRAIR 2924 Protocol 
 
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1. 
PROTOCOL SUMMARY 
1.1. 
Synopsis 
Name of Sponsor: 
The Surgeon General, Department of the Army 
Name of Investigational Product: 
A244, B.63521, and ALFQ 
Name of Active Ingredient: 
A244 consists of the gp120 envelope glycoprotein HIV-1 subtype CRF_01E A244 derived from the 
CM244 CRF_01AE. The A244 gp120 envelope has an 11 amino N-terminal deletion, similar to the 
A244 protein used in AIDSVAX B/E. 
B.63521 gp120 is a recombinant HIV-1 Env protein containing an 11 amino acid truncation at the N-
terminus of gp120 that enhances antigenicity and immunogenicity. 
ALFQ (Army Liposomal Formulation) is a liposomal adjuvant containing a synthetic 
monophosphoryl lipid A (MPLA) with the addition of QS-21. 
Protocol Number(s): RV 575 
Title of Study: A Phase 1, Randomized Study to Evaluate the Safety, Tolerability, and 
Immunogenicity of Ranging Doses of ALFQ Adjuvant in a Candidate HIV Vaccine Containing A244 
and B.63521  
Study Site(s): WRAIR Clinical Trials Center (CTC) 
Principal Investigator: 
MAJ Brennan R. Cebula, MD, MTM&H 
*Protocol Team Roster included as Attachment 1. 
Studied Period (years): 
Estimated date first participant enrolled: May 2022 Estimated 
date last participant completed: October2023 
Study duration: Specimen analysis and monitoring of 
medically attended adverse events (AEs) will continue 12 
months beyond the final participant vaccination. Anticipated 
end date is October 2028. 
Phase of Development: Phase I 
Objectives: 
Primary: 
To evaluate the safety and tolerability (including reactogenicity) of candidate vaccine A244/B.63521 
(300 μg each) with ALFQ adjuvant at the 200 μg, 100 μg, and 50 μg doses  
Secondary: 
To evaluate the effect of candidate vaccine A244/B.63521 (300 μg each) with ALFQ adjuvant at the 
200 μg, 100 μg, and 50 μg doses on cellular, and humoral immune responses 
Exploratory: 
To characterize B-cell functional specificities for each vaccination regimen 
To characterize innate immunity 
To assess the innate/gene expression induced across vaccination regimens 
To perform systems serology analyses 

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Name of Sponsor: 
The Surgeon General, Department of the Army 
To characterize the impact of vaccination with the ALFQ adjuvant on the permissiveness and 
expression to in vitro HIV infection of CD4+ T cells and differentiated macrophages 
 
Endpoints 
Primary 
Evaluate the occurrence and severity of solicited local and systemic AEs following candidate vaccine 
administration. 
Evaluate the occurrence, severity, and relationship to vaccination of unsolicited AEs after candidate 
vaccine administration. 
Evaluate the occurrence of SAEs, and new-onset medical conditions. 
Evaluate the occurrence of AEs of special interest (AESIs) following candidate vaccine 
administration. 
Secondary 
Compare plasma Immunoglobulin G (IgG) binding antibodies to gp120 in terms of magnitude, 
durability, and area under the curve among groups with differing ALFQ doses. 
Characterize and assess the magnitude of cell-mediated immune responses elicited across vaccination 
regimens including antigen -specific CD4 and CD8 T cell responses and polyfunctionality. 
Exploratory 
Characterize plasma IgG and Immunoglobulin A (IgA) binding antibodies to HIV gp120, 
neutralizing antibodies, and non-neutralizing effector functions such as Antibody-Dependent Cell-
Mediated Cytotoxicity (ADCC) and Antibody-Dependent Cell-Mediated Phagocytosis (ADCP) with 
emphasis on RV144 immune correlates of risk of HIV acquisition. 
Characterize B-cell functional specificities for each vaccination regimen by quantifying antigen-
specific responses through B-cell Enzyme-Linked Immunospot (ELISPOT), phenotyping the 
magnitude and activation status of B cell subsets via flow cytometry and isolation of monoclonal 
antibodies from selected vaccine recipients. 
Characterize innate immunity through quantification of soluble chemokines and cytokines and 
assessing the phenotype and function of cellular innate immune subsets such as Natural Killer Cell 
(NK), NKT, and dendritic cells. 
Characterize effects of host genetic polymorphisms on immune responses and characterize 
differentially expressed genes. 
Characterize the impact of vaccination with the ALFQ adjuvant on the permissiveness and expression 
to in vitro HIV infection of CD4+ T cells and differentiated macrophages through integrated HIV-
DNA PCR, cell associated-DNA and RNA, flow cytometry of various activation markers and HIV 
Gag protein, ELISA of HIV p24 in the cell culture supernatant, and production of 
cytokines/chemokines. 
Methodology 
This study is exploratory in nature. 
The purpose of this study is to optimize ALFQ dosing. We will also evaluate the B.63521 protein. 
Safety will be assessed through the frequency of the overall and specific post-vaccination reactions. 
Blood will be collected to assess humoral, cell-mediated, and innate immune responses. 
Healthy adults not living with HIV, who are available for 14 months will be enrolled. A total of 60 
participants will be enrolled in one of three arms, each comprised of 20 candidate vaccine recipients. 
Each arm will receive identical doses of A244 and B.63521 (300 micrograms of each protein). In 

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addition, Arm 1 will receive 200 micrograms of ALFQ. Arm 2 will receive 100 micrograms of 
ALFQ. Arm 3 will receive 50 micrograms of ALFQ.  The safety, reactogenicity, and immunogenicity 
will then be compared among the three arms to determine the optimal dose of ALFQ. 
All vaccinations will be split into 2 half doses, which will be administered intramuscularly (IM) into 
the same deltoid muscle. Vaccinations will occur at months 0, 1, and 2. The second vaccination will 
be administered into the contralateral deltoid at study month 1 compared to the first 
vaccination at study month 0. The third vaccination at study month 2 will be administered into 
the same deltoid as the first vaccination at study month 0. Participants will be followed for 12 
months following the last study vaccination. 
Number of Subjects: 
Maximum number of subjects to be screened: 150 
Maximum number of subjects to be enrolled: 60 
Inclusion Criteria 
Potential participants are required to meet all of the following criteria for enrollment into the study. 
1. Healthy adults between the ages 18-55 years (inclusive) 
2. Must be at low risk for HIV infection per investigator assessment and using the study risk 
assessment tool. 
3. Able and willing to provide written, informed consent 
4. Able and willing to comply with all research requirements, in the opinion of the Investigator 
5. Agreement to refrain from blood donation during the course of the study and within the 2 
months prior to study entry 
6. Minimum body weight of 110 pounds (lbs) (50kg) 
7. Laboratory Criteria within 45 days before enrollment: 
a. Hemoglobin ≥ 11.7 g/dL for women; ≥ 12.5 g/dL for men 
b. White Blood Cell count = 3,500-10,800 cells/mm3 
c. Platelets ≥140,000/mm3 and ≤ 450,000/mm3 
d. Alanine aminotransferase (ALT; SGPT) <1.25 x ULN 
e. Serum creatinine ≤ 1.25 x institutional upper limit of the reference range 
f. 
Negative HIV testing (HIV Ab / antigen 4th generation screen with reflex 
confirmatory RNA testing) 
g. Negative HBsAg and hepatitis C antibody testing 
Note: As above, Grade 1 lab abnormalities detected on screening may be repeated at PI discretion. 
Persistent Grade 1 abnormalities that are felt to represent the non-pathologic baseline for the subject 
will be documented before a subject is enrolled in the trial and are allowable per discretion of the PI. 
8. Birth control requirements: 
All participants assigned female at birth must meet one of the following two criteria: 
a. No reproductive potential due to post-menopausal status (12 months of natural 
[spontaneous] amenorrhea) or hysterectomy, bilateral oophorectomy, or tubal ligation 
b. People of childbearing potential should agree to practice highly effective contraception at 
least 30 days before enrollment and through 3 months post-last vaccination, using one of 
the following methods: condoms (male or female) with spermicide; diaphragm, or 
cervical cap with spermicide; intrauterine device; contraceptive pills, patch, injection, 

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Name of Sponsor: 
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intravaginal ring or other FDA-approved contraceptive method; male partner has 
previously undergone a vasectomy; abstinence  
c. All participants are encouraged to engage in safe sex practices to prevent HIV acquisition  
9. For all participants assigned female at birth, except those with a history of hysterectomy or 
bilateral oophorectomy, a negative β-HCG pregnancy test (urine) on the day of enrollment 
and each vaccination day is required. Because tubal ligations have a failure rate that is not 
insignificant, and because 12 months of spontaneous amenorrhea can be a result of 
polycystic ovarian syndrome and does not completely preclude pregnancy, a negative β-HCG 
pregnancy test at enrollment and on each vaccination day is also required for participants 
assigned female at birth with a history of either of these). 
10. No plans to travel outside the Washington DC metro area (DC, Maryland, and Virginia) that 
would prevent compliance with planned study visits 
11. Test of Understanding (TOU) (minimum passing score of 80% with 2 attempts permitted) 
Exclusion Criteria 
Potential participants meeting any of the following criteria will be excluded from enrollment. 
1. Receipt of any investigational HIV vaccine or investigational adjuvant 
2. Received an investigational product in the 30 days before enrollment, or planned to receive 
during the study period. This does not include products with emergency use 
authorization. 
3. Concurrent participation in another clinical research study 
4. Any serious medical illness or condition  
5. Receipt of immunoglobulins or blood products within 3 months before enrollment 
6. Any history of anaphylaxis or allergy to study product 
7. History of sickle cell trait or disease 
8. Pregnancy, lactation, or intention to become pregnant during the study 
9. History of active/recent cancer still within treatment or active surveillance follow-up (except 
basal cell carcinoma of the skin and cervical carcinoma in situ). Treated/resolved cancers 
with no likelihood of recurrence may be deemed acceptable at Principal Investigator (PI) 
discretion. 
10. History of autoimmune disease 
11. History of Potentially Immune-Mediated Medical Conditions (PIMMCs) 
12. Suspected or known current alcohol or drug abuse as defined by an alcohol intake of greater 
than 3 drinks a day on average for a man, and greater than 2 drinks a day on average for a 
woman 
13. Any other significant disease, disorder or finding which may significantly increase the risk to 
the volunteer because of participation in the study, affect the ability of the volunteer to give 
informed consent, participate in the study, or impair interpretation of the study data, in the 
opinion of the Investigator 
14. History of splenectomy 
15. History of confirmed or suspected immunodeficiency 
16. History of hereditary angioedema (HAE) acquired angioedema (AAE), or idiopathic forms of 
angioedema 

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17. History of asthma that is unstable or required emergent care, urgent care, hospitalization, or 
intubation during the past 2 years 
18. History of diabetes mellitus (type I or II), with the exception of gestational diabetes 
19. History of thyroid disease (except for well controlled hypothyroidism) 
20. History of idiopathic urticaria within the past year 
21. History of hypertension that is not well controlled by medication or that is persistently 
greater than 140/95 at screening 
22. History of bleeding disorder diagnosed by a doctor (e.g., factor deficiency, coagulopathy, or 
platelet disorder requiring special precautions) or significant bruising or bleeding difficulties 
with IM injections or blood draws 
23. History of chronic or active neurologic disease to include seizure disorder and chronic 
migraine headaches. Exceptions are: i) childhood febrile seizures, or ii) seizures secondary to 
alcohol withdrawal more than 3 years ago 
24. Subjects receiving any of the following substances: 
a. Systemic immunosuppressive medications or cytotoxic medications within 12 weeks 
before enrollment [with the exception of a short course of corticosteroids (≤ 14 days 
duration or a single injection) for a self-limited condition at least 2 weeks before 
enrollment; inhaled, intranasal or topical steroids are not considered exclusionary 
b. Treatment with known immunomodulators including allergy immunotherapy (other 
than nonsteroidal anti-inflammatory drugs [NSAIDs] or stable maintenance 
immunotherapy (doses not in the process of being increased), at the discretion of the 
Protocol Safety Review Team (PSRT)) for any reason 
c. Live attenuated vaccines within 30 days before initial study vaccine administration 
d. Medically indicated subunit, mRNA, or killed vaccines, e.g., influenza, 
pneumococcal, vaccines with QS-21 as an adjuvant, or allergy treatment with antigen 
injections, planned for administration 14 days before or after study vaccine 
administration 
25. History of arthritis diagnosis other than osteoarthritis 
26. History of other diagnosed rheumatoid disorders 
27. Has an acute illness or temperature ≥38.0 ºC/100.4 ºF on any study injection day or within 48 
hours of planned study injection. 
Note: Participants will not be excluded from further consideration for enrollment and study 
injections. Volunteers with fever or an acute illness on the day of study injection or in the 2 days 
before the study injection may be re-assessed by a study physician for resolution of the condition and 
enrolled and receive the study injection so long as the injection is within allowable windows. Military 
personnel will be excluded from participation in this study, regardless of leave status due to the 
potential for a false-positive HIV test result on mandatory HIV testing. This could have adverse 
effects on deployment status. 
Final evaluation of eligibility will be based on the medical judgment of the investigator based on 
his/her medical and research experience. 
 
 

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Investigational Product, Dosage, and Mode of Administration 
Arm 1: An injection containing A244/B.63521 with 200 μg of ALFQ adjuvant will be administered as 
an IM injection into the deltoid muscle at visits 1, 3, and 5 (corresponding to Day 1, Day 29, and Day 
57). 
Arm 2: An injection containing A244/B.63521 with 100 μg of ALFQ adjuvant will be administered as 
an IM injection into the deltoid muscle at visits 1, 3, and 5 (corresponding to Day 1, Day 29, and Day 
57). 
Arm 3: An injection containing A244/B.63521 with 50 μg of ALFQ adjuvant will be administered 
as an IM injection into the deltoid muscle at visits 1, 3, and 5 (corresponding to Day 1, Day 29, and 
Day 57). 
Investigational Products 
Products: A244, B.63521, and ALFQ 
Dosage: 200 μg, 100 μg, and 50 μg of ALFQ (Dose of MPLA/3D-PHAD® component), 300 μg 
A244, and 300 μg of B.63521 
Adjuvant: ALFQ (200 μg of MPLA/3D-PHAD® and 100 μg of QS-21 diluted in 0.5 mL Sorenson’s 
buffer)  
Total Volume: 1.8 mL 
Mode and Location of Administration: IM injection into the same deltoid muscle (same side) for all 
injections.  
Duration of Study Participation: Participants will receive IM administration of the 
investigational products at months 0, 1, and 2, after which they will be followed for 12 months 
after the 3rd injection for safety analyses. Total duration of participation will be 14 months. 
Criteria for Evaluation: 
Immunogenicity: 
Humoral responses will be assessed by HIV-specific binding antibody assays, HIV-specific 
neutralizing antibody assays, and non-neutralizing antibody function assays. 
HIV-specific cell-mediated responses will be assessed by intracellular cytokine staining, CD4 and 
CD8 T cell proliferation, and related assays. Innate immune responses and genetic responses will be 
quantified. 
Safety: 
Safety will be assessed both by direct physical examination and by diary cards, which serve as 
memory tools for better identification of reactions. Participants will be assessed at pre- vaccination 
baseline and will remain in clinic under direct observation for at least 30 minutes post vaccination. 
Participants will return to clinic post-vaccination to be assessed for symptoms of local and systemic 
reactogenicity. Results will then be compared among the three ALFQ dose arms. 
Participants will be followed for a total of 12 months post final vaccination. Serious Adverse Events 
and SAEs will be recorded at all visits along with timing and possible attribution to Investigational 
Product. Because this clinical trial involves an adjuvant, Adverse Events of Special Interest AESIs 
and Medically Attended Adverse Events (MAAE) will also be assessed. Safety laboratory analyses of 
complete blood count, liver function tests, urinalysis, and pregnancy test in females will also be 
performed. Safety results will then be compared among dose arms. 

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Statistical Methods: 
RV 575 is designed to optimize ALFQ dosing. The primary objective is to compare dose- dependent 
safety and reactogenicity of ALFQ. For safety analyses, the occurrence of local and systemic 
reactogenicity symptoms, AEs, and SAEs will be assessed as the proportions of participants 
experiencing such safety events and compared among the three study arms. The secondary objective 
is to assess immunogenicity. 

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1.2. 
Schema 
The study design is illustrated in Figure 1. 
Figure 1: 
Study Design 
Number of participants (N) = 60 
Vaccination schedule in months (days) 
 
Number of 
participants (N) 
Month 0 (Day 1) 
Month 1 (Day 29) 
Month 2 (Day 57) 
Arm 1 
20 
300μg A244 
300μg B.63521 200μg 
ALFQ 
300μg A244 
300μg B.63521 200μg 
ALFQ 
300μg A244 
300μg B.63521  
200μg ALFQ 
Arm 2 
20 
300μg A244 
300μg B.63521 100μg 
ALFQ 
300μg A244 
300μg B.63521 100μg 
ALFQ 
300μg A244 
300μg B.63521 
100 μg ALFQ  
Arm 3 
20 
300μg A244 
300μg B.63521 50μg 
ALFQ 
300μg A244 
300μg B.63521 50μg 
ALFQ 
300μg A244 
300μg B.63521 50μg 
ALFQ 

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1.3. 
Schedule of Activities 
The detailed schedule of study events is provided in Table 1 and a summary of laboratory assessments is provided in Table 2 . 
Table 1: 
Schedule of Evaluations (SOE) 
Visit  
Number 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
102/Exit
4 
Phone 
Follow-
up #1 
Phone 
Follow-
up #2 
Visit 
 Type 
Screen 
V1 
Safety &   
Immun 
V2 
Safety &   
Immun 
 
V3 
Safety & 
Immun 
Safety & 
Immun 
Safety & 
Immun 
Safety& 
Immun 
Safety & 
Immun 
MAAE 
follow-up 
MAAE 
follow-up 
Visit Day 
-45 to 
-3 
1 
15 
29 
43 
57 
58-59 
64 
71 
169 
337 
365 
393 
Window 
 
 
± 3d 
± 7d 
± 3d 
±7d 
 
± 3d 
± 3d 
± 7d 
± 7d 
± 7d 
± 7d 
Visit Week 
 
0 
2 
4 
6 
8 
8 
9 
10 
24 
48 
52 
56 
Clinical 
Briefing and 
Contact 
Information 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Informed 
Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Test of 
Understanding 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Enrollment and 
Randomization 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Vaccination 
 
X 
 
X 
 
X 
 
 
 
 
 
 
 
Vital Signs and 
Physical exam1 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
Medical History 
& Concomitant 
Medications 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 

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Visit  
Number 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
102/Exit
4 
Phone 
Follow-
up #1 
Phone 
Follow-
up #2 
Visit 
 Type 
Screen 
V1 
Safety &   
Immun 
V2 
Safety &   
Immun 
 
V3 
Safety & 
Immun 
Safety & 
Immun 
Safety & 
Immun 
Safety& 
Immun 
Safety & 
Immun 
MAAE 
follow-up 
MAAE 
follow-up 
Visit Day 
-45 to 
-3 
1 
15 
29 
43 
57 
58-59 
64 
71 
169 
337 
365 
393 
Window 
 
 
± 3d 
± 7d 
± 3d 
±7d 
 
± 3d 
± 3d 
± 7d 
± 7d 
± 7d 
± 7d 
Visit Week 
 
0 
2 
4 
6 
8 
8 
9 
10 
24 
48 
52 
56 
Adverse Event 
Documentation 
(AE, SAE, 
AESI)3 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
Medically 
Attended 
Adverse Event 
(MAAE) 
Documentation4 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Diary Card 
 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
 
HIV Risk 
Counseling 
X 
X 
X 
X 
X 
X 
 
 
X 
X 
X 
 
 
Pregnancy Test7 
& Pre/Post 
Counseling 
X 
X 
 
X 
 
X 
 
 
X 
X 
X 
 
 
CBC w/ diff 
4 
4 
4 
4 
4 
4 
 
 
4 
 
4 
 
 
Creatinine, ALT 
4 
4 
4 
4 
4 
4 
 
 
4 
 
4 
 
 
Hepatitis B and 
Hepatitis C 
Serology 
7.5 
 
 
 
 
 
 
 
 
 
 
 
 
HIV Testing* 
7.5 
 
7.5 
 
7.5 
 
 
 
7.5 
7.5 
7.5 
 
 

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Visit  
Number 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
102/Exit
4 
Phone 
Follow-
up #1 
Phone 
Follow-
up #2 
Visit 
 Type 
Screen 
V1 
Safety &   
Immun 
V2 
Safety &   
Immun 
 
V3 
Safety & 
Immun 
Safety & 
Immun 
Safety & 
Immun 
Safety& 
Immun 
Safety & 
Immun 
MAAE 
follow-up 
MAAE 
follow-up 
Visit Day 
-45 to 
-3 
1 
15 
29 
43 
57 
58-59 
64 
71 
169 
337 
365 
393 
Window 
 
 
± 3d 
± 7d 
± 3d 
±7d 
 
± 3d 
± 3d 
± 7d 
± 7d 
± 7d 
± 7d 
Visit Week 
 
0 
2 
4 
6 
8 
8 
9 
10 
24 
48 
52 
56 
Research5 
HIV Binding 
Antibody 
 
SP 
SP 
SP 
SP 
SP 
 
 
SP 
SP 
SP 
 
 
HIV 
Neutralizing 
Antibody 
Assays 
 
7.5 
7.5 
7.5 
7.5 
7.5 
 
 
7.5 
7.5 
7.5 
 
 
Functional 
Antibody Assays 
 
SP 
SP 
SP 
SP 
SP 
 
 
SP 
SP 
SP 
 
 
B-Cell Analysis 
 
17 
17 
17 
17 
17 
 
17 
17 
17 
17 
 
 
Multiparameter 
Flow 
Cytometry 
 
17 
17 
17 
17 
17 
 
17 
17 
17 
17 
 
 
Innate Cell 
Analysis 
 
17 
17 
 
17 
 
17 
17 
17 
17 
17 
 
 
Transcriptomics 
 
2.5 
2.5 
2.5 
2.5 
2.5 
2.5 
2.5 
2.5 
 
 
 
 
Additional 
Immunogenicity 
Testing 
 
42.5 
8.5 
8.5 
8.5 
8.5 
 
17 
25.5 
34 
34 
 
 

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Visit  
Number 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
102/Exit
4 
Phone 
Follow-
up #1 
Phone 
Follow-
up #2 
Visit 
 Type 
Screen 
V1 
Safety &   
Immun 
V2 
Safety &   
Immun 
 
V3 
Safety & 
Immun 
Safety & 
Immun 
Safety & 
Immun 
Safety& 
Immun 
Safety & 
Immun 
MAAE 
follow-up 
MAAE 
follow-up 
Visit Day 
-45 to 
-3 
1 
15 
29 
43 
57 
58-59 
64 
71 
169 
337 
365 
393 
Window 
 
 
± 3d 
± 7d 
± 3d 
±7d 
 
± 3d 
± 3d 
± 7d 
± 7d 
± 7d 
± 7d 
Visit Week 
 
0 
2 
4 
6 
8 
8 
9 
10 
24 
48 
52 
56 
Daily Volume 
(mL)6 
23 
111.5 
85 
60.5 
85 
60.5 
19.5 
70.5 
102 
100 
108 
 
 
Cumulative 
Volume (mL) 
23 
134.5 
219.5 
280 
365 
425.5 
445 
515.5 
617.5 
717.5 
825.5 
 
 
8-Week 
Cumulative 
Volume 
(mL) 
 
 
 
 
 
402.5 
422 
381 
483 
100 
108 
 
 
SP = Assay performed from stored plasma; no additional blood volume required 
1 Full physical examination at screening (visit S1) and targeted physical examination at all other visits; Vitals to be collected both pre- and at about 30 minutes post-
vaccination at Visit Day 1, 29, and 57. 
2 Participants will be contacted for the unblinding visit after the clinical data is locked. 
3This includes unsolicited AEs which will be collected until 28 days post each vaccination. 
4Medically Attended AEs will be followed through the end of the study, a full 12 months following the last vaccination. After clinic visits end, follow-up for MAAEs 
will occur by monthly phone calls for two months. 
For those participants who are unable to continue participation in the study, but who do not withdraw consent, an exit visit will be conducted. 
5 Research labs may be performed at any visit where sufficient stored samples are available  
6 Blood volumes for clinical lab assays may be adjusted as long as maximum daily blood volume is not exceeded. 
7 Pregnancy tests can be conducted at any visit if clinically indicated. 
8 Any clinically indicated safety labs can be obtained at investigator discretion 
*10 mL EDTA verification blood collection for HIV incident case resolution for enrolled participants. 
**Blood will be collected into vaccutainer tubes according to standard operational procedures.  Substitution of tube types may be made as long as the substitution does 
not interfere with the study performance.  A 10% variance in blood volume is acceptable, such as in the case of an error or a broken or clotted vial. 
 

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Table 2: 
Protocol-Required Safety Laboratory Assessments 
Laboratory Assessment 
Parameters a 
Hematology 
CBC with differential 
Clinical Chemistry 
Creatinine, ALT 
Laboratory Assay 
HIV 
a Acronyms 
 

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2. 
INTRODUCTION 
The investigational products to be used in this trial include A244, B.63521, and ALFQ.  
A244, a Duke University product, manufactured by Vetter Development Services USA, Inc. for 
NIH/NIAID, consists of the gp120 envelope glycoprotein HIV-1 subtype CRF_01E A244 
derived from the CM244 CRF_01E. The A244 gp120 envelope has an 11-amino N-terminal 
deletion, similar to the bivalent AIDSVAX® B/E protein used in RV1441, which has been safely 
administered to over 9,000 participants. 
B.63521Δ11mutc gp120 is a recombinant HIV-1 Env protein containing an 11 amino acid 
truncation at the N-terminus of gp120 that enhances antigenicity and immunogenicity. It is 
manufactured for NIH/NIAID by Vetter Skokie IL. 
ALFQ, a US Army product, is a liposomal adjuvant containing a synthetic monophosphoryl lipid 
A (MPLA/3D-PHAD®) with the addition of QS-21. ALF55 is manufactured by Avanti thru 
DAIDS-ABL contract. QS-21 is procured from Desert King thru DAIDS-ABL contract. ALF55 
and QS21 will be combined fill/finish at Walter Reed Pilot Bioproduction Facility to produce 
ALFQ. 
This trial will be conducted in compliance with the protocol, Good Clinical Practices (GCP), and 
the applicable regulatory requirements(s). 
2.1. 
Study Rationale 
Given the global burden of the human immunodeficiency virus (HIV) and specific burden to the 
Department of Defense (DoD), as the cost of treatment for its HIV-infected service members 
ranges from $21 to $54 million per year, HIV vaccine development would offer an immeasurable 
benefit. Models suggest that even a partially effective vaccine would make a substantial 
contribution to HIV incidence reduction. A study conducted by Harmon et al suggested that a 
vaccine with 70% efficacy could reduce disease incidence by 44% in the first decade, and up to 
78% in under 50 years.2 A model presented by Stover et al posited that even a vaccine with 50% 
efficacy could reduce the annual transmission rate by 34% over 15 years.3 Both studies 
concluded that a vaccine would be a cost- effective intervention. This prevention strategy would 
confer multiple advantages over other interventions in that it requires time-limited delivery, 
minimal maintenance, and offers long-term protection.  
Efforts to produce an HIV vaccine have primarily focused on the HIV-1 gp 120 subunit vaccines 
which incorporate outer envelope viral proteins. Protein based vaccines are commonly paired 
with an immunostimulatory adjuvant to improve durability of the immune response. ALFQ is an 
adjuvant containing both MPLA and saponin QS21 which has elicited a potent immune response 
in preclinical studies. Determining the optimal dose of ALFQ adjuvant may reduce the amount 
of material and cost necessary for future HIV vaccine production. 
Similarly, improving the rapidity of our response to new emerging infectious diseases will be 
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2.2. 
Background 
Although global initiatives have made great strides in controlling the HIV pandemic, HIV and 
acquired immunodeficiency disease syndrome (AIDS) continues to impact the lives and 
livelihoods of a significant portion of the population. In 2019, 38 million individuals were living 
with HIV worldwide and 690,000 died of AIDS-related causes.4 Despite extensive global efforts 
for disease control over the last 20 years, 1.7 million individuals contract HIV annually.4 Given 
these successes and challenges, the development of a safe and efficacious preventive HIV 
vaccine as part of a comprehensive prevention program remains among the highest global health 
priorities and the best long-term tool for the control of the HIV-1.5-7 To date, seven HIV vaccine 
efficacy trials have been conducted, including VAX0038, VAX0049 RV1441 , STEP10 , 
Phambili/HVTN50311 , HVTN50512 and HVTN70213. Of these, only the RV144 Phase 3 trial 
demonstrated efficacy. Healthy participants who received the 6-month vaccination regimen 
containing live recombinant ALVAC-HIV (vCP1521) and AIDSVAX B/E, adjuvanted with 
Rehydragel®, were 31% less likely to acquire HIV than participants who received placebo over a 
3.5-year follow-up period.1 In a post hoc analysis, efficacy was 60% at 12 months after initial 
vaccination, indicating that protective immunity may have waned rapidly14 . A follow-on study 
of immunologic correlates associated with HIV acquisition found that the magnitude of plasma 
immunoglobulin (Ig) G antibodies to variable regions 1 and 2 (V1/V2) of HIV-1 Env was 
inversely correlated with probability of HIV acquisition, and plasma IgA antibody titers to HIV-
1 gp120 Env were directly correlated with probability of HIV acquisition.15 Given these findings, 
the B/E envelope proteins were selected for this study, based on their similarity to proteins 
utilized in the RV144 trial.  
2.2.1. 
A244d11 gp120 
The A244 protein is an immunogen that consists of the gp120 envelope glycoprotein HIV-1 
subtype CRF_01AE. It has demonstrated its antigenicity and immunogenicity in human and non-
human primate (NHP) studies, as described below.16 
A244d11 gp 120 gD has been administered to thousands of clinical trial participants as a 
component of AIDSVAX® B/E. A phase 1/2 study performed in Thailand enrolled 133 
participants to test the safety and immunogenicity of the combination of ALVAC-HIV and 
AIDSVAX B/E. No serious vaccine-related events were reported.17 Only 4 AEs were determined 
to be vaccine related. These included myalgia, pruritis, and erythematous rash.17 The RV144 
trial, which enrolled 16,402 individuals, tested the efficacy of the combination of ALVAC and 
AIDSVAX B/E. Local reactogenicity was mild or moderate for the AIDSVAX component, and 
overall AEs were primarily mild or moderate in severity. As previously discussed, vaccine 
efficacy was 60% At 12 months, but decreased to 31% by 3.5 years. The RV306 trial assessed 
the immunologic effect of a late boost of either AIDSVAX B/E or the combination of 
AIDSVAX B/E and ALVAX-HIV at month 12, 15, or 18 after receiving the standard RV144 
series at months 0, 1, 3, and 6. There were no vaccine-related serious AEs recorded. Groups 
which received a late boost had increased peak plasma IgG-binding antibody levels against gp70 
V1V2, and those that received a late boost at 15 or 18 months, specifically, had improved gp 120 
responses. Additionally, groups with late boosts had increased functionality and 
polyfunctionality scores compared to the RV144 regimen only group.18 

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In addition to combination regimens with ALVAC-HIV, AIDSVAX B/E has also been 
administered as a stand-alone investigational product. A randomized, double-blind, placebo-
controlled trial assessing the efficacy of AIDSVAX B/E was performed in a population of more 
than 2500 injection drug users in Thailand. Although the product was well-tolerated, it did not 
prove efficacious. HIV acquisition rate was almost identical between the vaccine and placebo 
groups.8 AIDVAX was also administered alone in the AVEG 036 study, which, similarly, did not 
demonstrate vaccine efficacy. 
The A244-rpg 120 immunogen utilized in the AIDSVAX B/E product was created with an 11 
amino acid deletion at the gp120 N terminus and a herpes simplex virus gD peptide tag.19 The 
effects of these modifications were explored by comparing gp120 expression, antigenicity, and 
immunogenicity. The study demonstrated that the deletion of the N-terminal improves both 
envelope antigenicity and immunogenicity, and is sufficient to induce an enhanced immune 
response without the gD protein-derived tag.16 As a result, the A244d11 gp120 investigational 
product was formulated without the gD tag. This investigational product has not yet been used in 
humans, but is planned for use as a late boost for RV306 participants in the RV546 trial.  
2.2.2. 
B.63521 
The B.63521 gp120 is an investigational recombinant HIV- 1 Clade B T/F envelope protein 
containing an 11 amino acid truncation at the N-terminus of gp120. It has been shown to elicit 
HIV binding antibodies, neutralizing antibodies with wide antigenic breadth, and antibody 
dependent cellular cytotoxicity in animal models.20, 21 Although the B.63521 investigational 
product has not yet been used in humans, it is sufficiently similar to other HIV-1 gp120 proteins 
administered in multiple previous clinical trials, that it is thought to have a similar safety profile. 
There have been a large number of clinical trials with other HIV-1 Env gp120 proteins, and the 
most common adverse effects were local and resolving within 3 days. 
2.2.2.1. 
Non-Human Primate Study 
A non-human primate study was designed to augment the diversity of gp120 motifs in the 
immunogen utilized in the RV144 trial and to assess whether a broader antibody response could 
be elicited. Eighteen rhesus macaques received six doses of ALVAC–HIV delivered alone at 
doses 1 and 2 at weeks 0 and 4, respectively, and combined with intramuscular bivalent (B/E) or 
pentavalent (B/E/E/E/E) at doses 3-6 in weeks 13, 21, 47 and 88, respectively. A 100 mcg 
protein dose was administered for the first 3 protein immunizations at weeks 13, 21 and 47, and a 
300 mcg protein dose was administered for the 4th protein immunization at week 88. All doses 
were combined with GLA-SE adjuvant. The B.63521 gp120 was combined with CRF01 
AE.A244 fp120, and the pentavalent boost contained three additional subtype E gp120s 
(AE.AA058 / AE.AA104 / AE.AA107). Immunizations were well-tolerated and no AEs were 
observed. 
Both vaccine regimens induced Env-reactive plasma antibodies, with greater breadth in the 
monkeys in the pentavalent cohort. Additionally, all animals had detectable mucosal IgG 
antibodies against at least one of twelve tested gp120 antigens, and all but one animal in the 
pentavalent group had mucosal antibodies specific to B.63521gp120 Env. Tier-1 neutralizing 
antibodies were induced in both groups, and both could mediate antibody-dependent cellular 
phagocytosis of vaccine Env-coated targets, with weak phagocytosis of Tier-2 SHIV Env by 

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3 animals in the pentavalent group. Protection from SHIV challenge was achieved in 55% of the 
pentavalent cohort.22 
2.2.3. 
ALFQ 
Modulation of the response to vaccines with immunostimulatory adjuvants is one very promising 
approach to improve durability. The Army Liposome Formulation (ALF) family of adjuvants are 
ALFA (ALF adsorbed to aluminum salt), ALFQ (ALF mixed with the saponin QS21; US patent 
pending) and ALFQA (ALFQ adsorbed to aluminum salt). Both ALFA and ALFQ are potent 
adjuvants, non-pyrogenic, and nontoxic in rabbit pyrogenicity and toxicology studies.23 ALFQ is 
an alternative to the successful but proprietary adjuvant, AS01B (liposomes containing both 
MPLA and the saponin QS21). ALFQ is potent in preclinical studies and contains 4 times the 
amount of MPLA and twice the amount of QS21 when compared to AS01B. An ongoing NHP 
study comparing HIV gp120 A244 formulated with either ALFQ or with Aluminum hydroxide 
fluid gel (AHFG) has shown that animals that received ALFQ as the adjuvant compared to 
animals that received AHFG mounted significantly higher binding antibody responses to A244, 
gp70V1V2 A244, and V2 peptide (Haynes 2020, unpublished ). The responses were significantly 
higher after each immunization, as well as in ADCC responses, quantified using gp120 coated 
target cells. Given that antibody functions such as ADCC may have contributed to protection by 
non-neutralizing antibodies in RV144 vaccine recipients, ALFQ may contribute to increasing 
protective vaccine efficacy.  
ALFQ is currently being investigated in three clinical trials. The FMP013 and FMP014 
(NCT04268420 and NCT04296279) malaria investigational vaccines are adjuvanted with ALFQ 
at the 100mcg and 200mcg doses in the low dose and high dose arms, respectively. The FMP013 
and FMP014 groups were designed to enroll 10 participants each, which underwent 
investigational vaccine administration at months 0, 1, and 2, with 5 participants assigned to each 
of the high and low dose groups.  
In the FMP013 trial, 5 pariticipants received dose 1 and 2 in the low dose group and 4 
individuals recived dose 3. The participant who did not receive the final dose left the study for 
reasons unrelated to the study design. In the high dose group, 5 participants received product 
adminstration 1, 4 pariticpants received product admin istration 2, and 3 participants received 
product adminstration 3. One participant dropped out for reasons unrelated to the trial, and 
another participant was removed due to a likely pre-exisiting medical condition. In the low dose 
group, the majority of participants reported mild local and systemic symptoms, with a minority 
of partiicpants reporting moderate local and systemic symptoms (Figure 2). In the high dose 
group, participants reported mild to moderate local and systemic symptoms. The study product 
was well tolerated with no SAEs (Hutter, 2020). 
In the FMP014 trial, 10 out of 10 participants completed all 3 vaccinations. In the low dose 
group, the majority of pariticpants reported mild local and systemic symptoms, with a minority 
of participants reporting moderate local and systemic symptoms (Figure 2). In the high dose 
group, the majority of participants reported mild to moderate local and systemic symptoms 
(Figure 2). All severe systemic reactions were reported by a single individual who had 24 hours 
of flu like symptoms that resolved spontaneously. The study product was well-tolerated, with no 
SAEs (Hutter, 2021) (Hutter ASTMH 2020). 

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Figure 2: 
Reactogenicity Results for FMP013 and FMP014 
 
ALFQ has also been administered in the EID030 (SpFN) (IND27301, NCT04784767) trial. The 
spike ferritin nanoparticle was administered with 200 mcg of ALFQ at months 0, 1, and 6. In 
Arm 1, 24 partiticpants received dose 1 and 2, and 9 participants received dose 3. In Arm 2, 5 
participants received dose 1, 4 participants received dose 2, and 2 participants received dose 3. 
No SAEs were reported. A total of 8 unsolicited AEs occurring in 5 participants, including 5 
determined to be at least possibly related to study vaccination were reported. All unsolicited AEs 
reported were of mild severity. 
In Arm 1, a total of 14 participants (58.3%) reported at least one mild solicited event, and 7 
(29.2%) reported at least one moderate solicited event. Five participants reported moderate 
systemic events and 3 reported moderate local events. Sixteen participants reported mild 
injection site pain and 2 reported moderate injection site pain. Eight individuals reported mild 
myalgia and 1 reported moderate myalgia. Five participants reported mild fatigue and 2 reported 
moderate fatigue. Clinical safety laboratory assessments demonstrated that 12 participants had 
mild decreases in hemoglobin 8 days after product administration. (Personal communication with 
study PI) 
In Arm 2, 5 participants experienced Grade 1 solicited reactions, and one participant experienced 
a Grade 2 muscle pain at the injection site. Grade 1 change in hemoglobin was reported for two 
participants. One participant had a Grade 1 change in WBC and a Grade 3 absolute neutrophil 
count which was similar to baseline. (Personal communication with study PI) 
ALFQ will also be administered in one additional clinical trial which may initatie prior to the 
start of RV575.  In the upcoming RV546 trial (IND 27011, NCT04658667), participants in arms 3 
and 4 will receive a single IM injection of 200 mcg of ALFQ. These ALFQ doses refer to the 
MPLA/3DPHAD component of ALFQ. 
Manufacture of protein-based vaccine antigens can be technically complex and expensive, both 
of which can limit vaccine supply, particularly in times of high demand during an epidemic. 
Optimization to reduce vaccine dosing can help to alleviate these issues. RV 575 will perform a 
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immunogenicity. Even a similar, rather than superior, immunogenicity of decreased dosing 
would result in significant cost savings, thereby facilitating scale-up and delivery to populations 
at greatest need for a preventive HIV vaccine. 
2.3. 
Military Relevance 
Historically, infectious diseases have had a major impact on US Armed Forces. With an 
estimated 34 million infected individuals worldwide at the end of 2011, HIV poses a significant 
and persistent threat in terms of force readiness and protection and may act as a war-starter by 
affecting the stability and security of nation-states. 
HIV’s relevance the US Military has been recognized from the very beginning of the pandemic. 
In 1985, the US Military recognized the emerging HIV-1 epidemic as a new threat to US and 
allied forces worldwide. The United States Congress mandated the establishment of the US 
Military HIV Research Program to develop effective preventive measures to include prevention 
education, vaccine development and implementation of novel anti-viral therapies and clinical 
management tools for the US DoD and Allied Forces. In 2001, the Armed Forces Epidemiology 
Board identified HIV as a disease of military importance, and Army FOC 09-07, Global Casualty 
Prevention, requires detection, identification, and vaccination in order to protect US personnel 
against potential infectious disease (ID) threats. The 2001 DoD Report on Biological Warfare 
Defense Vaccine Research and Development identified HIV as the 4th greatest infectious disease 
threat to DoD forces. Department of Army Headquarters designated HIV vaccine development 
as an Army Technology Objective (ATO), a status reserves for the highest priority science and 
technology efforts. Furthermore, The National Security Strategy of the United States (2002, 
2006, and 2010) clearly identifies the threat of HIV/AIDS as a destabilizing force that threatens 
US National Security. Finally, in January 2011, The Department of the Army approved the HIV 
Vaccines Capability Development Document for HIV Vaccine, which addresses the need to 
provide service members with vaccine protection against HIV and AIDS. 
Medical care for HIV infection is extremely costly to the US Defense Health Program. The 
estimated lifetime cost of HIV infection is at least USD $400,000 and the estimated average 
yearly cost varies from $20,000 to $25,000, bringing the estimated lifetime healthcare cost of the 
5,000 HIV infected servicemen/women to $8B to $10B dollars. Total annual cost of treatment 
for HIV-infected service members ranges from $21 to $54 million per year. 
2.4. 
Benefit/Risk Assessment 
The potential risks and benefits based on the study design and what is known about the product 
are summarized in this section. More detailed information about the known and expected 
benefits and risks and reasonably expected AEs can be found in the Investigator’s Brochure (IB). 
2.4.1. 
Local Reactions 
Participants may exhibit post-vaccination reactions including local reactions at the injection site 
such as erythema, induration, pain/tenderness, swelling and limitation of arm movement. These 
reactions are generally of short duration and rarely require treatment. Should such reaction 
persist and require treatment, the participant will be referred to appropriate medical care services. 

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2.4.2. 
Systemic Reactions 
Participants may exhibit general signs and symptoms associated with administration of a vaccine 
injection, including fever, tiredness, chills, rash, myalgia, arthralgia, nausea, and headache. 
These side effects will be monitored but are generally of short duration and rarely require 
treatment. Should such reaction persist and require treatment, the participant will be referred to 
appropriate medical care services. As with all vaccines, an allergic reaction is possible and will 
be managed as per site standard operating procedure (SOP) for anaphylaxis. To mitigate this risk, 
participants will be observed in the clinic for at least 30 minutes post-injection. 
2.4.3. 
Risks to Study Personnel and the Environment 
The principal risk to personnel in the clinical setting is that associated with potential exposure to 
infectious agents (e.g., in the handling of needles that may be contaminated and the attendant 
risks, including exposure to hepatitis, human immunodeficiency virus, and other human 
pathogens). Adherence to SOPs for working with infectious agents and universal precautions 
will reduce the risk of exposure. 
This protocol poses no known risks to the environment other than those associated with the 
generation of biohazardous waste attendant to study procedures. All biohazardous waste will be 
disposed of and specimens collected as stipulated by local, state, and Federal regulations and in 
accordance with study site SOPs. 
2.4.4. 
Other Risks 
Venipuncture: Blood sampling carries a minimal risk of minor discomfort, the possibility of 
minor bruising at the site of the needle puncture, and, rarely, the possibility of infection at the 
needle puncture site. To minimize risk, blood will be drawn by trained health care providers.  
Changes at the needle puncture site will be tracked with the diary card and will be examined at 
the follow up visit. Participants with evidence of severe bruising or infection will be referred for 
appropriate medical care. 
Allergic Reaction: No matter what precautions are taken, the administration of any product 
always carries the risk of a serious, even life-threatening, allergic reaction. Medical emergency 
equipment is located at the study site. This is available to handle emergencies, such as 
anaphylaxis, angioedema, bronchospasm, and laryngospasm. 
Vaccine Induced Seroreactivity: As with every candidate HIV vaccine trial, there is a risk that 
participants will mount an antibody response to the vaccine, which may cause them to test 
positive on a standard clinical diagnostic assay for HIV. If a participant has an issue due to a 
false-positive HIV test result from receiving the vaccination, the PI will provide an explanatory 
letter (Appendix I). The study will provide re-testing for participants should they receive a 
positive HIV test elsewhere and require proof of vaccine induced seroreactivity.  Should a 
participant move out of the D.C. metro area, they will be provided with a contact number to call  
and ask to be connected to an MHRP study physician for further guidance on what to do if they 
need to receive HIV testing. 
Social Harm and Discrimination: In the unlikely event of HIV infection during this study, the 
primary concern is related to ascertaining and providing HIV diagnostic information and, in 

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particular, involuntary disclosure of HIV status to others. These disclosures may result in 
depression and rarely suicide among individuals learning that they are infected with the HIV 
virus. Furthermore, involuntary disclosure to others may result in prejudice by the community, 
family, employers, and psychosocial factors including stigma and discrimination. This risk will 
be minimized as study counselors will be trained in pre- and post-test counseling for HIV and 
will inform participants of all activities in the study and attendant risks and benefits. The 
candidate vaccines may also induce false positivity to standard HIV antibody tests and may 
result in problems when applying for life or health insurance, international travel, employment, 
or hospitalization. If a participant has VISP and becomes pregnant, the baby may also have 
VISP. If there are any problems related to the above situations, the investigators will provide 
HIV testing to confirm HIV status. They will provide this information to the persons involved. 
Currently, people who have received experimental HIV vaccines are deferred from blood 
donation even though they do not have HIV infection. The participant will not be able to donate 
blood if they have VISP. 
In addition to the risk of social harms resulting from HIV diagnosis, becoming aware of a 
positive test results for Hepatitis B and C or the discovery of incidental findings may also result 
in the emotional effects one may experience after becoming aware of a positive HIV test. The 
study staff will take appropriate action to assist participants with any discrimination they may 
experience by participating in this study. Such measures will be accompanied with appropriate 
counseling. 
Guillain–Barré Syndrome: Guillain-Barré syndrome (GBS) is a disorder in which the body’s 
immune system attacks part of the peripheral nervous system. It afflicts only about one person in 
100,000, and in rare instances vaccinations may increase the risk of GBS. The first symptoms of 
this disorder include varying degrees of weakness or tingling sensations in the legs. These 
symptoms can increase in intensity until certain muscles cannot be used at all. Most individuals 
recover from even the most severe cases of GBS, although some continue to have a certain 
degree of weakness.1 
2.4.5. 
Unknown Risks 
As with all research, there is the remote possibility of risks that are unknown or that cannot be 
foreseen based on current information. This would include late-onset effects. 
Risks to unborn babies are unknown at this time; the effect of the candidate HIV vaccines on 
pregnancy and fetus are unknown as they have not been specifically studied in pregnant people. 
Participants who are pregnant or plan to become pregnant during the first 3 months post- 
vaccination will be excluded from enrollment. Female participants must use an adequate form of 
birth control including condoms (male or female) with spermicide, diaphragm, or cervical cap 
with spermicide, intrauterine device, contraceptive pills, patch, injection, intravaginal ring or 
other FDA-approved contraceptive method, male partner has previously undergone a vasectomy, 
or abstinence for 30 days prior to study injection until at least 3 months after receipt of the final 
study injection. A pregnancy test will be performed at screening, prior to the study injection 
(same day), and prior to any of the invasive procedures. Should a participant become pregnant 
within the 3 months post-study injection, she will be followed for safety to term.  Partners of 
male participants who become pregnant will not be followed. 

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All participants are encouraged to engage in safe sex practices to prevent HIV acquisition. 
2.4.6. 
Alternative to this Product or Study 
The alternative is not to participate in this protocol. 
2.4.7. 
Benefits 
Study participants will receive no direct benefit from study participation. HIV and other sexually 
transmitted infection (STI) testing may reveal HIV or STI transmission. In this case, the 
participant would be referred for counseling and care. Findings of medical concern will be 
referred for appropriate care and treatment. Others may benefit from knowledge gained in this 
study that may contribute to the development of an HIV vaccine. 
3. 
OBJECTIVES AND ENDPOINTS 
The objectives and associated endpoints are provided in Table 3. 
Table 3: 
Objectives and Endpoints 
Objective 
Endpoint 
Primary: 
To evaluate the safety and tolerability (including 
reactogenicity) of candidate vaccine 
A244/B.63521 with ALFQ adjuvant at the 200 
μg, 100 μg, and 50 μg doses 
Evaluate the occurrence and severity of solicited 
local and systemic AEs following candidate 
vaccine administration. 
Evaluate the occurrence, severity, and relationship 
to vaccination of unsolicited AEs after candidate 
vaccine administration. 
Evaluate the occurrence of SAEs, and new-onset 
medical conditions. 
Evaluate the occurrence of AEs of special interest 
(AESIs) following candidate vaccine 
administration. 
Secondary: 
To evaluate the effect of candidate vaccine 
A244/B.63521 with ALFQ adjuvant at the 200 
μg, 100 μg, and 50 μg doses on cellular, and 
humoral immune responses 
Compare plasma IgG binding antibodies to gp120 
in terms of magnitude, durability, and area under 
the curve among groups with differing ALFQ 
doses. 
Characterize and assess the magnitude of cell-
mediated immune responses elicited across 
vaccination regimens including antigen-specific 
CD4 and CD8 T cell responses and 
polyfunctionality. 
Exploratory: 
To characterize B-cell functional specificities for 
each vaccination regimen 
Characterize plasma IgG and IgA binding 
antibodies to HIV gp120, neutralizing antibodies, 

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Objective 
Endpoint 
Primary: 
To characterize innate immunity 
To assess the innate/gene expression induced 
across vaccination regimens 
To perform systems serology analyses 
To characterize the impact of vaccination with the 
ALFQ adjuvant on the permissiveness and 
expression to in vitro HIV infection of CD4+ T 
cells and differentiated macrophages 
and non- neutralizing effector functions such as 
ADCC and ADCP with emphasis on RV144 
immune correlates of risk of HIV acquisition. 
Characterize B-cell functional specificities for 
each vaccination regimen by quantifying antigen-
specific responses through B-cell ELISPOT, 
phenotyping the magnitude and activation status 
of B cell subsets via flow cytometry and isolation 
of monoclonal antibodies from selected vaccine 
recipients. 
Characterize innate immunity through 
quantification of soluble chemokines and 
cytokines and assessing the phenotype and 
function of cellular innate immune subsets such as 
NK, NKT, and dendritic cells. 
Characterize effects of host genetic 
polymorphisms on immune responses and 
characterize differentially expressed genes. 
Characterize the impact of vaccination with the 
ALFQ adjuvant on the permissiveness and 
expression to in vitro HIV infection of CD4+ T 
cells and differentiated macrophages through 
integrated HIV-DNA PCR, cell associated-DNA 
and RNA, flow cytometry of various activation 
markers and HIV Gag protein, ELISA of HIV p24 
in the cell culture supernatant, and production of 
cytokines/chemokines. 
4. 
STUDY DESIGN 
4.1. 
Overall Study Design 
The purpose of this Phase I randomized, double-blind clinical trial is to optimize ALFQ dosing. 
Safety will be assessed through the frequency of the overall and specific post-vaccination 
reactions. Blood will be collected to assess humoral, cell-mediated, and innate immune 
responses. 
Healthy adults not living with HIV who are available for 14 months will be enrolled. A total of 
60 participants will be enrolled to one of three arms, each comprised of 20 candidate vaccine 
recipients. Each arm will receive identical doses of A244 and B.63521 (300 micrograms of 
each). In addition, Arm 1 will receive 200 micrograms of ALFQ. Arm 2 will receive 100 
micrograms of ALFQ. Arm 3 will receive 50 micrograms of ALFQ. The safety, reactogenicity, 
and immunogenicity will then be compared among the three arms to determine the optimal dose 
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All vaccinations will be split into 2 half doses which will both be administered intramuscularly 
(IM) into the same deltoid muscle. Vaccinations will occur at months 0, 1, and 2. The second 
vaccination will be administered into the contralateral deltoid at study month 1 compared to the 
first vaccination at study month 0. The third vaccination at study month 2 will be administered 
into the same deltoid as the first vaccination at study month 0. Participants will be followed for 
12 months following the last study vaccination. Safety and tolerability will be assessed with both 
clinical and laboratory monitoring. Vaccine- related reactions will be observed and solicited for 
30 minutes post-vaccination and with the aid of a diary card and interview of participants during 
the 14 days post vaccination. 
The information gained from the review of the diary card and the interview with the participants 
will be documented in the clinical study chart. In addition, AEs will be documented at each 
clinical encounter. AEs will be graded for seriousness, severity, and relationship to the 
investigational product. 
4.2. 
Number of Participants 
The site will screen approximately 150 participants to achieve a total of 60 enrolled. 
4.3. 
Sentinel Group Allocation 
A sentinel group including 6 participants will be enrolled. Four participants in the sentinel group 
will  be randomized to the 200 mcg ALFQ group (Arm 1), one participant  will be randomized to 
the 100 mcg ALFQ group (Arm 2), and one participant will be randomized to the 50 mcg ALFQ 
group (Arm 3). Safety data for seven days following vaccination one will be reviewed by a 
PSRT. If it is determined to be necessary by the PSRT, the MHRP SMC will be consulted to 
review unblinded data to provide a recommendation on continuing enrollment.  Additionally, the  
PSRT will consult the SMC if either of the following conditions are met:  
1) One (or more) participants experience an SAE that is assessed as related to a study agent, or 
2) Two (or more) participants experience grade 3 or 4 AEs of the same type (e.g., elevated ALT) 
related to a study agent.  This would exclude monitoring lab values if a clear alternate etiology is 
identified.  Additionally, CBC differential counts would not contribute. 
After safety has been confirmed by PSRT review, the remaining 54 participants will be enrolled. 
4.4. 
Scientific Rational for Study Design 
To date there have been no comparative studies regarding optimal dosing of adjuvant ALFQ. 
The ALFQ doses in Arm 1, 2, and 3 are based on the MPLA/(3D-PHAD®) component of 
ALFQ. ALFQ has been demonstrated to elicit potent antibody binding responses in animal 
models. 
ALFQ adjuvant dose optimization has the potential to limit AEs and improve efficiency and cost 
effectiveness of vaccine administration while maximizing immunogenicity. As a result, RV575 
will perform a dose de-escalation study.  

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4.5. 
Justification for Dose 
The A244 protein was included in the AIDSVAX vaccine at the 300 microgram dose. This dose 
was found to be safe and well tolerated.24  
B.63521has been tested in animal trails at doses up to 300 micrograms. It has not yet been 
utilized in human trials. B.63521 is very similar to the MN protein utilized in AIDSVAX which 
was administered at the 300 microgram dose in RV 144.10 
ALFQ had already demonstrated safety and immunogenicity at the 200 microgram and 100 
microgram dose.(Hutter, ASTMH, 2020) 
This study will contribute information about safety and immunogenicity of ALFQ at the 50 
microgram dose and perform a direct dose comparison for both safety and immunogenicity. This 
data is essential to optimizing efficiency of adjuvant inclusion in future vaccine trials. 
4.6. 
Method of Treatment Assignment 
Randomization will be conducted using a randomization block scheme described in a separate 
randomization plan held by the protocol statistician. The randomization list will be generated by 
the protocol statistician and provided and trained on with the site delegated un-blinded study 
staff/nurse. Once a randomization number has been assigned, it will not be re-assigned. 
4.7. 
End of Study Definition 
The end of the study primary completion date is defined as the last subject last visit (LSLV). 
Each participant will be followed for 12 months after the final vaccination. Follow up includes 
10 months of clinic visits and two additional telephone follow up visits at 1-month intervals for a 
total of 12 months. The completion of laboratory assays and data analysis is anticipated to take 
up to 5 years from the primary completion date. 
5. 
STUDY POPULATION 
5.1. 
Participant Recruitment 
The study population will consist of 60 healthy adults not living with HIV between the ages of 
18 and 55 years at the time of study enrollment at the WRAIR CTC. 
Healthy adult volunteers, male and female, will be recruited by noncoercive means through the 
WRAIR CTC according to applicable US Army regulations. Participants will be recruited from 
the Baltimore-Washington, D.C. metro area through the WRAIR CTC according to applicable 
U.S. Army regulations (i.e., 32 CFR 219 and 21 CFR 50, 54, 56, and 312). Adult men and 
women will be recruited by the use of WRAIR IRB approved advertisements using multiple 
media formats, to include, but not limited to: newspapers, flyers, emails, the WRAIR CTC 
website, postings on public listservs, social media (such as Facebook or other apps/websites), 
posters, bus ads, generic radio and television ads, word of mouth and referrals from volunteers 
currently in studies. Email announcements will include information on ad posters excluding any 
photos unless attached as complete flyer. See Appendix F below for the recruitment script. 
Recruitment may also include oral presentations (using standardized audio script) at events and 

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meetings. All advertisements, both general and specific to this study, will have been reviewed 
and approved by the WRAIR IRB prior to their use. All recruiting methods will direct the 
volunteer to contact the CTC via e-mail or phone, or to the CTC web site where the same contact 
information will be given. 
In the case of oral presentations and other meeting-related recruitment, potential subjects may be 
given the opportunity to sign up directly for a briefing and screening appointment. Any subject 
who so desires will be asked to provide basic contact information and an appointment made. 
Upon calling the recruitment phone number, a recruitment script will be used by study personnel 
to inform potential subjects of the details of the study. If the potential subject desires, some basic 
contact information will be obtained and an appointment for a formal briefing and screening will 
be made at that time. 
Upon contacting the CTC via e-mail, potential participants will be provided an electronic version 
of the recruitment script to inform them of the details of the study. If the potential subject 
desires, some basic contact information will be obtained and an appointment for a formal 
briefing and screening will then be made (via further e-mail or phone conversations). 
Given the value of the information in this recruitment script, a version of the script without 
compensation figures will be submitted as a separate “information flier” to be used on the 
WRAIR CTC web site, email correspondence (as noted above), and other electronic or social 
networking platforms. 
Participants who have participated in studies at the WRAIR CTC will be given the opportunity to 
receive compensation for referring additional subjects. Subjects will be asked to direct interested 
persons to contact the CTC. Subjects will receive $50 for each referred person who then attends 
a screening session and meets all inclusion and none of the exclusion criteria. 
Compensation will be independent of the referred person’s decision to enroll. Final authority 
over dispensation of referral compensation will lie with the CTC Director. 
Volunteers can also be recruited via WRAIR 2038, which is a generic screening protocol 
managed by the WRAIR CTC. Under this screening protocol, volunteers will undergo a generic 
screening visit with medical history, exam, vitals, lab tests, along with other potential screening 
tests. If the volunteer is interested in learning about this clinical trial, they will need to attend a 
briefing and sign this study’s informed consent form. If the volunteer consents, the screening 
documentation from WRAIR 2038 can be shared with this study to fulfill screening 
requirements, and a copy of said documents can be added to the volunteer’s study file for this 
clinical trial. 
5.2. 
Participant Identification Numbers 
All participants consented will be assigned a Participant Identification Number (PIN), which is a 
7-digit number. The first three digits are the protocol number (575), followed by a 4-digit 
number which, will be assigned sequentially. 
The PIN will be used to label all source documents, Case Report Forms (CRFs) and specimens. 
Source documents, CRFs and specimens are never labeled with participant names or national 
identification numbers. 

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5.3. 
Participant Eligibility 
Screening for eligible participants will be performed within 45 days prior to administration of the 
investigational vaccine. Participants who have passed the TOU and have given written informed 
consent will undergo a complete medical history, physical examination, and screening laboratory 
assessments to determine eligibility for study participation. Screening assessments (found in the 
SOE) will be completed after the informed consent process has been completed. 
Screening evaluations for specific eligibility criteria (see Section 5.3.1 and Section 5.3.2) must 
be completed within the screening visit window specified (Table 1) prior to enrollment. 
Counseling for HIV risk reduction and the potential risks of becoming pregnant during 
participation will be provided. The PI or designee will make the final decision regarding 
eligibility. Clinical data and specimens collected during screening from participants subsequently 
found to be ineligible for participation in the study will become part of the study records and 
specimens will be evaluated in accordance with protocol procedures. One re-screening attempt 
will be allowed if the there was an elevation and laboratory values on the first screening attempt 
which is suspected to be temporary, and or insignificant by the study personnel. Only eligible 
participants will be given the investigational product at the enrollment visit. A participant is 
considered enrolled in the study upon receipt of the first study vaccination. 
5.3.1. 
Inclusion Criteria 
Potential participants are required to meet all of the following criteria for enrollment into the 
study. 
1. Healthy adults between the ages 18-55 years (inclusive) 
2. Must be at low risk for HIV infection per investigator assessment and using the study risk 
assessment tool. 
3. Able and willing to provide written, informed consent 
4. Able and willing to comply with all research requirements, in the opinion of the Investigator 
5. Agreement to refrain from blood donation during the course of the study and within the 2 
months prior to study entry 
6. Minimum body weight of 110 pounds (lbs) (50kg) 
7. Laboratory Criteria within 45 days before enrollment: 
a. Hemoglobin ≥ 11.7g/dL for women; ≥ 12.5 g/dL for men 
b. White Blood Cell count = 3,500-10,800 cells/mm3 
c. Platelets ≥140,000/mm3 and ≤ 450,000/mm3 
d. Alanine aminotransferase (ALT; SGPT) <1.25 x ULN 
e. Serum creatinine ≤  1.25 x institutional upper limit of the reference range 

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f. Negative HIV testing (HIV Ab / antigen 4th generation screen with reflex confirmatory 
RNA testing) 
g. Negative HBsAg and hepatitis C antibody testing 
Note: As above, Grade 1 lab abnormalities detected on screening may be repeated at PI 
discretion. Persistent Grade 1 abnormalities that are felt to represent the non-pathologic baseline 
for the subject will be documented before a subject is enrolled in the trial and are allowable per 
discretion of the PI. 
8. Birth control requirements: 
All participants assigned female at birth must meet one of the following 2 criteria: 
a. No reproductive potential due to post-menopausal status (12 months of natural 
[spontaneous] amenorrhea) or hysterectomy, bilateral oophorectomy, or tubal ligation 
b. People of childbearing potential should agree to practice highly effective contraception 
at least 30 days before enrollment and through 3 months post-last vaccination, using one 
of the following methods: condoms (male or female) with spermicide; diaphragm, or 
cervical cap with spermicide; intrauterine device; contraceptive pills, patch, injection, 
intravaginal ring or other FDA-approved contraceptive method; male partner has 
previously undergone a vasectomy; abstinence  
c. All participants are encouraged to engage in safe sex practices to prevent HIV 
acquisition  
9. For all participants assigned female at birth, except those with a history of 
hysterectomy or bilateral oophorectomy, a negative β-HCG pregnancy test (urine) 
on the day of enrollment and each vaccination day is required. Because tubal 
ligations have a failure rate that is not insignificant, and 12 months of spontaneous 
amenorrhea can be a result of polycystic ovarian syndrome which does not 
completely preclude pregnancy, a negative β-HCG pregnancy test at enrollment and 
on each vaccination day is also required for all participants assigned female at birth 
with a history of either of these. 
10. No plans to travel outside the Washington DC metro area (DC, Maryland, and 
Virginia) that would prevent compliance with planned study visits 
11. Test of Understanding (TOU) (minimum passing score of 80% with 2 attempts permitted) 
5.3.2. 
 Exclusion Criteria 
Potential participants meeting any of the following criteria will be excluded from enrollment. 
1. Receipt of any investigational HIV vaccine or investigational adjuvant 
2. Received an investigational product or vaccine in the 30 days before enrollment. 
This does not include products with emergency use authorization. 
3. Concurrent participation in another clinical research study 
4. Any serious medical illness or condition  

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5. Receipt of immunoglobulins or blood products within 3 months before enrollment 
6. Any history of anaphylaxis or allergy to study product 
7. History of sickle cell trait or disease 
8. Pregnancy, lactation, or intention to become pregnant during the study 
9. History of active/recent cancer still within treatment or active surveillance follow-
up (except basal cell carcinoma of the skin and cervical carcinoma in situ). 
Treated/resolved cancers with no likelihood of recurrence may be deemed 
acceptable at PI discretion 
10. History of autoimmune disease 
11. History of PIMMCs 
12. Suspected or known current alcohol or drug abuse as defined by an alcohol intake 
of greater than 3 drinks a day on average for a man, and greater than 2 drinks a 
day on average for a woman 
13. Any other significant disease, disorder or finding which may significantly 
increase the risk to the participant because of participation in the study, affect the 
ability of the participant to give informed consent, participate in the study, or 
impair interpretation of the study data, in the opinion of the Investigator 
14. History of splenectomy 
15. History of confirmed or suspected immunodeficiency 
16. History of Hereditary angioedema (HAE) acquired angioedema (AAE), or 
idiopathic forms of angioedema 
17. History of asthma that is unstable or required emergent care, urgent care, 
hospitalization, or intubation during the past 2 years 
18. History of diabetes mellitus (type I or II), with the exception of gestational 
diabetes 
19. History of thyroid disease (except for well controlled hypothyroidism) 
20. History of idiopathic urticaria within the past year 
21. History of hypertension that is not well controlled by medication or that is 
persistently greater than 140/95 at screening 
22. History of bleeding disorder diagnosed by a doctor (e.g., factor deficiency, 
coagulopathy, or platelet disorder requiring special precautions) or significant 
bruising or bleeding difficulties with IM injections or blood draws 
23. History of chronic or active neurologic disease to include seizure disorder and 
chronic migraine headaches. Exceptions are: i) childhood febrile seizures, or ii) 
seizures secondary to alcohol withdrawal more than 3 years ago 
24. Participants receiving any of the following substances 

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a. Systemic immunosuppressive medications or cytotoxic medications within 12 weeks 
before enrollment [with the exception of a short course of corticosteroids (≤ 14 days 
duration or a single injection) for a self-limited condition at least 2 weeks before 
enrollment; inhaled, intranasal or topical steroids are not considered exclusionary 
b. Treatment with known immunomodulators including allergy immunotherapy (other 
than NSAIDs) for any reason 
c. Live attenuated vaccines within 30 days before initial study vaccine administration 
d. Medically indicated subunit, mRNA, or killed vaccines, e.g., influenza, 
pneumococcal, vaccines with QS-21 as an adjuvant, or allergy treatment with antigen 
injections, planned for administration 14 days before or after study vaccine 
administration 
25. History of arthritis diagnosis other than osteoarthritis 
26. History of other diagnosed rheumatoid disorders 
27. Has an acute illness or temperature ≥38.0 ºC/100.4 ºF on any study injection day 
or within 48 hours of planned study injection. 
28. Note: Participants will not be excluded from further consideration for enrollment 
and study injections. Volunteers with fever or an acute illness on the day of study 
injection or in the 2 days before the study injection may be re-assessed by a study 
physician for resolution of the condition and enrolled and receive the study 
injection so long as the injection is within allowable windows. Military personnel 
will be excluded from participation in this study, regardless of leave status due to 
the potential for a false-positive HIV test result on mandatory HIV testing. This 
could have adverse effects on deployment status. 
Final evaluation of eligibility will be based on the medical judgment of the investigator based on 
his/her medical and research experience. 
5.4. 
Lifestyle and Reproduction Restrictions 
Only restrictions to contraception and pregnancy are required. 
5.4.1. 
Contraception and Pregnancy 
Risks to unborn babies and nursing infants are unknown at this time. Therefore, participants 
childbearing potential must agree to follow contraceptive guidance and must agree not to 
breastfeed while in the study and for 2 months after the study. 
5.4.1.1. 
Individuals of Childbearing Potential 
Participants are not considered of childbearing potential if they meet any of the following 
criteria: 
x Premenopausal with 1 of the following: 
 Documented hysterectomy 

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 Documented bilateral salpingectomy 
x Documented bilateral oophorectomy 
x Premenarchal 
x Postmenopausal 
 Defined as no menses for 12 months without an alternative medical cause (A 
high follicle stimulating hormone (FSH) level in the postmenopausal range 
may be used to confirm a postmenopausal state in women not using 
hormonal contraception or hormonal replacement therapy (HRT). However, 
in the absence of 12 months of amenorrhea, a single FSH measurement is 
insufficient.) 
 People assigned female sex at birth on HRT and whose menopausal status is 
in doubt will be required to use one of the nonhormonal highly effective 
contraception methods if they wish to continue their HRT during the study, 
or they must discontinue HRT to allow confirmation of postmenopausal 
status before protocol enrollment 
5.4.1.2. 
Participants of Childbearing Potential 
Participants of reproductive potential are eligible to participate if they agree to use a highly 
effective method of contraception as described the section below. 
5.4.1.3. 
Effective Methods of Contraception 
Highly effective contraceptive methods that have a failure rate of <1% per year when used 
consistently and correctly: 
x User Dependent Methods: 
 Oral, intravaginal, or transdermal combined (estrogen- and 
progestogen- containing) hormonal contraception associated with 
inhibition of ovulation 
 Oral or injectable progestogen-only hormonal contraception associated 
with inhibition of ovulation 
x User Independent Methods: 
 Implantable progestogen-only hormonal contraception associated with 
inhibition of ovulation 
 Intrauterine device (IUD) 
 Intrauterine hormone-releasing system (IUS) 
 Bilateral tubal occlusion 

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x Vasectomized partner 
x Sexual abstinence 
5.4.1.4. 
Pregnancy Testing 
Participants of childbearing potential will be included in the study only after a negative 
pregnancy test. Additional pregnancy testing will be performed according to Table 1 Schedule of 
Evaluations and whenever a menstrual cycle is missed or when pregnancy is otherwise 
suspected. 
6. 
STUDY INTERVENTION 
6.1. 
Treatments Administered 
All products to be administered in this study are described in Table 4. Investigational product is 
defined as any investigational treatment or a product/device being used for an indication that has 
not been approved/cleared by the U.S. Food and Drug Administration (FDA). 
Table 4: 
Products Administered 
 
Investigational 
Product 
Investigational 
Product 
Investigational Product 
Compound 
A244 
B.63521 
ALFQ 
Dosage formulation: 
0.5 mg/mL of the 
A244 gp120 protein, 
20 mM Tris, 123 
mM 
NaCl pH 7.4 
0.60 mg/mL of 
the 
B.63521gp120 
protein, 20 mM 
sodium 
phosphate, 150 
mM NaCl, 0.02% 
polysorbate 
80(PS80), pH 
6.5. 
The concentration of DMPC is 
25.8+/-3.9 mM. The concentration 
of DMPG 2.9 +/- 0.4 mM. The 
synthetic cholesterol 
concentration is 34.9+/1 5.2mM 
Unit dose 
strength(s)/dos age 
level(s): 
300 ug 
300 ug 
Arm 1 will contain 200ug 
MPLA/3D-PHAD®, Arm 2 will 
contain 100 ug MPLA/3D-
PHAD®, and Arm 3 will contain 
50ug MPLA/3D-PHAD®. ALFQ 
also contains 100ug of QS-21. 
Route of 
administration: 
IM 
IM 
IM 
Manufacturer: 
Vetter Development 
Services USA Inc. 
For: NIH/NIAID 
Vetter Skokie, IL 
for NIH/HIAID 
Walter Reed Pilot Bioproduction 
Facility 

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6.2. 
Preparation, Handling, Storage, and Accountability 
6.2.1. 
Acquisition Accountability 
The sponsor’s representative is responsible for distributing the investigational product to the 
study site and has ultimate responsibility for product accountability. 
Responsibility for investigational product (IP) accountability at the trial site rests with the 
investigator/institution. 
The sponsor’s representative may release the product to other clinical study sites. 
After the investigational product is distributed, the site pharmacy personnel are responsible for 
maintaining logs of investigational product receipt, storage, reconstitution, accountability by 
subject, and remaining before final disposition within the site. The investigational product should 
be stored as specified by the sponsor. The PI may delegate, in writing, this responsibility to 
another individual, but the PI is ultimately responsible for the investigational product and its 
proper storage upon receipt at the study site until it is transferred back to the sponsor’s 
representative, or designee, or is destroyed, as directed by the sponsor’s representative. 
The empty vials and the unused portion of the vial should be discarded in a biohazard 
containment bag and incinerated after verification from the monitor. Any unopened vials (past 
the re-test date or otherwise) that remain will be destroyed at the discretion of the sponsor in 
accordance with local site SOP as directed by the Sponsor’s representative and as stipulated by 
local, state, and Federal regulations. Partially used vials or expired prepared doses will not be 
administered to other participants or used for in vitro experimental studies. They will be disposed 
of in accordance with institutional pharmacy policy. Retest dates shall be updated as per 
analytical analyses of the IP. No IP shall be administered to a participant beyond the current 
retest date. The decision for final disposition of the investigational product rests with the 
sponsor. 
Any temperature excursions during product storage must be reported to the sponsor and the 
affected IP must be placed in quarantine status. The sponsor will determine the disposition on the 
affected product. 
Further guidance and information for the final disposition of unused study treatment are provided 
in the pharmacy section of the Manual of Operations (MOP). 
6.2.2. 
Product Storage and Stability 
All study products will be transported in accordance with the labeled storage conditions. 
Once at the study site, all study products will be stored in a secure, environmentally controlled, 
and monitored (manual or automated) area in accordance with the labeled storage conditions 
with access limited to the investigator and authorized site staff. Only participants enrolled in the 
protocol may receive study injections, and only authorized site staff may supply or administer 
study injections. 
The investigator or designee will confirm that appropriate temperature conditions have been 
maintained during transit for all study products received and that any discrepancies are reported 
and resolved before use of the study products. 

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6.2.3. 
Product Preparation 
All investigational product preparation will be performed by the unblinded study nurse on the 
day of vaccine administration, after confirmation by the clinical staff that the participant is 
present, is eligible for enrollment into the study, and has received their randomization 
assignment. The unblinded study nurse will perform all manipulations of investigational products 
in accordance with GCP guidelines. The unblinded study nurse will wear a lab coat and gloves 
for all manipulations. The center seals of the vials will be removed and the vial stoppers wiped 
with an alcohol wipe prior to insertion of a sterile needle attached to a sterile syringe. The 
Pharmacy section of the MOP outlines specific product preparations. 
6.3. 
Study Intervention Compliance 
All IP is administered by the study team and documented on the Study Vaccine Administration 
CRF. A study vaccination outside of a participant’s vaccination window may occur at the 
discretion of the PSRT. 
6.4. 
Measures to Minimize Bias: Randomization and Blinding 
The PI, study staff, and participants will be blinded as to group allocation as it pertains to the 
dose of ALFQ. Since the vaccines are not identical in appearance, to preserve blinding the 
material inside the syringe will be masked. In addition, the delegated un-blinded study 
staff/nurse preparing the vaccine syringes will not be involved in the clinical assessment of 
participants and will be instructed not to comment on the appearance of experimental agent to 
study staff. For all participants, the volume of injection will be consistent. 
Delegated un-blinded study staff/nurse staff will be trained in GCP and instructed not to discuss 
the vaccine randomization lists, codes, or participant assignments with study personnel. 
Unblinded study staff will be the only person(s) on site who will know the randomization 
assignment. The randomization assignment will not appear on any label or source document. 
Samples will be labeled on site using labels containing participant study numbers, protocol 
number and visit number. The samples will be accompanied by a specimen tracking form that 
records the study number, date and time of sample collection. 
Any request for unblinding, with its rationale, must be forwarded through the PI. The PI will 
evaluate the request and will notify the Protocol Chair/Sponsor's Medical Expert for the Trial 
and HIV Vaccine Product Manager. They will evaluate the request and will advise the United 
States Army Medical Research and Development Command (USAMRDC) Office of Regulated 
Activities (ORA) Product Safety Surveillance Office (PSSO) regarding a course of action. A 
PVG physician from the USAMRDC ORA PSSO will decide whether to approve the request for 
unblinding. 
In the case of a decision to unblind, the Sponsor will authorize the independent statistician to 
provide this information to the PI, who must notify the WRAIR IRB (see Table 7) and provide 
the study assignment to the site physician. It should be noted that there are very few 
circumstances in which unblinding will be essential to the medical management of a vaccine 
recipient. The site investigator will report episodes of accidental unblinding, with an explanation, 
to Protocol Chair/Sponsor's Medical Expert for the Trial, the Institution Review Boards (IRBs) 

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and the USAMRDC ORA PSSO. Follow-up of unblinded participants will continue through the 
duration of the trial. 
After completion of the study and database lock, the Sponsor will authorize the independent 
statistician to prepare a list containing each participant’s unblinded treatment assignment. 
Participants will be contacted to return to clinic or will be notified via phone or letter 
(Appendix A ) of their unblinded treatment assignment. 
6.5. 
Concomitant Therapy 
Concomitant medications will be recorded from 45 days prior to first vaccination until 10 months 
after the final vaccination. An initial review of past medical history will include a review of 
medications. Information pertaining to receipt of non-study vaccines, research agents, 
immunoglobulin preparations, immunosuppressive medication, antiretroviral drugs, and any 
blood products will be elicited at study visits and recorded in source documents. 
To ensure appropriate medical follow-up for study participants, information regarding 
concomitant medications used in association with an AE will be collected and recorded in the 
source documents. 
Study participants can receive medications such as acetaminophen, non-steroidal anti-
inflammatory drugs (NSAIDs), or antihistamines as required, although they must be 
documented. 
6.6. 
Dose Modification 
Excluding the aforementioned study arm dose-escalation of ALFQ, no modification of dosage 
for any of the vaccine products will be allowed in this study. 
6.7. 
Treatment After the End of the Study 
There is no plan to continue with study injections after the end of the study. 
7. 
DISCONTINUATION CRITERIA 
Each participant has the right to withdraw from the study at any time for any reason without 
penalty. In those cases where a “withdrawal of consent” is requested by the participant, 
documentation of the withdrawal of consent and the reason(s) for the request will be captured in 
the participant’s clinical research records and/or on the status change CRF. Although the 
participant is not obligated to give reason(s) for withdrawing, the investigator should make a 
reasonable effort to ascertain the reason(s) while fully respecting the participant’s rights. The 
participant will also be asked if she/he will agree to complete the clinical assessments listed for 
the study termination/exit visit. 
7.1. 
Screening Failures 
Screen failures are defined as subjects who consent to participate in the clinical study but either 
are deemed not eligible or are not subsequently randomized. The investigator will ensure that the 

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following information is collected for each screen failure—demographic information, and the 
reason for the screen failure. 
7.2. 
Discontinuation of Study Intervention 
Under certain circumstances, a participant will be terminated from participating in further 
vaccinations. These specific experiences include: 
o Investigator’s clinical judgment is that it is in the best interest of the subject 
o Confirmed diagnosis of HIV 
o Pregnancy 
o Clinically significant Type 1 hypersensitivity associated with vaccination 
o Serious concomitant illness that is not expected to resolve prior to the next 
scheduled vaccination or will require contact isolation during the next 
scheduled vaccination.  
o Treatment with chronic systemic glucocorticoids (e.g., prednisone) or other 
immunomodulators other than NSAIDs for greater than 10 days. Inhaled and 
topical steroid use is not exclusionary 
o Need for concomitant vaccine that requires discontinuation 
o After initial vaccination, missing any subsequent vaccination by more than 15 days 
Participant’s request Immediate Early Discontinuation will occur: 
1. Upon learning of pregnancy; 
2. Due to an SAE that is believed to be related to study vaccination, including:  
a. Any Grade 3 systemic adverse reaction that is assessed as related to the study agent,  
b. Any Grade 4 AE assessed as related to the study agent, or  
3. If a participant develops infection with HIV-1.  
Each case will be reviewed by the Protocol Safety Review Team (PSRT) to assess impact on 
study conduct overall and continuation of individual participants with referral to the U.S. FDA 
and WRAIR IRB, in consultation with the study sponsor. 
7.2.1. 
PSRT 
The PSRT will include the following: 
x Sponsor’s Medical Expert or designee 
x HIV Vaccine Product Manager or designee 
x ORA Pharmacovigilance (PVG) Physician 
x PI or designee 
x Independent Safety Monitor (ISM) 
Additional PSRT participants may include, as needed, the following: 
x Associate Investigators 
x Clinical research nursing staff 
x Laboratory directors 

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x Data management and regulatory staff 
x DAIDS Medical Officer 
PSRT conference calls or meetings will require, at minimum, the participation of the Sponsor’s 
Medical Expert or designee, ISM, and the PI or designee. The WRAIR IRB will be notified of 
the decisions taken by the PSRT if they involve suspensions, clinical holds (voluntary or 
involuntary), or terminations of this research in accordance with Table 8. The USAMRDC ORA 
PSSO will be responsible for safety reporting to the U.S. FDA. 
The DAIDS Medical Officer will serve in a consultant role on the PSRT but is not required for 
PSRT or any other protocol team decisions.  
Participants who are discontinued from additional study vaccinations will be discussed with the 
PSRT to determine the duration, if any, of  follow-up visits to further evaluate safety and monitor 
adverse experiences. 
Wherever possible, the tests and evaluations listed for the termination visit should be carried out 
if the subject refuses follow-up according to the protocol visit schedule. 
7.2.2. 
Pregnancy 
Subjects who become pregnant post investigational product administration will not receive any 
additional investigational product. The subject will be followed for safety to term by phone 
interview and laboratory assessment. 
7.3. 
Participant Discontinuation/Withdraw from the Study 
A participant may withdraw from the study at any time at his/her own request, or may be 
withdrawn at any time at the discretion of the investigator for safety, behavioral, or 
administrative reasons. Reasons may include the following: 
x Repeated failure to comply with protocol requirements 
x Recommended withdrawal by the study investigator, e.g., because of worsening health 
status, intercurrent illness, or AEs interfering with study assessments as determined by 
the investigator 
x Participant requests withdrawal. 
If the participant withdraws consent for disclosure of future information, the sponsor’s 
representative may retain and continue to use any data collected before such a withdrawal of 
consent. If a participant withdraws from the study, he/she may request destruction of any 
samples taken and not tested, and the investigator must document this in the site study records. 
Participants who acquire HIV will be discontinued from study vaccinations, and additional 
research blood draws will not be performed. However, the participant will continue with safety 
assessments: pregnancy test, CBC with differential, and Creatinine/ALT.  
Refer to the schedule of activities in Table 1 for data to be collected at the time of study 
discontinuation and follow-up and for any further evaluations that need to be completed. 

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7.3.1. 
Replacement of Withdrawn Participants 
Participants who withdraw or are withdrawn from study vaccinations or the study completely, or 
are lost to follow-up after signing the ICF and administration of the study product will be 
replaced if enrollment is still open. If a participant withdraws from the study or discontinues 
vaccination before all slots have been filled, the vacated randomization slot will be filled with the 
next available participant. Participants who did not receive the study injection will not count 
toward the total participants enrolled in the study. 
7.3.2. 
Follow-up for Participants Who Withdraw Consent 
After withdrawal of consent, the study team will engage in no further communication with the 
participant except as directed by an IRB and/or USAMRDC regarding information concerning 
participant safety. 
7.4. 
Lost to Follow-up 
A participant will be considered lost to follow-up if he or she repeatedly fails to return for 
scheduled visits and is unable to be contacted by the study site. 
The following actions must be taken if a participant fails to return to the clinic for a required 
study visit. 
The site must attempt to contact the participant and reschedule the missed visit as soon as 
possible and counsel the participant on the importance of maintaining the assigned visit schedule 
and ascertain whether or not the participant wishes to and/or should continue in the study. 
x In cases in which the participant is deemed lost to follow up, the investigator or 
designee must make every effort to regain contact with the participant (where 
possible, three (3) telephone calls and, if necessary, a certified letter to the 
participant’s last known mailing address or local equivalent methods). These contact 
attempts should be documented in the participant’s medical record. 
x Should the participant continue to be unreachable for 3 months, he/she will be 
considered to have withdrawn from the study with a primary reason of lost to follow-
up. 
8. 
STUDY ASSESSMENTS AND PROCEDURES 
Study procedures and their timing are summarized in Section 1.3, Schedule of Activities. 
All screening evaluations will be completed and reviewed to confirm that potential subjects meet 
all eligibility criteria. The investigator will maintain a screening log to record details of all 
subjects screened and to confirm eligibility or record reasons for screening failure, as applicable. 
The maximum amount of blood collected from each subject during the study will not exceed 550 
mL per 8 weeks. Blood will be collected into vacutainer tubes according to standard operating 
procedures. Substitution of tube types may be made as long as the substitution does not interfere 
with the study performance. A 10% variance in blood volume is acceptable, in case of an error or 
a broken or clotted vial. 

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8.1. 
Safety Assessments 
8.1.1. 
Demographic/Medical History 
Age, sex at birth, gender identity, date of birth, birthplace, level of education, occupation and 
baseline medical history of participants will be recorded. Participant’s medical conditions, 
previous procedures, medications, and history of abnormal laboratory values, if available, will be 
recorded in the participant’s medical history. 
8.1.2. 
Physical Examination 
For the purpose of assessing AEs, symptoms and directed medical examination will be 
performed based on the medical judgment of the investigator. In addition, participants will be 
asked to record their temperature and complete a diary card at home in the evening after the 
vaccination and each evening thereafter for a total of 14 days. Study staff will provide 
thermometers and participants will be trained how to complete the diary card, use the 
thermometer, and to measure injection site induration and redness using the ruler after each 
vaccination. 
Investigators will pay special attention to clinical signs related to previous serious illnesses. 
8.1.3. 
Vital Signs 
Body temperature, pulse, respiratory rate, and blood pressure will be measured prior to study 
vaccination and again at 30 minutes post-vaccination. Blood pressure and pulse measurements 
will be assessed in the seated position with legs uncrossed with a completely automated device. 
Manual techniques will be used only if an automated device is not available. Blood pressure and 
pulse measurements will be preceded by at least 5 minutes of rest for the subject in a quiet 
setting without distractions (e.g., television, cell phones). 
8.1.4. 
Clinical Laboratory Assessments 
Clinical laboratory assessments will be performed according to standard operating procedures by 
the laboratory listed in the General Information of this protocol. The clinical laboratory tests to 
be performed and the timing and frequency is provided in Section 1.3, Schedule of Activities. 
Screening labs may be repeated one time. 
Laboratory values will be graded and their clinical significance evaluated according to the 
following procedures: 
x All abnormal laboratory values included in the Toxicity Grading Scale are 
graded by the investigator per Appendix A. 
x The investigator will exercise his or her medical and scientific judgment in 
deciding whether an abnormal laboratory finding or other abnormal assessment 
is clinically significant; however abnormal laboratory results will be judged by 
investigators to be clinically significant if they meet one or both of the following 
criteria: 

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 The abnormality suggests a disease and/or organ toxicity that is new 
or has worsened from baseline. 
 The abnormality is of a degree that requires additional active management 
(e.g., change of dose, discontinuation of the drug, close observation, more 
frequent follow-up assessments, or further diagnostic investigation). 
x Abnormal laboratory findings or other abnormal assessments judged by the 
investigator to be clinically significant will be recorded as AEs or SAEs if 
appropriate. 
x All events (Grade 1 or higher) must be recorded, regardless of suspected causal 
relationship or clinical significance (e.g., including those determined by the 
physician investigator as non-clinically significant). 
x Clinically significant abnormal laboratory findings or other abnormal assessments 
that are detected before the first administration of investigational product will be 
documented on medical history as pre-existing conditions and will be reported as 
AEs or SAEs only if the pre-existing conditions are worse following the vaccination. 
x Clinically significant abnormal laboratory findings or other abnormal assessments 
that are detected after the first administration of investigational product during the 
study will be reported as AEs or SAEs. 
x Chronic clinically significant laboratory abnormalities will be followed until 
they return to normal or to baseline, stabilize with the probability of becoming 
chronic, or a satisfactory explanation is provided by the investigator. 
8.1.4.1. 
Hematology 
Complete blood cell count (with differential) will be performed on whole blood as per the SOE 
(Table 1). 
8.1.4.2. 
Blood Chemistry 
Creatinine and alanine amino-transferase will be tested on plasma or serum according to the 
SOE. 
8.1.4.3. 
Laboratory assays 
HIV EIA and HIV Western blot or immunochromatographic test U.S. FDA-approved kits maybe 
used. 
HIV nucleic acid tests (NAAT): serum and/or plasma collected in a suitable anti-coagulant 
(EDTA or ACD) will be assayed for HIV NAAT using at least one nucleic acid test platform. 
Supplemental nucleic acid tests may also be incorporated. 
Samples that are Western Blot or immunochromatographic test indeterminate may also be 
subject to supplemental Nucleic Acid testing. 

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8.1.4.4. 
Pregnancy Screen 
Pregnancy tests will be performed at screening, and prior to the vaccination as indicated in the 
SOE. No vaccine procedures will be performed if the pregnancy test is positive. 
8.2. 
Adverse Events 
The following terms, as defined by 21 CFR 312.32, apply to Investigational New Drug (IND) 
application safety reporting. 
8.2.1. 
Definitions 
8.2.1.1. 
Adverse Event and Suspected Adverse Reaction 
An adverse event: 
x Is any untoward medical occurrence associated with the use of a drug in 
humans, whether or not it is considered drug related 
x Including any unfavorable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal (investigational) product 
A suspected adverse reaction: 
x Is any AE for which there is a reasonable possibility that the 
investigational product caused the adverse event, with “reasonable 
possibility” meaning there is evidence to suggest a causal relationship 
between the investigational product and the AE 
x Implies a lesser degree of certainty about causality than adverse reaction, 
which means any AE caused by an investigational product 
8.2.1.2. 
Solicited Adverse Event 
A solicited adverse event: 
x Is a pre-determined event identified in the IB that may reflect safety concerns 
related to the investigational product 
x Is specifically asked about (orally or in writing) during subject questioning 
The solicited AEs for this protocol include: 
Local 
x Pain/tenderness 
x Itching 
x Warmth 
x Redness/erythema measurement of largest dimension 

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x Induration measurement of largest dimension 
Systemic 
x Fever 
x Myalgia 
x Arthralgia 
x Headache 
x Fatigue 
x Chills 
x Rash 
x Nausea 
x Dizziness 
Solicited adverse events will be recorded daily by the participant, starting the evening of each IP 
administration and each evening thereafter for a total of 14 additional days following each IP 
administration. They will use a diary card, for recording any reactions they experience, which 
will be provided to participants at the time of each IP administration.  The diary card will act as a 
memory tool for the participant and may be used as a source document. Participants must 
complete the full diary card prior to the administration of any subsequent vaccine. 
8.2.1.3. 
Serious Adverse Event 
An AE or suspected adverse reaction is considered serious if it results in any of the following 
outcomes: 
x Death 
x Life-threatening AE 
 In the view of either the investigator or sponsor, its occurrence 
places the participant at immediate risk of death 
 It does not include an AE or suspected adverse reaction that, had it 
occurred in a more severe form, might have caused death 
x Inpatient hospitalization or prolongation of existing hospitalization 
x Persistent or significant incapacity or substantial disruption of the ability to 
conduct normal life functions 
x Congenital anomaly/birth defect 

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x Important medical events that may not result in death, be life-threatening, or 
require hospitalization may be considered serious when, based upon 
appropriate medical judgment: 
 They may jeopardize the participant and may require medical or 
surgical intervention to prevent one of the outcomes listed above 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 
Planned hospitalizations or surgical procedures for pre-existing conditions are not to be 
considered SAEs unless the condition worsens or requires additional interventions post 
investigational product administration. Pre-existing conditions, in general, are not to be 
considered SAEs unless the condition worsens (in frequency, severity, or intensity) or requires 
additional interventions post-investigational product administration. 
8.2.1.4. 
HIV Diagnostic Algorithm 
Diagnostic HIV testing will utilize a series of validated tests that will differentiate between 
vaccine induced seroreactivity and true HIV infection. HIV test information provided to the 
study staff at each vaccine trial site will not include the test results of individual tests, but rather 
will be provided with an aggregate interpretation as HIV "infected" or "not infected", or that 
repeat testing is needed (as in the case of need for a verification specimen). Report of results will 
be delayed to at least 10 days from blood collection so that the timing of HIV test reporting does 
not compromise the double-blind nature of the trial, as a result returned immediately after EIA 
testing could signal the clinical team that this specimen did not require the Western blot or 
immunochromatographic testing which would be needed if vaccine had induced an antibody 
response. 
In the case of suspected HIV infection with a positive nucleic acid test, a verification specimen 
may be requested immediately.  If HIV infection is confirmed, results may be returned to the 
participant immediately. 
8.2.1.5. 
Medically Attended Adverse Events (MAAE) 
The FDA requests that clinical trials of preventive vaccines with adjuvants such as ALFQ 
provide for collection and analysis of data relating to MAAEs among subjects in all treatment 
groups through 12 months or longer following the last study vaccination, due to the theoretical 
potential for induction of autoimmune or auto-inflammatory diseases. 
MAAEs are defined as AEs with medically attended visits including hospital, emergency room, 
urgent care clinic, or other visits to or from medical personnel for any reason. Adverse events 
(e.g., abnormal vitals) identified at a routine study visit will not be considered MAAEs. MAAEs 
include, but are not limited to, PIMMCs. 

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8.2.1.6. 
Unexpected Adverse Event or Unexpected Suspected Adverse Reaction 
An AE or suspected adverse reaction is considered unexpected if: 
x It is not listed in the IB or is not listed at the specificity or severity that has 
been observed 
x If an IB is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current 
application, as amended 
x It is mentioned in the IB as occurring with a class of drugs or as anticipated from 
the pharmacological properties of the drug but is not specifically mentioned as 
occurring with the particular drug under investigation 
For example: 
x Hepatic necrosis would be unexpected (by virtue of greater severity) if 
the Investigator’s Brochure referred only to elevated hepatic enzymes or 
hepatitis 
x Cerebral thromboembolism and cerebral vasculitis would be unexpected (by 
virtue of greater specificity) if the IB listed only cerebral vascular accidents. 
8.2.2. 
Timeframe for Collecting Adverse Event Information 
All AEs and SAEs will be collected from the start of treatment until the end of the protocol-
defined follow-up period, at the time points specified in Section 1.3, Schedule of Activities. 
Adverse events and SAEs will be assessed at all study visits, documented in the source records, 
and recorded on the CRFs using accepted medical terms and/or the diagnoses that accurately 
characterize the event. All unsolicited AEs will also be collected from at least 28 days after each 
vaccination.  Medical occurrences that begin before the start of study treatment but after 
obtaining informed consent will be recorded on the Medical History/Current Medical Conditions 
section of the CRF. 
Unsolicited adverse events (AEs) and Medically Attended Adverse Events (MAAEs) will be 
collected from the day of IP administration and for the next 28 days.  The investigator will also 
document SAEs, MAAEs and AESIs (including PIMMCs) for at least 12 months after subjects 
receive their last study vaccination. 
8.2.3. 
Methods for Obtaining AE and SAE Data 
Adverse events will be reported by the subject or detected by the investigator or designee during 
examination or assessment. 
The investigator and any designees are responsible for detecting, documenting, and reporting 
events that meet the definition of an AE or SAE and remain responsible for following up AEs 
that are serious, considered related to the study treatment or the study, or that caused the subject 
to discontinue the study treatment or the study (Section 7.3). Participants will be asked for their 
consent for photography if the clinician wishes to document an unusual or unexpected finding. 
Participants will not be able to be identified by the photographs, which will be kept indefinitely. 

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When an AE or SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) related to the 
event. 
8.2.4. 
Recording Adverse Event and Serious Adverse Event Data 
When a diagnosis is known, the AE term recorded on the case report form will be the diagnosis 
rather than a constellation of symptoms. The investigator will assess all AEs for seriousness 
(Section 0), severity (Section 8.2.5), relationship to investigational product (Section 8.2.6), and 
other possible etiologies. When an event has not resolved by study closure, it will be documented 
on the AE case report form as “not recovered/not resolved.” The AE verbatim (in the clinical 
database) and the SAE verbatim (in the safety database) should be recorded consistently and that 
any updates to the event should be made in both places. 
Serious adverse events will be recorded on the SAE report form as follows: 
x Start date - date the AE met serious criteria. 
x Resolution date - date the event returned to baseline status or the condition 
stabilized or was determined chronic 
x If not resolved by study closure, the event will be documented on the SAE 
report form as “not recovered/not resolved.” 
8.2.5. 
Severity Assessment 
All AEs (laboratory and clinical symptoms) will be graded for severity and assessed for 
relationship to study product using the DAIDS Table for Grading the Severity of Adult and 
Pediatric Adverse Events (DAIDS AE Grading Table), Version 2.1 dated July 2017 that is found 
on the website https://rsc.niaid.nih.gov/clinical-research-sites/daids-adverse-event-grading-
tables. AEs 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. Changes in the severity of an AE will be documented to allow an 
assessment of the duration of the event at each level of intensity. Adverse events not identified in 
the grading table, follow the guidelines provided at the beginning of the table.  
All other events will be graded according to the scale in Table 5. The case report form for AEs 
will reflect only the highest severity for continuous days an event occurred. 
8.2.6. 
Estimating Severity Grade for Parameters Not Identified in the Grading Table 
The functional table below should be used to grade the severity of an AE that is not specifically 
identified in the grading table. In addition, all deaths related to an AE are to be classified as 
Grade 5. 

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Table 5: 
DAIDS Adverse Event Grading Scale 
PARAMETER 
GRADE 1 
MILD 
GRADE 2 
MODERATE 
GRADE 3 
SEVERE 
GRADE 4 
POTENTIALLY 
LIFE-
THREATENING 
Clinical adverse 
event NOT 
identified 
elsewhere in the 
grading table 
Mild symptoms 
causing no or 
minimal 
interference with 
usual social & 
functional 
activities with 
intervention not 
indicated 
Moderate 
symptoms 
causing greater 
than minimal 
interference with 
usual social & 
functional 
activities with 
intervention 
indicated 
Severe symptoms 
causing inability 
to perform usual 
social & 
functional 
activities with 
intervention or 
hospitalization 
indicated 
Potentially life-
threatening 
symptoms causing 
inability to perform 
basic self-care 
functions with 
intervention 
indicated to prevent 
permanent 
impairment, 
persistent disability, 
or death 
8.2.7. 
Exceptions to the Grading Table 
8.2.7.1. 
Creatinine 
Creatinine is required to be reported as an AE only if it is gradable per the increase from local 
lab upper limit of normal (ULN) parameter. Do not grade elevated creatinine based on the 
change from the baseline parameter.  
Do not grade creatinine clearance or eGFR based on the change from the baseline parameter. Do 
not grade on the basis of eGFR if there is clinical concern for kidney injury. 
8.2.7.2. 
Injection Site Erythema and Induration 
Injection Site Erythema and Injection Site Induration will not consider surface area and 
interference with usual social and functional activities such that: 
x Grade 1 is: 2.5 to < 5 cm in diameter; 
x Grade 2 is: ≥ 5 to < 10 cm in diameter; 
x Grade 3 is: ≥ 10 cm in diameter OR ulceration OR secondary infection OR phlebitis 
OR sterile abscess OR drainage; 
x Grade 4 is: Potentially life-threatening consequences (e.g., abscess, exfoliative 
dermatitis, necrosis involving dermis or deeper tissue 
8.2.7.3. 
Unintentional Weight Loss 
Unintentional weight loss is required to be reported as an AE only if it is considered to be 
potentially deleterious to the participant’s health. 

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8.2.8. 
Causality Assessment 
A physician must assign a relationship of each AE to the receipt of the investigational product 
using the relationship categories in Table 6. 
Table 6: 
Causality Relationships 
Relationship 
Description 
Not related 
No relationship to investigational product. Applies to those events for 
which evidence exists that there is an alternate etiology. 
Unlikely 
Likely unrelated to the investigational product. Likely to be related to 
factors other than the investigational product but cannot be ruled out with 
certainty. 
Possible 
An association between the event and the administration of investigational 
product cannot be ruled out. There is a reasonable temporal association, but 
there may also be an alternative etiology, such as the subject’s clinical 
status or underlying factors (including other therapy). 
Probable 
There is a high degree of certainty that a relationship to the investigational 
product exists. There is a reasonable temporal association, and the event 
cannot be explained by known characteristics of the subject’s clinical state 
or factors including other therapy. 
Definite 
An association exists between the receipt of investigational product and the 
event. An association to other factors has been ruled out. 
8.2.8.1. 
Serious Adverse Events 
The investigator will report SAEs to the ORA PSSO according to the requirements in Table 8. A 
summary of the required information is provided in Table 9. When reporting via email, the 
subject lines should be formatted as follows: 
Initial: SAFETY REPORT – IND # xxx  
, Sponsor Study #S-xx-xx  
, Subject# 
 
, Event term:  
 
Follow-Up: SAE Case Number provided by PSSO (e.g., US-000123), SAFETY 
REPORT – IND # xxx  
, Sponsor Study #S-xx-xx  
, 
Subject#  
, Event term:  
 
Investigators must also follow all relevant requirements regarding the timely reporting of AEs to 
the WRAIR IRB and the USAMRDC Office of Human and Animal Research  Oversight 
(OHARO). 
There may be instances when copies of medical records for certain cases are requested by the 
sponsor’s safety office. In this case, all subject identifiers, with the exception of the subject 
number, must be redacted on the copies of the medical records before submission to the 
sponsor’s safety office. 
8.2.9. 
Sponsor Independent Safety Monitor 
Independent Safety Monitor (ISM): The Sponsor Independent Safety Monitor (ISM) is a 
physician with relevant expertise whose primary responsibility is to provide independent safety 
monitoring in a timely fashion. The ISM is required to review and provide an unbiased written 
report for all SAEs and subject deaths to the ORA PSSO (Safety Office) within 24 hours of their 

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awareness of the event. The report provided must include, at a minimum, a brief summary of the 
ISMs review of the event and event outcome, relationship of the event to the investigational 
product/device and alternate etiology if the event is deemed unrelated to the investigational 
product/device.   
In addition to the responsibilities above, the ISM maybe required to attend safety meetings and 
providing assessments on subject withdrawals, sentinel subject data reviews, study halting 
criteria, etc., and must be clearly defined in the sponsor approved clinical trial protocol. 
8.2.10. 
Reporting Serious and Unexpected Adverse Events 
Contact information for reporting SAEs is provided in Table 7. 
Table 7: 
Study Contacts for Reporting Serious and Unexpected Adverse Events 
Sponsor’s Safety Office 
US Army Medical Research & Development Command Office 
of Regulated Activities 
Product Safety Surveillance Office ATTN: FCMR-ORA 
1430 Veterans Drive 
Fort Detrick, MD 21702-5009 
Fax: 301-619-7790 
Phone Number: +1 301 619 1005 
Email: usarmy.detrick.medcom-usamrmc.mbx.sae- 
reporting@health.mil 
Institutional Review Board 
Walter Reed Army Institute of Research IRB 503 Robert Grant 
Avenue 
Silver Spring, MD 20910, USA  
Phone Number: 301-319-9940 
Email: usarmy.detrick.medcom-wrair.mbx.hspb@health.mil 
USAMRDC Office of Human and 
Animal  Research Oversight 
Office of Human and Animal Research Oversight (OHARO) 
Office of Human Research Oversight (OHRO) 
US Army Medical Research and Development Command, 
ATTN: FCMR-RPH 
504 Scott Street 
Fort Detrick, MD 21702-5012 Fax: +1 301 619 7803 
Phone Number: 301-619-2165 
Email: usarmy.detrick.medcom-usamrmc.other.hrpo@health.mil 
Independent Safety Monitor 
Melanie McCauley, MD 
Research Physician | HJF 
in support of the Emerging Infectious Diseases Branch (EIDB) 
Walter Reed Army Institute of Research (WRAIR) 
6720A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
Phone Number: 301- 500-3636 

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8.2.10.1. Reporting to WRAIR IRB 
All SAEs related to participation in the study, and all deaths will be promptly (within 48 hours) 
reported to the WRAIR IRB and written reports will be submitted to the WRAIR IRB within 10 
working days. The WRAIR Human Subjects Protection Branch (HSPB) will report all related 
SAEs and all participant deaths to USAMRDC Office of Human and Animal Research  
Oversight (OHARO) Office of Human Research Oversight (OHRO) as per USAMRDC 
Command Policy 21 Follow up reports will be submitted as additional information becomes 
available. 
A summary of the non-serious AEs and SAEs (both related and unrelated) that occurred during 
the reporting period will be included in the continuing review report (CRR) to the WRAIR IRB. 
Investigators are required to forward safety information provided by the sponsor’s representative 
to the WRAIR IRB in a timely manner.  
8.2.11. 
Additional Immediately Reportable Events 
8.2.11.1. Potentially Immune-Mediated Medical Conditions (PIMMCs) 
PIMMCs (Appendix L ) constitute a group of AEs that includes diseases which are clearly 
autoimmune in etiology and other inflammatory and/or neurologic disorders which may or may 
not have autoimmune etiologies. 
Sponsor’s Safety Office 
US Army Medical Research & Development Command Office 
of Regulated Activities 
Product Safety Surveillance Office ATTN: FCMR-ORA 
1430 Veterans Drive 
Fort Detrick, MD 21702-5009 
Fax: 301-619-7790 
Phone Number: +1 301 619 1005 
Email: usarmy.detrick.medcom-usamrmc.mbx.sae- 
reporting@health.mil 
Email: mmccauley@eidresearch.org 
Alternate Independent Safety 
Monitor 
Roger Ying, MD  
Research Physician | HJF 
U.S. Military HIV Research Program (MHRP) 
6720A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
Email:RYing@global-id.org 
 

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8.2.11.2. Unanticipated Problems Involving Risks to Subjects or Others  
Unanticipated problems involving risks to subjects or others (UPIRTSO) are any incident, 
experience, or outcome that meets all of the following criteria: 
x Is unexpected (in terms of nature, severity, or frequency) given 
 the procedures that are described in the protocol, IB, or informed consent 
document; and 
 the characteristics of the subject population 
x Is related or possibly related to participation in the study 
x Suggests that the protocol places participants or others at a greater risk of harm than 
was previously known or recognized 
These events encompass a broader category of events than SAEs and may include issues such as 
problems with loss of control of participant data or the investigational product; adverse 
psychological reactions; or breach of confidentiality. 
The PI will determine whether a given incident, experience or outcome constitutes a UPIRTSO 
and will promptly (within 48 hours) report UPIRTSOs to the WRAIR IRB and to the sponsor. 
The PI will then submit a written report within 10 working days to the WRAIR IRB. 
Follow up reports should be submitted as soon as additional information becomes available. A 
summary of UPIRTSOs will also be included in the continuing review report submitted to the 
WRAIR IRB. The WRAIR HSPB will report UPIRTSOs to the USAMRDC OHARO OHRO as 
per USAMRDC Command Policy 21. 
8.2.11.3. Study Pause or Termination 
The PSRT will closely monitor and analyze study data as they become available and will make 
determinations regarding the presence, severity, and seriousness of AEs. The administration of 
study injections and new enrollments will be paused, and the Sponsor will be promptly notified 
regarding a safety issue according to the following criteria: 
x One (or more) participant experiences an SAE that is assessed as related to study 
agent, or  
x Two (or more) participants experience grade 3 or 4 AEs of the same type related to a 
study agent. This would exclude monitoring lab values if a clear alternate etiology is 
identified.  Additionally, CBC differential counts would not contribute. 
The WRAIR IRB will be immediately notified of any suspensions, clinical holds (voluntary or 
involuntary), or terminations of this research by any of the WRAIR IRB, the PSRT, the 
institution, the sponsor, or regulatory agencies. WRAIR HSPB will forward the notification to 
USAMRDC OHARO OHRO as per USAMRDC Command Policy 21. 
8.2.11.4. Participant Status Change: Incarceration 
Participation of prisoners is not planned, and any participant will be suspended from study visits 
while incarcerated, except visits or telephone contact for ensuring their safety. No study product 

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will be administered to a participant who is incarcerated. The IRB will be notified of the period 
of incarceration. Data and samples that were collected before the period of incarceration may 
still be stored and used for analyses. After release from incarceration, either the participant may 
return to participation in study visits according to the SOE or study participation may be 
terminated at the discretion of the PI. 
Any participant who is incarcerated will be re-consented before rejoining the study. 
When a previously enrolled participant becomes a prisoner and the relevant research protocol 
was not reviewed and approved by the IRB in accordance with the requirements of Department 
of Defense Instruction (DODI) 3216.02 subparagraphs 7.b (1) and (2), to include prisoners as 
research participants, the PI shall promptly notify the WRAIR IRB. WRAIR HSPB will forward 
the notification to USAMRDC OHARO OHRO as per USAMRDC Command Policy 21. 
8.2.11.5. AE-Related Withdrawal of Consent 
Any AE-related withdrawal of consent during the study must be reported by email or fax to 
USAMRDC ORA PSSO within 24 hours of event awareness. The WRAIR IRB will be notified 
of the withdrawal in a follow-up report as per Table 8. 
8.2.12. 
Pending Inspections/Issuance of Reports 
The knowledge of any pending compliance inspection/visit by the FDA, Office for Human 
Research Protections (Department of Health and Human Services), or other government agency 
concerning clinical investigation or research, the issuance of Inspection Reports, FDA Form 483, 
warning letters, or actions taken by any regulatory agency including legal or medical actions and 
any instances of serious or continuing noncompliance with the regulations or requirements will 
be reported immediately to the WRAIR IRB and the sponsor’s representative. The WRAIR 
HSPB will report knowledge of any pending inspections/audits by regulatory agencies to the 
USAMRDC  OHARO OHRO as per USAMRDC Command Policy 21. 
8.2.13. 
IND Annual Report to the FDA 
The sponsor’s representative will notify the PI of the due date of the annual report with sufficient 
time for the PI to assemble and submit all of the required clinical study information to the 
sponsor’s representative. 
The Sponsor and Sponsor’s representative (USAMRDC) will be responsible for the preparation 
of the detailed annual synopsis of clinical activity, including AEs, for submission to the US 
FDA. Each annual report will summarize IND activity for one year beginning approximately 3 
months before the IND US FDA anniversary date.  
8.2.14. 
Final Report 
A final study report will be prepared in accordance with “Guidance for Industry: Submission of 
Abbreviated Reports and Synopses in Support of Marketing Applications” and ICH E3 Guideline 
“Structure and Content of Clinical Study Reports” and provided to the sponsor’s representative 
for review and approval. The sponsor’s representative will use this report to prepare the final 
clinical study report for submission to the U.S. FDA. 

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Table 8: 
Sponsor’s Adverse Event Reporting Requirements 
Event 
Report 
Notification Time 
Notification 
Method 
Serious adverse 
events 
SAE Report 
Within 24 hours of event 
awareness a 
Email: 
usarmy.detrick.me
dcom- 
usamrmc.mbx.sae- 
reporting@health.
mil 
If email is not 
available, the 
information can be 
faxed or called in: 
Fax: 301-619-
0197 
Phone: 301-619-
1005 
Independent Safety 
Monitor Report 
Within 24 hours of event 
awareness 
Pregnancy 
Pregnancy Report 
Within 24 hours of event 
awareness 
Events of Special 
Interest (including 
PIMMCs) 
SAE/AESI) Report 
Within 24 hours of event 
awareness a 
Independent Safety 
Monitor Report 
Within 24 hours of event 
awareness 
Adverse Event 
Related Withdrawals 
Contact PSSO  
Within 24 hours of awareness of 
event awareness 
Safety-related 
Deviations 
Contact PSSO 
Within 5 business days of event 
awareness 
 
a Submission of the reports to the PSSO must not be delayed. Do not wait to receive the independent safety 
monitor’s report. 
Table 9: 
Summary of Information Required When Reporting Adverse Events to the 
Sponsor 
Information to Include on the Serious Adverse Event Report Form includes, but is not limited to: 
Investigational new drug application number, sponsor study number 
Name of the investigational product 
Investigator name and contact number  
Subject identification number 
Serious adverse event term, description 
Onset date 
Date(s) of investigational product administration 
Severity 
Relationship to the investigational product 
Subject’s current status 
Admission/Discharge Summary 
Consultations 
Medical record progress notes including pertinent laboratory/diagnostic test results 
8.2.15. 
Pregnancy Reporting 
Each pregnancy must be reported within 24 hours of identification by completing and submitting 
the Pregnancy Report Form by email or fax to the sponsor's safety office (ORA, Product Safety 

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Surveillance Branch (PSSO)). Pregnancies must be reported within 48 hours of becoming aware 
of the event to the IRB. WRAIR HSPB will forward the notification to USAMRDC OHARO 
OHRO as per USAMRDC Command Policy 21. 
The following information will be reported on the Pregnancy Report: type and date of delivery, 
Apgar scores, and health status of the mother and child including the child's sex, head 
circumference, gestational age at delivery, length, and weight. Complications and or 
abnormalities should be reported including any premature terminations. A pregnancy is reported 
as an AE or SAE only when there is suspicion that the investigational product may have 
interfered with the effectiveness of contraception or there was a serious complication in the 
pregnancy including a spontaneous abortion or an elective termination for medical rationale. 
Otherwise, pregnancy will not be categorized as an AE. 
8.2.16. 
Hepatitis B and C and HIV Reporting to the Local Health Authorities 
Hepatitis B and C, and Human Immunodeficiency Virus (HIV) test results are required by law to 
be reported to the local health authorities. The test results reported to local health authorities will 
contain the participant’s name, contact information, including address and telephone numbers, 
and the type of testing conducted. 
8.2.17. 
Serious Adverse Event Follow-up 
After the initial /SAE report, the investigator will proactively follow each subject at subsequent 
visits or contacts via phone or in person. All SAEs, and events of special interest, if applicable, 
will be followed until resolution, even if this extends beyond the prescribed reporting period. 
Resolution is the return to baseline status or stabilization of the condition with the probability 
that it will become chronic. The SAE outcomes will be reported to the ISM and ORA PSSO 
using the SAE Report Form. The investigator will submit any updated SAE data to the sponsor 
and ISM within 24 hours of receipt of the information. 
If a subject dies during participation in the study or during a recognized follow-up period, the 
investigator will need to provide a copy of any post-mortem findings, including histopathology, 
if available. 
Investigators are not obligated to actively seek SAEs in former subjects; however, if a SAE that 
is considered to be related to the investigational product occurs in a former subject and is brought 
to the attention of the investigator at any time following completion of the study, the event will 
be reported to the sponsor’s representative (Table 8). 
8.2.18. 
 Safety Monitoring Committee 
An independent safety monitoring committee (SMC) will be convened to review safety data and 
provide recommendations to the sponsor’s representative. The SMC for this study will be 
comprised of an independent group of experts who will review cumulative safety data during 
scheduled in-process and ad hoc reviews to evaluate safety, study conduct, and scientific validity 
and integrity of the trial. The SMC will meet annually. After each review of study data, the SMC 
will make a recommendation to the sponsor’s representative. The SMC recommendations will be 
documented and provided in writing (via the SMC Chair Recommendation Form) to the ORA 
PSSO. The sponsor’s representative will then decide whether to continue, modify, or suspend the 

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study. The ORA PSSO will communicate the final decision to the investigator, who in turn will 
notify the IRB as appropriate. The ORA regulatory affairs scientist will communicate the final 
decision to the U.S. FDA, as appropriate. A further description of the SMC roles and 
responsibilities, functions, reporting requirements, meeting frequency, and the study 
stopping/continuation criteria is included in the SMC charter. 
8.3. 
Immunogenicity Assessments 
8.3.1. 
Specification of Immunogenicity Endpoints 
Table 10: 
Immunology Assays 
Humoral Assays 
Serum or plasma Function Measurement 
ADCC, ADCP, and other non- 
neutralizing antibody functions 
Frozen Plasma/ 
Serum 
Measures lysis of HIV expressing targets 
mediated by HIV specific antibodies 
HIV-specific binding 
Frozen Plasma/ 
Serum 
Binding antibody to vaccine antigens 
HIV-specific neutralizing 
antibodies 
Frozen Serum 
Neutralizing activity against luciferase 
reporter gene expression 
Cellular and Innate Assays 
Serum or plasma Function Measurement 
Cellular response by cytokines 
such as IFN-J and IL2 after 
stimulation with HIV-specific 
antigens 
Frozen PBMC 
CD4+ and CD8+ antigen-specific 
response 
Lymphocyte proliferation 
Frozen PBMC 
Characterize the function of proliferating 
cells in response to HIV antigens 
B-cell ELISPOT 
Frozen PBMC 
Measures cytokine secretion from B cells 
in response to HIV antigens 
Flow cytometry for innate 
immune cell phenotyping and a 
cytokine array assay 
Frozen Plasma/ 
Serum 
Phenotype NK and other innate cells and 
characterize the cytokines elicited by the 
different vaccine regimens 
DNA Microarray: gene 
expression to vaccine antigens 
Frozen PBMC 
Host gene expression profile and 
signature to vaccine antigens 
RNA sequencing: gene 
transcription to vaccine antigens 
Frozen PBMC 
Host gene transcription profile and 
signature to vaccine antigens 
8.3.2. 
Immunogenicity Endpoints 
Immunogenicity will be assessed using the following assays at time points defined in the SOE. 

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8.3.3. 
Humoral Immune Responses 
HIV-specific Binding Antibody ELISA Assays will be performed to detect serum or plasma 
IgG and IgA binding antibodies to HIV-1 gp120, in particular to the V1V2 loops. Capture 
antigens will include V1V2 sequences from both subtype AE and Subtype B HIV-1 Env 
(gp70V1V2 92TH023 and gp70V1V2 Case A2), and HIV-1 Env gp120 proteins matched to 
sequences in AIDSVAX® B/E without the gD tag and with an 11 amino acid N-terminal 
deletion, represented as gp120 A244gD- D11 and gp120 MNgD- D11. Durability of IgG 
response will be assessed for each participant by estimating the decline in log10 IgG from peak 
to 6 and 12 months post final vaccination. 
Neutralizing Antibody Assays will be measured as a function of reductions in luciferase (Luc) 
reporter gene expression after a single round of infection in TZM-bl cells using high throughput 
analysis. Stocks of molecularly cloned envelope-pseudotyped viruses have been prepared by 
transfection in 293T/17 cells (American Type Culture Collection) and titrated in TZM-bl cells. 
This assay has been formally optimized and validated and was performed in compliance with 
Good Clinical Laboratory Practices (GCLP), including participation in a formal proficiency 
testing program. Tier 2 neutralization was assessed using a panel of 11 pseudoviruses, and a 
global panel. 
Antibody Profiling will be conducted to assess antibody breadth and correlation of Fc receptor 
usage with antibody function. The current capacity for Antibody Profiling is the assessment of 
over 300 samples in one experiment. The beads presenting the HIV antigens will be distributed 
robotically in a high-throughput manner into 384 well plates. Similarly, the samples will be 
diluted robotically and incubated with the beads. Detection reagents will be added to the wells 
after washing. These detection reagents are inclusive of subclass, isotype and Fc receptor usage. 
Therefore, in one well, we will be able to capture 900 to 1000 data points from a single sample, 
and over 250,000 data points per experiment that will allow evaluation of the humoral immune 
response to these vaccine strategies. 
Antibody-dependent cell-mediated cytotoxicity and antibody-dependent cellular 
phagocytosis will evaluated using an amine reactive staining (Aqua) to identify killed target 
cells compare to loss of carboxyfluorescein succinimidyl ester (CFSE) in the traditional rapid 
and fluorometric ADCC (RFADCC). This Aqua-based assay is high throughput, has a better 
signal to noise ratio and a lower coefficient of variation than the conventional CFSE-based 
RFADCC assay. The ADCP phagocytic score will also be calculated and compared with 
Antibody profiling data to evaluate the impacts on IgG subclass and Fc receptor usage on non- 
neutralizing function between groups. 
8.3.4. 
Cellular Immune Responses 
Intra-cellular cytokine staining (ICS) assay. Cryopreserved peripheral blood mononuclear cell 
(PBMC) will be stimulated with HIV-specific antigens and gated on cell surface markers such as 
CD4 and CD8 to determine antigen-specific cellular responses by individual cytokines, such as 
interferon gamma (IFNγ) and interleukin 2 (IL2), as well as a composite, calculated functional 
and polyfunctional response using the COMPASS analysis method. 

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Cellular Proliferation. The proliferative responses of participant’s PBMC to HIV-specific 
antigens and mitogens will be measured by the CFSE assay to characterize the function of 
proliferating cells. 
B-cell Responses. A highly sensitive Env-specific B-cell ELISPOT system was developed, 
which allows the enumeration of antibody-secreting cells from diverse anatomical compartments 
directed against different structural determinants of HIV envelope. Both plasmablast and 
memory B cell responses will be quantified to detect gp120 A244-specific IgG producing 
plasmablasts and long-lived memory B cells using cryopreserved PBMC. 
8.3.5. 
Innate Immunity 
Characterization of Innate Immunity. Flow cytometric panels will be used to phenotype the 
different types of NK cells and functional ICS to characterize their function. Luminex® multiplex 
assays will be performed to compare soluble cytokines and other factors in plasma/serum across 
vaccine regimens, to further characterize innate biologic pathways engaged. 
Innate and Early Adaptive Responses by Gene Activation Assessment. DNA microarrays 
will explore the innate and early adaptive immune responses to vaccination by screening 
genomic expression profile in PBMCs. 
Transcriptomics. Transcriptome data will be generated using RNASeq to investigate gene 
expression that correlates with increased immunogenicity to particular vaccine combination. 
8.4. 
Specimen Archiving and Transfer 
Biological samples such as serum, plasma, PBMCs, whole blood, remaining after all assays 
described in this protocol have been completed will be bar coded and archived using electronic 
specimen storage and tracking system. PBMC will be stored at -125oC or lower and plasma/sera 
will be stored at -70oC or lower. Samples will be archived at  the WRAIR Diagnostics and 
Countermeasures Branch (DCB) Specimen Processing Laboratory and Biorepository (SPL/BioR) 
indefinitely. If the study team determines to destroy these specimens, they will be destroyed per 
applicable SOPs of the archiving institutions. The PI, the Sponsor and authorized regulatory 
bodies may have access to the data regarding the archived specimens.  
Archived samples will only be used for research and will not be sold; nor will study participants 
receive payment, should samples lead to the development of new products in the future. Any 
future study requesting these samples must first be reviewed and approved by the IRB of each 
applicable institute. With concurrence from these parties, archived specimens will be shipped to 
the U.S. Military HIV Research Program (MHRP) and collaborating laboratories for assays. 
8.5. 
Future Use and Storage of Data and Biological Samples 
Each participant will be asked to voluntarily consent to their data and blood samples to be stored 
for other research studies that may be done after this study is complete. Future testing may 
involve genetic tests. In case the participant is unwilling to have their data and/or biological 
samples stored for future use, they can consent to participate in this study only, without having 
their data and blood samples stored for future testing. In this case, their data and blood samples 
will be destroyed within one year of receiving notification from the PI that all protocol testing is 

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complete. Samples remaining in the biorepository will be destroyed by laboratory staff following 
approved laboratory procedures. 
All data and samples for which consent has been obtained and for which additional material is 
available after study specified testing is complete will be stored indefinitely for future testing. 
However, WRAIR IRB approval will be sought before any such data and/or samples are used for 
analysis not specified in the protocol or a protocol amendment approved by the IRB. Data and 
samples may be shared with external collaborators but only after WRAIR IRB approval is 
received. 
During the study, blood samples for safety testing will be tested as soon as possible and within 
the allowable sample stability timeframe for each test.  
9. 
STATISTICS 
This study intends to descriptively characterize safety and immune responses following 
administration of candidate vaccine A244/B.63521 with ALFQ adjuvant in healthy adults across 
three doses of ALFQ: 200 μg, 100 μg and 50 μg, at baseline, month 1 and month 2. Statistical 
analysis will be performed on the following three groups: 
x Arm 1: A244/B.63521 + 200 μg ALFQ, N=20 
x Arm 2: A244/B.63521 + 100 μg ALFQ, N=20 
x Arm 3: A244/B.63521 + 50 μg ALFQ, N=20 
Of specific interest is determining the safety of the candidate vaccine A244/B.63521 with ALFQ 
regimens as well as whether immunologic differences can be detected between the 3 study arms. 
Response rates, and, where applicable, titer level distributions with appropriate summary 
statistics will be estimated for each immunologic assay performed. The study will evaluate a 
series of cellular, humoral, and innate immune assays. 
Due to limited sample size, rapid enrollment of subjects and short period of vaccination delivery 
over two months, no seasonal and other temporal effects are expected. 
9.1. 
Statistical Criteria for the Termination of the Trial 
There are no statistical criteria for the purpose of study termination in this clinical trial. 
9.2. 
Procedures for Reporting Deviations from the Statistical Plan 
Any deviation(s) from the statistical plan as indicated in the protocol will be described in an 
amendment to the protocol or the clinical study report. The protocol amendment will be 
submitted to the WRAIR IRB as per Section 10.1.1. 
9.3. 
Accounting for Missing, Unused, and Spurious Data 
Non-analyzable data will be documented in the deviations. 

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9.4. 
Sample size and Power calculation 
The safety, reactogenicity, and tolerability of the vaccine regimens are the primary endpoints for 
this study. The table below demonstrates effect size with a power of 80%, calculated at a variety 
of sample sizes. 
In Table 11, the sample size calculation for the antibody titer (secondary outcome) will be based 
on the published data from Pitisuttithum P et.al18 . For the analysis of this experiment we plan on 
observing a single measurement of peak antibody level per participant. Thus, the study is 
designed around a 1-way ANOVA model. Tukey's method will be used in the pairwise 
comparison test. In the analysis we looked at the size of the error bars on the peak of the log 
antibody titer and based upon our visual inspection we assumed that the residual standard 
deviation is = 0.2. A sample size of approximately 20 participants per arm was selected to 
provide ≥80% power to detect the effect size of ≥ 0.202 between study arms. Statistical 
significance will be set at p=0.05. 
Table 12 depicts SAE or AESI rates and associated 95% confidence intervals in each potential 
grouping of interest given 0-5 participants with observed events. If we were to observe zero 
events within a single active study group (n=20), the upper limit of a 2-sided exact 95% CI 
would be 13.9% and true events above this could be ruled out at the α=0.025 level. 
Table 11: 
Sample size per arm and minimum effect size with 80% power, assuming 
residual standard deviation of 0.2 
 
Part A: 3 Active Arms 
n per arm Minimum effect size 
Power 
5 
0.448 
0.801 
6 
0.398 
0.801 
7 
0.362 
0.801 
8 
0.334 
0.800 
9 
0.312 
0.800 
10 
0.294 
0.801 
11 
0.280 
0.804 
12 
0.266 
0.802 
13 
0.254 
0.800 
14 
0.244 
0.800 
15 
0.236 
0.803 
16 
0.228 
0.804 
17 
0.220 
0.801 
18 
0.214 
0.803 
19 
0.208 
0.803 

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Part A: 3 Active Arms 
20 
0.202 
0.802 
21 
0.198 
0.806 
22 
0.192 
0.801 
23 
0.188 
0.803 
24 
0.184 
0.804 
25 
0.180 
0.803 
26 
0.176 
0.802 
27 
0.174 
0.808 
 
Table 12: 
Exact 95% Clopper-Pearson CI for the SAE Rate in Each Study Group 
Number of 
Participants with 
Event 
All Active Groups 
(n=60) 
Any Active Group 
(n=20) 
Arm 1  
(n=22) 
Arms 2 or 3 
(n=19) 
0 
(0%, 4.9%) 
(0%, 13.9%) 
(0%, 12.7%) 
(0%, 14.6%) 
1 
(<0.1%, 8.9%) 
(0.1%, 24.9%) 
(0.1%, 22.8%) 
(0.1%, 26.0%) 
2 
(0.4%, 11.5%) 
(1.2%, 31.7%) 
(1.1%, 29.2%) 
(1.3%, 33.1%) 
3 
(1.0%, 13.9%) 
(3.2%, 37.9%) 
(2.9%, 34.9%) 
(3.3%, 39.6%) 
4 
(1.8%, 16.2%) 
(5.7%, 43.7%) 
(5.2%, 40.3%) 
(6.0%, 45.5%) 
5 
(2.8%, 18.4%) 
(8.7%, 49.1%) 
(7.8%, 45.4%) 
(9.1%, 51.2%) 
9.5. 
Populations for Analyses 
Analysis populations are defined in Table 13. 
Table 13: 
Analysis Populations 
Population 
Description 
Enrolled 
All participants who receive the first study vaccination 
Randomized 
All participants who are randomized to a study arm 
Evaluable 
All participants who receive the first study vaccination 
Safety 
All randomized participants who take at least 1 dose of study treatment. 
Participants will be analyzed according to the treatment they actually received 
Immunogenicity 
The immunogenicity population will include all participants who received at 
least one dose of vaccination and have at least one post-vaccination blood 
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9.6. 
Statistical Analyses 
Immunologic measurements and AEs including solicited and unsolicited events and clinically 
significant abnormal laboratory values are the main database for RV575. The statistical analysis 
plan will be developed and finalized before database lock and will describe the selection of 
participants to be included in the analyses, and procedures for accounting for missing, unused, 
and spurious data. 
Below is a summary of planned statistical analyses of the primary and secondary endpoints. 
9.6.1. 
Demographics and Baseline Characteristics 
Participant demographics and baseline characteristics will be summarized and may be presented 
in table form. The number of participants who enroll in the trial and the number and percentage 
of participants who complete each assessment will be presented. The percentage of patients who 
withdraw from the trial or discontinue the study drug and reasons for withdrawal will be 
summarized. 
9.6.2. 
Safety Analysis 
The primary endpoints of this Phase 1 study are related to the safety, reactogenicity, and 
tolerability of the candidate vaccine A244/B.63521 with varying doses of ALFQ adjuvant. The 
occurrence and severity of SAEs, AEs, AESIs, and unsolicited AEs will be tabulated by study 
group and compared between and among groups. 
The occurrence of local and systemic reactogenicity symptoms will be assessed as the 
proportions of participants experiencing such safety events along with 95% confidence intervals. 
Pairwise, between group comparisons will be performed. These comparisons may be performed 
at a two-sided α=0.05 level, using the normal approximation to the binomial distribution or an 
alternate testing procedure as appropriate (e.g. Fisher’s Exact test for smaller samples). 
9.6.3. 
Immunogenicity Analysis 
The primary immunogenicity endpoint is the magnitude of immune response to the endpoint titer 
of plasma IgG. Immune response over time (durability) will be assessed by the positive 
incremental area under the curve (AUC) based on a graph with log endpoint titer on the y-axis 
and visit week on the x-axis. Summary statistics at specific time points of interest will also be 
provided using the mean and standard deviation (or median and interquartile range (IQR) as 
appropriate). Primary comparisons may use a 2-sample t-test with an unadjusted alpha of 0.05. 
When differences are found, pairwise comparisons between vaccination groups will be made 
using the Holm-Bonferroni method and a 5% significance threshold for the adjusted p-values. 
Further exploratory comparisons may be performed and adjusted for multiplicity as appropriate. 
Secondary immunogenicity endpoints will entail primarily descriptive analyses. In general, 
continuous outcomes will be assessed using summary statistics such as the mean, median, 
standard deviation and IQR. Binary responses will be described using frequencies and 
percentages. Changes from baseline may be assessed within groups using paired t-tests (or 
Wilcoxon signed rank as appropriate) or McNemar’s test for binary outcomes. Changes over 

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time between groups may be evaluated using longitudinal models with unstructured covariance 
matrices where possible or covariance strictures selected as appropriate for the observed data. 
9.6.4. 
Missing Values and Outliers 
All attempts will be made to collect all data per protocol. Frequencies of missingness for primary 
outcomes will be calculated overall and within each study group. Less than 10% missing data 
that are missing completely at random will be considered ignorable missingness and complete 
case analysis will be utilized. No imputation will be performed for missing values. Data 
visualizations such as boxplots, histograms, or scatter plots will be used to identify outliers. 
Single outliers may be identified if values are outside of 1.5x the interquartile range depending 
on normality assumptions. Outliers will not be excluded from the primary analyses. Outliers 
identified during the analysis will be discussed in the analysis report. 
9.6.5. 
Clinical Laboratory Data Analyses 
Clinical laboratory values, including change from baseline, will be summarized for each 
participant according to time and study arm. The values will be graded according to the DAIDS 
grading scale in Appendix A and, if clinically significant, reported as AEs. 
Descriptive summary statistics (mean, standard deviation (SD), median, minimum, and 
maximum) for clinical laboratory data at admission and each applicable post-dosing visit, 
including changes from the baseline value collected at admission, will be calculated. If multiple 
baseline values are obtained, only the most recent value will be analyzed. For change-from- 
screening summaries, participants with an undefined change from screening, due to missing data, 
will be excluded. Clinical significance of abnormalities will be indicated in the listings. 
9.6.6. 
 Planned Interim Analyses 
No formal interim safety analyses are planned but may be performed if requested by the safety 
monitoring committee as outlined in Section 8.2.18.  
Interim immunogenicity analyses will be conducted after all enrolled participants have reached at 
least week 10 (20.8% of follow-up accrual) and may be conducted at additional time points if 
requested by the safety monitoring committee. The main goal of planned interim analyses 
through week 10 is to assess short-term effect of the candidate vaccine A244/B.63521 (300 μg 
each) with ALFQ adjuvant at the 200, 100, and 50 μg doses on cellular and humoral immune 
responses. This immunogenicity group analysis will not compromise the integrity of the trial in 
terms of the maintenance of the study blind, participant retention, or safety or immunogenicity 
endpoint assessments. All participant IDs will be replaced with alternate IDs by the unblinded 
study staff and only the minimum necessary summary level of information will be shared with 
blinded study staff. Results from planned interim immunogenicity analyses will be used to 
inform the design of a subsequent clinical trial.  
To minimize the risk of type-I error, an alpha-spending function, such as O’Brien-Fleming 
boundary will be used to monitor primary and secondary endpoints using an overall, two-sided 
alpha of 0.05, with the overall goal being to maximize the amount of alpha reserved for the final 
study analysis. Specific details of alpha-spending methods will be included in the statistical 
analysis plan. 

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The Principal Investigator will ensure concurrence from USAMRDC ORA as outlined in 
Section Error! Reference source not found. prior to presentation or publication of data from 
interim analyses. Any analyses conducted prior to the end of the scheduled follow-up visits will 
not compromise the integrity of the trial in terms of participant retention or safety or 
immunogenicity endpoint assessments. 
10. 
ETHICS 
10.1. 
Ethics Review 
The protocol is based on adequately performed laboratory and animal experimentation and will 
be conducted under a protocol reviewed by the IRB (see Table 7) and USAMRDC OHARO 
OHRO. The study is to be conducted by scientifically and medically qualified persons. The 
WRAIR IRB will determine whether the benefits of the protocol are in proportion to the risks. 
The rights and welfare of the participants will be respected, the physicians conducting the study 
will ensure that the hazards do not outweigh the potential benefits, the results to be reported will 
be accurate, participants will give their informed consent and will be competent to do so and not 
under duress, and all study staff will comply with the ethical principles in 21 CFR Part 50 and 
the Belmont Principles. 
10.1.1. 
Protocol Modifications 
All modifications to the protocol and supporting documents (informed consent, protocol-specific 
procedures, standard operating procedures, recruitment materials, etc.) will be reviewed and 
approved prior to implementation: 
1. Integrated product team review and approval 
2. Scientific Review Board review and approval (if applicable) 
3. Protocol Review Board (sponsor’s representative) review and approval 
4. Institutional Review Board review and approval 
5. U.S. FDA submission 
Any protocol amendment will be agreed upon and approved by the sponsor’s representative, 
through the PRB prior to submission to the IRB and approved by the IRB prior to 
implementation of said change or modification. 
Any modification that could potentially increase risk to participants will be submitted to the US 
FDA prior to implementation. Substantive modifications to the protocol and any modifications 
that could potentially increase risk to participants must also be submitted to the USAMRDC 
OHARO OHRO for approval prior to issuance of the WRAIR Commander Approval 
Authorization. The USAMRDC OHARO OHRO defines a substantive modification as a change 
in PI, change or addition of an institution, elimination or alteration of the consent process, 
change in the IRB of Record, change to the study population that has regulatory implications 
(e.g., adding children, adding active duty population, etc.), significant change in study design 
(i.e., would prompt additional scientific review) or a change that could potentially increase risks 

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to participants. Any changes of the IRB used to review and approve the research will be 
promptly reported to the USAMRDC OHARO OHRO and ORA PRB. The WRAIR HSPB will 
submit protocol amendments and modifications to the USAMRDC OHARO OHRO as per 
USAMRDC Command Policy 21. 
The Informed Consent Form must be revised to concur with any significant amendment that 
directly affects participants and must also be reviewed and approved with the amendment. New 
participants enrolled in the study will be consented with the most recent approved consent form. 
Any participant already enrolled in the protocol will be informed about the revision and asked to 
sign the revised informed consent document if the modification directly affects the individual’s 
participation in the protocol. A copy of the revised, signed, and dated informed consent 
document will be given to the participant. All original versions of the informed consent 
document will be retained in the protocol regulatory file.  
Modifications or updates to the IBs will also be submitted as protocol amendments to the 
WRAIR IRB for review and approval. 
10.1.2. 
Protocol Deviations 
All subject-specific deviations from the protocol (e.g., failure to return for follow-up visits or 
blood collection within the time indicated in the protocol) are to be documented. These are 
considered non-emergent or minor deviations. The PI or designee will be responsible for 
identifying and reporting all deviations, which are defined as isolated occurrences involving a 
procedure that did not follow the protocol or protocol-specific procedure. Non- emergent/minor 
deviations will be reported annually in the continuing review report to the IRB and, if 
appropriate, in the final study report. Action taken in response to the deviation and the impact of 
the deviation will be assessed by the PI or subinvestigator and recorded as significant or 
nonsignificant. 
For any protocol deviation that adversely affects the safety or rights of a subject or scientific 
integrity of the study (emergent deviation), the deviation will be reported within 24 hours to the 
sponsor’s representative’s clinical trial monitor or clinical trial manager and the IRB. 
Emergent/significant deviations should be reported promptly (within 48 hours) to the IRBs and 
the sponsor, upon becoming aware of the event, by telephone or email. A written report is 
required to be submitted by the PI to the WRAIR IRB and sponsor within 10 working days of 
knowledge of the significant deviation. Deviations will be reported by the WRAIR HSPB to the 
USAMRDC OHARO OHRO as per USAMRDC Command Policy 21. 
Waivers and exemptions to protocol procedures are not allowed with the exception of immediate 
safety concerns and these will be discussed with the sponsor immediately upon occurrence or 
awareness to determine whether the subject should continue or discontinue study treatment. 
10.2. 
Ethical Conduct of the Study 
This study will be conducted in accordance with all applicable Federal and DoD human research 
protections requirements and the Belmont Principles of respect for persons, beneficence, and 
justice. The procedures set out in this protocol are designed to ensure that the sponsor’s 
representative and all study personnel abide by the principles of the ICH GCP Guideline and the 
CFR. The PI confirms this by signing this protocol and FDA Form 1572. 

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Clinical research supported (directly or indirectly) by all HHS agencies, including the NIH, must 
comply with 45 CFR 46 (See 45 CFR 46.116), the OHRP regulations, in addition to all other 
applicable requirements. 
10.3. 
Confidentiality 
HIPAA requires that researchers obtain the subject’s permission (HIPAA authorization) to use 
and disclose health information about the subject that is either created by or used in connection 
with this protocol. The information includes the entire research record and supporting 
information from the subject’s medical records, results of laboratory tests, and both clinical and 
research observations made during the individual’s participation in the protocol. In this protocol, 
the subject’s health information will be collected and used to conduct the study; to monitor the 
subject’s health status; to measure effects of the investigational product; to determine research 
results; and possibly to develop new tests, procedures, and commercial products. Health 
information is used to report results of research to the sponsor’s representative and Federal 
regulators and may be reviewed during study audits for compliance with study plans, regulations, 
and research policies. After the study ends, each subject has the right to see and receive a copy of 
his/her information. 
Release of personally identifiable genetic information may pose a risk of discrimination or 
increased difficulty in obtaining certain types of insurance for participants and their family 
members. However, every measure will be taken to keep the information confidential. An added 
protection of privacy is the Genetic Information Nondiscrimination Act of 2008 (Pub. L. 110-
233), also known as "GINA”, which is a federal law that prohibits discrimination in health 
insurance coverage and employment based on genetic information. However, GINA does not 
apply to employers with fewer than 15 employees. GINA's protections in employment do not 
extend to the US military, nor does it apply to health insurance through the TRICARE military 
health system, the Indian Health Service, the Veterans Health Administration, or the Federal 
Employees Health Benefits Program. Lastly, the law does not cover long term care insurance, 
life insurance, or disability insurance. 
The Surgeon General ( as the IND sponsor) the sponsor’s representative, the representatives of 
USAMRDC OHARO OHRO, WRAIR IRB, the U.S. FDA, the U.S. DoD, and other regulatory 
agencies, as part of their responsibilities for insuring the protection of research participants (the 
DoD, and the FDA), are eligible to photocopy and review records related to this protocol as a 
part of their responsibility to protect the subjects of this protocol. In addition, representatives 
from these organizations may witness study procedures to ensure the safety of subjects. 
When study information is communicated or published to support this protocol, no personal 
identifier will be used. The subject’s identification number will be used in the event it becomes 
necessary to identify data specific to a single subject. 
10.4. 
Compensation for Participation 
Volunteers will be compensated $200 for their time, blood draws, and any inconvenience during 
each scheduled vaccination visit, $130 for other follow-up visits, and $50 for each screening 
visit. Unscheduled visits may be compensated up to $100 each if the PI deems appropriate. 
Participants who are ineligible to continue with the study and are subsequently withdrawn, will 

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be compensated $130 at their exit visit. This totals a maximum of $1,910 if the subject returns 
for each visit, and meets all study requirements for Diary Card return, and end of study 
completion fee of $350.  The two phone call follow-ups, conducted after clinical study visits 
have concluded, are not compensated but must be completed in order to receive the study 
completion fee. 
By regulation, federal employees can be compensated only for visits in which blood draws occur, 
and then only $50 per visit, unless the visits occur during off-duty hours or when they are on 
leave. If the volunteer is off-duty or on leave, he or she will be paid the same as non-federal 
personnel. 
The total amount of compensation may vary depending on the number of visits completed. 
Civilian (non-Federal employee) volunteers who undergo injections may receive approximately 
$1,910 in compensation. Subjects who are federal employees who are part of a vaccine group are 
expected to receive approximately $900 in compensation, unless they are on approved leave or 
participating outside of normal duty hours. 
Compensation for visits that are not specifically planned (scheduled) in the protocol, such as may 
be required to repeat labs to verify/clarify results or labs drawn to better evaluate abnormal lab 
values or AEs may be compensated at the discretion of the PI relative to the severity of the event 
and scope of evaluations. Compensation for transportation to the clinic for study visits may be 
provided at the discretion of the PI. 
Participants will receive $50 for each referred person who then attends a screening session and 
meets all inclusion and none of the exclusion criteria. 
Other than the payments discussed in this section, there is no other compensation available for 
your participation in this research. For all volunteers receiving more than $600.00, an IRS Form 
1099 will be issued. 
10.5. 
Medical Care for Research-Related Injury 
All nonexempt research involving human subjects shall, at a minimum, meet the requirement of 
32 CFR 219.116(a)(6). 
If a subject is injured because of participation in this protocol and is a DoD healthcare 
beneficiary (e.g., military spouse or dependent), the subject is entitled to medical care for that 
injury within the DoD healthcare system, as long as the subject remains a DoD healthcare 
beneficiary. This care includes, but is not limited to, free medical care at Army hospitals or 
clinics. 
If a subject is injured because of participation in this protocol and is not a DoD healthcare 
beneficiary, the subject is entitled to medical care for that injury at an Army hospital or clinic; 
medical care charges for care at an Army hospital or clinic will be waived. The subject is also 
entitled to care for that injury at other DoD (non-Army) hospitals or clinics, but such care for that 
injury at non-Army hospitals or clinics may be limited by time, and the subject’s insurance may 
be billed. It cannot be determined in advance which Army or DoD hospital or clinic will provide 
care. If the subject obtains care for research-related injuries outside of an Army or DoD hospital 
or clinic, the subject or the subject’s insurance will be responsible for medical expenses. 

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Participants who experience illness or injury arising from participation in the study should 
contact the study site immediately. An emergency contact card will be provided to study 
participants for this purpose. The site will either treat and care for the participant directly or refer 
the participant to an appropriate medical facility for care and treatment. Other than medical care 
and other payments as stated in this protocol, there is no other compensation available to the 
study participant from this research study. 
10.6. 
COVID-19 Information 
In light of the circulating severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) and 
the associated coronavirus disease (COVID-19) pandemic, the clinic will provide to the 
participant at time of consent information regarding proposed modifications put in place to 
safeguard the health and well-being of participants. The modifications provide flexibility for 
conducting study visits and procedures when needed to monitor participant safety. These 
modifications are expected to be time-limited in relation to the COVID-19 pandemic. In 
consultation with the Sponsor and MHRP, the study team will determine when, in the future, the 
guidance is no longer applicable. WRAIR will be notified when such a determination is made. 
Study personnel will follow the CTC SOP for conducting clinical trials in the setting of COVID-
19 for the duration of the pandemic in order to ensure that proper precautions are in place for the 
minimization of COVID-19 exposure. 
10.7. 
Source Documents 
The primary source document for this study will be the participant’s study folder. If separate 
research records are maintained by the investigator(s), the medical record and the research 
records will be considered the source documents for the purposes of auditing the study. The 
source documents will be retained at each site. During the study, all hard copy data collection 
forms will be locked in cabinets accessible only by appropriate study staff when not in use. 
For this study, an electronic data capture (EDC) database system will be used for the collection 
of the study data in an electronic format. The EDC database system will be designed based on 
the protocol requirements, the approved eCRF layouts and specifications, and in accordance with 
21 CFR Part 11. The eCRF layouts and specifications define and identify the applicable source 
data that will be collected and captured into the EDC database system. The applicable source 
data will be electronically transcribed by the site designee onto the electronic Case Report Forms 
(eCRF) (data entry screens) in the EDC database system. No source data will be recorded 
directly in the eCRF without a prior written record of the data. The investigator is ultimately 
responsible for the accuracy of the data transcribed on the eCRF. Data monitoring and 
management will be performed in the EDC database system by the study clinical monitor and 
DCAC. After database lock, data will be transferred from DCAC to the PI and Sponsor in a 
secure manner. 
A detailed data management plan will be written and approved by the study team and the PI prior 
to study start, with approval by the sponsor’s data manager in the USAMRDC ORA. All updates 
to the data management plan must be approved before study close-out and database lock. 

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10.8. 
Study and Site Closure 
The PI, the sponsor’s representative, the IRB, the USAMRDC Office of Research Protections, or 
the U.S. FDA, may stop or suspend the use of this product at any time. Otherwise, the study will 
be closed after all study procedures are completed. 
At study closure, all electronic source documents will be filed in the sponsor’s electronic Trial 
Master File (eTMF) and all hard copy documents will be delivered to USAMRDC ORA for 
archiving. Hard copy documents should be shipped to: 
USAMRDC ORA 
ATTN: Document Control Group 1430 Veterans Drive 
Fort Detrick, MD 21702-5009 
10.9. 
Written Informed Consent 
The PI (or designated staff) will obtain PRB and IRB approval of the informed consent 
document prior to initiation of the study. The informed consent will be administered individually 
with strict respect for confidentiality. The investigators or their designees will present the 
protocol in lay terms to individual participants. 
The PI or designee will solicit questions from participants on the purpose of the protocol, 
protocol procedures, and risks and allow the participant time to review the document. The person 
conducting consent will ensure that the participant understands the following principles of 
consent 
x Participation is entirely voluntary 
x The participant may withdraw from participation at any time 
x Refusal to participate involves no penalty 
x The individual is free to ask any questions that will allow him/her to understand the 
nature of the protocol 
Participants will be asked to indicate on the consent how their samples and/or data may be used 
in future studies. 
The designated individuals will obtain a copy of the consent form signed by each participant 
before any study-related procedures are initiated for that participant. The participant will also be 
given a copy of the participant event schedule. This signed and dated consent document will be 
retained as part of the study records. Each participant will receive a copy of the signed informed 
consent document. 
Should the protocol be modified, the informed consent document must be reviewed and revised 
to reflect the changes to the protocol, if applicable. If a previously enrolled participant is directly 
affected by the change, the approved revision must be read, signed, and dated by the participant. 
A copy of the signed and dated consent must be provided to the participant. 
The participant will be informed that a description of this clinical trial will be available on 
http://www.ClinicalTrials.gov, as required by US law. 

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11. 
ADMINISTRATIVE 
11.1. 
Study Monitoring  
Study monitoring will be the responsibility of the USAMRDC ORA. Upon successful approval 
of the protocol and establishment of the regulatory file, the clinical monitor will establish a 
clinical monitoring plan. To ensure that the investigator and the study staff understand and 
accept their defined responsibilities, the clinical monitor will maintain regular correspondence 
with the site and may be present during the course of the study to verify the acceptability of the 
facilities, compliance with the investigational plan and relevant regulations, and the maintenance 
of complete records. As needed, the clinical monitor may witness the informed consent process 
or other applicable study procedures to assure the safety of participants and the investigators’ 
compliance with the protocol and GCPs. 
Monitoring visits conducted by a sponsor’s representative-designated clinical monitor will be 
scheduled to take place at the initiation of the study, during the study at appropriate intervals, and 
after the last participant has completed the study. A report of monitoring observations will be 
provided to the PI (for corrective actions), USAMRDC ORA, and the product manager. A copy 
of the study monitoring report will be provided to the WRAIR IRB, upon request. 
11.2. 
Data Quality Assurance 
All data management activities will be conducted according to the approved study-specific data 
management plan. 
All participant data relating to the study will be recorded on printed or electronic case report 
forms unless transmitted to the sponsor or designee electronically (e.g., laboratory data). 
The investigator is responsible for: 
x Verifying that data entries are accurate and correct by physically or electronically 
signing the case report forms 
x Maintaining accurate documentation (source data) that supports the information 
entered in the case report forms. 
x Permitting study-related monitoring, audits, IRB review, and regulatory agency 
inspections and providing direct access to source data documents. 
The sponsor or designee is responsible for the data management of this study, including quality 
checking of the data. 
Study monitors will: 
x Perform ongoing source data verification based on the study-specific monitoring plan 
to confirm that data entered into the case report forms by authorized site personnel are 
accurate, complete, and verifiable from source documents 
x Verify that the safety and rights of participants are being protected 

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x Verify that the study is being conducted in accordance with the currently approved 
protocol and any other study agreements, International Conference on Harmonisation 
Good Clinical Practice, and all applicable regulatory requirements  
11.3. 
Record Retention 
Records and documents, including signed informed consent forms, pertaining to the conduct of 
this study will be retained by the investigator for 5 years after study completion 
No records may be destroyed during the retention period without the written approval of the 
sponsor. 
No records may be transferred to another location or party without written notification to the 
sponsor. 
11.4. 
Pending Inspections/Issuance of Reports 
The knowledge of any of the following will be reported immediately to the [IRB and/or the 
USAMRDC Office of Research Protections] and the sponsor’s representative through 
USAMRDC Office of Regulated Activities Quality and Compliance Office: 
x Pending compliance inspection/visit by the U.S. FDA, Office for Human Research 
Protections (Department of Health and Human Services), or other government agency 
concerning clinical investigation or research 
x The issuance of Inspection Reports, FDA Form 483, warning letters, or actions taken 
by any regulatory agency, including legal or medical actions 
x Any instances of serious or continuing noncompliance with the regulations or 
requirements 
11.5. 
Reports and Publications 
11.5.1. 
Annual Report to the FDA 
The PI will be responsible for the preparation of a detailed annual synopsis of clinical activity, 
including AEs, for submission to USAMRDC ORA. Each annual report will summarize IND 
activity for 1 year beginning approximately 3 months before the IND FDA anniversary date. 
USAMRDC ORA will notify the PI of the due date with sufficient time for the PI to assemble 
the required information. 
11.5.2. 
Final Clinical Study Report 
A final clinical study report will be prepared in accordance with the following guidelines will be 
provided to the USAMRDC ORA for review and approval: 
x Guidance for Industry: Submission of Abbreviated Reports and Synopses in Support 
of Marketing Applications 
x ICH E3 Guideline “Structure and Content of Clinical Study Reports” 
x The Comprehensive Table of Contents Headings and Hierarchy 

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x All applicable electronic Common Technical Document standards 
The USAMRDC ORA will use this report to prepare/review the final clinical study report for 
submission to the FDA. 
11.5.3. 
Continuing Review and Closeout Reports 
The PI is responsible for submitting the required CRRs and associated documents to the WRAIR 
IRB for review and approval, allowing sufficient time for review and continuation determination 
prior to the established continuing review date. Summaries of enrollment and safety reports will 
be provided in the CRR, as they are made available by the sponsor. 
After all study related activities, including data analysis are completed, a closeout report will be 
submitted as required to the same bodies. A closeout report will be submitted after 5 years or 
upon completion of the study, whichever occurs first. The WRAIR HSPB will forward CRRs 
and closeout report to the USAMRDC OHARO OHRO as per USAMRDC Command Policy 21. 
11.5.4. 
Publication Policy 
All data collected during this study will be used to support this investigational new drug 
application. Data may be published in the open medical or military literature with the identity of 
the participants protected. Anyone desiring to publish or present data obtained during the 
conduct of the study will forward the publication to the USAMRDC Office of Regulated 
Activities prior to submission. 
11.6. 
Roles and Responsibilities 
A list of study personnel will be maintained by the PI, or designee. The list must be available for 
review by clinical study monitors and representatives of the sponsor (ORA). Study roles and 
responsibilities are described in Appendix B. 

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12. 
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21. Liao HX, Tsao CY, Alam SM, et al. Antigenicity and immunogenicity of 
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12 
22. Bradley T, Pollara J, Santra S, et al. Pentavalent HIV-1 vaccine protects against simian-
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doi:10.1038/ncomms15711 
23. Cawlfield A, Genito CJ, Beck Z, et al. Safety, toxicity and immunogenicity of a malaria 
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formulation containing QS21 (ALFQ). Vaccine. 06 2019;37(29):3793-3803. 
doi:10.1016/j.vaccine.2019.05.059 

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13. 
APPENDICES 

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APPENDIX A. DAIDS TABLE FOR GRADING THE SEVERITY OF 
ADVERSE EVENTS 

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APPENDIX B. ROLES AND RESPONSIBILITIES 
x Principal Investigator 
 Supervises sub investigators and protocol nurse coordinator 
 Is responsible for completing informed consent process 
 Trains study staff 
 Screens individuals and reviews all of the screening tests to determine 
eligibility of an individual to participate in the study 
 Makes the final participant eligibility decisions or appoints a qualified designee 
 Permits auditing by the site quality unit, the US Army MRDC Quality 
Unit, and the ORA Quality and Compliance Office 
 Assures adequate resources, time, and participant population to 
meet study requirements 
 Provides medical care for AEs 
 Documents protocol deviations 
 Reports all SAEs to (WRAIR IRB, MRDC ) and sponsor’s safety office 
(US Army Medical Research and Development Command) Product Safety 
Surveillance Office (PSSO), WRAIR IRB 
 Delegates investigational product accountability and administration 
 May reconstitute vaccine 
 May administer investigational product to study participants 
 Evaluates AEs for diagnosis, relationship, and severity 
 Ensures documentation and safety reporting 
x Sub-investigator 
 Assists the PI in conduct of the study and may include (as indicated on the 
Delegation of Responsibilities Log) the following duties: 
 Is responsible for completing informed consent process 
 Assists in supervising study staff 
 Reviews all of the screening tests to determine eligibility of an 
individual to participate in the study 

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 Screens individuals for protocol participation (physical examination and 
medical history) 
 Can make final eligibility decisions 
 May reconstitute vaccine as designated by the PI 
 May administer investigational product to study participants 
 Evaluates AEs 
 If designated by the PI, to assist the PI in protocol adherence and execution, 
management and reporting, and any other duties described under PI 
responsibilities 
x Protocol Nurse/Study Coordinator 
 May be responsible for investigational vaccine accountability 
 May be responsible for completing informed consent process 
 Reviews all of the screening tests to determine eligibility of an 
individual to participate in the study 
 May reconstitute vaccine as designated by the PI 
 May administer investigational product to study participants 
 Schedules follow-up appointments 
 Follows up on all moderate and SAEs 
 Coordinates all study activity to include all participant visits and any non-
compliant participants 
 Completes case report forms 
 Trains staff on protocols/protocol changes 
 Documents protocol deviations 
x Protocol Support Staff 
 Screens study participants 
 Collects pre- and post-vaccination vital signs 
 Contacts study participants to collect AE data 
 Ensures efficient clinic flow 
x Clinical Monitor 

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 Conducts qualification, initiation, interim, and closeout study site visits 
 Monitors protocol adherence – reviews informed consent, standard 
operating procedures, regulations, and Good Clinical Practices 
 Monitors study and regulatory documents 
 Performs source data verification of case report form data 
 Checks investigational product accountability 
x Statistical Support Team 
 Assist in the review of statistical analysis 
 Assist with the review of/support the writing of the final Clinical Study Report 
x Data Manager 
 Provides study database end-user training to users requiring access to the 
study database 
 Oversees the data entry and data discrepancy management activities 
 Ensures data consistency, quality, and integrity 
 Performs Medical Dictionary for Regulatory Affairs (MedDRA) and 
World Health Organization (WHO) coding 
 Manages and maintains the study database 
x Statistician 
 Analyzes the verified data according to the protocol, statistical analysis 
plan, and any amendments in place 
 Provides documentation of statistical findings to the PI for 
incorporation into required reports 
x Sponsor’s Safety Office 
 Receives safety events, reviews, follows until resolution and performs 
SAE case processing 
 Performs medical evaluation of SAEs and provides the final 
determination on relatedness and expectedness and determines 
expedited reporting 
 Queries site for additional information as appropriate 
 Reports expedited SAEs to regulatory authorities 

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x DAIDS Medical Officer 
 Responsible for protocol development and liaison for the submission to the 
DAIDS Protocol Sciences Review Committee (PSRC) for review.  The DAIDS 
Medical Officer (MO) will be consulted on safety issues, if necessary, but will 
not have contact with study participants or identifiers. 

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APPENDIX C. PARTICIPANT VISIT SCHEDULE 

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APPENDIX D. INFORMED CONSENT FORM 

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APPENDIX E. TEST OF UNDERSTANDING 

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APPENDIX F. RECRUITMENT SCRIPT 

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APPENDIX G. INFORMATION SHEET REGARDING 
COMPENSATION TO FEDERAL PERSONNEL WHEN 
THEY PARTICIPATE IN RESEARCH 

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APPENDIX H. VISP LETTER 

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APPENDIX I. UNBLINDING LETTER 

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APPENDIX J. EMERGENCY CONTACT CARD 

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APPENDIX K. BRIEFING SLIDES 

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APPENDIX L. LIST OF POTENTIALLY IMMUNE-MEDIATED 
MEDICAL CONDITIONS 
Gastrointestinal 
x Celiac disease 
 
x Crohn’s disease 
 
x Ulcerative colitis 
 
x Ulcerative proctitis 
 
Liver 
 
x Autoimmune cholangitis 
 
x Autoimmune hepatitis 
 
x Primary biliary cirrhosis 
 
x Primary sclerosing cholangitis 
 
x Metabolic diseases 
 
x Addison’s disease 
 
x Autoimmune thyroiditis (including Hashimoto 
thyroiditis) 
 
x Diabetes mellitus type I 
 
x Grave's or Basedow’s disease 
 
Musculoskeletal 
 
x Dermatomyositis, Polymyositis and Antisynthetase 
Syndrome 
 
x Juvenile chronic arthritis (including Still’s disease) 
 
x Mixed connective tissue disorder 
 
x Polymyalgia rheumatic 
 
x Psoriatic arthropathy 
 
x Relapsing polychondritis 
 
x Rheumatoid arthritis 
 
x Scleroderma, including diffuse systemic form and 
CREST syndrome 
 
x Spondyloarthritis, including ankylosing spondylitis, 
reactive arthritis (Reiter's Syndrome) and 
undifferentiated spondylarthritis 
 
x Systemic lupus erythematosus 
 

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x Systemic sclerosis 
 
Neuroinflammatory: 
 
x Acute disseminated encephalomyelitis, including site 
specific variants (e.g., non-infectious encephalitis, 
encephalomyelitis, myelitis, radiculomyelitis) 
 
x Cranial nerve disorders, including paralyses/paresis 
(e.g., Bell’s palsy) 
 
x Guillain-Barré syndrome, including Miller Fisher 
syndrome and other variants 
 
x Immune-mediated peripheral neuropathies and 
plexopathies, including chronic inflammatory 
demyelinating polyneuropathy, multifocal motor 
neuropathy and polyneuropathies associated with 
monoclonal gammopathy 
 
x Multiple sclerosis 
 
x Narcolepsy 
 
x Optic neuritis 
 
x Transverse myelitis 
 
x Myasthenia gravis, including Eaton-Lambert syndrome 
 
Skin: 
 
x Alopecia areata 
 
x Autoimmune bullous skin diseases, including 
pemphigus, pemphigoid and dermatitis herpetiformis 
 
x Cutaneous lupus erythematosus 
 
x Erythema nodosum 
 
x Morphea 
 
x Lichen planus 
 
x Psoriasis 
 
x Rosacea 
 
x Sweet’s syndrome 
 
x Vitiligo 
 
Vasculitides: 
 
x Large vessels vasculitis including: giant cell arteritis 
such as Takayasu's arteritis and temporal arteritis 
 

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x Medium sized and/or small vessels vasculitis including: 
polyarteritis nodosa, Kawasaki's disease, microscopic 
polyangiitis, Wegener's granulomatosis, Churg–Strauss 
syndrome (allergic granulomatous angiitis), Buerger’s 
disease thromboangiitis obliterans, necrotizing 
vasculitis and anti-neutrophil cytoplasmic antibody 
(ANCA) positive vasculitis (type unspecified), Henoch- 
Schonlein purpura, Behcet's syndrome, leukocytoclastic 
vasculitis 
Other: 
x Antiphospholipid syndrome 
 
x Asthma and other immune-based lung diseases such as 
idiopathic pulmonary fibrosis, BOOP 
 
x Autoimmune hemolytic anemia 
 
x Autoimmune glomerulonephritis (including IgA 
nephropathy, glomerulonephritis rapidly progressive, 
membranous glomerulonephritis, 
membranoproliferative glomerulonephritis, and 
mesangioproliferative glomerulonephritis) 
 
x Autoimmune myocarditis/cardiomyopathy 
 
x Autoimmune thrombocytopenia 
 
x Goodpasture syndrome 
 
x Pernicious anemia 
 
x Raynaud’s phenomenon* note: Up to 10% of healthy 
female population has this without ever developing an 
autoimmune disease 
 
x Sarcoidosis 
 
x Sjögren’s syndrome 
 
x Stevens-Johnson syndrome 
 
x Uveitis 

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APPENDIX M. RISK ASSESSMENT TOOL  
 
 

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ATTACHMENT 1: PROTOCOL TEAM ROSTER 
Principal Investigator 
Brennan R. Cebula, MD, MTM&H 
MAJ, MC, USA 
U.S. Military HIV Research Program (MHRP)  
Walter Reed Army Institute of Research (WRAIR)  
6720A Rockledge Drive 
Bethesda, MD 20817  
bcebula@hivresearch.org 
814-233-0841 
 
Associate Investigators 
Julie A. Ake, MD, MSc COL, MC, USA 
Director 
US Military HIV Research Program (MHRP)  
Walter Reed Army Institute of Research (WRAIR)  
Office Phone: 301-319-7154 
Mobile Phone: 240-506-1915 E-mail: jake@hivresearch.org 
 
Sandhya Vasan, MD 
U.S. Military HIV Research Program (MHRP)  
Walter Reed Army Institute of Research (WRAIR)  
6720A Rockledge Drive 
Bethesda, MD. 20817  
Phone Number: 301-500-3622 
Email: svasan@hivresearch.org 
 
Elisavet Serti Chrisos, MSc, PhD 
U.S. Military HIV Research Program (MHRP)/Henry M. Jackson Foundation for the 
Advancement of Military Medicine (HJF) 
6720 Rockledge Drive 
Bethesda, MD 20817 
Phone: 301-500-3789 
Email: esertichrisos@hivresearch.org 
 
MAJ Trevor Wellington 
Walter Reed Army Institute of Research Center for Enabling Capabilities 
503 Robert Grant Ave.  
Silver Spring, MD 20910  
Phone Number: 301-319-3095 
Email: trevor.r.wellington.mil@health.mil 
 
 
 
 
 

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Paul Robben, MD/PhD, FACP 
LTC MC FS 
Chief, Clinical Investigations 
Malaria Biologics Branch, WRAIR 
503 Robert Grant Ave.  
Silver Spring, MD 20910 
Office: 301-319-9034 
Cell: 301-651-0351 
Email: paul.m.robben.mil@health.mil 
 
Trevor A. Crowell, MD, PhD 
Associate Director, Department of Epidemiology and Threat Assessment (DETA) 
Henry M. Jackson Foundation   
In support of the U.S. Military HIV Research Program (MHRP) 
6720A Rockledge Drive, Suite 400 
Bethesda, MD 20817 
Office: 301-500-3990 
Cell: 301-275-6047 
Email: tcrowell@hivresearch.org 
 
James E. Moon, MD, FACP, FIDSA 
COL, MC, USA 
Director, Center for Enabling Capabilities (CEC)  
Walter Reed Army Institute of Research (WRAIR) 
503 Robert Grant Avenue 
Rm 2S34A 
Silver Spring, MD, 20910-7500 
Office (301)-319-9176 
Work Cell (240)-406-6497 
james.e.moon.mil@health.mil 
 
Anna Stein, MD 
MAJ, MC, USA 
Walter Reed Army Institute of Research Center for Enabling Capabilities 
503 Robert Grant Ave.  
Silver Spring, MD 20910  
Email: anna.e.stein.mil@health.mil 
 
WRAIR Laboratory Investigators 
 
Sheila A Peel, MSPH, Ph.D. 
Director, Diagnostics and Countermeasures Branch (DCB) 
Laboratory Director, HIV Diagnostics and Reference Laboratory (HDRL); 
Leishmania Diagnostics Laboratory (LDL) 
Walter Reed Army Institute of Research (WRAIR) 
503 Robert Grant Ave 

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Silver Spring, MD 20910 
Tel: 301-319-2297 
Email: Sheila.a.peel2.civ@health.mil 
 
Janice Darden, MS 
Chief of Operations 
Diagnostics and Countermeasures Branch (DCB) 
Walter Reed Army Institute of Research (WRAIR) 
9100 Brookville Road, Building 508 
Silver Spring, MD 20910 
Cell: 301-275-1190 
Email: jdarden@hivresearch.org  
 
Anais Valencia-Ruiz, MS 
Laboratory Director  
Specimen Processing Laboratory and Biorepository (SPL/BioR) | HJF 
Diagnostics and Countermeasures Branch (DCB) 
Walter Reed Army Institute of Research (WRAIR) 
13 Taft Court, Suite 100 
Rockville, MD 20850 
Office: 301-251-3032  
Cell: 301-742-5787  
Email: avalencia-ruiz@hivresearch.org 
 
Jennifer Malia, DrPH, MS, MT (ASCP)  
Deputy Director, Diagnostics and Countermeasures Branch (DCB)  
Associate Laboratory Director, HIV Diagnostics and Reference Laboratory (HDRL); 
Leishmania Diagnostics Laboratory (LDL) 
Walter Reed Army Institute of Research (WRAIR) 
503 Robert Grant Ave, 1A34 
Silver Spring, MD 20910 
Phone: 301-319-3510  
Cell: 301-273-8015 
Email: jmalia@hivresearch.org  
 
Kristina Peachman, Ph.D. 
Research, Assessment and Development (RAD) 
Diagnostics and Countermeasures Branch (DCB) 
Walter Reed Army Institute of Research (WRAIR) 
13 Taft Court, Suite 100 
Rockville, MD 20850 
Office: 301-319-7513 
Cell: 240-463-7879 
Email: kpeachman@hivresearch.org 
 

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DAIDS Medical Officer 
Laura Polakowski, MD, MSPH 
Medical Officer 
Vaccine Clinical Research Branch 
Vaccine Research Program 
Division of AIDS, NIAID, NIH 
5601 Fishers Lane, 9D11B 
Rockville, MD 20852 
Phone Number: 301-627-3040 

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ATTACHMENT 2: TABLE OF EXTERNAL COLLABORATORS 
(When external collaborators are identified, they will be added here.)