A Phase-1 Open-label Clinical Trial to Evaluate the Safety and Immunogenicity of Synthetic DNAs Encoding NP-GT8 and IL-12, With or Without a TLR-agonist- Adjuvanted HIV Env Trimer 4571 Boost, in Adults Without HIV
Phase: PHASE1
Conditions: HIV Infections
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

 
 
PROTOCOL 
HVTN 305 
A phase-1 open-label clinical trial to evaluate the safety 
and immunogenicity of synthetic DNAs encoding NP-
GT8 and IL-12, with or without a TLR-agonist–
adjuvanted HIV Env Trimer 4571 boost, in adults without 
HIV 
DAIDS DOCUMENT ID 38886 
IND 028962 HELD BY DAIDS 
CLINICAL TRIAL SPONSORED BY 
Division of AIDS (DAIDS) 
National Institute of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
Department of Health and Human Services (DHHS) 
Bethesda, Maryland, USA 
STUDY PRODUCTS PROVIDED BY 
 The Wistar Institute, Philadelphia, Pennsylvania, USA 
The Betty and Dale Bumpers Vaccine Research Center (VRC) 
Bethesda, Maryland, USA 
Inovio Pharmaceutical Inc. 
Plymouth Meeting, Pennsylvania, USA 
Access to Advanced Health Institute (AAHI) 
Seattle, Washington, USA 
Division of AIDS (DAIDS) 
Bethesda, Maryland, USA 
Jan 06, 2023 
Final 
HVTN 305 
Version 2.0 

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Protocol Signature Page 
 
A phase-1 open-label clinical trial to evaluate the safety and immunogenicity of 
synthetic DNAs encoding NP-GT8 and IL-12, with or without a TLR-agonist–
adjuvanted HIV Env Trimer 4571 boost, in adults without HIV 
I will conduct the study in accordance with the provisions of this protocol and all 
applicable protocol-related documents. I agree to conduct this study in 
compliance with United States (US) Health and Human Service regulations (45 
CFR 46); applicable US Food and Drug Administration regulations; standards of 
the International Conference on Harmonization Guideline for Good Clinical 
Practice (E6); Institutional Review Board/Ethics Committee determinations; all 
applicable in-country, state, and local laws and regulations; and other applicable 
requirements (eg, US National Institutes of Health [NIH], Division of AIDS 
[DAIDS]) and institutional policies. 
 
 
 
 
 
 
 
 
 
Investigator of Record Name (print) 
 
Investigator of Record Signature 
 
Date 
 
 
 
DAIDS Protocol Number: HVTN 305 
DAIDS Protocol Version: Version 2.0 
Protocol Date: Jan 06, 2023 

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Acronyms and abbreviations 
AAHI 
 
Access to Advanced Health Institute 
Ab 
 
antibody 
ADCC 
 
antibody-dependent cell-mediated cytotoxicity 
ADCP 
 
antibody-dependent cellular phagocytosis 
AE 
 
adverse event 
AESI 
 
adverse event of special interest 
AF 
 
aqueous formulation 
AoU 
 
assessment of understanding 
ALT 
 
alanine transaminase 
Alum 
 
aluminum hydroxide suspension 
Anti-HCV  
anti-Hepatitis C antibody 
AUC 
 
area under the curve 
β-HCG 
 
beta human chorionic gonadotropin 
BCR 
 
B-cell receptor 
BMI 
 
body mass index 
bnAb 
 
broadly neutralizing antibody 
BP 
 
blood pressure 
CAB 
 
Community Advisory Board 
CBC 
 
complete blood count 
CI 
 
confidence interval 
CMIA 
 
chemiluminescent microparticle immunoassay 
CRF 
 
case report form 
CRPMC 
 
NIAID Clinical Research Products Management Center 
CRS 
 
clinical research site 
CSS 
 
clinical safety specialist 
DAERS 
 
DAIDS Adverse Experience Reporting System 
DAIDS 
 
Division of AIDS 
DM 
 
diabetes mellitus 
EAE 
 
expedited adverse event 
EC 
 
Ethics Committee 
ECLIA 
 
electrochemiluminescence 
eGFR 
 
estimated glomerular filtration rate 
EIA 
 
enzyme immunoassay 
EMPEM  
electron microscopy-polyclonal epitope mapping 
Env 
 
HIV/HIV-1 envelope protein 
EP 
 
electroporation 
FDA 
 
US Food and Drug Administration 
FNA 
 
fine-needle aspiration 
GCP E6 (R2) 
ICH Guideline for Good Clinical Practice 

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GINA 
 
Genetic Information Nondiscrimination Act 
GPP 
 
Good Participatory Practices 
HSV2 
 
herpes simplex virus type 2 
HVTN 
 
HIV Vaccine Trials Network 
IB 
 
Investigator’s Brochure 
IBC 
 
Institutional Biosafety Committee 
ICH 
 
International Council for Harmonization 
ICS 
 
intracellular cytokine staining 
ID 
 
intradermal 
IFN-gamma 
interferon-gamma 
IL-12 
 
Interleukin 12 
IM 
 
intramuscular 
IND 
 
investigational new drug (application) 
INR 
 
international normalized ratio 
IRB 
 
Institutional Review Board 
IUD 
 
intrauterine device 
LABA 
 
long-acting beta agonist 
MAAE 
 
medically attended adverse event 
MBL 
 
mannose-binding lectin 
MedDRA  
Medical Dictionary for Regulatory Activities 
MO 
 
medical officer 
MOP 
 
Manual of Procedures 
MSD 
 
Meso Scale Discovery 
MSM 
 
person born male with partner(s) born male 
nAb 
 
neutralizing antibody 
NAT 
 
HCV nucleic acid test 
NHP 
 
nonhuman primate 
NIAID 
 
National Institute of Allergy and Infectious Diseases 
NIH 
 
US National Institutes of Health 
NLT 
 
native-like trimer 
NSAID 
 
nonsteroidal anti-inflammatory drug 
OHRP 
 
US Department of Health and Human Services Office for 
 
 
Human Research Protections 
PAB 
 
DAIDS Pharmaceutical Affairs Branch 
PAGE 
 
polyacrylamide gel electrophoresis 
PBMC 
 
peripheral blood mononuclear cell 
PEF 
 
peak expiratory flow 
PI 
 
principal investigator 
PID 
 
pelvic inflammatory disease 
POPIA 
 
Protection of Personal Information Act 

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PrEP 
 
HIV pre-exposure prophylaxis 
PSRT 
 
Protocol Safety Review Team 
PV 
 
PENNVAX-B DNA vaccine 
RAB 
 
DAIDS Regulatory Affairs Branch 
RBD 
 
receptor binding domain 
RE 
 
Regulatory Entity 
RM 
 
rhesus macaque 
RML 
 
Regional Medical Liaison 
RSC 
 
DAIDS Regulatory Support Center 
SAE 
 
serious adverse event 
SAP 
 
statistical analysis plan 
sD-NP-GT8 
synthetic-DNA–encoded nanoparticle GT8 
SDMC 
 
statistics and data management center 
SHIV 
 
simian-human immunodeficiency virus 
SICF 
 
sample informed consent form 
SMB 
 
Safety Monitoring Board 
SOP 
 
Standard Operating Procedures 
SPT 
 
DAIDS Safety and Pharmacovigilance Team 
SSP 
 
Study-specific Procedures 
SUSAR 
 
suspected unexpected serious adverse reaction 
TDS 
 
Ichor Medical Systems TriGrid Delivery System 
TEM 
 
transmission electron microscopy 
TLR 
 
toll-like receptor 
ULN 
 
upper limit of normal 
VISP 
 
vaccine-induced seropositivity 
VRC 
 
Dale and Betty Bumpers Vaccine Research Center 
VSV 
 
vesicular stomatitis virus 
WBC 
 
white blood cell 
SWFI 
 
sterile water for injection 

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Contents 
1 Executive summary ........................................................................................................9 
1.1 
Title ................................................................................................................9 
1.2 
Study products ................................................................................................9 
1.3 
Study devices ................................................................................................10 
1.4 
Study population ..........................................................................................10 
1.5 
Study plan and schema table ........................................................................10 
1.6 
Duration per participant ...............................................................................12 
1.7 
Estimated total study duration ......................................................................12 
1.8 
Study sites ....................................................................................................12 
2 Introduction ..................................................................................................................13 
2.1 
Rationale for evaluation of sD-NP-GT8 ......................................................14 
2.2 
Preclinical data: mice, guinea pigs and macaques .......................................14 
2.3 
Clinical experience with related DNA vaccines encoding HIV Env 
immunogens with and without IL-12 adjuvants ...........................................18 
2.4 
Safety of HIV Envelope proteins adjuvanted with 3M-052-AF + Alum: 
Clinical Experience ......................................................................................21 
2.5 
Rationale for evaluation of 3M-052-AF + Alum adjuvant ..........................23 
2.6 
Rationale for schedule ..................................................................................23 
2.7 
Rationale for evaluation of boost with trimeric protein ...............................24 
2.8 
Rationale for targeting the same draining lymph node with both DNA and 
protein trimer ................................................................................................24 
2.9 
Rationale for dose of Trimer 4571 ...............................................................26 
2.10 Rationale for dose of 3M-052-AF adjuvant .................................................27 
2.11 Rationale for FNA ........................................................................................28 
2.12 Rationale for leukapheresis ..........................................................................29 
2.13 Risks and benefits .........................................................................................30 
3 Objectives and endpoints .............................................................................................33 
3.1 
Primary objectives and endpoints ................................................................33 
3.2 
Secondary objectives and endpoints ............................................................33 
3.3 
Exploratory objectives ..................................................................................33 
4 Laboratory strategy ......................................................................................................35 
5 Study design .................................................................................................................36 
5.1 
Study population ..........................................................................................36 
5.2 
Participant departure from vaccination schedule or withdrawal ..................41 
6 Statistical considerations ..............................................................................................45 
6.1 
Sample size justification and accrual ...........................................................46 
6.2 
Randomization .............................................................................................50 
6.3 
Blinding ........................................................................................................50 
6.4 
Statistical analyses ........................................................................................51 
7 Study vaccine preparation, storage, and administration ..............................................53 
7.1 
Vaccine regimen ...........................................................................................53 
7.2 
Study product formulation and storage ........................................................53 
7.3 
Product preparation ......................................................................................54 
7.4 
Study vaccine administration .......................................................................57 

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7.5 
Acquisition of study products ......................................................................58 
7.6 
Study vaccine accountability ........................................................................58 
7.7 
Final disposition of study product ................................................................58 
8 Clinical procedures ......................................................................................................60 
8.1 
Screening ......................................................................................................60 
8.2 
Definition of “study day” and “study visit” .................................................60 
8.3 
Reactogenicity assessments ..........................................................................60 
8.4 
EP injection site assessment .........................................................................61 
8.5 
Lymph node FNA .........................................................................................62 
8.6 
Leukapheresis ...............................................................................................62 
8.7 
Visit procedures, schedule, windows, and missed visits ..............................63 
8.8 
Monitoring for acquisition of HIV ...............................................................63 
8.9 
Early termination visit ..................................................................................64 
8.10 AESI contact ................................................................................................64 
9 Safety and AEs .............................................................................................................66 
9.1 
AEs ...............................................................................................................66 
9.2 
SAEs .............................................................................................................67 
9.3 
Safety monitoring .........................................................................................68 
9.4 
Total blood volume ......................................................................................69 
9.5 
Initial safety review ......................................................................................69 
9.6 
Safety pause and prompt PSRT AE review..................................................69 
9.7 
Study termination .........................................................................................71 
9.8 
Pregnancy .....................................................................................................71 
10 Protocol conduct and informed consent .......................................................................72 
10.1 Protocol conduct ...........................................................................................72 
10.2 Compliance with NIH guidelines for research involving products containing 
recombinant or synthetic nucleic acid molecules .........................................72 
10.3 Informed consent ..........................................................................................73 
11 Exploratory studies ......................................................................................................74 
11.1 Specimen storage and other use of specimens .............................................74 
12 Literature cited .............................................................................................................75 
Appendix A Schedule of procedures: Participants undergoing FNA ...............................80 
Appendix B Schedule of procedures: Participants not undergoing FNA .........................82 
Appendix C Visit windows ...............................................................................................84 
Appendix D Sample informed consent form ....................................................................85 
Appendix E Sample consent form for use of samples and information in other studies 113 
Appendix F Low-risk guidelines for US ........................................................................118 
Appendix G Low-risk guidelines for South Africa .........................................................122 
Appendix H Approved birth control methods ................................................................123 
Appendix I Adverse events of special interest (AESIs) ................................................125 
Appendix J Protocol team ..............................................................................................126 

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Appendix K Version history ...........................................................................................128 

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1 
Executive summary 
1.1 
Title 
A phase-1 open-label clinical trial to evaluate the safety and immunogenicity of 
synthetic DNAs encoding NP-GT8 and IL-12, with or without a TLR-agonist–
adjuvanted HIV Env Trimer 4571 boost, in adults without HIV. 
This is a randomized trial to examine safety and immunogenicity of  INO-6172 
(synthetic DNAs encoding the GT8 nanoparticle and Interleukin-12), alone and 
with HIV envelope protein (Env) Trimer 4571 adjuvanted with 3M-052-AF + 
Alum. The primary hypothesis is that INO-6172 will elicit VRC01-class B-cell 
responses as well as antigen-specific T-cell responses. 
1.2 
Study products 
• INO-6172 (sD-NP-GT8 coformulated with IL-12 DNA (pGX6001)): 
Synthetic-DNA–encoded nanoparticle GT8 (sD-NP-GT8) was developed by 
the Wistar Institute and consists of a single plasmid, pGX1072 (in pGX0001 
vector backbone), encoding the soluble self-assembling nanoparticle vaccine, 
which is decorated with a germline-targeting, B-cell precursor activating 
immunogen GT8. pGX6001, developed by Inovio Pharmaceuticals, is derived 
from a pGX0003 backbone. pGX6001is a single plasmid, which contains a 
dual promoter system for the expression of both the IL-12 p35 and p40 genes, 
necessary for production of the active heterodimeric IL-12 protein. The 
plasmid ratio is 4:1 sD-NP-GT8: IL-12 DNA (0.8 mg pGX1072/0.2 mg 
pGX6001 or 0.2 mg pGX1072/0.05 mg pGX6001) per 0.1-mL injection. 
• Trimer 4571: HIV-1 Env Trimer 4571 (VRC-HIVRGP096-00-VP) is a 
soluble protein that consists of BG505 DS-SOSIP.664 gp140 Env and is 
supplied as a sterile, aqueous, buffered solution filled into single-dose vials at 
a concentration of 500 mcg/mL and a volume of 1.2 mL in 3-mL glass vials. 
Trimer 4571 is provided by the Dale and Betty Bumpers Vaccine Research 
Center (VRC) and will be used at a dose of 100 mcg. 
• 3M-052-AF: This adjuvant is an aqueous formulation (AF) of the small 
molecule imidazoquinoline, which works as a toll-like receptor (TLR) 7/8 
agonist. To be administered at 5 mcg admixed with 100 mcg Trimer 4571 and 
500 mcg Alum. 
• Aluminum Hydroxide Suspension (Alum) adjuvant: Alhydrogel at 500 mcg 
will be admixed with 5 mcg 3M-052-AF along with 100 mcg Trimer 4571. 
 
 
 

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1.3 
Study devices 
Electroporation device:  The Inovio CELLECTRA Adaptive Constant Current 
Electroporation (EP) Device is a portable, battery-powered medical device 
designed to facilitate the introduction of DNA into skin through EP. The Inovio 
CELLECTRA 2000 will be used for intradermal (ID) delivery following Mantoux 
injection of the DNA vaccine, which is provided by Inovio Pharmaceuticals. 
1.4 
Study population 
Forty-five healthy volunteers without HIV-1 aged 18 to 55 years, inclusive. 
1.5 
Study plan and schema table 
Participants will receive INO-6172 (sD-NP-GT8 + IL-12 DNA) vaccinations at 
months 0, 1, and 3, with a total dose of 0.5 mg (Group 1) or 2 mg (Groups 2 and 
3). For Group 1, sD-NP-GT8 DNA will be administered at a dose of 0.4 mg along 
with IL-12 DNA at a dose of 0.1 mg (0.5 mg total). For Groups 2 and 3, sD-NP-
GT8 DNA will be administered at a dose of 1.6 mg along with 0.4 mg of IL-12 
DNA (2 mg total). These will be administered intradermally via EP of the skin. 
Due to volume limitations of the device, the dose of the DNA will be 
administered as 2 separate intradermal (ID) injections. Each of 2 sites will receive 
0.1 mL of study product (either the 1.0-mg/0.1-mL concentration or the 0.25-
mg/0.1-mL concentration) via ID injection (Mantoux injection) bilaterally 1 on 
each upper arm. Following ID injections, EP will be performed with the Inovio 
CELLECTRA 2000 EP device. 
A total of 18 out of 45 participants (Group 3) will also be administered Trimer 
4571 at a total dose of 100 mcg at months 3 and 6. The Trimer dose is split over 2 
injection sites. The injections are delivered intramuscularly (IM) via needle and 
syringe, 1 in each deltoid muscle. Participants will be evaluated for safety and 
immune responses through blood collection, lymph node fine-needle aspiration 
(FNA), and leukapheresis at specified timepoints throughout the study. The study 
schema is presented below in Table 1-1: 

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Table 1-1 Schema 
 
 
 
 
Injection schedule in months (days) 
Group 
N 
Product/Dose 
Route 
Month 0 
(Day 1) 
Month 1 
(Day 29) 
Month 3 
(Day 85) 
Month 6 
(Day 169) 
1** 
9 
 
INO-6172 
(0.5 mg) 
ID EP  
(2 
injections; 1 
into skin of 
each upper 
arm) 
X 
X 
X 
-- 
2§ 
18 
 
INO-6172 
(2 mg) 
ID EP  
(2 
injections; 1 
into skin of 
each upper 
arm) 
X 
X 
X 
-- 
3§ 
18 
 
INO-6172 
(2 mg) 
ID EP  
(2 
injections; 1 
into skin of 
each upper 
arm) 
X 
X 
X† 
-- 
Trimer 4571 
(100 mcg) + 
3M-052 AF 
(5 mcg) / 
Alum (500 
mcg) 
IM  
(2 
injections; 1 
into each 
deltoid) 
-- 
-- 
X† 
X 
Total 
45* 
 
 
 
 
 
 
 
Notes: 
*Up to 9 additional participants may be enrolled (for a total of up to 54), if 
needed, to have approximately 45 participants contribute to immunogenicity 
analyses. Specific scenarios that could necessitate enrollment of additional 
participants in order to prevent loss of statistical power of the study include, but 
are not limited to, the following: loss of participants due to moving, withdrawal of 
consent, missing vaccine visits, inability to complete FNA, or variations in the 
clinical care due to unpredictable events. Participants will not be replaced after 
completion of visit 6, the first FNA visit. 
** Initial enrollment will be restricted to 1 participant per day for the first 5 
participants in Group 1, and enrollment will pause after the first 5 participants are 
enrolled in Group 1. The Protocol Safety Review Team (PSRT) will review 
cumulative safety information for these first 5 participants recorded through the 
visit scheduled 2 weeks post–first vaccination and will determine whether it is 
safe to proceed with full enrollment in all groups. 
†For the first five participants in Group 3, the Month 3 vaccination will be limited 
to 1 participant per day. 

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§FNA will not be conducted for Group 1 but may be optional for Groups 2 and 3 
and will be offered to participants during screening. A minimum target of 9 
participants in these groups will be asked to provide consent for receiving the 
FNA. 
1.6 
Duration per participant 
The total duration per participant is 18 months, including 12 months of scheduled 
clinic visits (main study) followed by an adverse event of special interest (AESI) 
health contact at month 18. 
1.7 
Estimated total study duration 
At least 22 months (includes enrollment, planned safety holds, follow-up, and 
AESI health contact). 
1.8 
Study sites 
HIV Vaccine Trials Network (HVTN) clinical research sites (CRSs) in the US 
and South Africa, to be specified in the Site Announcement Memo. 

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2 
Introduction 
This first-in-human study will evaluate the safety and immunogenicity of a 
synthetic-DNA–encoded, self-assembling nanoparticle (sD-NP-GT8). NP-GT8 is 
an engineered outer domain of gp120 containing the CD4bs with an optimized 
human IgE-leader, which is displayed on a lumazine synthase scaffold to form a 
self-assembling 60-mer nanoparticle. NP-GT8 targets VRC01-class germline cells 
(1, 2). The immunization series will include priming with a synthetic DNA 
plasmid (pGX1072/sD-NP-GT8) encoding the nanoparticle displaying GT8, 
adjuvanted with a plasmid encoding human IL-12 (pGX6001) and boosting with 
sD-NP-GT8/pGX6001 alone and in combination with an HIV envelope soluble 
protein trimer, Trimer 4571, adjuvanted with 3M-052-AF + Alum. The DNAs 
will be delivered via ID EP, and the protein boost with Trimer 4571 will be 
administered via IM injection. 
Preclinical data show that self-assembling nanoparticle vaccines enhance the 
kinetics of particle trafficking to the draining lymph nodes, and the uptake by the 
dendritic cells and macrophages (3-5). Multivalent antigen display on self-
assembling nanoparticles can also enhance B-cell activation, thereby boosting the 
magnitude, affinity, and durability of humoral responses (3, 6, 7). 
The long-term utility of this platform will be to employ sD-NP-GT8 as both a 
prime and boost with engineered booster immunogens to produce broadly 
neutralizing antibodies (bnAbs), to employ sD-NP primes and heterologous 
boosts for alternative epitopes, or potentially to employ sD-NP cocktails to 
provide diverse HIV isolates and generate broader responses. 
Self-assembling antigens encoded into a DNA plasmid can be delivered in vivo, 
bypassing laborious in vitro protein purification steps. DNA is easily mass 
produced, temperature stable, easy to deliver clinically using the new ID EP 
technologies, and can be rapidly reformulated for new vaccine immunogens, 
making it an attractive vaccine technology for iterative HIV-1 vaccine studies (8, 
9). 
Advantages of the synthetic DNA vaccine platform with adjuvant IL-12 DNA 
(pGX6001), especially when administered in combination with native-like trimer 
(NLT) protein boosts, include: 
• Potential to elicit more robust immune responses than DNA alone; 
• Potential dose sparing of DNA vaccine required in the regimen; and 
• Unique induction of cellular responses to act synergistically with induced 
humoral responses. 
This clinical trial will inform future precursor-activating/germline-
targeting/lineage B-cell–focused nanoparticles and DNA clinical studies. It will 
be important to discover the immune responses (both B cell and T cell) to the self-

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assembling nanoparticles delivered by this advanced DNA platform, and to 
compare the responses to other studies using alternate methods of antigen 
delivery. In addition, this study can help answer the question in human clinical 
HIV-1 vaccinology of whether activation of bnAb precursors can be 
accomplished by repeated boosts with multiple engineered Envs delivered via 
DNA vaccination, or if a wild-type HIV-1 Env administered as a protein is 
capable of maturing and broadening desired antibody (Ab) responses. 
Clinical evaluation of sD-NP-GT8 will allow examination of important goals in 
the HIV vaccine field, such as: (1) activation of CD4bs bnAb precursors by a self-
assembling nanoparticle that is produced in vivo and can be trafficked in a 
mannose-binding lectin (MBL)–independent manner and (2) evaluation of 
advanced nucleic-acid–delivered self-assembling vaccines, which could change 
the clinical landscape for many future studies. 
2.1 
Rationale for evaluation of sD-NP-GT8 
Single immunization of adjuvanted GT8 has been shown to be capable of priming 
germline VRC01-class responses. One example is the eOD-GT8-60mer, a self-
assembling nanoparticle targeting VRC01-class precursors (4, 11-19). Preliminary 
results from a proof-of-concept trial (IAVI G001; clinicaltrials.gov identifier: 
NCT03547245) using vaccination with a recombinant eOD-GT8-60mer protein 
showed that VRC01-class precursor Abs were elicited (10), and these data 
strongly support the goals of this current protocol. We built upon this concept by 
developing DNA-launched nanoparticles encoding the GT8 immunogen (sD-NP-
GT8). Vaccination with the sD-NP-GT8 nanoparticle elicits strong humoral and 
cellular responses. As with the recombinant protein, sD-NPs can induce similar 
levels of immunogenicity and develop human Ab responses in human Ab 
repertoire mice ((4) and unpublished). 
2.2 
Preclinical data: mice, guinea pigs and macaques 
In a recent study in mice (4), we developed a synthetic-DNA–based nanoparticle 
platform (sD-NP) and observed in vivo assembly of nanoparticles as measured by 
muscle immunofluorescence, native polyacrylamide gel electrophoresis (PAGE) 
of muscle homogenates, transmission electron microscopy (TEM) of thin muscle 
sections and multimeric binding of MBL (see Figure 2-1) (4). This is the first time 
that in vivo assembly of a designed, self-assembling nanoparticle delivered by 
DNA has been demonstrated. sD-NP provides rapid seroconversion and is 
significantly more immunogenic than monomeric antigens in both the phase 
immediately postvaccination and the long-term timepoints. We also observed 
large dose-sparing effects, as a 25-fold lower dose of sD-NP can produce 10 times 
higher Ab titers compared to monomeric antigens. The sD-NP platform also 
produced strong CD8+ T cells as compared to recombinant protein with a number 
of distinct adjuvants. 

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The strong immunogenicity of the sD-NP platform was also demonstrated in 
guinea pigs (Figure 2-2) (4). Additionally, we observed that the sD-NP platform 
can function independently of the MBL-complement pathway, unlike HIV Env-
decorated protein self-assembling nanoparticles. This observation is likely of 
interest for clinical translation of such vaccines, as approximately 5% to 20% of 
human populations have MBL deficiency (plasma MBL < 100 ng mL–1) (11). In 
published macaque studies by other groups, DNA/Env-protein coadministration at 
the same site during each vaccination can improve induced binding and 
neutralizing Ab (nAb) responses (12) and can also improve challenge outcomes 
(13). 

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Figure 2-1 In vitro and in vivo assembly of sD-NP-GT8. (A) Nanoparticle structural model (left); 
size-exclusion chromatography and multiangle light scattering data with protein molecular weight 
on right y-axis (middle); negative stain EM imaging of NP-GT8 (right). (B) TEM images of muscle 
sections from mice injected with control or sD-NP-GT8 seven days postinjection that were 
immunolabeled with VRC01 and gold antihuman IgG. (C) Pseudonative PAGE analysis comparing 
migration of in vivo–produced sD-GT8 and sD-NP-GT8 to in vitro–produced SEC-purified 
recombinant GT8-monomer (labeled as STD mono) and eOD-GT8-60mer (labeled as STD nano) 
protein standards. 
 

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Figure 2-2 Enhanced immunogenicity of sD-NP-GT8. (A) Serology for 4 animal models 
immunized with sD-NP-GT8 compared to monomeric GT8 delivered by DNA. (B) Dose-sparing 
study showing that sD-NP-GT8 given at one-tenth of the dose induces > 1 log higher endpoint 
titers. (C) sD-NP-GT8 induces stronger CD4+ and CD8+ T cells than monomeric GT8. 
 

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2.3 
Clinical experience with related DNA vaccines encoding HIV Env 
immunogens with and without IL-12 adjuvants 
Table 2-1 Clinical trials with related DNA vaccines encoding HIV Env immunogens 
Protocol 
Products 
N 
Route of 
administration 
Safety  
HVTN 070 / 
NCT00528489 
PENNVAX-B DNA 
vaccine: A mixture of 3 
expression plasmids 
encoding HIV-1 clade B 
Env, Gag, and Pol 
IL-12 plasmid DNA: 
GENEVAX-IL-12-4532 
expressing human IL-12 
proteins p35 and p40 under 
separate regulatory control, 
formulated with 
bupivacaine 
IL-15 DNA 
120 
IM 
administration 
with needle and 
syringe 
No severe systemic 
reactogenicity 
No related severe 
adverse events (AEs) 
One discontinuation 
due to possibly 
related AE (cervical 
radiculopathy) (14) 
HVTN 080 / 
NCT00991354 
PENNVAX-B DNA 
vaccine: A mixture of 3 
expression plasmids 
encoding HIV-1 clade B 
Env, Gag, and Pol 
IL-12 plasmid DNA: 
GENEVAX-IL-12-4532, 
formulated with 
bupivacaine 
48 
IM 
administration 
with EP using 
VGX 
CELLECTRA 
EP system 
No severe systemic 
reactogenicity 
No related severe 
AEs 
Two discontinuations 
due to injection-site 
pain (14) 
HVTN 087 / 
NCT01578889 
HIV multiantigen DNA: 
Consists of 2 plasmid 
DNA expression vectors, 
HIV-1 Gag/Pol and HIV-1 
Nef/Tat/Vif, Env 
IL-12 plasmid DNA: 
GENEVAX IL-12-4532, 
formulated with 
bupivacaine 
VSV-HIV Gag 
100 
IM 
administration 
with EP using 
Ichor Medical 
Systems TriGrid 
Delivery System 
(TDS) EP device 
No severe systemic 
reactogenicity 
No severe related 
AEs (15) (16) 
HVTN 098 / 
NCT02431767 
PENNVAX-GP DNA 
vaccine: Admixture of 2 
plasmids encoding 
consensus clade A and C 
HIV-1 Env and 2 plasmids 
containing HIV-1 
multiclade (A, B, C, and 
D) consensus Pol and Gag 
IL-12 plasmid DNA: 
Inovio INO-9012 
expressing human IL-12 
p35 and p40 
94 
IM and ID 
administration 
with EP using 
CELLECTRA 
3P EP for ID 
delivery, and 
CELLECTRA 
5P EP for IM 
delivery 
No severe systemic 
reactogenicity 
No severe related 
AEs 
One participant 
discontinued due to 
pain, anxiety, and 
presyncopal episode 
(17, 18) 
 

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2.3.1 
Safety of DNA Vaccines Encoding HIV Antigens Administered Via 
Electroporation 
In general, DNA plasmids encoding HIV antigens coadministered with IL-12 
plasmids via EP have a well- established track record for safety. The 4 most 
applicable trials are summarized in Table 2-1 above and include data from over 
350 individuals. DNA vaccines were well tolerated, with no severe systemic 
reactogenicity and no severe related adverse events (AEs). Two participants who 
received IM injections discontinued further vaccinations due to pain at the 
injection site. It is important to note that there are differences between IM 
injection and ID injections followed by EP, with ID administration generally 
being less associated with injection-site pain. 
Edupuganti et al reported on the safety, tolerability, and acceptability of IM and 
ID EP of the HIV-1 PENNVAX-GP DNA vaccine and IL-12 DNA in HVTN 098 
(17). Systemic reactogenicity symptoms with IM injection of DNA vaccines and 
with ID injection of DNA vaccines did not differ (17). Injections were generally 
well tolerated but skin lesions (flat scars) occurred in 49% of participants with ID 
injections. These changes were found to be acceptable in 96% of participants and 
did not lead to vaccination discontinuation for any participant. Based on data from 
HVTN 098, there is no reason to suggest a different outcome. Accumulating 
safety data on trimer protein immunogens suggest no safety differences from 
monomers. 
When the injection and EP are given in the skin, the needles from the EP device 
may leave marks, such as red bumps or scabs. The marks may heal later but may 
still leave light or dark spots or small scars. For some people, these marks have 
lasted 9 months or more (17). These marks tend to be more noticeable on darker 
skin. 
In an ongoing phase 2/3 clinical trial (NCT04642638), a SARS-CoV-2 DNA 
vaccine is being administered with the aid of the Cellectra 2000 EP device. So far, 
over 3,200 participants have received over 9,300 active, blinded, or placebo 
product administrations via ID EP, across multiple countries. A few people have 
noticed skin discolorations after getting the vaccines, but so far, no health 
concerns have been reported. For more details, please see the Investigator’s 
Brochure (IB). 
2.3.2 
Immunogenicity of DNA Vaccines Delivered Via EP 
DNA vaccines can be efficiently introduced into cells via EP, as the transfection 
of DNA by in vivo EP allows entry of the macromolecules via a brief electrical 
field pulse. Previous approaches have used IM EP, but more recent approaches 
have delivered the DNA vaccine into the dermis (14, 15, 19). The dermis is an 
immunologic milieu rich in antigen-presenting cells (20). 
Experience from the immunogenicity results from several prior clinical studies 
has informed the design of the current study. HVTN 070 and HVTN 080 tested 
the same PENNVAX-B DNA vaccine (PV), but with (HVTN 080) and without 

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(HVTN 070) IM EP (14). Comparing similar vaccine regimens between these 
trials (PV + IL-12) after the third vaccination, the HIV-specific CD4+ T-cell 
response rate increased from 19.2% to 80.8% and the CD8+ T-cell response rate 
increased from 6.9% to 51.9% when administered by EP. These results clearly 
demonstrated the markedly enhanced T-cell immunogenicity with EP, especially 
notable for CD8+ T cells that previously had not been induced well by DNA (21). 
The responses were primarily directed to Gag and Pol, and not to Env. Because of 
this, in HVTN 098, the Env DNA vaccine was modified, including 2 Env 
plasmids encoding clade A and C Env immunogens, and including a substitution 
of an optimized IgE leader sequence as well as the deletion of their cytoplasmic 
tail to improve surface expression (18). The dose of the Env plasmids was also 
increased relative to the Gag and Pol plasmids. 
ID delivery is dose sparing. HVTN 098 compared IM and ID EP, with the EP 
administration at one-fifth the dose of the IM. CD4+ and CD8+ HIV-specific T-
cell responses were equivalent for ID and IM, demonstrating the dose-sparing 
effect of ID EP, with the highest responses to Env. Unlike prior trials, Env-
specific binding Ab responses were induced in the majority of vaccine recipients 
for both the ID and IM groups, with higher response rates and/or magnitudes for 
the ID + IL-12 group for some Env antigens. IgG-binding Abs to V1V2 Env 
antigens were detected in up to 56% of participants, with the highest responses 
from the ID + IL-12 group. Neutralizing Abs were mainly detected only to a tier-
1A viral isolate. Only a few participants developed detectable Ab-dependent 
cellular cytotoxicity (ADCC), and these were mainly in the ID + IL-12 group. 
Thus, HVTN 098 demonstrated equivalent or sometimes superior immune 
responses for ID relative to IM (both including IL-12), and that the ID route was 
dose sparing. 
Inclusion of IL-12 plasmid DNA. IL-12 induces the production of interferon-
gamma (IFN-gamma), favoring the differentiation of T-helper cells (22, 23). 
Biologically active IL-12 has pleotropic effects but is generally considered to be a 
proinflammatory cytokine that biases the CD4+ T-cell response toward a Th1 
phenotype. Previous studies have demonstrated that the inclusion of IL-12 
(GENEVAX IL-12-4532) with HIV PENNVAX-B DNA was dose sparing (14). 
The inclusion of IL-12 did not affect the tolerability of an HIV DNA vaccine (24). 
Regarding the benefit of IL-12, in HVTN 080, the sample size of the group 
without IL-12 was small, likely reducing the ability to detect significant 
differences between the responses of the group with IL-12 and those of the group 
without IL-12. Although not statistically significant, the response rates including 
IL-12 were higher than those without IL-12 (CD4+ HIV-specific T-cell response 
rates increased from 44.4% to 80.8% and CD8+ increased from 33.3% to 51.9% 
after the third dose). In HVTN 098, the CD4+ HIV-specific T-cell response rate 
increased from 56.3% to 96.4% comparing the ID groups with and without IL-12 
after the fourth dose. The binding Ab responses to Env for the ID groups with IL-
12 and without IL-12 were similar after the fourth vaccination, but tended to be 
lower after the third vaccination for the group without IL-12, suggesting that the 
inclusion of IL-12 accelerated the response achieving near maximal Ab binding 
after the third rather than the fourth vaccination. The effect of IL-12 was 
examined in another HVTN study, HVTN 087, which tested increasing doses of 

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IL-12 included with DNA administered intramuscularly with EP for 3 doses 
followed by a vesicular stomatitis virus (VSV) boost (16). HIV-specific CD8+ T-
cell response rates were generally increased when IL-12 was included, and the 
magnitude for these responses was significantly higher after the boost for the 
highest IL-12 dose group compared to the group without IL-12. Unexpectedly, the 
addition of IL-12 led to significantly decreased CD4+ HIV-specific T-cell 
response rates after the prime for the low- and medium-dose groups. 
2.4 
Safety of HIV Envelope proteins adjuvanted with 3M-052-AF + Alum: 
Clinical Experience 
HVTN 137 (NCT04177355): HVTN 137 Part A is a first-in-human, double-
blinded, dose-escalation study testing the combination of 100 mcg BG505 
SOSIP.664 gp140 with 3M-052-AF at 2 doses, including 1 mcg (Group 1) and 5 
mcg (Group 2), both combined with 500 mcg Alum. The study is ongoing and 
remains blinded to within-group treatment assignment. As of April 2022, both the 
1-mcg 3M-052-AF group (6 participants total; 5 receiving protein/adjuvant, 1 
receiving placebo) and the 5-mcg 3M-052-AF group (11 participants total; 10 
receiving protein/adjuvant, 1 receiving placebo) have completed enrollment. Part 
A of the initial protocol specified that participants in Groups 1 and 2 would 
receive 2 total doses, one at month 0 and the other at month 2. 
Five out of 6 participants received 2 doses of the 1-mcg dose (1 participant 
discontinued for reasons unrelated to vaccination) and 10 out of 11 participants 
received 2 doses of the 5-mcg dose. One participant in the 5-mcg dose group 
(Group 2) decided to discontinue vaccination due to Grade-3 induration/erythema 
first noted on Day 8 postvaccination. The maximum size of both the induration 
and erythema was measured at 17 x 17 cm on Day 8 (Grade 3). The erythema 
resolved over 3 days and the induration subsided to less than 5 cm on Day 11 
(Grade 1). The induration was measured at 2 x 2 cm on Day 14 but did not 
completely resolve until 41 days postinjection. At no point was the 
pain/tenderness greater than mild, and the participant continued to work. It is 
unknown at present whether this participant received placebo or study product. 
Another participant in Group 2 experienced 2 days of Grade-3 induration and 
erythema from Days 6-7 postvaccination that resolved completely by Day 8. In 
consultation with the PSRT, the participant did receive the second dose, which 
was uneventful. In addition, 4 participants reported Grade-3 systemic 
reactogenicity events. All 4 participants experienced severe fatigue and myalgia 
which lasted for 1-2 days. Additionally, 2 participants experienced severe 
myalgia, arthralgia, chills and headache that lasted for a day.  
The protocol was later amended and participants in Groups 1 and 2 were given the 
option to receive a third dose for 3 total doses. Nine (out of 17) participants 
elected to receive a third dose. There were no unsolicited Grade-3 or -4 AEs and 
no related serious adverse events (SAEs), AESIs, or deaths in either group after 
this third dose. Both local and systemic reactogenicity were similar between all 3 
doses in participants who received 3 doses. 

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HVTN 137 Part B is evaluating the safety and immunogenicity of 100 mcg 
BG505 SOSIP.664 gp140 in combination with 3 TLR agonists, including 5 mcg 
3M-052-AF + 500 mcg Alum, and Alum alone. As of April 17, 2022, enrollment 
is complete and 88 participants have been enrolled. Of these 88 participants, 20 
have been randomized to the 3M-052-AF group. All 88 participants have received 
at least two doses and 63 have received the third dose. Given that the trial remains 
blinded across 4 treatment arms, interpretation of the blinded safety data from 
HVTN 137 Part B is challenging, but there have been no related SAEs, AESIs, 
deaths, or unplanned study pauses. No additional grade 3 local reactogenicity 
events have been observed. 
Overall, the BG505 SOSIP.664 with 3M-052-AF was generally well tolerated, 
with no unsolicited Grade-3 or -4 AEs, no related SAEs, AESIs, or deaths, and no 
unplanned study pauses. Other than the persistent erythema in 1 of the HVTN 137 
Part A Group 2 participants described above, all reactogenicity symptoms 
resolved within 14 days and generally within 7 days. For more detailed 
information, please see the IB. 
HVTN 300 (NCT04915768): This study is a first-in-human, unblinded trial 
testing a 300-mcg dose of a CH505 TF chTrimer (a stabilized, chimeric SOSIP 
Env trimer) in combination with 5 mcg 3M-052-AF + 500 mcg Alum delivered 
via split injection into both the right and left deltoids. The planned schedule is 5 
total vaccinations at months 0, 2, 4, 8, and 12. As of September 7, 2022, the study 
is ongoing and all 13 participants have received the first vaccination, 10 
participants have received the second vaccination, 9 participants have received the 
third vaccination, 9 participants have received the fourth vaccination, and 6 
participants have received the fifth vaccination. Five (5) participants have 
discontinued further vaccinations, 1 due to a panic attack after the first injection 
(this participant had a history of panic attacks before being part of the study), 3 
due to reactogenicity events and 1 was lost to follow-up. Out of these 5, 2 
terminated from the study early (panic attack and lost to follow-up) and 3 have 
remained in the study for follow-up. 
All participants experienced at least some local reactogenicity during the trial, 
mostly mild to moderate. One (1) participant experienced severe pain/tenderness 
in both the right and left injection sites 3 days following the fourth vaccination, 
though it lasted only one day. 
Nearly all participants reported some systemic reactogenicity during the course of 
the trial to date, mostly mild to moderate. For example, 11 of 13 participants 
experienced systemic reactogenicity after the first dose. Five (5) participants 
reported Grade 3 (severe) systemic reactogenicity during the trial through 
September 7, 2022. One (1) of those participants received a subsequent 
vaccination without any Grade 3 reactogenicity, one participant experienced an 
additional repeat Grade 3 systemic reactogenicity symptom after a subsequent 
vaccination, one participant has not yet received a subsequent vaccination, and 
two (2) participants declined further vaccinations after their first Grade 3 systemic 
reactogenicity event.  

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All Grade 3 events resolved within the 7-day reactogenicity period; the longest 
duration for severe reactogenicity was 2 days. 
There have been no related SAEs, AESIs, or deaths as of September 7, 2022. 
2.5 
Rationale for evaluation of 3M-052-AF + Alum adjuvant 
3M-052-AF contains 3M-052 and an emulsifier in an aqueous formulation. 3M-
052 is a TLR7 and TLR8 agonist (TLR7/8 agonist) that induces production of 
cytokines in vitro from immune cells, such as dendritic cells, macrophages, and 
monocytes (25-28). 
Nonclinical studies in guinea pigs, rats, rabbits, and rhesus macaques comparing 
the immunogenicity of the related SOSIP protein BG505 SOSIP.664 formulated 
with a diverse range of adjuvants have consistently shown that the protein 
adjuvanted with 3M-052-AF elicits neutralizing and binding Abs more quickly, to 
higher-peak magnitudes, and with greater durability than any of the other protein-
adjuvant combinations tested (29). 
In rhesus macaques among 8 adjuvants combined with an Env gp140 immunogen, 
the Alum/TLR7 was the most potent in assays characterizing both Ab and cellular 
responses. This hierarchy of potency was sustained throughout the longitudinal 
follow-up. The overall effect of the Alum/TLR combination was to boost Env 
binding Ab titers 3-to-10-fold compared to Alum alone (30). 
The 3M-052-AF adjuvant is a TLR7/8 agonist added to vaccine antigens to 
generate strong Ab responses in several nonhuman primate (NHP) models, and is 
currently being evaluated in HVTN 137 and HVTN 300 clinical trials (see Section 
2.4). Combining 3M-052-AF with Alum results in a synergistic increase in 
immune responses to vaccine antigens in preclinical models, including Ab titers, 
CD4+ T-cell responses, infiltrating monocytes and dendritic cells, and B-cell 
activation when compared to either Alum or 3M-052-AF alone (25). In NHPs 
immunized with SARS-CoV-2, the receptor binding domain (RBD) formulated 
with 3M-052-AF combined with Alum and increased Ab titers, RBD binding and 
effector Abs, Th1 biased CD4+ T cells, and protection from SARS-CoV-2 viral 
challenge when compared to Alum alone (31). 
For additional information, see the IB. 
2.6 
Rationale for schedule 
Previous trials of DNA vaccines for HIV-1 Env immunogens have used a 0-, 1-, 
3-, and 6-month vaccination schedule, exactly as proposed in this study. Those 
regimens were well tolerated and immunogenic (14, 15). Specifically for ID 
delivery, both Ab and T-cell responses were induced (17, 18). In HVTN 098, the 
fourth vaccination boosted the magnitude of total Ab response for all treatment 
groups against gp140 antigens from clades A and B as compared to the magnitude 

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of total Ab response after the third dose (18). By including a boost of Trimer 4571 
at month 6 in Group 3, we can assess whether additional doses of protein and 
DNA lead to enhanced immunogenicity compared to DNA alone in this study 
(Group 2) and previous studies. By maintaining a standard schedule, data will be 
directly comparable to previous DNA vaccine studies. 
2.7 
Rationale for evaluation of boost with trimeric protein 
Trimer 4571 is a stabilized NLT derived from the BG505 isolate in a gp140 
format, modified with a different set of mutations to prevent CD4-induced 
conformational triggering (32-34). In proof-of-concept preclinical studies in 
guinea pigs, Trimer 4571, adjuvanted with Alum, induced autologous nAbs to 
BG505.W6M.C2 after the second administration, which increased further after the 
third administration. Rhesus macaques immunized via IM injection with research-
grade Trimer 4571 produced nAbs to BG505.W6M.C2.T332N (see the IB for 
more details on these BG505 variants). 
2.8 
Rationale for targeting the same draining lymph node with both DNA 
and protein trimer 
Direct targeting of the draining lymph node with a multipronged immune stimulus 
consisting of both a DNA vaccine and an adjuvanted protein vaccine led to 
improved outcomes in a variety of several preclinical models (12, 35-37). The 
most directly relevant study is that of Felber et al in rhesus macaques (37). This 
study compared vaccination with monomeric HIV envelope administered via both 
DNA and an adjuvanted protein delivered into the same limb or into contralateral 
limbs. This method allowed for a direct comparison of 2 injections with the same 
regimen (protein and DNA) at the same time in the same limb versus spacing 
simultaneous injections into contralateral limbs. Vaccination targeting the same 
draining lymph nodes led to improvement in a variety of immune outcomes, 
including functional Ab responses (increased ADCC activity and improved 
FcRIIIa binding) and cellular responses (CD4+ and CD8+). The proposed 
mechanism involves simultaneous priming of both CD4+ T cells (mostly induced 
by DNA) and Env-specific B cells (mainly induced by the adjuvanted protein 
component) in the same lymph node when injections were given in the same limb. 
Most importantly, vaccination targeting the same draining lymph nodes resulted 
in superior protection against the simian-human immunodeficiency virus (SHIV) 
challenge (see Figure 2-3). 

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Figure 2-3 Coadministration group shows significant protection from SHIV.CH505 infection. 
(A) Schematic representation of vaccine delivery of the 2 components (DNA and protein) in the 2 
vaccination regimens, “coadministration” in the same anatomical sites and “separate 
administration” in contralateral sites. The coadministration group received DNA delivered via IM 
EP, followed immediately by IM injection of the adjuvanted protein. The separate administration 
group received the vaccine components in different anatomical sites, with DNA delivered by IM EP 
in the left site and protein IM in the right site. The same vaccine components and the same total 
vaccine dose were used for both regimens. (B) Vaccination schedule indicating the sequentially 
isolated CH505 immunogens used. Five weeks after the last vaccination, the animals were 
exposed weekly to repeated low-dose vaginal challenges using SHIV.CH505. (C) Kaplan-Meier 
curves show the viral acquisition rate after repeated low-dose SHIV.CH505 challenges of the 2 
vaccine groups (n = 18 and n = 17, respectively) and of the control group (n = 20). The RMs were 
exposed to 15 weekly intravaginal challenges. Infection was defined by 2 consecutive positive 
plasma viral load (VL) measurements. No RMs were censored. P value, exact log-rank test (37). 
In keeping with the well tolerated safety profile of simultaneous delivery of HIV 
immunogens via both DNA+ envelope protein administration strategies (38) (39), 
no AEs were noted with either strategy (separation versus targeting the same 
draining lymph node), including assessments via clinical chemistry, complete 

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blood counts (CBCs), body weight, temperature, and regular physical exams by 
veterinarians. Additionally, theoretical concerns regarding increased local 
reactogenicity are mitigated by the observation that the DNA vector does not 
disperse from the site of injection (40) and that the DNA will be delivered 
intradermally, whereas the protein will be delivered intramuscularly. 
2.9 
Rationale for dose of Trimer 4571 
The HIV-1 Env Trimer 4571 (VRC-HIVRGP096-00-VP) variant vaccine BG505 
DS-SOSIP.664 consists of purified, formulated HIV-1 recombinant glycoprotein 
(gp140) as a soluble homotrimeric assembly of gp120-gp41 heterodimers, derived 
from the clade A HIV-1 strain BG505 and produced in a Chinese Hamster Ovary 
(CHO) DG44 cell line. Specifically, the BG505 DS-SOSIP.664 (Trimer 4571) 
possesses an engineered stabilizing disulfide bond between residues 201 and 433 
(DS), maintaining the truncation at residue 664 in gp41, producing a soluble, 
cleaved Env trimer in a prefusion closed conformation. The ongoing and 
completed clinical trials with Trimer 4571 are listed in Table 2-2. Data on the 
safety and immunogenicity of Trimer 4571 + Alum from VRC 018 are available. 
No related SAEs were reported. The immunologic assays and analysis for this 
study are ongoing. Trimer 4571–specific Ab titers in serum samples were 
measured by electrochemiluminescence (ECLIA) using a Meso Scale Discovery 
(MSD) platform at baseline and at 2 weeks after the third product administration. 
Both the 100-mcg (n = 3) and 500-mcg (n = 5) doses elicited Trimer 4571–
specific Abs, with geometric mean areas under the curve (AUCs) 8-fold and 40-
fold over background, respectively. No substantial difference was observed 
between the 100-mcg and the 500-mcg dose, so the 100-mcg dose will be tested 
in this trial. 
For additional information, see the IB. 

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Table 2-2 Clinical trials with Trimer 4571 
Protocol 
Study 
status 
N 
receiving 
Trimer 
4571 + 
Alum 
Route of 
administration 
and dose 
Schedule 
Safety data 
(clinicaltrials.gov) 
VRC 018 / 
NCT03783130 
 
Completed 
16 
IM or 
subcutaneous 
(SC) at 100 
mcg or 500 
mcg Trimer 
with 500 mcg 
Alum  
0, 2, 5 
months 
 As of August 2021, 
final study data: no 
related SAEs, related 
grade AEs, or related 
MAAEs were reported 
or other unexpected 
reactions, and no study 
pause criteria were met 
at any time. The clinical 
experience with Trimer 
4571 will be updated in 
the IB Version 4.0 
expected in September 
2022 
NIAID 19-I-
0069 / 
NCT03878121 
 
Enrolling  
Max 100 
IM at 500 mcg 
Trimer with 
500 mcg Alum  
Boost post 
adenovirus 
vector 
prime 
 As of Jan 5, 2022, 12 
participants have 
received study product. 
No SAEs, no SUSARs 
NCT04985760 
Enrolling 
24 
IM at 100 mcg 
or 500 mcg 
Trimer with 
500 mcg Alum 
0, 2 and 5 
months 
As of January 01, 
2022, 5 participants 
have received either 
the 4571/alum or 
control. 2 participants 
have received two 
doses of study 
product and the 
remaining 3 have 
received a single 
dose. No related 
SAEs, no SUSARS 
2.10 
Rationale for dose of 3M-052-AF adjuvant 
The 5-mcg dose of 3M-052-AF adjuvant has been selected based on preclinical 
data and clinical experience with other recombinant HIV envelope proteins in 
HVTN 137 and HVTN 300. The 500-mcg dose of Alum (ie, aluminum content by 
weight) adjuvant also matches the regimen in HVTN 137 and HVTN 300. 
HVTN 137 has now completed enrollment in Part A and Part B. Based upon the 
available evidence, the HVTN 137 study team decided to proceed with a 5-mcg 
dose over a 1-mcg dose. 

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2.11 
Rationale for FNA  
To improve assessment of the effects of sD-NP-GT8 on B-cell lineage 
development, this protocol will employ FNA of an axillary lymph node proximal 
to the sites of vaccine administration. To assess the frequency of VRC01-class B 
cells from lymph nodes, FNA will be performed in a subset of participants in 
Groups 2 and 3 (at minimum 9 per group, half of each group). FNA will be 
performed approximately 3 weeks after both the second dose and the third dose of 
the sD-NP-GT8 immunogen. The germinal center response, which occurs 
exclusively in the lymphatic tissue, is critical for directing B-cell lineage 
development. Havenar-Daughton et al have employed repeated direct probing of 
germinal center responses by fine needle lymph node aspirates in NHP studies 
and found that the induction of nAbs against tier-2 autologous HIV strains best 
correlated with germinal center B-cell magnitude and Env-specific CD4+ Tfh 
cells (41). Similar sampling strategies are under evaluation in licensed vaccine 
and early-phase human HIV vaccine studies (42). 
Fine needle aspirate of the axillary lymph node is a safe procedure with multiple 
studies reporting no significant complications with either FNA or core biopsy (a 
more invasive procedure than FNA) (43). The protocol team has developed 
eligibility requirements for the proposed procedures based upon guidelines from 
the American Society of Anesthesiologists, the American Board of Internal 
Medicine, the American Red Cross, the Society of Interventional Radiology, and 
a review of the scientific literature.  
Specifically, it is a considered decision to include participants with obesity. The 
rate of obesity varies substantially by self-reported race with higher rates in 
populations identifying as Hispanic or non-Hispanic Black (44), and eligibility 
criteria related to obesity have been cited as a specific trial design barrier 
impeding the participation of women and underrepresented minorities (45). An 
extremely low rate of significant adverse events precludes identification of risk 
factors for poor outcomes, but we could find no evidence that Body Mass Index 
(BMI) is associated with increased complications with FNA. Additionally, 
diagnostic yields are higher in participants with obesity undergoing clinical FNA 
(46), an observation attributed to enhanced contrast between adipose and 
lymphatic tissue. 
The team has also considered whether to include temporal limits on 
platelets/International Normalized Ratio (INR), and a temporal requirement to 
abstain from Non-steroidal anti-inflammatory drug (NSAID)/aspirin use prior to 
procedures. Routine measurement of INR has demonstrated no ability to identify 
patients who are at risk of periprocedural bleeding, even with higher risk 
procedures such as placement of central venous catheters (47). Additionally, 
platelets are assessed throughout the trial (including within 30 days of FNA). 
Hence the team has decided not to include INR measurement as a restriction. We 
also note that the trial eligibility criteria exclude persons with bleeding disorders 
(eg, factor deficiency, coagulopathy, or platelet disorder requiring special 
precautions [see Section 5.1.2]). Interventional radiology guidelines state that 
aspirin and/or NSAIDS are not a contraindication to FNA (48).  

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2.12 
Rationale for leukapheresis 
To assess the expansion of rare precursor B-cell populations that occur at a 
frequency of approximately 1 out of 20 million cells, a large number of peripheral 
blood mononuclear cells (PBMCs) is required. Leukapheresis can provide 6 to 10 
billion PBMCs without depleting red blood cells or other blood components. 
There are approximately 1 million PBMCs per milliliter of blood; therefore, 
peripheral blood sampling at volumes associated with typical blood donation 
(~500 mL) would not provide sufficient PBMC numbers. Furthermore, 
leukapheresis is associated with fewer AEs when compared to blood donation. 
Hence, this protocol will employ leukapheresis in all study participants to ensure 
adequate specimens for analysis of rare precursors. Leukapheresis will be 
performed at screening and after the third dose of sD-NP-GT8 has been 
administered. The rationale for performing leukapheresis at 2 weeks after the third 
dose is based on previous studies that have assessed potentially rare VRC01-class 
memory B cells in the periphery (49). 
Leukapheresis is a safe and well tolerated procedure with several studies reporting 
no significant late effects attributable to the procedure (50, 51). The protocol team 
has developed eligibility requirements for the proposed procedures (see Section 
5.1) and leukapheresis eligibility criteria (see Section 8.6) as well as accounting 
for guidelines from the American Society for Apheresis, the American Society of 
Hematology, and review of the scientific literature.  
The team has also considered whether to include temporal limits on platelets/INR, 
and a temporal requirement to abstain from NSAID/aspirin use prior to 
leukapheresis and has decided not to include these as restrictions (48). We note 
that the trial eligibility criteria exclude persons with bleeding disorders (eg, factor 
deficiency, coagulopathy, or platelet disorder requiring special precautions (see 
Section 5.1.2). Assessment of patients post leukapheresis found the most frequent 
side effect (39%) was paresthesia due to citrate-related hypocalcemia, managed 
with oral calcium supplements and/or slower rates. No bleeding occurred (52). 
The study team conclude that adverse reactions with leukapheresis are similar to 
those reported for conventional PBMC collections. Routine measurement of INR 
has demonstrated no ability to identify patients who are at risk of periprocedural 
bleeding even with higher risk procedures such as placement of central venous 
catheter (47). 

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2.13 
Risks and benefits 
2.13.1 
Potential risks 
Table 2-3 Summary of potential risks of study products and administration 
Risk type 
Summary 
Common 
• 
Mild to moderate injection-site pain, tenderness, erythema, or 
swelling/induration/edema 
• 
Malaise/fatigue, myalgia, or headache in the first few days following 
injection 
• 
A vaccine-induced positive HIV Ab test result 
• 
Visible lesion(s) at the injection site, such as erythematous papules with 
eschar, hypopigmentation, hyperpigmentation, or scar due to EP 
Less 
common 
• 
Severe injection-site pain or tenderness 
• 
Fever, chills, flu-like syndrome, arthralgia, rash, nausea, or dizziness in 
the first few days following injection 
• 
Vasovagal reaction/lightheadedness/dizziness related to the injection 
procedure 
• 
Transient changes in clinical laboratory values 
• 
Injection-site hematoma, bruising/ecchymosis, other transient lesions, 
itching, or bleeding related to the injection procedure 
Uncommon 
or rare 
• 
Severe localized injection-site reaction, such as sterile abscess or 
secondary bacterial infection 
• 
Allergic reaction, including rash, urticaria, angioedema, bronchospasm, or 
anaphylaxis 
Theoretical 
• 
Muscle damage at the injection site 
• 
Electrical injury with EP 
• 
Disruption of implanted electrical devices 
• 
Exacerbation of cardiac arrythmia with EP 
• 
Effects on a participant’s response to an approved HIV vaccine 
administered in the future 
• 
Effects on susceptibility to HIV if the participant is exposed to HIV 
• 
Effects on the course of HIV disease if the participant acquires HIV 
• 
Effects on the fetus and on pregnancy if the participant becomes pregnant 
In addition to risks from ID injections and EP of DNA vaccines (see Section 2.3), 
frequent potential side effects resulting from subcutaneous (local anesthetic prior 

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to FNA) injections include stinging, discomfort, redness of skin, and mild 
bruising at injection site. 
Blood drawing: May cause pain, bruising, fainting, and, rarely, infection at the 
site where the blood is taken.  
Leukapheresis: Can  cause hypocalcemia, pain, bruising, and rarely, infection. 
Additional risks may depend on local leukapheresis procedures. Fainting can also 
occur but occurs less frequently than with blood drawing. 
FNA of lymph nodes: Can sometimes cause pain, bruising, and bleeding at the 
site where the needle is inserted. Similar to any procedure where a needle is 
inserted into the skin (such as vaccination, as described above), there is a rare risk 
of localized infection. Rarely, the local anesthetic can cause arm numbness or 
weakness. 
Risks during pregnancy: Because possible effects of the study vaccine on a fetus 
or nursing infant are unknown, persons assigned female sex at birth who have 
reproductive potential will be tested for pregnancy at screening and prior to 
administration of each dose of the study vaccine. Such persons will be asked to 
notify the site immediately if they suspect or learn they are pregnant during this 
study. In case of pregnancy, participants will continue to be followed for safety 
and the participant will not receive any additional vaccinations. The participant 
will be contacted about the outcome of a pregnancy that begins during the study. 
Other risks: The medical tests performed as part of this research protocol may 
result in new diagnoses or abnormal values without clinical significance (“false 
positives”). Depending on the medical findings and consequences of being 
provided with the results of these tests, the study participant may view this as 
either a risk or a benefit. Any such information will be shared and discussed with 
the participant and, if requested by the participant, may be forwarded to their 
primary health care provider for further workup and management. 
Participants in this study risk experiencing discrimination or other personal 
problems that may result from study participation itself: these types of risks are 
known collectively as “negative social impacts.” The HVTN CRS is obliged to 
provide advocacy for and assistance to participants regarding these negative social 
impacts associated with the vaccine trial. If HVTN CRS staff have questions 
regarding ways to assist a participant dealing with a negative social impact, a 
designated HVTN Core representative can be contacted. 
2.13.2 
Benefits 
Study participants will not receive direct health benefits from study participation. 
Others may benefit from knowledge gained in this study that may aid in the 
development of an HIV vaccine. The investigational vaccines are not expected to 
provide protection from HIV. 

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Participants may benefit from more frequent counseling, laboratory tests, and 
physical exams while enrolled in the study. Participants may also experience 
positive social impacts as a benefit of study participation. When asked, 
participants say that being in a study made them feel good about helping others, 
increased their knowledge about HIV, and improved their self-esteem. 

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3 
Objectives and endpoints 
3.1 
Primary objectives and endpoints 
 
Objectives 
Endpoints 
1) To evaluate the safety 
and tolerability of 3 doses of 
sD-NP-GT8 + IL-12 DNA 
adjuvant with or without 2 
doses of Trimer 4571 + 3M-
052-AF/Alum adjuvant  
a) Local and systemic reactogenicity signs and symptoms will 
be collected for a minimum of 2 weeks following receipt of 
any study vaccine 
b) SAEs, medically attended adverse events (MAAEs), AESIs, 
and AEs leading to early participant withdrawal or permanent 
discontinuation will be collected throughout the study and for 
12 months following any receipt of study products. 
Additionally, all AEs will be collected for 30 days after any 
receipt of study vaccination.  
 
2) To evaluate the 
immunogenicity of 3 doses 
of sD-NP-GT8 + IL-12 
DNA adjuvant with or 
without 2 doses of Trimer 
4571 + 3M-052-AF/Alum 
adjuvant 
a) Response rate and magnitude of HIV-1–specific binding Ab 
responses to eOD-GT8-60mer, eOD-GT8 monomer, or Trimer 
4571 as assessed by multiplex assay 2 weeks following the 
third (Groups 1 and 2) and fourth (Group 3) vaccinations 
b) Response rate and magnitude of CD4+ and CD8+ T-cell 
responses, measured by flow cytometry, to HIV-1–specific Env 
peptide pools 2 weeks following the third (Groups 1 and 2) and 
fourth (Group 3) vaccinations 
3) To evaluate the induction 
of VRC01-class B cells 
a) Frequency of VRC01-class B cells from lymph nodes and 
periphery following the second and third vaccinations 
3.2 
Secondary objectives and endpoints 
 
Objectives 
Endpoints 
1) To further evaluate the 
immunogenicity of 3 doses of 
sD-NP-GT8 + IL-12 DNA 
adjuvant with or without 2 
doses of Trimer 4571 + 3M-
052-AF/Alum adjuvant 
a) eOD-GT8-60mer and monomer Ab response rate and 
magnitude measured by multiplex assay at 2 weeks 
following the first, second, and third immunizations 
between Group 1 and Group 2 to assess dose-sparing 
effects of self-assembling nanoparticle vaccines 
b) Quantify and compare antigen-specific germinal center 
Tfh and B-cell responses across groups at 3 weeks after 
the second and third vaccinations 
c) Evaluate nAb magnitude and breadth against tier-2 
HIV-1 isolates after third and fourth vaccinations 
3.3 
Exploratory objectives 
1. To clinically evaluate EP-injection–related skin changes for 6 months after the 
last study product administration and subjective assessment by participant of 
tolerability at 12 months after the last study product administration   

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2. Evaluate ADCC and antibody-dependent cellular phagocytosis (ADCP) after the 
last vaccination 
3. Describe the binding specificities of sera using electron microscopy-polyclonal 
epitope mapping (EMPEM) at 2 weeks post–third and fourth vaccinations, if Env 
binding is observed 
4. Evaluate Abs for cross-reactive binding to eOD-GT8 and Trimer 4571 6 weeks 
after the fourth vaccination and after 6 months post–fourth vaccination for Group 
3 
5. To conduct analyses related to furthering the understanding of HIV, immunology, 
vaccines, and clinical trial conduct, including but not exclusive to B-cell 
repertoire analysis (including analysis of rare B-cell lineages associated with 
bnAb precursors), and assessment of lymph node aspirate for germinal center 
activity, including cellular phenotyping and mutational frequency analysis 
suggestive of somatic hypermutation and affinity maturation following 
immunization. 

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4 
Laboratory strategy 
The primary immunogenicity endpoint assay will be a combination of 
measurements describing whether VRC01-like B-cell lineages are elicited and 
whether Env-specific CD4+ and CD8+ T cells are also elicited. Intracellular 
cytokine staining (ICS) assays will be performed at the Fred Hutch HVTN 
laboratory. 
The detailed laboratory strategy and the technical details are described in the 
Central Assay Plan and will be updated as new reagents and techniques are 
incorporated into assay planning. This document will be available on the HVTN 
305 protocol webpage. Descriptions of the standard HVTN laboratory assays can 
be found online at https://www.hvtn.org/en/science.html. 

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5 
Study design 
This is an open-label study to examine the safety and immunogenicity of synthetic 
DNAs encoding NP-GT8 and IL-12 with or without a TLR-agonist–adjuvanted 
Env Trimer 4571 boost in adults without HIV. The primary hypothesis is that 
vaccination with a recombinant DNA vaccine encoding a germline-targeting 
epitope followed by a trimeric protein boost will be safe and immunogenic. 
5.1 
Study population 
All inclusion and exclusion criteria must be met for eligibility. Screening 
procedures to determine eligibility must be performed within 56 days prior to 
enrollment. 
Investigators should always use good clinical judgment when considering a 
volunteer’s overall fitness for trial participation. Some volunteers may not be 
appropriate for enrollment even if they meet all inclusion/exclusion criteria. 
Medical, psychiatric, occupational, or other conditions may make evaluation of 
safety and/or immunogenicity difficult, and some volunteers may be poor 
candidates for retention. 
5.1.1 
Inclusion criteria 
1. Able and willing to complete the informed consent process, including an 
Assessment of Understanding (AoU): volunteer demonstrates understanding of 
this study; completes a questionnaire prior to first vaccination with verbal 
demonstration of understanding of all questionnaire items answered incorrectly. 
2. 18 to 55 years old, inclusive, on day of enrollment. 
3. Available for clinic follow-up through the last clinic visit and willing to be 
contacted at least 12 months after the last vaccine administration. 
4. Willing to undergo leukapheresis (see Appendix A and Appendix B). 
5. Agrees not to enroll in another study of an investigational agent during 
participation in the trial. 
6. In good general health according to the clinical judgement of the site investigator. 
7. Physical examination and laboratory results without clinically significant findings 
that would interfere with assessment of safety or reactogenicity in the clinical 
judgement of the site investigator. 
8. Assessed as low risk for HIV acquisition per low-risk guidelines (see Appendix F 
and Appendix G), agrees to discuss HIV-infection risks, agrees to risk-reduction 
counseling, and agrees to avoid behaviors associated with high risk of HIV 

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exposure through the final study visit. Low risk may include persons stably taking 
HIV pre-exposure prophylaxis (PrEP) as prescribed for 6 months or longer. 
9. Hemoglobin: 
• ≥ 11.0 g/dL for volunteers who were assigned female sex at birth. 
• ≥ 13.0 g/dL for volunteers who were assigned male sex at birth and 
transgender males who have been on hormone therapy for more than 6 
consecutive months. 
• ≥ 12.0 g/dL for transgender females who have been on hormone therapy for 
more than 6 consecutive months. 
• For transgender participants who have been on hormone therapy for less than 
6 consecutive months, determine hemoglobin eligibility based on the sex 
assigned at birth.  
10. Platelets = 125,000-550,000/mm3. 
11. White blood cell (WBC) count = 2,500-12,000/mm3 (not exclusionary: if count 
greater than 12,000 with investigation showing general good health and PSRT 
approval). The Leukapheresis Center may impose a higher lower limit of 
3,500/mm3 
12. Alanine aminotransferase (ALT) < 2.5 x upper limit of normal (ULN) based on 
the institutional normal range. 
13. Serum creatinine ≤ 1.1 x ULN based on the institutional normal range. 
14. Corrected total serum calcium level of > 8.5 mg/dL  
15. Blood pressure in the range of 90 to < 140 mmHg systolic and 50 to < 90 mmHg 
diastolic. 
16. Negative results for HIV infection by a US Food and Drug Administration 
(FDA)-approved enzyme immunoassay (EIA) or chemiluminescent microparticle 
immunoassay (CMIA). 
17. Negative for anti-Hepatitis C Abs (anti-HCV), or negative HCV nucleic acid test 
(NAT) if anti-HCV Abs are detected. 
18. Negative for Hepatitis B surface antigen. 
19. For a volunteer capable of becoming pregnant: 
• Volunteers who were assigned female sex at birth and are of reproductive 
potential must agree to use effective means of birth control from at least 21 

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days prior to enrollment through 8 weeks after their last vaccination timepoint 
(see Appendix H). 
• Has negative beta human chorionic gonadotropin (β-HCG) pregnancy test 
(urine or serum) on day of enrollment. 
5.1.2 
Exclusion criteria 
1. Volunteer who is breast-feeding or pregnant. 
2. Morbid obesity. Enrollment of individuals with body mass index (BMI) that is 
≥ 40, whom the site investigator assesses are in good health, may be 
considered by PSRT on a case-by-case basis. 
3. Diabetes mellitus (DM). Type 2 DM, controlled with diet alone, or a history 
of isolated gestational diabetes are not exclusionary. Enrollment of individuals 
with Type 2 DM that is well-controlled on hypoglycemic agent(s) may be 
considered, provided the HgbA1c is ≤ 8% within the last 6 months (sites may 
draw these at screening). 
4. Previous or current recipient of an investigational HIV vaccine (previous 
placebo recipients are not excluded). 
5. Systemic glucocorticoid use equal to or greater than prednisone 10 mg/day 
within 3 months prior to enrollment, congenital or acquired 
immunodeficiency, or other systemic medication use likely to impair immune 
response to vaccine in the opinion of the site investigator. 
6. Blood products or immunoglobulin within 16 weeks prior to enrollment; 
receipt of immunoglobulin within 16 weeks prior to enrollment requires PSRT 
approval. 
7. Receipt of any live attenuated vaccine within 4 weeks prior to enrollment. 
(Note: ACAM2000 vaccine for Mpox (previously known as Monkeypox) 
received within 30 days prior to enrollment or receipt of study vaccine, or if 
ACAM2000 received greater than 30 days prior to enrollment, or prior to 
receipt of study vaccine and vaccination scab still present; or planned 
administration within 30 days after enrollment or receipt of study vaccine). 
8. Receipt of any vaccines that are not live attenuated within 14 days prior to 
enrollment; replication incompetent vaccines such as the Jynneos vaccine for 
the prevention of Mpox disease are not considered to be live vaccines.  
9. Non-HIV experimental vaccine(s) received within the last 1 year in a prior 
vaccine trial. Exceptions may be made by the HVTN 305 PSRT for vaccines 
that have subsequently undergone licensure or Emergency Use Authorization 
by the FDA or, if outside the United States, equivalent authorization by the 
national regulatory authority.  

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10. Initiation of antigen-based immunotherapy for allergies within the previous 
year (stable immunotherapy is not exclusionary); inclusion of participants 
who initiated immunotherapy within the previous year requires PSRT 
approval.  
11. Receipt of investigational research agents with a half-life of 7 or fewer days 
within 4 weeks prior to enrollment. If a potential participant has received 
investigational agents with a half-life greater than 7 days (or unknown half-
life) within the past year, PSRT approval is required for enrollment.  
12. Serious reactions to vaccines that preclude receipt of study injections as 
determined by the principal investigator (PI) or designee, including history of 
serious reaction (eg, hypersensitivity, anaphylaxis) to any or any component 
of the study vaccine.  
13. Hereditary angioedema, acquired angioedema, or idiopathic forms of 
angioedema.  
14. Idiopathic urticaria within the past year.  
15. Bleeding disorder diagnosed by a doctor (eg, factor deficiency, coagulopathy, 
or platelet disorder requiring special precautions).  
16. Seizure disorder; febrile seizures as a child or seizures secondary to alcohol 
withdrawal more than 5 years ago are not exclusionary.  
17. Asplenia or functional asplenia.  
18. Active duty and reserve US military personnel.  
19. Any other chronic or clinically significant condition that in the clinical 
judgement of the investigator would jeopardize the safety or rights of the 
study participant, including, but not limited to: clinically significant forms of 
drug or alcohol abuse, serious psychiatric disorders, persons with any suicide 
attempt within the past one year (if between 1-2 years, consult PSRT) or 
cancer that, in the clinical judgment of the site investigator, has a potential for 
recurrence (excluding basal cell carcinoma).  
20. Asthma is excluded if the participant has ANY of the following: 
• Required either oral or parenteral corticosteroids for an exacerbation 
two or more times within the past year; OR 
• Required either oral or parenteral corticosteroids for an exacerbation 2 
or more times within the past year; OR 
• Needed emergency care, urgent care, hospitalization, or intubation for 
an acute asthma exacerbation within the past year (eg, would NOT 
exclude individuals with asthma who meet all other criteria but sought 

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urgent/emergent care solely for asthma medication refills or coexisting 
conditions unrelated to asthma); OR 
• Uses a short-acting rescue inhaler more than 2 days per week for acute 
asthma symptoms (ie, not for preventive treatment prior to athletic 
activity); OR 
• Uses medium-to-high-dose inhaled corticosteroids (greater than 250 
mcg fluticasone or therapeutic equivalent per day), whether in single-
therapy or dual-therapy inhalers (ie, with a long-acting beta agonist 
[LABA]); OR 
• Uses more than 1 medication for maintenance therapy daily. Inclusion 
of anyone on a stable dose of more than 1 medication for maintenance 
therapy daily for greater than 2 years requires PSRT approval.  
21. A participant with a history of an immune-mediated disease, either active or 
remote. Specific examples are listed in Appendix I (AESI index). Not 
exclusionary: 1) remote history of Bell’s palsy (>2 years ago) not associated 
with other neurologic symptoms, 2) mild psoriasis that does not require 
ongoing systemic treatment.  
22. History of allergy to local anesthetic (Novocaine, Lidocaine).   
23. Investigator concern for difficulty with venous access based upon clinical 
history and physical examination. For example, history of IV drug abuse or 
substantial difficulty with previous blood draws.  
24. Presence of implanted electronic medical device (eg, pacemaker, implantable 
cardioverter defibrillator).  
25. Presence of surgical or traumatic metal implant in either upper arm and/or 
upper torso.  
26. History of cardiac arrhythmia (eg, supraventricular tachycardia, atrial 
fibrillation) (Not excluded: sinus arrhythmia).  
27. Tattoo overlying the injection sites preventing assessment of reactogenicity in 
the view of the investigator or skin condition at the injection sites.  
28. History or presence of keloid scar formation or hypertrophic scar 

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5.2 
Participant departure from vaccination schedule or withdrawal 
5.2.1 
Delaying vaccinations for a participant 
Under certain circumstances, a participant’s scheduled vaccination will be 
delayed. The factors to be considered in such a decision include but are not 
limited to the following: 
• Intercurrent illness or prevaccination abnormal vital signs or clinical 
symptoms that may mask assessment of vaccine reaction. 
• Treatment with systemic glucocorticoids (eg, prednisone or other 
glucocorticoid), immune targeting monoclonal Abs or other 
immunomodulators (other than nonsteroidal anti-inflammatory drugs 
[NSAIDs]), with the exception that study injection may continue per PI 
discretion if the next study injection occurs at least 2 weeks following 
completion of glucocorticoid treatment. 
• Receipt of any live attenuated vaccines within 4 weeks prior to study vaccine 
administration. (Note: ACAM2000 vaccine for Mpox received within 30 days 
prior to enrollment or receipt of study vaccine, or if ACAM2000 received 
greater than 30 days prior to enrollment, or prior to receipt of study vaccine 
and vaccination scab still present; or planned administration within 30 days 
after enrollment or receipt of study vaccine).  
• Receipt of any vaccines that are not live attenuated vaccines within 14 days 
prior to study vaccine administration. Replication incompetent vaccines such 
as the Jynneos vaccine for the prevention of Mpox disease are not considered 
to be live vaccines. 
Vaccinations should not be administered outside the visit window period specified 
in Appendix C without PSRT approval. Any vaccination(s) administered outside 
the visit window period specified in Appendix C without PRST approval should 
be reported as protocol deviations. 
5.2.2 
Discontinuation of study vaccine administration 
Under certain circumstances, an individual participant’s vaccinations will be 
permanently discontinued. Specific events that will result in stopping a 
participant’s vaccination schedule include: 
• SAE that is subsequently considered to be related to vaccination 
• Pregnancy (regardless of outcome) 
• HIV acquisition 
• Grade 3 AE assessed as related to study vaccine with the exception of fever 
and subjective local and systemic symptoms. For grade 3 injection site 

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erythema and/or induration, upon review, the PSRT may allow continuation of 
vaccination. 
• Grade 4 AE assessed as related to study vaccine 
• Clinically significant type-1 hypersensitivity associated with study vaccine 
For ease of reference and review, the clinically significant type-1 
hypersensitivity definition, per the Second National Institute of Allergy and 
Infectious Disease/Food Allergy and Anaphylaxis Network symposium 
criteria for anaphylaxis (19), is provided below: 
“Anaphylaxis is highly likely when any one of the following criteria are 
fulfilled: 
1. Acute onset of an illness (minutes to several hours) with 
involvement of the skin, mucosal tissue, or both (eg, generalized hives, 
pruritus or flushing, swollen lips-tongue-uvula)  
AND AT LEAST ONE OF THE FOLLOWING:  
a. Respiratory compromise (eg, dyspnea, wheeze-bronchospasm, 
stridor, reduced peak expiratory flow (PEF), hypoxemia)  
b. Reduced blood pressure (BP) or associated symptoms of end-organ 
dysfunction (eg, hypotonia [collapse], syncope, incontinence)  
2. Two or more of the following that occur rapidly after exposure to a 
likely allergen for that patient [participant] (minutes to several hours): 
a. Involvement of the skin-mucosal tissue (eg, generalized hives, itch-
flush, swollen lips-tongue-uvula) 
b. Respiratory compromise (eg, dyspnea, wheeze-bronchospasm, 
stridor, reduced PEF, hypoxemia)  
c. Reduced BP or associated symptoms (eg, hypotonia [collapse], 
syncope, incontinence)  
d. Persistent gastrointestinal symptoms (eg, crampy abdominal pain, 
vomiting)  
3. Reduced BP after exposure to known allergen for that patient 
[participant] (minutes to several hours). Adults: systolic BP of less 
than 90 mm Hg or greater than 30% decrease from that person’s 
baseline” 
• PI assessment that it is not in the best interest of the participant to continue 
receiving study vaccine 

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• Newly disclosed AESI (see Appendix I) 
• Coenrollment in a study with an investigational research agent (rare 
exceptions allowing for the continuation of vaccinations may be granted with 
the unanimous consent of the HVTN 305 PSRT) 
• Participants discontinuing study vaccine for reasons other than HIV 
acquisition should be encouraged to participate in follow-up visits and 
procedures per the protocol for 12 months following their last study vaccine 
administration. At the discretion of the CRS clinician and the PSRT (for 
composition of PSRT, see Section 9.3), some clinic procedures and sample 
collections may be modified or discontinued. 
• If a participant acquires HIV during the course of the study, no additional 
study vaccine will be administered. Participants will be encouraged to 
continue scheduled study visits for 12 months following their last study 
vaccine administration. At postdiagnosis follow-up visits, only samples for 
protocol-specified clinical labs (with the exception of HIV diagnostic testing) 
will be collected. In addition, some clinic procedures may be modified or 
discontinued. 
5.2.3 
Participant departure from vaccine schedule 
If a participant misses a scheduled vaccination, they are eligible for future 
vaccinations (see Section 5.2.1). The site investigator can petition the PSRT for 
an out of window vaccination. 
5.2.4 
Discontinuation of study participation 
A participant may be discontinued from protocol participation for the following 
reasons: 
• Participant voluntarily withdraws; 
• CRS determines the participant is lost to follow-up; 
• The investigational new drug (IND) application sponsor or regulatory 
authorities stop the study; or, 
• PI assessment that it is not in the best interest of the participant to continue 
participation in the study or that the participant’s compliance with the study is 
not sufficient. 
If a participant terminates participation in the study early for any reason, the site 
PI should consider if the following assessments are appropriate: end-of-study HIV 
test, complete blood count (CBC with differential, serum chemistry [ALT and 
creatinine], physical examination, and if indicated, a pregnancy test [see 
Appendix A and Appendix B]). For participants living with HIV, please see 

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Section 8.8. If the site PI has questions regarding a termination visit, they should 
consult with the PSRT. 

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6 
Statistical considerations 
This phase-1 study will target recruiting a total of 45 adult participants without 
HIV aged 18 to 55 years. 
In Group 1, a total of approximately 9 participants will receive 3 administrations 
of sD-NP-GT8 DNA at a dose of 0.4 mg, coformulated with IL-12 DNA at a dose 
of 0.1 mg at days 1, 29, and 85. Study products will be administered intradermally 
via EP of the skin on each upper arm. 
In Group 2, a total of approximately 18 participants will receive 3 administrations 
of sD-NP-GT8 DNA at a dose of 1.6 mg, coformulated with IL-12 DNA at a dose 
of 0.4 mg at days 1, 29, and 85. Study products will be administered intradermally 
via EP of the skin on each upper arm. 
In Group 3, a total of approximately 18 participants will receive 3 administrations 
of sD-NP-GT8 DNA at a dose of 1.6 mg, coformulated with IL-12 DNA at a dose 
of 0.4 mg at days 1, 29, and 85. These doses will be administered intradermally 
via EP of the skin on each upper arm. All participants in Group 3 will also receive 
2 administrations of Trimer 4571 at a dose of 100 mcg adjuvanted with 5 mcg of 
3M-052-AF + 500 mcg Alum via IM injections into the deltoid muscle at days 85 
and 169. 
The first 5 participants enrolled in the study will be assigned to Group 1. The 
PSRT will convene to review safety data recorded from these 5 participants 
through the visit scheduled 2 weeks post–first vaccination. If the PSRT 
determines that it is safe to proceed with enrollment, the remaining participants 
(approximately 40) will be randomized between Groups 1, 2, and 3. 
Participants will be monitored for up to 18 months postenrollment. We anticipate 
that the study will take at least 22 months to be completed. 
Participants will be evaluated for safety and immune responses through blood 
collection, lymph node FNA, and leukapheresis at specified timepoints 
throughout the study. 
The study will target enrolling at least 50% of participants willing to undergo 
FNA in Groups 2 and 3. Participants enrolled in Group 1 will not undergo FNA. 
See Sections 1.5 and 2.11 for details.  
Primary and secondary objectives will seek to characterize and compare 
immunogenicity endpoints between treatment groups. Since allocation ratios used 
to enroll participants may vary between groups during enrollment (eg, participants 
will be preferentially assigned to Groups 2 and 3 relative to Group 1 after Group 1 
has enrolled its first 5 participants), group comparisons will be cautiously 
interpreted. Moreover, in order to mitigate potential biases, statistical analyses 
may adjust for potential confounders (eg, age) when comparing endpoints 
between groups. 

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6.1 
Sample size justification and accrual 
The protocol team will convene after the final enrolled participant has completed 
Visit 6 (first FNA visit) to determine if further participants should be enrolled to 
ensure adequate samples for immunogenicity analysis. Examples of reasons that 
might necessitate enrollment of additional participants are provided in Section 
1.5. If some participants of the original 45 enrolled will not contribute data to 
some of the immunogenicity analyses, the protocol team may enroll up to 9 
additional fully evaluable participants, or a total of 54 participants enrolled, with 
the goal of 45 contributing to the final immunogenicity analyses. Participants will 
not be replaced after completion of visit 6, the first FNA visit. 
Every study participant will be followed up for an AESI contact at month 18. 
Details about the pace of enrollment can be found in Section 1.5. 
Since enrollment is concurrent with receiving the first vaccination, all participants 
will provide some safety data. However, for immunogenicity analyses, data may 
be missing for a variety of reasons, including participants terminating from the 
study early, problems in shipping specimens, or high assay background. 
Immunogenicity data from 17 phase 1, 2, and phase 2a HVTN vaccine trials, 
which began enrolling after June 2005 (data as of September 2014), indicate that 
17% is a reasonable estimate for the rate of missing data. Therefore, the sample-
size calculations in Section 6.1.1 below account for about 17% of enrolled 
participants having missing data. 
6.1.1 
Power calculations for immunogenicity 
The primary goal of the statistical immunogenicity analyses is to evaluate 
vaccine-induced immune responses in each treatment group by timepoint. The 
primary immunogenicity endpoints will be the response rate and magnitude of 
HIV-1–specific binding Ab responses, as measured by a multiplex assay; the 
response rate and magnitude of CD4+ and CD8+ T-cell responses to HIV-1–
specific Env peptide pools, as measured by flow cytometry; and the frequency of 
VRC01-class B cells from lymph nodes and periphery following the second and 
third vaccinations, as measured by B-cell receptor (BCR) sequencing. 
The primary and secondary statistical analyses of immunogenicity data will 
include the following comparisons: 
• Group k postbaseline versus Group k at baseline to assess vaccine-induced 
immune responses within Group k, where k = 1, 2, 3. Groups 2 and 3 may be 
combined when comparing immune responses measured between baseline and 
Day 85 and those measured at baseline because participants in these 2 groups 
will have received the same products until Day 85. 
• Group 1 versus Groups 2 and 3 combined to compare immune responses in 
the low- and high-dose groups (without administration of Trimer 4571) 
between Day 1 and Day 85. 

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• Group 1 versus Group 2 to compare immune responses in the low- and high-
dose groups (without administration of Trimer 4571) post–Day 85. 
• Group 2 versus Group 3 to compare immune responses with and without 
administration of Trimer 4571 post–Day 85. 
Response magnitudes may be summarized using sample means and their two-
sided 95% confidence intervals (CIs). Response rates will be estimated using 
empirical proportions; their two-sided 95% CIs will be constructed using the 
score method described in Agresti and Coull (1998) (53). 
Comparisons of response magnitudes between timepoints within individual 
treatment groups or pooled treatment groups (eg, Groups 2 and 3 postbaseline 
versus baseline) will be carried out using Wilcoxon signed-rank tests. 
Comparisons of response magnitudes between 2 independent groups (eg, Group 2 
versus Group 3) within timepoints will be done using Wilcoxon rank-sum tests. 
The power achieved by the Wilcoxon rank-sum and the Wilcoxon signed-rank 
tests was computed as a function of the effect size for various sample sizes: n = 9, 
15, 18, 30, 36, 45, including cases when no data are missing and cases when 
~17% of the data are missing. These calculations also describe the power 
achieved by statistical comparisons of data measured in FNA samples if only 50% 
of participants enrolled in Groups 2 and 3 (ie, n = 9 participants per group or n = 
18 in Groups 2 and 3 combined) consent to providing FNA samples. The 
calculations assumed that observations are normally distributed and that the 
number of observations in both groups or timepoints is identical. 
The power function of the tests under these scenarios is presented in Figure 6-1. 
For instance, when comparing 2 independent groups of equal sizes n = 15 (eg, 
Groups 2 or 3 with about 17% loss to follow-up), the study achieves about 80% 
power to detect a significant difference at the 5% significance level using a two-
sided Wilcoxon rank-sum test if the average increase in the standardized 
(Cohen’s) effect size is about 1.1. For example, if the proportion of VRC01-class 
B cells two weeks after the second vaccination was 1.0% in Group 1 and 2.1% in 
Group 2, with a standard deviation of about 1.0% in both groups, the standardized 
effect size would be 1.1 = (2.1%-1.0%)/1.0%, and the study would have 
approximately 80% to detect a significant difference between the two groups 
using a two-sided Wilcoxon rank-sum test at the 5% significance level. With a 
sample size of n = 18 per group (eg, Groups 2 or 3 with no loss to follow-up), the 
test has about 80% power to detect a significant difference if the standardized 
effect size is 1.0. Thus, if the proportion of VRC01-class B cells two weeks after 
the second vaccination was 1.0% in Group 1 and 2.0% in Group 2, with a 
standard deviation of about 1.0% in both groups, the standardized effect size 
would be 1.0 = (2.0%-1.0%)/1.0%, and the study would have approximately 80% 
to detect a significant difference between the two groups using a two-sided 
Wilcoxon rank-sum test at the 5% significance level. 

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Figure 6-1 Left panel: Power of a two-sided Wilcoxon rank-sum test comparing 2 
independent groups. Right panel: Power of a two-sided Wilcoxon signed-rank test 
comparing 2 groups of paired observations (eg, the frequency of VRC01-class B cells 
postvaccination and at baseline within a single group). The power was calculated as a function 
of (1) the standardized effect size, defined as the average difference between the 2 group means 
divided by the common group standard deviation (left-hand side) or as the mean difference divided 
by the standard deviation of the difference (right-hand side) and (2) the sample size (n = 9, 15, 18, 
30, 36, and 45). 
6.1.2 
Power calculations for safety 
The goal of the safety evaluation for this study is to identify safety concerns 
associated with vaccine administration. The ability of the study to detect SAEs 
(see Section 9.2) can be expressed by the true event rate above which at least 1 
SAE would likely be observed and the true event rate below which no events 
would likely be observed. Specifically, for each treatment group of size n = 18 
(Groups 2 or 3), there is at least a 90% chance of observing at least 1 event if the 
true rate of such an event is 12.5% or more, and there is at least a 90% chance of 
observing no events if the true rate is 0.53% or less. When pooling Groups 2 and 
3 together (n = 36), there is at least a 90% chance of observing at least 1 event if 
the true rate of such an event is 6.5% or more, and there is at least a 90% chance 
of observing no events if the true rate is 0.29% or less. Safety data will be 
summarized as described in Section 6.4.3 and evaluated using historical controls. 
As a reference, in HVTN vaccine trials conducted in the US from April 2008 
through March 2018, about 1% of participants who received placebos experienced 
an SAE. 
Binomial probabilities of observing 0 events, 1 or more events, and 2 or more 
events among 9 (or 18 or 36 or 45) participants receiving the study vaccine are 
presented in Table 6-1 for a range of possible true AE rates. These calculations 
provide a more complete picture of the sensitivity of this study design to identify 
potential safety problems with the vaccine. 

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Table 6-1 Probability of observing 0 events, 1 or more events, and 2 or more events among a 
group of 9, 18, 36, or 45 study participants for different true event rates 
True 
event 
rate 
(%) 
Arm 
size 
0 
events 
1+ events 
2+ events 
1 
9 
0.91 
0.09 
0.00 
4 
9 
0.69 
0.31 
0.05 
10 
9 
0.39 
0.61 
0.23 
20 
9 
0.13 
0.87 
0.56 
30 
9 
0.04 
0.96 
0.80 
1 
18 
0.83 
0.17 
0.01 
4 
18 
0.48 
0.52 
0.16 
10 
18 
0.15 
0.85 
0.55 
20 
18 
0.02 
0.98 
0.90 
30 
18 
0.00 
1.00 
0.99 
1 
36 
0.70 
0.30 
0.05 
4 
36 
0.23 
0.77 
0.42 
10 
36 
0.02 
0.98 
0.89 
20 
36 
0.00 
1.00 
1.00 
30 
36 
0.00 
1.00 
1.00 
1 
45 
0.64 
0.36 
0.07 
4 
45 
0.16 
0.84 
0.54 
10 
45 
0.01 
0.99 
0.95 
20 
45 
0.00 
1.00 
1.00 
30 
45 
0.00 
1.00 
1.00 
An alternative way of describing the statistical properties of the study design is in 
terms of the 95% CI for the true rate of an AE based on the observed data. Table 
6-2 shows the two-sided 95% CIs for the probability of an event based on a 
particular observed rate. Calculations are done using the score test method for CIs 
described in Agresti and Coull formula 2 (53). If none of the 18 participants 
receiving the study vaccine in Groups 2 or 3 experience a safety event, the 95% 
two-sided upper confidence bound for the true rate of such events in the total 
vaccinated population is 17.6%. Table 6-2 also includes CIs for groups of size n = 
9 (Group 1 alone), n = 36 (Groups 2 and 3 pooled together), and n = 45 (all 3 
treatment groups combined). 

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Table 6-2 Two-sided 95% CIs for the probability of observing a safety event based on 
observing a particular rate of safety endpoints in a group of 18, 36, or 54 study participants 
Observed 
event rate 
95% CI (%) 
0/9 
[0 ; 29.9] 
1/9 
[2.0 ; 43.5] 
2/9 
[6.3 ; 54.7] 
0/18 
[0 ; 17.6] 
1/18 
[1.0 ; 25.8] 
2/18 
[3.1 ; 32.8] 
0/36 
[0 ; 9.6] 
1/36 
[0.5 ; 14.2] 
2/36 
[1.5 ; 18.1] 
0/45 
[0 ; 6.6] 
1/45 
[0.3 ; 9.8] 
2/45 
[1.0 ; 12.5] 
6.2 
Randomization 
A participant’s randomization assignment will be computer generated and 
provided to the HVTN CRS pharmacist through a Web-based randomization 
system. Pause rules discussed in Section 9.6 will be accounted for in this process. 
At each institution, the pharmacist with primary responsibility for dispensing 
study products is charged with maintaining security of the treatment assignments 
(except in emergency situations as specified in the HVTN MOP). 
Enrollment will be restricted to 1 participant per day for the first 5 participants in 
Group 1 and pause once the first 5 participants are enrolled in Group 1. If the 
PSRT determines that it is safe to complete enrollment in all three groups, the 
remaining participants (approximately 40: n=4 in Group 1, n=18 in Group 2, and 
n=18 in Group 3) will be block randomized. Block size and allocation ratio will 
be determined based on group sizes.  
The study will aim to ensure balanced representation with respect to sex assigned 
at birth. Sites will therefore be encouraged to enroll at least approximately 40% of 
each sex assigned at birth in the study. 
6.3 
Blinding 
This is an open-label study. Participants and site staff will be unblinded to 
participants’ group assignments. Laboratory program staff will be blinded to 
participants’ group assignments during assay analysis, whenever feasible. 

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6.4 
Statistical analyses 
This section outlines the final study analysis, unblinded as to treatment arm 
assignment. All data from enrolled participants will be analyzed regardless of how 
many vaccinations each participant received. All analyses will be performed using 
SAS or R. More details about the statistical analyses will be described in the 
Statistical Analysis Plan (SAP). 
6.4.1 
Baseline demographics 
Participants’ baseline characteristics will be summarized using descriptive 
statistics. 
6.4.2 
Immunogenicity analyses 
For the statistical analysis of immunogenicity endpoints, data from enrolled 
participants will be used according to the initial randomization assignment, 
regardless of how many injections each participant received. Additional analyses 
may be performed, limited to participants who received all scheduled injections 
per protocol. Assay results that are unreliable, or are from participants living with 
HIV postdiagnosis, are excluded. Since the exact date of HIV acquisition is 
unknown, any assay data from blood draws 4 weeks prior to a participant living 
with HIV’s last seronegative sample and thereafter may be excluded. If a 
participant living with HIV does not have a seronegative sample postenrollment, 
then all data from that participant may be excluded from the analysis. 
The goals of the primary and secondary immunogenicity analyses, including 
group comparisons, are presented in Section 3. 
Discrete categorical immunogenicity endpoints (eg, response rates) will be 
analyzed by tabulating the frequency of positive response by timepoint and 
treatment group. Crude response rates will be presented with their corresponding 
95% CI estimated based on the score test method (42). Barnard’s exact tests may 
be employed to compare the response rates of certain immune responses between 
2 treatment groups. 
Quantitative assay endpoints (eg, frequencies of CD4-bs B cells as measured by 
flow cytometry) will be summarized by timepoint within treatment groups by 
tabulating univariate summary statistics (eg, mean, median, standard deviation, 
range) and plotting observations using boxplots. Comparisons of continuous 
outcome variables measured in 2 independent groups will be performed using the 
Wilcoxon rank-sum test. Comparisons of paired continuous outcome variables 
measured in 1 group will be performed using the Wilcoxon signed-rank test. 
6.4.3 
Safety analyses 
Reactogenicity: The number and percentage of subjects experiencing each type 
of reactogenicity sign or symptom will be tabulated by severity. For a given sign 

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or symptom, each subject’s reactogenicity will be counted once under the 
maximum severity for all assessments. 
AEs: AEs will be coded into Medical Dictionary for Regulatory Activities 
(MedDRA)–preferred terms. The number and percentage of subjects experiencing 
each specific AE will be tabulated by severity and relationship to study vaccine. 
For the calculations in these tables, each subject’s AE will be counted once under 
the maximum severity or strongest recorded causal relationship to treatment. A 
complete listing of AEs for each subject will provide details including severity, 
relationship to treatment, onset, duration, and outcome. 

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7 
Study vaccine preparation, storage, and 
administration 
7.1 
Vaccine regimen 
The schedule of vaccinations is shown in Section 1 and additional information is 
given below. 
Group 1 
Treatment 1 (T1): 
INO-6172, 0.5 mg to be administered as 2 separate ID injections (0.1 mL each) 
followed by EP using the CELLECTRA 2000 EP device at months 0, 1, and 3. 
Group 2  
Treatment 2 (T2):  
INO-6172, 2 mg to be administered as 2 separate ID injections (0.1 mL each) 
followed by EP using the CELLECTRA 2000 EP device at months 0, 1, and 3. 
Group 3 
Treatment 3 (T3):  
INO-6172, 2 mg to be administered as 2 separate ID injections (0.1 mL each) 
followed by EP using the CELLECTRA 2000 EP system at months 0, 1, and 3. 
Trimer 4571, 100 mcg admixed with 3M-052-AF, 5 mcg and Alum, 500 mcg to 
be administered as 2 separate IM injections (0.2 mL each) at months 3 and 6. 
7.2 
Study product formulation and storage 
7.2.1 
INO-6172 
INO-6172 Drug Product is sD-NP-GT8 coformulated with IL-12 DNA at a 
plasmid ratio of 4:1 (0.8 mg sD-NP-GT8 and 0.2 mg IL-12 DNA per 0.1 mL). 
The formulation is supplied in Water for Injection (WFI) at a concentration of 10 
mg/mL, and a labeled volume of 0.4 mL in 2-mL glass vials. Vials may contain a 
volume greater than the labeled volume.  INO-6172 is a clear, colorless solution 
essentially free of visible particulate matter. Store INO-6172 frozen at -25ºC to -
15ºC. The study product is described in further detail in the IB. 

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7.2.2 
HIV-1 Trimer 4571 (VRC-HIVRGP096-00-VP) 
Trimer 4571 will be provided as 3-mL, single-use glass vials with a labeled 
volume of 1.2-mL and a concentration of 500 mcg/mL. Each vial contains a 
sterile, aqueous, preservative-free, buffered solution that is clear and colorless. 
Some small, white, or translucent particles may be present. 
Store Trimer 4571 frozen at -35ºC to -15ºC. Vials should not be refrozen after 
thaw. Thawed vials can be stored at 2ºC to 8ºC for up to 48 hours or at 15ºC to 
27ºC for up to 24 hours. Vials are intended for single use only and do not contain 
preservatives. A single vial may not be used for multiple dose preparations. The 
study product is described in further detail in the IB. 
7.2.3 
3M-052-AF (labeled as AP 60-702) 
3M-052-AF will be provided in 2-mL Type-1 glass vials with a rubber stopper 
and flip-off aluminum seal. Each vial contains a fill volume of 0.4 mL at a 
concentration of 50 mcg/mL. 3M-052-AF is a clear to slightly hazy, colorless 
liquid. The product is stored at 2-8° C. Do not freeze. The study product is 
described in further detail in the IB. 
7.2.4 
Aluminum Hydroxide Suspension (Alum) 
The Alum adjuvant is composed of Alhydrogel 2% (Brenntag Biosector, 
Frederikssund, Denmark). The Alum adjuvant is a sterile, pyrogen-free, off-white 
suspension filled into 3-mL glass vials with a labeled volume of 0.7 mL, diluted 
with water for injection to an aluminum concentration of 5 mg/mL. 
Store Alum adjuvant refrigerated at 2°C to 8°C. Do not freeze. Vials are intended 
for single use only and do not contain preservatives. A single vial may not be used 
for multiple dose preparations. The study product is described in further detail in 
the IB. 
7.3 
Product preparation 
Pharmacists must follow appropriate aseptic techniques and sterile preparation 
procedures/guidance as outlined in USP <797> [medium risk], utilizing a 
pharmacy biosafety cabinet/isolator or better. Local regulations and site 
institutional policies and procedures regarding use of personal protective 
equipment, such as gloves, gowns, masks, and safety glasses, must be followed. 
Pharmacists should follow the requirements of their country, their institution, and 
their pharmacy regulatory authority regarding these procedures. 
Any unused portion of study product will not be used for another participant. 
Empty vials, unused portions of entered vials, and unused prepared study product 
should be discarded in a biohazard container and disposed of in accordance with 
institutional or pharmacy policy. 

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7.3.1 
INO-6172, 0.5 mg (Group 1) 
INO-6172, 10 mg/mL will be further diluted to a final concentration of INO-6172, 
2.5 mg/mL for Group 1 dosing. 
1. Remove 1 vial of INO-6172, 10 mg/mL from the freezer and record this as the 
preparation start time on the Investigational Product Preparation Sheet. 
2. Once contents of vial are thawed, withdraw 0.4 mL from the INO-6172 vial 
and inject into the sterile empty glass mixing vial. Note: Thawed vials of INO-
6172 will appear clear and colorless, essentially free of visible particulate 
matter.  
3. Withdraw 1.2 mL from the SWFI USP vial and inject into the sterile glass 
mixing vial, which contains 0.4 mL of INO-6172. 
Note: The final total volume in the sterile glass mixing vial should be 1.6 mL, 
for a final concentration of INO-6172, 2.5 mg/mL. 
4. Gently swirl the INO-6172, 2.5 mg/mL in the sterile glass mixing vial. 
5. Insert the sterile BD ½ mL Tuberculin Syringes with permanently attached 
needle, 27G x ½ inch (BD Reference Number 305620) into the sterile glass 
mixing vial containing INO-6172, 2.5 mg/mL and withdraw 0.1 mL. Place the 
needle cap back onto the needle. Repeat this step for a total of 2 syringes, each 
containing INO-6172, 2.5 mg/mL, 0.1 mL each. 
6. Label the syringes according to Section 7.3.4 and include on the label “syringe 
1 of 2” for the first syringe prepared and “syringe 2 of 2” for the second 
syringe prepared. 
The prepared syringe(s) may be stored at room temperature and used within 4 
hours of study product preparation time. 
7.3.2 
INO-6172, 2 mg (Group 2 and Group 3) 
1. Remove 1 vial of INO-6172, 10 mg/mL from the freezer and record this as the 
preparation start time on the Investigational Product Preparation Sheet. 
2. Once contents of vial are thawed, insert the sterile BD 0.5 mL Tuberculin 
Syringes with permanently attached needle, 27G x ½ inch (BD Reference 
Number 305620) into the sterile glass vial containing INO-6172, 10mg/mL 
and withdraw 0.1 mL. Place the needle cap back onto the needle. Repeat this 
step for a total of 2 syringes, each containing INO-6172, 10 mg/mL, 0.1 mL 
each. Note: Thawed vials of INO-6172 will appear clear and colorless, 
essentially free of visible particulate matter. 
3. Label the syringes according to Section 7.3.4 and include on the label “syringe 
1 of 2” for the first syringe prepared and “syringe 2 of 2” for the second 
syringe prepared. 

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The prepared syringe(s) may be stored at room temperature and used within 4 
hours of study product preparation start time. 
7.3.3 
Trimer 4571, 100 mcg admixed with 3M-052-AF, 5 mcg and Alum, 500 
mcg (Group 3) 
1. Thaw 1 vial of Trimer 4571 at ambient temperature (15˚C to 27˚C) for a 
minimum of 30 minutes. Vial should not be moved directly from a freezer to a 
refrigerator to thaw. Record this as the preparation start time. While vial is 
thawing proceed to the next steps.  
2. Remove 1 vial of Alum adjuvant from the refrigerator and equilibrate at 
ambient temperature (15˚C to 27˚C) for a minimum of 15 minutes. Mix the 
vial of Alum adjuvant by gently inverting the vial 5 times. 
3. Remove 1 vial of 3M-052-AF from the refrigerator and equilibrate at ambient 
temperature (15˚C to 27˚C) for a minimum of 15 minutes. Mix the vial of 3M-
052-AF by gently inverting the vial 5 times.   
4. Withdraw 0.2 mL of 3M-052-AF and inject into the sterile empty glass 
mixing vial. 
5. Withdraw 0.2 mL of Alum and inject into the sterile glass mixing vial, which 
contains 0.2 mL of 3M-052-AF.  
6. Mix and wait for 30 minutes. 
7. Swirl the thawed, equilibrated Trimer 4571 vial for about 30 seconds with 
sufficient force to mix the solution while avoiding foaming. Do not shake the 
vial. If some white to translucent particles are observed, vials may be used for 
the preparation for IM administration. 
8. Withdraw 0.4 mL of Trimer 4571 and inject into the sterile glass mixing vial, 
which contains 0.2 mL of 3M-052-AF, and 0.2 mL of Alum. The final volume 
in the sterile mixing vial will be 0.8 mL. 
9. Invert mixing vial gently 5 times to mix. Withdraw 0.2 mL from the sterile 
glass mixing vial. Pull back on the syringe plunger to ensure all product is in 
the syringe.  Discard preparation needle and either cap the syringe or attach a 
needle for IM administration, per institutional procedure. Repeat this step for 
a total of 2 syringes containing 0.2 mL each. 
10. Label the syringes according to Section 7.3.4 and include on the label “syringe 
1 of 2” for the first syringe prepared and “syringe 2 of 2” for the second 
syringe prepared. Invert each syringe gently 10 times to mix immediately 
prior to administration. 

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The prepared syringes may be stored for up to 8 hours at 2°C to 8°C and/or up to 
4 hours at ambient temperature (15°C to 27°C), including dose administration 
time.  
7.3.4 
Labeling 
Label the study product as follows: 
• Participant identifier(s) 
• Study product name 
• Final volume (mL) 
• Route (IM or ID) 
• Beyond-use date and time 
• Any additional information required by jurisdiction 
7.4 
Study vaccine administration 
7.4.1 
INO-6172 
INO-6172, 0.5 mg and INO-6172, 2 mg will be administered as 2 separate 0.1-mL 
injections intradermally, bilaterally 1 on each arm at months 0, 1, and 3. 
Following ID injections of INO-6172, EP will be performed using the Inovio 
CELLECTRA 2000 EP Device. 
The ID injections will be administered into the skin overlying the deltoid area of 
the arm. The needle will be inserted into the skin at a 5- to 15-degree angle to the 
skin and bevel side up until the bevel is seen to be fully under the skin. The 
syringe contents will be injected to form a small bleb. One injection should be 
completed (including EP) prior to administering the second injection.  
7.4.2 
Trimer 4571, 100 mcg admixed with 3M-052-AF, 5 mcg and Alum, 500 
mcg  
Trimer 4571, 100 mcg admixed with 3M-052-AF, 5 mcg and Alum, 500 mcg will 
be administered to Group 3 participants following administration of INO-6172 
and at months 3and 6, as 2 separate 0.2-mL IM injections (1 in each arm) 
intramuscularly into the deltoid muscles by needle and syringe. The injection site 
for the protein injection (Trimer 4571) should be at least 2.5 centimeters (1 inch) 
from the ipsilateral EP injection site of INO-6172 (see 7.4.1).  Immediately prior 
to administration, gently invert each syringe 10 times.  
If an ID or IM injection needs to be administered to an alternate body site (eg, 
lateral thigh) due to a medical contraindication, it should not be administered in 

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the same deltoid as the injection site where the other half of the dose is 
administered, but rather in the thigh. If no injection can be administered in either 
deltoid muscle, the injections should be administered in separate alternate body 
sites. The appropriate study staff should document this clearly. Under this 
circumstance, this is NOT a protocol violation. 
When preparing a dose in a syringe and administering the dose, consideration 
should be given to the volume of solution in the needle before and after the dose 
is administered. Particularly, if the needle used to withdraw the product is 
replaced prior to vaccine administration, consideration should be given to 
conserving the full dose of product. The pharmacy and clinic staff members are 
encouraged to work together to administer the dose specified in the protocol. 
7.5 
Acquisition of study products 
 sD-NP-GT8 and IL-12 DNA are manufactured under quality controls and 
production protocols developed and tech transferred by Inovio Pharmaceuticals. 
INO-6172, the coformulation of sD-NP-GT8 with IL-12 DNA, is formulated 
under guidance provided by Inovio under subcontract to IDT and is being 
provided to HVTN to support this program under an IPCAVD grant to The Wistar 
Institute (Philadelphia, PA USA). 
Trimer 4571, and Alum adjuvant are manufactured by Leidos Biomed (Vaccine 
Research Center, Frederick, MD US). Trimer 4571 will be provided by VRC and 
Alum will be provided by DAIDS, NIAID, and NIH (Rockville, MD USA) 
3M-052-AF (labeled as AP 60-702) is manufactured and provided by Access to 
Advanced Health Institute (AAHI) 
Once an HVTN CRS is protocol registered, the pharmacist can obtain study 
products from the NIAID Clinical Research Products Management Center 
(CRPMC) by following the ordering procedures given in Pharmacy Guidelines 
and Instructions for DAIDS Clinical Trials Networks. 
Syringes, needles, SWFI, and other ancillary supplies will be locally sourced by 
the site. 
7.6 
Study vaccine accountability 
The HVTN CRS pharmacist is required to maintain complete records of all study 
products. 
7.7 
Final disposition of study product 
For US CRSs, all unused study products must be returned to the CRPMC after the 
study is completed or terminated unless otherwise instructed by the study sponsor. 

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For non-US CRSs, all unused study products must be destroyed after the study is 
completed or terminated unless otherwise instructed by the study sponsor. The 
procedures are included in the Pharmacy Guidelines and Instructions for DAIDS 
Clinical Trials Networks. 

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8 
Clinical procedures 
8.1 
Screening 
Screening for eligibility will be performed after informed consent has been 
obtained and properly documented before enrollment. Screening evaluations and 
sample collection include medical history review, physical exam, and any clinical 
laboratory tests, as detailed in the schedule of procedures (Appendix A and 
Appendix B), needed to confirm eligibility. Persons assigned female sex at birth 
who are of reproductive potential will be given a pregnancy test. Additional 
assessments of health may be conducted at screening based on clinical judgment. 
An AoU will be completed prior to enrollment. Records will be kept documenting 
the reason that screened participants did not enroll. 
Leukapheresis specimens for baseline comparison will be drawn during the 
screening period if a participant meets all additional eligibility criteria and wishes 
to join the study (Sections 5.1.1 and 8.6). 
8.2 
Definition of “study day” and “study visit” 
“Study Day 1” is defined as the day of the first vaccination. A study visit may be 
conducted remotely, such as via phone, text message, email, or other electronic 
means in lieu of, or in combination with, in-person visits. As long as they are 
completed within the visit window (see Appendix C), procedures for a study visit 
can be completed over multiple days. 
8.3 
Reactogenicity assessments 
Reactogenicity definition: Signs and symptoms considered to represent 
reactogenicity from the vaccine include systemic events of increased body 
temperature, fatigue, generalized myalgia, generalized arthralgia, headache, chills, 
nausea, and local events at the injection site, including pain/tenderness, 
induration, and erythema.  
The reactogenicity assessment period is the day of vaccination and 7 full days 
following vaccine administration for INO-6172 alone and 14 full days following 
vaccine administration for INO-6172 along with Trimer 4571 adjuvanted with 
3M-052-AF/Alum and Trimer 4571 alone, adjuvanted with 3M-052-AF/Alum. 
Clinicians will follow and collect resolution information for any reactogenicity 
signs and symptoms that have not resolved within the reactogenicity assessment 
period. 
Prevaccine administration: Medical history and evaluations, including vital 
signs and planned injection-site evaluations, are performed as described in 
Appendix A and Appendix B  

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Postvaccine administration in-clinic evaluation: Following the first vaccine 
administration of sD-NP-GT8, participants will be observed for a minimum of 60 
minutes postadministration. For all subsequent injections, participants will be 
observed for a minimum of 30 minutes with the following exception: for Group 3, 
participants will be observed for 60 minutes postadministration of Trimer 4571 + 
3M-052-AF + Alum. During this time, vital signs will be recorded, the injection 
site will be inspected for evidence of local reaction, and any evidence of systemic 
symptoms will be assessed. 
Postvaccine administration contact with participant: Remote or in-person 
contact between the participant and the site staff should take place at least once on 
the third or fourth day following vaccination. Any postvaccination reaction that is 
Grade 2 or higher will be assessed by a clinician within 48 hours after onset 
unless the reaction is improving and/or has completely resolved. Additionally, 
other clinical concerns may prompt a study visit based on the judgment of a study 
clinician. 
Guidance for participants scheduling licensed vaccines or allergy treatments: 
In order to prevent interference with reactogenicity assessments, participants who 
plan to receive licensed vaccines or allergy treatments should be counseled to 
avoid scheduling receipt of these substances, when possible, within the 2 week 
interval after study product administration. To avoid unnecessary delays in study 
product administration, participants should be counseled to avoid scheduling 
receipt of these substances before study product administration (see Section 
5.2.1).  
Symptom diaries: Participants are asked to record symptoms on a daily basis 
using an electronic participant diary (paper alternative is available). All 
participants will be given a thermometer for oral temperature measurement and a 
ruler, and will be provided access to the electronic diary (paper alternative 
available). All participants will be provided training on diary completion, proper 
thermometer usage, and how to use of the measuring device to measure any 
injection-site induration and/or erythema. Participants will use the diary to record 
daily their highest temperature as well as local and systemic signs and symptoms. 
For Groups 1 and 2, symptoms are recorded for the day of vaccination and for 7 
full days following each vaccination. For Group 3 only, symptoms are recorded 
for the day of vaccination, for 7 full days following the first two vaccinations, and 
for 14 full days following the third and fourth vaccinations. Participant diaries 
will be reviewed by a clinician and reconciled for accuracy and completeness. 
Attribution assessment will be performed and recorded on a case report form 
(CRF) for systemic reactogenicity events reported in the participant diary after 
additional evaluation of the participant by clinician. Clinicians may photograph 
the injection site in order to document injection-site reactogenicity. 
8.4 
EP injection site assessment 
To document the appearance of injection sites over time, the area to be injected 
will be assessed prior to receipt of an injection, and at subsequent scheduled 

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visits. The purpose of this assessment is to evaluate the appearance of the 
injection sites after time has allowed for healing. A description including type and 
size of any skin changes related to vaccination that are not described in Section 
8.3 will be recorded. Clinicians may photograph the injection site in order to 
document EP injection-site skin changes. An abbreviated questionnaire to assess 
EP-related skin changes and tolerability will be asked at the AESI contact 
(Section 8.10) 
8.5 
Lymph node FNA 
Tissue sampling of an axillary lymph node will be carried out percutaneously by 
FNA for participants who have opted for this procedure. A minimum of half of 
the participants in Groups 2 and 3 will undergo this procedure. This procedure 
will be performed in accordance with the standard practices of the participating 
provider and/or facility. The procedure involves tissue retrieval with a needle via 
a small skin incision under sonographic guidance. Approximately 2 to 4 passes 
will be made to retrieve cytologic material. 
Postprocedural safety assessments will be performed in accordance with the 
standard practices of the participating provider and/or facility. The participant will 
be advised to contact the study site if they experience severe pain, fever 
(postprocedural body temperature ≥ 38.0°C or 100.4°F), other evidence of 
infection (inflammation and/or pus) at the aspiration site, or arm numbness or 
weakness. 
Eligibility for lymph node FNA: 
• Participant may not be taking warfarin, oral antithrombin equivalents 
(including, but not limited to, apixiban, rivaroxiban, dabigatran), enoxaparin 
injections, or nonsteroidal anti-inflammatory drugs (NSAID), aspirin, or other 
medications at doses that would increase the risk of bleeding, as assessed by 
the clinician performing the procedure. No evidence of localized infection 
directly superior to aspiration site 
• No other contraindication to procedure as assessed by the clinician performing 
the procedure 
• Participants who can become pregnant must have a negative urine pregnancy 
test within 48 hours prior to the procedure 
8.6 
Leukapheresis 
Collection of PBMCs via leukapheresis will be performed in accordance with the 
standard practices of the participating apheresis center. Typically, in this 
procedure, approximately 3 liters of blood will be processed using peripheral 
veins for venous access. The blood will be anticoagulated in accordance with 
standard practice of the apheresis center. 

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Postprocedural safety assessments will be performed in accordance with the 
standard practices of the participating apheresis center. Additionally, the 
participant will be advised to contact the study site if they experience any AEs 
following the procedure. 
Eligibility for leukapheresis: 
• Prior to leukapheresis, participant must meet all apheresis center requirements 
for this procedure 
• Participants who can become pregnant must have a negative urine pregnancy 
test within 48 hours prior to the procedure 
8.7 
Visit procedures, schedule, windows, and missed visits 
The schedules of visits and evaluations performed at each visit are shown in 
Appendix A (for participants undergoing lymph node FNA) and Appendix B (for 
participants not undergoing lymph node FNA). Visit windows are shown in 
Appendix C. The procedures for documenting missed visits and out-of-window 
visits are described in the HVTN 305 Study-specific Procedures (SSP). If the 
missed visit is one that required safety assessments or local safety labs, HVTN 
CRS staff should attempt to bring the participant in for an interim visit as soon as 
possible. 
8.8 
Monitoring for acquisition of HIV  
Study participants will be tested for presence of HIV periodically throughout the 
study, as indicated in the schedules included in Appendix A and Appendix B. The 
Laboratory Program (or approved diagnostic laboratory) will follow the HVTN 
HIV testing algorithm (see HVTN Laboratory Center Manual of Procedures 
(MOP), which is able to distinguish vaccine-induced Ab responses from the 
presence of actual HIV. Participants will be promptly informed and counseled if 
they acquire HIV during the study and will be referred for treatment (see Section 
5.2.2). 
Study participants will receive regularly scheduled counseling regarding 
avoidance of HIV acquisition in accordance with the most recent Centers for 
Disease Control and Prevention HIV-counseling guidelines. 
Although the study vaccine will not cause HIV acquisition, it may induce Abs 
detectable by standard HIV screening techniques. This is referred to as vaccine-
induced seropositivity (VISP). The following steps will be taken to protect 
participants from adverse consequences associated with VISP: 
• Participants will be counseled to avoid HIV Ab testing outside of the HVTN 
CRS during study participation. 

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• Participants can receive HIV diagnostic testing from the CRS following their 
last scheduled visit until they are told they do not have VISP. 
• Participants with VISP will be periodically offered free-of-charge poststudy 
HIV diagnostic testing (per the HVTN poststudy HIV testing algorithm) as 
medically/socially indicated (approximately every 6 months) unless or until 
HIV Ab testing is no longer standard in clinical settings. 
• Unless participants request that their names be removed, the names of all 
participants in HVTN studies are entered into a secure VISP registry in order 
to verify that an individual received an HIV vaccine (and therefore has the 
potential for VISP) and to qualify former participants for poststudy HIV 
testing to distinguish between VISP and HIV acquisition. Information in the 
VISP registry is not used for research. 
8.9 
Early termination visit 
If a participant terminates participation in the study early for any reason, the site 
PI should consider if the following assessments are appropriate: end-of-study HIV 
test, CBC with differential, serum chemistry (ALT and creatinine), physical 
examination, social-impact assessment, and, if indicated, a pregnancy test (see 
Appendix A and Appendix B). For participants living with HIV, please see 
Section 5.2.2. If the site PI has questions regarding a termination visit, they 
should consult with the PSRT. 
8.10 
AESI contact 
CRS staff will contact study participants at month 18 to collect the information 
listed below and will ask an abbreviated set of EP-injection-site related skin 
changes and tolerability assessment questions that can be conducted over the 
phone. If indicated, the participant may be asked to come in for a clinical 
assessment, which may also include referrals for an AESI assessment. AESIs are 
described further in Appendix I. 
• Confirmation of vital status; if deceased, attempt to learn cause and date of 
death 
• If participant is alive, record the participant’s responses to the following: 
SAEs, EAEs; 
AESIs (a sample list of AESI is provided in Appendix I). AESIs are reported 
regardless of relationship to study product(s); 
Medically attended adverse events (MAAEs), defined as any AE leading to an 
unscheduled visit to a healthcare professional. MAAEs are reported regardless of 
relationship to study product(s); 

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New diagnosis of HIV acquisition; and 
Pregnancies and outcomes, including congenital anomalies/birth defects. 
All such events will be recorded and AEs will be assessed for relationship to 
study vaccines. 

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9 
Safety and AEs 
9.1 
AEs 
AEs will be collected over a period of 30 days after each vaccination. All 
collected AEs are captured in the clinical database on the appropriate CRF. Clinic 
staff should evaluate every AE to determine if (1) the AE meets the requirements 
for expedited reporting (see Section 9.2.1), (2) the AE meets the criteria for a 
safety pause/prompt AE review (see Section 9.6), (3) the AE meets the criteria for 
an MAAE, and (4) the AE is a potential immune-mediated disease that may be 
listed as an AESI. A sample list of AESIs is provided in Appendix I. 
In addition, a limited set of AEs will be collected and reported until (but not 
including) the month 18 visit: 
• SAEs/expedited adverse events (EAEs), 
• AESIs, 
• MAAEs, and 
• AEs leading to early participant withdrawal or early discontinuation of study 
vaccine(s) administration. 
AEs will be graded according to the DAIDS Table for Grading the Severity of 
Adult and Pediatric Adverse Events, Corrected Version 2.1 
(https://rsc.niaid.nih.gov/clinical-research-sites/daids-adverse-event-grading-
tables), with the following exceptions:  
• 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 (see HVTN 
305 SSP). 
• Do not grade creatinine clearance or estimated glomerular filtration rate 
(eGFR) based on the change from the baseline parameter. Do not grade on the 
basis of eGFR if there is clinical concern for acute kidney injury. 
• Injection-site erythema or redness and injection-site induration or swelling 
will not consider surface area and interference with usual social and functional 
activities, such that: 
Grade 1 is: 2.5 to < 5 cm in diameter; 
Grade 2 is: ≥ 5 to < 10 cm in diameter; 
Grade 3 is: ≥ 10 cm in diameter OR ulceration OR secondary infection OR 
phlebitis OR sterile abscess OR drainage; 

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Grade 4 is: potentially life-threatening consequences (eg, abscess, exfoliative 
dermatitis, necrosis involving dermis or deeper tissue). 
9.2 
SAEs 
The term “Serious Adverse Event” (SAE) is defined in 21 CFR 312.32 as follows:  
“An adverse event or suspected adverse reaction is considered serious if, in the 
view of either the investigator or the sponsor, it results in any of the following 
outcomes: 
• death, 
• a life-threatening adverse event, inpatient hospitalization or prolongation of 
existing hospitalization, 
• a persistent or significant incapacity or substantial disruption of the ability to 
conduct normal life functions, 
• congenital anomaly/birth defect. 
Important medical events that may not result in death, be life-threatening, or 
require hospitalization may be considered serious when, based upon appropriate 
medical judgment, they may jeopardize the patient or subject and may require 
medical or surgical intervention to prevent one of the outcomes listed in this 
definition. Examples of such medical events include allergic bronchospasm 
requiring intensive treatment in an emergency room or at home, blood dyscrasias 
or convulsions that do not result in inpatient hospitalization, or the development 
of drug dependency or drug abuse.” 
“Life-threatening” refers to an AE that, at occurrence, represents an immediate 
risk of death to the subject. Similarly, a hospital admission for an elective 
procedure is not considered an SAE. 
9.2.1 
Expedited reporting of adverse events to DAIDS 
Requirements, definitions, and methods for expedited reporting of AEs are 
outlined in Version 2.0 (January 2010) of the Manual for Expedited Reporting of 
Adverse Events to DAIDS (DAIDS EAE Manual), which is available on the 
DAIDS Regulatory Support Center (RSC) website at 
https://rsc.niaid.nih.gov/clinical-research-sites/manual-expedited-reporting-
adverse-events-daids. 
The internet-based DAIDS Adverse Experience Reporting System (DAERS) must 
be used for EAE reporting to DAIDS. In the event of system outages or technical 
difficulties, EAEs may be submitted via the DAIDS EAE form at: 
https://rsc.niaid.nih.gov/clinical-research-sites/paper-eae-reporting. 

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For questions about DAERS, please contact CRMSsupport@niaid.nih.gov or 
from within the DAERS application itself. 
For questions about EAE reporting, please contact the DAIDS RSC Safety Office 
at DAIDSRSCSafetyOffice@tech-res.com. 
The SAE reporting category will be used throughout the study. After completion 
of the study, the Suspected Unexpected Serious Adverse Reaction (SUSAR) 
reporting category will be used if clinical staff becomes aware of an event on a 
passive basis. 
The study products for which expedited reporting are required are:  
• 
INO-6172, 
• 
[Trimer 4571 + 3M-052-AF + Alum], and 
• 
EP device.  
There is no placebo product administered in this protocol. The NIAID/DAIDS 
will report all unexpected SAEs related to the study products observed in this 
clinical trial to the FDA, in accordance with 21 CFR 312.32 (IND Safety 
Reports). 
9.3 
Safety monitoring 
9.3.1 
PSRT 
The PSRT comprises: DAIDS Medical Officer (MO) representative, Protocol 
Chair and Cochair, Protocol Team Leader (PTL), Clinical Safety Specialist 
(CSS), and Regional Medical Liaison (RML). The protocol team’s clinic 
coordinator, clinical data manager, vaccine developer representative, clinical trial 
manager, and others may also be included in HVTN 305 PSRT meetings. The 
PSRT will review study safety information on a weekly basis through 2 weeks 
after the last participant receives the final study injection. Less frequent safety 
reviews will be conducted at the discretion of the PSRT. 
9.3.2 
HVTN Safety Monitoring Board 
The Safety Monitoring Board (SMB) is a multidisciplinary group consisting of 
biostatisticians, clinicians, and experts in HIV vaccine research that, collectively, 
has experience in the conduct and monitoring of vaccine trials. Members of the 
SMB are not directly affiliated with the protocols under review. 
The SMB reviews safety data (including cumulative reactogenicity events, AEs, 
laboratory safety data, and individual SAE reports) approximately every 4 
months. The SMB conducts additional special reviews at the request of the HVTN 
305 PSRT. 
Study sites will receive SMB summary minutes and are responsible for 
forwarding them to their Institutional Review Board (IRB)/Ethics Committee 
(EC) and any applicable Regulatory Entity (RE). 

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9.4 
Total blood volume 
Required blood volumes per visit are shown in Appendix A and Appendix B. Not 
shown is any additional blood volume that would be required if a safety lab needs 
to be repeated or if a serum pregnancy test needs to be performed. The additional 
blood volume would likely be minimal. The total blood volume drawn for each 
participant will not exceed 500 mL in any 56-day (8-week) period, per American 
Red Cross guidelines for blood donation (https://www.redcrossblood.org/donate-
blood/how-to-donate/eligibility-requirements.html). 
9.5 
Initial safety review 
The trial will begin with enrollment in Group 1 only. Enrollment will be restricted 
to a maximum of 1 participant per day for the first 5 participants in Group 1. 
Enrollment will then be held. However, participants who have been enrolled will 
continue to receive vaccinations during the planned hold. The PSRT will review 
cumulative safety data reported through the visit scheduled 2 weeks post–
enrollment. Upon PSRT determination that it is safe to proceed, full and 
unrestricted enrollments in all Groups may commence without restriction. 
9.6 
Safety pause and prompt PSRT AE review 
The PSRT (see Section 9.3) will closely monitor participant safety. The trial can 
be paused at any time for any reason by the PSRT. When a trial is placed on 
safety pause, all enrollment and vaccination will be held until further notice. The 
AEs that will lead to a safety pause or prompt HVTN 305 PSRT AE review are 
summarized in Table 9-1. Vaccinations may be suspended for safety concerns 
other than those described in the table or before pause rules are met if, in the 
judgment of the HVTN 305 PSRT, participant safety may be threatened. Criteria 
for an individual participant’s departure from the schedule of vaccinations are 
listed in Section 5.2.3. 

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Table 9-1 Pause rules 
Event and 
relationship to 
study vaccine 
Severity 
Grade 
HVTN Site Actions 
HVTN Core 
Action 
SAE, related 
5 or 4 
Phone 24/7 Safety Phone immediately 
Email vtn.clin.safety.spec@hvtn.org 
Submit CRFs immediately 
Immediate pause 
SAE, related 
3, 2, or 1 
Email CSS/RML 
Submit CRFs immediately 
Prompt PSRT 
AE review to 
consider a pause 
AE, related 
(see Grade 3 
exceptions below) 
4 or 3 
Email CSS/RML 
Submit CRFs immediately 
Prompt PSRT 
AE review to 
consider a pause  
Exceptions to related Grade 3 AEs (for Grade-3 subjective reactogenicity 
events): 
• Injection-site pain/tenderness 
• Fatigue 
• Generalized myalgia 
• Generalized arthralgia 
• Chills 
• Headache 
• Nausea (unless IV rehydration required) 
Unrelated participant death: Sites will call the CSS/RML office phone upon 
learning of any unrelated participant deaths. The site will also email the 
CSS/RML and immediately submit CRFs. The CSS/RML will then promptly 
notify the PSRT. 
If you need to contact the CSS/RML, refer to phone numbers and email addresses 
found on the HVTN 305 protocol home page on the HVTN Members’ site at 
https://members.hvtn.org/protocols/hvtn305. 
9.6.1 
Plan for review of pause rules 
For all safety pauses, HVTN Core notifies the HVTN 305 PSRT, HVTN 
Regulatory Affairs, DAIDS Pharmaceutical Affairs Branch (PAB), DAIDS 
Regulatory Affairs Branch (RAB), DAIDS Safety and Pharmacovigilance Team 

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(SPT), and participating HVTN CRSs. When an immediate safety pause is 
triggered, HVTN Core notifies the SMB. 
Once a trial is paused, the HVTN 305 PSRT reviews safety data and decides 
whether the pause can be lifted or a permanent discontinuation of vaccination is 
appropriate, consulting the SMB if necessary. HVTN Core notifies the 
participating HVTN CRSs, HVTN Regulatory Affairs, DAIDS PAB, DAIDS 
RAB, and DAIDS SPT of the decision regarding resumption or discontinuation of 
study vaccinations. Based on the HVTN 305 PSRT assessment, DAIDS RAB 
notifies the FDA as needed. 
If an immediate HVTN 305 PSRT notification or prompt HVTN 305 PSRT AE 
review is triggered, HVTN Core notifies the HVTN 305 PSRT as soon as possible 
during working hours (local time) or, if the information was received during off 
hours, by the morning of the next workday. If a prompt HVTN 305 PSRT AE 
review cannot be completed within 72 hours of notification (excluding weekends 
and US federal holidays), an automatic safety pause occurs. 
The HVTN requires that each CRS submit to its IRB/EC and any applicable RE 
protocol-related safety information (such as IND safety reports, notification of 
vaccine holds due to the pause rules, unanticipated problems involving risks to 
participants or others, and notification of other unplanned safety pauses). CRSs 
must also follow all applicable RE reporting requirements. 
9.7 
Study termination 
This study may be terminated early by the determination of the HVTN 305 PSRT, 
the NIH, the United States Department of Health and Human Services Office for 
Human Research Protections (OHRP), the FDA, or study product developers. In 
addition, the conduct of this study at an individual HVTN CRS may be terminated 
by the determination of the IRB/EC and any applicable RE. 
9.8 
Pregnancy 
If a participant becomes pregnant during the course of the study, no more 
injections of study product will be given but remaining visits and study 
procedures, including AESI health contact, should be completed unless medically 
contraindicated or applicable regulations require termination from the study. 
During follow-up of persons who are confirmed pregnant, pregnancy testing is not 
required unless clinically indicated. If the participant terminates from the study 
prior to the pregnancy outcome, the site should make every effort to keep in touch 
with the participant in order to ascertain the pregnancy outcome. Pregnancies and 
pregnancy outcomes will be reported as described in the HVTN 305 SSP. If the 
participant is no longer pregnant, refer to Section 5.2.2. 

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10 
Protocol conduct and informed consent 
10.1 
Protocol conduct 
This research study will be conducted in the US and countries outside of the US, 
with funding from the NIH, among others. Due to this, the trial is subject to both 
US and local regulations and guidelines on the protection of human research 
subjects and ethical research conduct. Where there is a conflict in regulations or 
guidelines, the HVTN strives toward maximum protection of human research 
participants. 
The study will comply with the International Council for Harmonization (ICH) 
Guideline for Good Clinical Practice (GCP E6 [R2]), HVTN and DAIDS policies 
and procedures as specified in the HVTN Manual of Operations and DAIDS 
Clinical Research Policies and Standard Procedures Documents, and all 
applicable regulatory requirements. These policies and procedures include 
protocol monitoring (on-site and remote) and compliance. DAIDS and HVTN 
policies and procedures are available for review by any IRB/EC/RE upon request. 
Any policies or procedures that vary from DAIDS and HVTN standards or require 
additional instructions (eg, instructions for randomization specific to this study) 
will be described in the HVTN 305 SSP. Regarding protocol registration, sites 
should follow procedures outlined in the current version of the DAIDS Protocol 
Registration Manual. 
HVTN scientists and operational staff are committed to substantive community 
input into the planning, conduct, and follow-up of its research, ensuring that 
locally appropriate cultural and linguistic needs of study populations are met. 
Community Advisory Boards (CABs) are required by DAIDS and supported at all 
HVTN research sites to ensure community input in accordance with Good 
Participatory Practices (GPP). 
10.2 
Compliance with NIH guidelines for research involving products 
containing recombinant or synthetic nucleic acid molecules 
Because this study is evaluating products containing recombinant or synthetic 
nucleic acid molecules and is funded by the NIH, it must comply with regulations 
set forth in the NIH’s Guidelines for Research Involving Recombinant or 
Synthetic Nucleic Acid Molecules (April 2019). Information about the study must 
be submitted to the Institutional Biosafety Committee (IBC) for each CRS. 
Investigators at each CRS are responsible for obtaining IBC approval per NIH 
guideline Section IV-B7-a-(1). IBC review and approval must be documented by 
the investigator and submitted as part of DAIDS’s initial protocol registration for 
this trial before participants are enrolled at the CRS. If this protocol is amended, 
investigators should follow the requirements of their respective IBC. 

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10.3 
Informed consent 
The sample informed consent form (SICF) in Appendix D describes the 
investigational vaccine and all aspects involved in study participation. 
Documentation of appropriate informed consent must be in place prior to 
conducting study procedures with participants. Periodic assessment of 
participants’ continued understanding of key study concepts and informed consent 
must also be documented. Study sites are strongly encouraged to have their local 
CABs review their site-specific consent forms. This review should include, but 
should not be limited to, issues of cultural competence, local language 
considerations, and the SICF’s level of comprehensibility. 
If any new information is learned that might affect the participants’ decisions to 
stay in the trial, this information will be shared with them. If necessary, 
participants will be asked to sign revised informed consent forms. 
An HVTN CRS may employ recruitment efforts prior to the participant 
consenting. For example, some HVTN CRSs use a telephone script to prescreen 
people before they come to the clinic for a full screening visit. Participants must 
sign a screening or protocol-specific consent before any procedures are performed 
to determine eligibility. HVTN CRSs must submit recruitment and prescreening 
materials to their IRB/EC and any applicable RE for human subjects protection 
review and approval. 
10.3.1 
Screening consent form 
Without a general screening consent, screening for a specific study cannot take 
place until the site receives protocol registration from the DAIDS RSC Protocol 
Registration Office. Sites should follow procedures outlined in the current version 
of the DAIDS Protocol Registration Manual. 
Some HVTN CRSs have approval from their IRB/EC and any applicable RE to 
use a general screening consent form that allows screening for an unspecified HIV 
vaccine trial. In this way, HVTN CRS staff can continually screen potential 
participants and, when needed, proceed quickly to obtain protocol-specific 
enrollment consent. Sites conducting general screening or prescreening approved 
by their IRB/EC and any applicable RE may use the results from this screening to 
determine eligibility for this protocol, provided that the tests are conducted within 
the time periods specified in the eligibility criteria. 

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11 
Exploratory studies 
Samples may be used for other testing and research related to furthering the 
understanding of HIV immunology or vaccines. In addition, cryopreserved 
samples may be used to perform additional assays to support standardization and 
validation of existing or newly developed methods. 
Only genetic testing that is in accordance with the language in the SICF 
(Appendix D) may be performed on samples. 
11.1 
Specimen storage and other use of specimens 
The HVTN stores specimens from all study participants indefinitely unless a 
participant requests that specimens be destroyed or if destruction or a time limit 
for storage is required by IRB/EC or RE. 
“Other use” of specimens is defined as “studies not covered by the protocol or the 
informed consent form for the main study” (see Appendix D). 
This research may relate to HIV, vaccines, the immune system, or other diseases. 
This could include genetic testing and, potentially, genome-wide studies. This 
research is done only to the extent authorized in each study site’s informed 
consent form or as otherwise authorized under applicable law. Other research on 
specimens (“other use”) will occur only after review and approval by the HVTN, 
the IRB/EC of the researcher requesting the specimens, and the IRBs/ECs/REs of 
the CRSs, if required. 
As part of consenting for the study, participants will document their initial 
decision to allow or not allow their specimens to be used in other research, and 
they may change their decision at any time. The participant’s initial decision 
about other use of their specimens, and any later change to that decision, is 
recorded by their CRS in an internet-based tool that documents participants’ 
current decisions for other use of their specimens. The HVTN will only allow 
other research to be done on specimens from participants who allow such use. 
CRSs must notify HVTN Regulatory Affairs if institutional or local governmental 
requirements pose a conflict with or impose restrictions on specimen storage or 
other use of specimens. 

HVTN 305 Version 2.0 / Jan 06, 2023 
HVTN305_v2.0_Final_06Jan23 / Page 75 of 129 
12 
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30. Francica JR, Zak DE, Linde C, Siena E, Johnson C, Juraska M, et al. Innate 
transcriptional effects by adjuvants on the magnitude, quality, and durability 
of HIV envelope responses in NHPs. Blood Adv. 2017;1(25):2329-42. 
31. Pino M, Abid T, Pereira Ribeiro S, Edara VV, Floyd K, Smith JC, et al. A 
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32. Chuang GY, Geng H, Pancera M, Xu K, Cheng C, Acharya P, et al. 
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33. Kwon YD, Pancera M, Acharya P, Georgiev IS, Crooks ET, Gorman J, et al. 
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34. Sanders RW, Derking R, Cupo A, Julien JP, Yasmeen A, de Val N, et al. A 
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macaques. Hum Vaccin Immunother. 2012;8(11):1620-9. 
37. Felber BK, Lu Z, Hu X, Valentin A, Rosati M, Remmel CAL, et al. Co-
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38. Hosseinipour M, Innes C, Naidoo S, Tomaras G, Morris L, DeCamp A, et al. 
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to a Prime‐Boost Approach, but Needle‐Free Injection of DNA Shows No 
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39. Rouphael N, Morgan C, Li S, Jensen R, Sanchez B, Swann E, et al. DNA and 
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CD4+ T‐cell Responses as Traditional Prime/Boost Regimens (HVTN 105). 
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40. Sardesai NY, Weiner DB. Electroporation delivery of DNA vaccines: 
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41. Havenar-Daughton C, Carnathan DG, Torrents de la Pena A, Pauthner M, 
Briney B, Reiss SM, et al. Direct Probing of Germinal Center Responses 
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Responses to HIV Env Trimer. Cell Rep. 2016;17(9):2195-209. 
42. Jardine JG, Ota T, Sok D, Pauthner M, Kulp DW, Kalyuzhniy O, et al. HIV-1 
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Appendix A 
Schedule of procedures: Participants undergoing FNA 
 
 
Green shading: Vaccination visit for all groups; Blue shading: Vaccination visit for Group 3 only. 
Visit Number 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
AESI18 
Study Week 
 
0 
2 
4 
6 
7 
12 
14 
15 
24 
26 
52 
78 
Study Month 
 
0 
0.5 
1 
1.5 
1.75 
3 
3.5 
3.75 
6 
6.5 
12 
18 
Study Day 
-56 to 0 
1 
15 
29 
43 
50 
85 
99 
106 
169 
183 
365 
547 
Procedure 
Screen1 
Vac 1 
 
Vac 2 
 
FNA 
Vac 3 
 
FNA 
Vac 417 
 
 
 
Study procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Assessment of Understanding 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Informed consent 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history2 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Physical exam3 
 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Contraception status assessment4 
 
√ 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
Social impact assessment 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Social impact questionnaire 
 
 
 
 
√ 
 
 
√ 
 
 
 
 
√ 
 
Risk-reduction counseling5 
 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Concomitant medications6 
 
√ 
√ 
√ 
√ 
√7 
√ 
√ 
√7 
√ 
√ 
√ 
√ 
 
Adverse Events (AEs) 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
 
 
AESIs/MAAEs/SAEs 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
√ 
Vaccination8 
 
 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
Reactogenicity assessment9 
 
 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
EP injection site assessment 
 
 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√18 
Clinical labs 
Tube 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy test (urine or serum) 10,11 
 
√ 
√ 
 
√ 
 
√ 
√ 
√ 
√ 
√ 
 
 
 
HBsAg/anti-HCV11 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
HIV screening test11,12 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
Calcium11 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
CBC/Differential11 
EDTA 
5 
 
5 
 
5 
 
 
5 
 
 
5 
 
 
ALT & Creatinine11 
SST 
5 
 
5 
 
5 
 
 
5 
 
 
5 
 
 
HIV diagnostic test 
EDTA 
 
 
 
 
 
 
10 
 
 
 
10 
20 
 
Research samples13 
 
 
 
 
 
 
 
 
 
 
 
 
 
PBMC for assays & storage 
ACD 
 
 
42.5 
 
136 
 
13614 
 
 
 
136 
178.5 
 
Serum for assays and storage 
SST 
34 
 
34 
 
34 
 
 
34 
 
 
34 
34 
 
Leukapheresis15 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
Fine Needle Aspiration16 
 
 
 
 
 
 
√ 
 
 
√ 
 
 
 
 
Daily volume (mL) 
59 
0 
86.5 
0 
180 
0 
146 
44 
0 
0 
190 
232.5 
 
56-day total volume (mL) 
 
 
145.5 
145.5 
325.5 
325.5 
326 
370 
370 
0 
190 
232.5 
 

HVTN 305 Version 2.0 / Jan 06, 2023 
HVTN305_v2.0_Final_06Jan23 / Page 81 of 129 
 
1 Screening evaluations at Visit 01 are performed no more than 56 days before Day 1. 
2 Medical history: A complete medical history is performed during screening. At enrollment and at subsequent visits, an interim medical history may be performed. 
3 A complete physical exam is performed at screening and last clinic visit to include height, weight, and vital signs, and clinical assessments of head, ears, eyes, nose, and throat; neck and lymph 
nodes; heart; chest; abdomen; extremities; neurological function; and skin. At other visits, a targeted physical exam will be performed as needed, based on participant reports or indications of 
illness. 
4 Contraception status assessment is required only for participants who were assigned female sex at birth and are capable of becoming pregnant. Participants will also be provided with counselling 
about contraception. 
5 Risk-reduction counseling per CRS Standard Operating Procedures (SOP). 
6 Concomitant medications, including prescription and nonprescription drugs, vitamins, topical products, alternative/complementary medicines, recreational drugs, vaccinations, and allergy shots, 
are recorded during screening, at enrollment, and at each subsequent clinic visit. 
7 During concomitant medications check-in participant is counseled to avoid medications that would increase the risk of bleeding for 7 days prior to FNA (see Section 8.5).  
8 Vaccination (in-clinic assessments): At least 60 minutes after the first vaccination and after the fourth vaccination, and 30 minutes after for all other vaccinations. Prior to clinic discharge, the 
injection site and systemic symptoms will be assessed, and participants will have vital signs taken. 
9 Reactogenicity: Clinic staff will follow new or unresolved reactogenicity symptoms present at the final reactogenicity day (see Section 8.3 and HVTN 305 SSP)  
10 Pregnancy test: For participants assigned female sex at birth. Pregnancy test may be performed on blood specimens. Persons who are NOT capable of becoming pregnant due to total hysterectomy 
or bilateral oophorectomy (verified by medical records) or menopause (no menses for ≥ 1 year) are not required to undergo pregnancy testing. For persons who are confirmed pregnant, pregnancy 
testing is not required unless clinically indicated. 
11 Local labs may assign appropriate alternative tube types for locally performed tests. 
12 HIV screening test: See Section 5.1.1. 
13 Research samples: Blood-draw volumes for each tube type shown. Alternate tube types may be used under certain conditions (eg, product shortages) upon approval of the HVTN Laboratory 
Center. Refer to the Specimen Collection SSP for more information. 
14 Blood draw will be performed before vaccination. 
15 Leukapheresis: May be performed prior to enrollment visit once the participant has been found to have met all protocol enrollment criteria (see Section 5.1.1 and 5.1.2) and leukapheresis eligibility 
criteria (see Section 8.6). 
16 Fine Needle Aspiration: Target for FNA is 3 weeks post–second and –third vaccinations. 
17 Vaccination 4/Visit 10 will be conducted only for participants assigned to Group 3. 
18 AESI contact: CRS staff will contact study participants at month 18 to collect the information listed in Section 8.10. The EP-injection–site assessment will be an abbreviated questionnaire that can 
be conducted over phone. 

HVTN 305 Version 2.0 / Jan 06, 2023 
HVTN305_v2.0_Final_06Jan23 / Page 82 of 129 
Appendix B 
Schedule of procedures: Participants not undergoing FNA 
Green shading: Vaccination visit for all groups; Blue shading: Vaccination visit for Group 3 only; Gray shading: FNA visit not applicable to this subgroup. 
 
Visit Number 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
AESI16 
Study Week 
 
0 
2 
4 
6 
7 
12 
14 
15 
24 
26 
52 
78 
Study Month 
 
0 
0.5 
1 
1.5 
1.75 
3 
3.5 
3.75 
6 
6.5 
12 
18 
Study Day 
-56 to 0 
1 
15 
29 
43 
50 
85 
99 
106 
169 
183 
365 
547 
Procedure 
Screen1 
Vac 1 
 
Vac 2 
 
 
Vac 3 
 
 
Vac 415 
 
 
 
Study procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Assessment of Understanding 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Informed consent 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history2 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Physical exam3 
 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Contraception status assessment4 
 
√ 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
Social impact assessment 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Social impact questionnaire 
 
 
 
 
√ 
 
 
√ 
 
 
 
 
√ 
 
Risk-reduction counseling5 
 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Concomitant medications6 
 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Adverse Events (AEs) 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
 
 
AESIs/MAAEs/SAEs 
 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
√ 
Vaccination7 
 
 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
Reactogenicity assessment8 
 
 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
EP injection site assessment 
 
 
 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
√16 
Clinical labs 
Tube 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy test (urine or serum) 9,10 
 
√ 
√ 
 
√ 
 
 
√ 
√ 
 
√ 
 
 
 
HBsAg/anti-HCV10 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
HIV screening test, 10,11 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
Calcium11 
SST 
5 
 
 
 
 
 
 
 
 
 
 
 
 
CBC/Differential10 
EDTA 
5 
 
5 
 
5 
 
 
5 
 
 
5 
 
 
ALT & Creatinine10 
SST 
5 
 
5 
 
5 
 
 
5 
 
 
5 
 
 
HIV diagnostic test 
EDTA 
 
 
 
 
 
 
10 
 
 
 
10 
20 
 
Research samples12 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
PBMC for assays & storage 
ACD 
 
 
42.5 
 
136 
 
13613 
 
 
 
136 
178.5 
 
Serum for assays and storage 
SST 
34 
 
34 
 
34 
 
 
34 
 
 
34 
34 
 
Leukapheresis14 
 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
Daily volume (mL) 
59 
0 
86.5 
0 
180 
 
146 
44 
 
0 
190 
232.5 
 
56-day total volume (mL) 
 
 
145.5 
145.5 
325.5 
 
326 
370 
 
0 
190 
232.5 
 

HVTN 305 Version 2.0 / Jan 06, 2023 
HVTN305_v2.0_Final_06Jan23 / Page 83 of 129 
1 Screening evaluations at Visit 01 are performed no more than 56 days before Day 1. 
2 Medical history: A complete medical history is performed during screening. At enrollment and at subsequent visits, an interim medical history may be performed. 
3 A complete physical exam is performed at screening and last clinic visit to include height, weight, and vital signs, and clinical assessments of head, ears, eyes, nose, and throat; neck and lymph 
nodes; heart; chest; abdomen; extremities; neurological function; and skin. At other visits, a targeted physical exam will be performed as needed, based on participant reports or indications of 
illness. 
4 Contraception status assessment is required only for participants who were assigned female sex at birth and are capable of becoming pregnant. Participants will also be provided with counselling 
about contraception. 
5 Risk-reduction counseling per CRS Standard Operating Procedures (SOP). 
6 Concomitant medications, including prescription and nonprescription drugs, vitamins, topical products, alternative/complementary medicines, recreational drugs, vaccinations, and allergy shots, 
are recorded during screening, at enrollment, and at each subsequent clinic visit. 
7 Vaccination (in-clinic assessments): At least 60 minutes after the first vaccination and after the fourth vaccination, and 30 minutes after for all other vaccinations. Prior to clinic discharge, the 
injection site and systemic symptoms will be assessed and participants will have vital signs taken. 
8 Reactogenicity: Clinic staff will follow new or unresolved reactogenicity symptoms present at the final reactogenicity Day (see Section 8.3 and HVTN 305 SSP)  
9 Pregnancy test: For participants assigned female sex at birth. Pregnancy test may be performed on blood specimens. Persons who are NOT capable of becoming pregnant due to total hysterectomy 
or bilateral oophorectomy (verified by medical records) or menopause (no menses for ≥ 1 year) are not required to undergo pregnancy testing. For persons who are confirmed pregnant, pregnancy 
testing is not required unless clinically indicated. 
10 Local labs may assign appropriate alternative tube types for locally performed tests. 
11 HIV screening test: See Section 5.1.1. 
12 Research samples: Blood-draw volumes for each tube type shown. Alternate tube types may be used under certain conditions (eg, product shortages) upon approval of the HVTN Laboratory 
Center. Refer to the Specimen Collection SSP for more information. 
13 Blood draw will be performed before vaccination.  
14 Leukapheresis: May be performed prior to enrollment visit once the participant has been found to have met all protocol enrollment criteria (see Section 5.1.1 and 5.1.2) and leukapheresis eligibility 
criteria (see Section 8.6). 
15 Vaccination 4/Visit 10 will be conducted only for participants assigned to Group 3. 
16 AESI contact: CRS staff will contact study participants at month 18 to collect the information listed in Section 8.10. The EP-injection–site assessment will be an abbreviated questionnaire that can 
be conducted over the phone. 

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Appendix C 
Visit windows 
Visit # 
Visit Type 
Lower 
allowable 
window (-) 
Lower 
target 
window (-) 
Target day* 
Upper target 
window (+) 
Upper 
allowable 
window 
(+)up 
01 
Screening 
Leukapheresis 
-56 
- 
0 
- 
+0 
02 
Enrollment / 
Vaccination 1 
- 
- 
1 
- 
- 
03 
2 weeks postvaccination 1 
- 
-4 
15 
+4 
+7 
04 
Vaccination 2 
- 
- 
29 
+9 
+28 
05 
2 weeks postvaccination 2 
- 
-4 
43  
+4 
+7 
06* 
FNA postvaccination 2 
- 
-4 
50  
+4 
+10 
07 
Vaccination 3 
- 
- 
85  
+14 
+28 
08 
2 weeks postvaccination 3 
Leukapheresis 
Primary immunogenicity  
- 
-4 
 99 
+4 
+7 
09* 
FNA postvaccination 3 
- 
-4 
 106 
+4 
+10 
10 
Vaccination 4 (Group 3 
only) 
- 
-7 
169 
+14 
+28 
11 
2 weeks postvaccination 4 
for Group 3; follow-up 
visit for Groups 1 and 2 
- 
-4 
183 
+4 
+7 
12 
Final clinic visit 
-28 
-14 
365 
+14 
+28 
AESI** 
AESI contact 
-21 
-14 
547 
+14 
+28 
All target dates are relative to Day 1, with the exception of visits, 3, 5, 8, and 11 (for 
Group 3 only) which are relative to the vaccination immediately preceding the visit. 
*Visits 06 and 09 are only applicable to participants providing lymph node FNA 
collections. The FNA visits are relative to the preceding vaccination.  
**This contact must be at least 12 months post last vaccination

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Appendix D 
Sample informed consent form 
Title: A phase 1 open-label clinical trial to evaluate the safety and 
immunogenicity of synthetic DNAs encoding NP-GT8 and IL-12, with or without 
a TLR-agonist-adjuvanted HIV Env Trimer 4571 boost, in adults without HIV 
HVTN protocol number: HVTN 305 
Site: [Insert site name] 
Thank you for your interest in our research study. Please read this consent form or 
ask someone to read it to you. If you decide to join the study, we will ask you to 
sign or make your mark on this form. We will offer you a copy to keep. We will 
ask you questions to see if we have explained everything clearly. You can also 
ask us questions about the study. 
Research is not the same as treatment or medical care. The purpose of a research 
study is to answer scientific questions. 
Key information 
• This is the first study in which one of the study vaccines will be given to 
people. This is also the first time these study products will be given together 
to people.  
• Being in this research study is voluntary. It is your choice. 
• You are being asked to take part in this study because you are aged 18 to 55, 
HIV negative, and healthy. 
• The purpose of this study is to see if the study vaccines are safe, and how a 
person’s immune system responds to them. 
• You will be in this study for up to 12 months of clinic visits, with a follow-up 
contact at least 1 year after your last vaccination, to check on your health. 
• Procedures will include blood draws and injections of study vaccine, as well 
as the collection of white blood cells. White blood cells will be collected by a 
procedure called leukapheresis. In this procedure, some blood will be drawn 
out of your body and passed through a device that will separate and save the 
white blood cells before returning the rest of the blood back into your body. 
You may also be asked if we can collect cells from your lymph nodes. Lymph 
node cells will be collected by a doctor using a very thin needle guided by 
ultrasound. We will tell you more about these procedures later in this consent 
form. 
• One of the study vaccines will be given with electroporation (EP). This 
procedure uses a handheld device to give an electrical pulse into the skin 
where the injection is given using 3 short needles. The pulse briefly opens tiny 

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pores in the cells where the injection is given. We will tell you more about this 
procedure later in this consent form. 
• There are risks from participating: 
The most common risks of taking blood and giving injections that we know about are 
bruising, pain, fainting, soreness, redness, swelling, itching, a sore, and bleeding. 
Collection of white blood cells (leukapheresis) and collecting cells from your lymph node 
can cause pain, bruising, and rarely, an infection. 
Because one of the vaccines has not been given to people before, and the other vaccine has 
only been given to a few people, we do not know what all of the risks may be. 
We will tell you more information about risks later in this consent form. 
The EP device can cause pain ranging from mild to severe that goes away quickly. It can also 
cause soreness, bruising, redness, swelling, itching, or hardness/stiffness in the arm where 
you got the EP. There is also a risk of marks on the skin, such as red bumps, scabs, and 
changes in skin color that may last for months. 
• We do not expect the study vaccines to benefit you in any way. 
About the study 
The HIV Vaccine Trials Network (HVTN) and [Insert site name]  are doing a 
study to test 2 HIV vaccines. HIV is the virus that causes AIDS. 
Up to 54 people will take part in this study. The researcher in charge of this study 
at this clinic is [Insert name of site PI]. The US National Institutes of Health 
(NIH) is paying for the study. 
1. We are doing this study to answer several critical questions. 
• Are the study vaccines safe to give to people? 
• Are people able to take the study vaccines without becoming too 
uncomfortable? 
• How do people’s immune systems respond to the study vaccine? (Your 
immune system protects you from infections and disease). 
• After getting the vaccines, do people develop a certain type of cell in their 
lymph nodes that researchers think are important for protecting against HIV? 
2. The study vaccines cannot give you HIV. 
The study vaccines are not made from HIV. It is impossible for the study vaccines 
to give you HIV. Also, they cannot cause you to give HIV to someone else. 

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We do not know if the study vaccines will decrease, increase, or not change your 
risk of getting HIV if you are exposed to the virus. 
3. The study vaccines are experimental. 
We are testing 2 study vaccines.  
One is INO-6172. We will call it the DNA vaccine. It is made up of two different 
pieces of DNA that were made in a laboratory. DNA is a natural substance found 
in all living things, including people and viruses. DNA tells cells to make 
proteins.  
The second vaccine is called Trimer 4571. We will call it the protein vaccine. It is 
made with an adjuvant called 3M-052-AF + Alum. Adjuvants are products that 
help alert the immune system to have a stronger response. 
Everyone in this study will get the DNA vaccine. It will tell your body to make 
small amounts of two different proteins. One of them is a protein that is found in 
HIV and the second protein is called IL-12 which is naturally found in people. 
Your body’s immune system might recognize the HIV protein and prepare itself 
to fight HIV by making antibodies and T cells. IL-12 helps the immune system to 
make antibodies and T cells more efficiently. Antibodies and T cells are parts of 
an immune response that defend the body against diseases. DNA tells cells to 
make proteins. The DNA vaccine is very much like natural DNA, but it was made 
in a laboratory. You cannot get HIV or AIDS from the DNA vaccine or from the 
proteins the body makes in response to it. 
Some people in the study will also get the protein vaccine. The protein vaccine is 
a small, lab-made piece of protein that looks like a part of a protein found on the 
surface of the HIV virus. The immune system might be able to see this protein 
piece and learn how to recognize HIV if you are ever exposed to it in the future.  
The protein vaccine will be given with an adjuvant called 3M-052-AF + Alum. 
3M-052 was originally developed by 3M Corporation. 3M-052 is one of many 
similar products developed by 3M Corporation to treat skin conditions and 
tumors, and to make vaccines more effective. It is designed to stimulate parts of 
the immune system that recognize invaders like viruses. But in this study, we will 
use the improved 3M-052-AF adjuvant. 
The protein vaccine and the 3M-052-AF adjuvant have not been given together to 
people before. In this study, 3M-052-AF is dissolved in water and is mixed with 
Alum. Alum is made from Aluminum Hydroxide. Alum is an adjuvant with a 
long standing safety record that has been used in approved vaccines for more than 
90 years 
Where do the study products come from? 

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The DNA vaccine was developed by the Wistar Institute and Inovio 
Pharmaceuticals. It is provided for this study by the Division of AIDS (DAIDS) at 
the US National Institutes of Health (NIH) in Rockville, Maryland.  
The protein vaccine was developed and is provided by the Dale and Betty 
Bumpers Vaccine Research Center (VRC) in Bethesda, Maryland, which is part 
of the NIH  
3M-052 was developed by 3M Corporation in St. Paul, Minnesota USA. In this 
study, the 3M-052 is dissolved in water and this mixture is being provided by the 
Access to Advanced Health Institute (AAHI)in Seattle, Washington USA. 
The Alum was manufactured by Brenntag Biosector in Frederikssund, Denmark. 
It is being provided by DAIDS at the US National Institutes of Health in 
Rockville, Maryland, USA. 
What do we know about the study vaccines from other studies? 
The DNA vaccine has not been given to people before. Similar HIV DNA 
vaccines with the same IL-12 adjuvant have been given to more than 350 people 
in other studies. In general, people who got the DNA vaccine in those studies 
were not too uncomfortable, and did not have any serious health problems related 
to the vaccine. Three participants had pain with the injection of the DNA vaccine, 
and decided not to get any more injections. 
The protein vaccine combined with just Alum was given to people in 3 different 
ongoing studies. In the first study, VRC081, 16 people got the same protein 
vaccine mixed with the same Alum adjuvant that we will use in this study. Each 
person got 3 injections of the vaccine at the same dose or higher than the dose we 
will use in this study. The injections did not cause any serious health problems. 
Most people in that study had at least 1 of the side effects listed in the “Risks of 
Vaccines” described below. The participants said their side effects were mild, and 
they went away within about a week.  
In the second study, NIAID 19-I-0069, 3 people got a single dose of this protein 
vaccine combined with Alum at a higher dose than the one we will use in this 
study. The injections did not cause any serious health problems. This study is still 
ongoing and plans to enroll up to 100 people.  
In the third study, NCT04985760, 5 people got injections that had either, a) the 
protein vaccine mixed with Alum at the same dose or higher than the dose we will 
use in this study or, b) just the Alum. We do not yet know who got which 
products. Out of the 5 people, 2 received 2 injections, and 3 received 1 injection. 
The injections have not caused any serious health problems. This study is still 
ongoing and plans to enroll 32 people out of which 24 people will get the protein 
vaccine mixed with Alum. 
Another protein vaccine and 3M-052-AF + Alum adjuvant combination is being 
tested for the first time in people in two other studies called HVTN 137 and 

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HVTN 300. As of September 7, 2022, 48 people in these 2 studies have had at 
least 1 study injection and at least 35 people have had 2 injections. These 
injections have not caused any serious health problems. 
As of September 7, 2022, HVTN 300 has enrolled 13 people, all of whom 
received at least 1 injection. Ten (10) people received at least 2 injections, nine 
(9) received at least 4 injections, and six (6) have received all 5 injections.  
All people in the study had at least some injection site reactions during the trial, 
mostly mild to moderate. One person had severe pain/tenderness in both the right 
and left injection sites 3 days following the fourth vaccination, though it lasted 
only one day. 
 All people had some side effects that they described as mild to moderate. Five (5) 
people had severe side effects, including chills, headache, muscle aches, and 
generally feeling unwell. Two (2) of these 5 people decided to stop getting study 
injections because they felt generally unwell after injections, and it was affecting 
their daily lives. Another 2 of the 5 people who had severe side effects got more 
injections; one of them had more severe side effects, while the other did not. And 
1 of the 5 people who had severe side effects has not yet received another 
injection.  
Two more people also decided to stop getting the study injections and dropped 
out of the study. One stopped because they experienced a panic attack after the 
first injection. However, this person had a history of panic attacks before being 
part of this study. The other did not return for their next injection visit and is no 
longer in the study. We do not know the reason for this.  
HVTN 137 gives some people a study vaccine and some people a placebo which 
does not contain any study vaccine. The study has two parts, Part A and Part B. 
As of April 2022, 17 people in Part A have had 2 injections (15 people got the 
study vaccine and 2 people got a placebo). These injections have not caused any 
serious health problems. In HVTN 137 Part A, two people had redness and 
swelling over a large area on their arm where they got the injection. The swelling 
started about 1 week after the injection. The redness lasted for about 2 to 3 days. 
For one of these people, the swelling went away within 2 to 3 days. The second 
person had significant swelling for about 3 days. The swelling went down but it 
took about 5 weeks to completely go away. They had mild pain which did not 
prevent them from going to work. Four other people had symptoms that did 
prevent them from going to work. All 4 felt severely tired and generally unwell 
for up to 2 days. Two of them also had severe muscle and joint pain, severe 
headache and chills for a day. The study is ongoing and we do not know if these 
people got the study vaccine or placebo. 
HVTN 137 was later updated to offer people in Part A, a third injection, and 9 out 
of the 17 people agreed to it. In Part B of HVTN 137, 88 people received 
injections of either the study vaccine and adjuvant or the placebo. These 
injections have not caused any serious health problems.  
 

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Given the side effects experienced in HVTN 300 and HVTN 137, we can expect 
that many people in this study who receive the 3M-052-AF + Alum adjuvant will 
have some side effects. The side effects we expect would be similar to the Risks of 
vaccines listed below. 
These are experimental HIV vaccines, which means we do not know if the 
vaccines will be safe to use in people, or if they will work to prevent HIV 
infection. These vaccines are used only in research studies. 

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Risks of vaccines: 
All vaccines can cause fever, chills, headaches, rash, aches, pains, nausea, 
dizziness, and feeling tired. Vaccines can also cause tenderness, swelling, redness 
or itching where you got the injection. Most side effects do not interfere with 
daily activities or make a person visit the doctor. 
Rarely, a vaccine can cause an allergic reaction, including a rash, hives, or trouble 
breathing. Allergic reactions can be life threatening. Tell us if you have ever had a 
bad reaction to an injection or vaccine. 
Risks of DNA vaccines: 
More than 1,000 people have been given DNA vaccines being tested against HIV, 
and thousands of people have received other experimental DNA vaccines for 
other diseases. These other DNA vaccines have not caused serious health 
problems. We expect the risks of the DNA vaccine in this study to be similar to 
those of other DNA vaccines. 
Risks of the protein vaccine: 
This protein vaccine has been given to a small number of people so far and no 
serious health concerns have been reported. Because of this, we do not know all 
the side effects. There may be side effects, even serious or life-threatening ones, 
that we do not know about yet. 
Risks of the study adjuvant: 
The combination of the protein vaccine and adjuvant used in this study has not 
been given to people before, so we do not know all the risks. However, the 
adjuvant in combination with other similar protein vaccines is being given to 
people for the first time in other studies. A small number of people have received 
these vaccinations so far. A few of them have reported having some of the side 
effects described in ‘Risks of vaccine’, but no serious health concerns have been 
reported so far. 
These are the side effects that we know about. There may be others that we don’t 
know about. We will tell you if we learn about new side effects that could affect 
your willingness to stay in the study. 
4. The DNA vaccine is given using electroporation (EP). 
Besides adjuvants, another way to improve immune responses to DNA vaccines is 
to use an EP device when giving the vaccine. EP gives an electric pulse using 3 
short needles into the skin. The pulse briefly opens tiny pores in the cells where 
the injection is given. The DNA can enter the cells through these pores. Previous 
studies have shown that EP helped to get DNA into cells better. One study also 
showed increased immune responses to another experimental DNA vaccine. 

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Using EP in people is an experimental procedure, and the EP device is an 
experimental device. The EP device is only used in people in research studies. 
EP is done in this study using a device called the CELLECTRA 2000 EP System. 
It was developed and is being provided for this study by Inovio Pharmaceuticals, 
Inc.  
The EP device has been used with injections into the skin in at least 350 people. 
In another study, a DNA vaccine given together with an IL-12 DNA adjuvant was 
given into the skin with EP to 55 people. EP did not cause any serious health 
problems in these participants. 
In a previous study using the EP device in healthy volunteers, some people said 
that they felt only a little discomfort, while others said it was very painful, even 
after the injections were over. However, only 3 out of 48 volunteers left the study 
because of pain and discomfort. 
Another large study being done in multiple countries is currently giving EP with a 
DNA vaccine for SARS-CoV-2 (the virus that causes COVID-19). The study 
began in November 2020, and so far over 3,200 people have received more than 
9300 injections with an EP device. A few people have noticed marks on the skin 
after getting the vaccines with EP, but no health concerns have been reported so 
far. 
Risks of EP: 
EP causes brief muscle contractions during the procedure. In previous studies 
using EP, people felt initial pain that ranged from mild to severe. For most people, 
the pain eased quickly. EP can also cause soreness, bruising, redness, swelling, 
itching, or hardness/stiffness in the upper arm where you got the injection. When 
the injection and EP are given to the skin, the needles may leave marks, such as 
red bumps or scabs. Later, the marks may heal, but may leave light or dark spots 
on the skin. In some people, these marks lasted 12 months after their last 
injection. We do not know if the marks went away later, because we did not 
follow the participants beyond that time. On darker skin, these marks tend to 
remain visible for longer, with less fading. We do not know how the skin at the 
injection site will change in appearance after EP with these study products 
because they have not been given with EP before. In addition, every person’s skin 
can react differently. We will show you photos of people who got EP in a 
previous study to give you examples of what these injection marks can look like 
over time. 
Site: Sites must review the sample photos of EP injection marks with potential 
participants during the study informed consent process. 
On rare occasions, the EP device may cause infection at the part of your body 
where you got the injection. 
Having the procedure or thinking about it may cause some stress and anxiety. If 
you feel anxious, please tell us and we will try to help you. In a previous study 

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where a DNA vaccine was given into the skin with EP, over 99% of the 
participants said they would recommend EP with an effective HIV vaccine to 
their family or friends if it felt the same as their study experience. Of these 
participants, 96% said that the appearance at injection sites was acceptable. 
We do not know if EP will change the risks for the DNA vaccine with the IL-12 
adjuvant. We do not know all the risks of EP because it has only been used in a 
limited number of people before this study and has not been used with these study 
products. 
Joining the study 
5. It is up to you whether or not to join the study. 
Take your time. Talk to people you trust. If you decide not to join this study or if 
you leave after you have joined, it will not affect your other care at this clinic nor 
the benefits or rights you would normally have. 
You cannot be in this study while you are in another study where you get a study 
product. If you do not join this study, you may be able to join another study. 
Site: The boxed text is for sites in South Africa. 
During the study, you should not donate blood or tissue. 
Site: Remove item 6 if you use a separate screening consent that covers these 
procedures. 
6. If you want to join the study, we will screen you to see if you are eligible. 
Screening involves a physical exam, HIV test, and health history. A physical 
exam may include, but is not limited to: 
• Checking your weight, temperature, and blood pressure 
• Looking in your mouth and throat 
• Listening to your heart and lungs 
• Feeling your abdomen (stomach and liver) 
We will also do blood tests. These tell us about the health of your kidneys and 
liver. We will ask you about medicines you are taking, including HIV pre-
We check to make sure that you are not in more than one study by taking your 
fingerprint on an electronic system. This information is only accessed by a 
few members of the study team using a secure password. 

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exposure prophylaxis (PrEP). We will ask you about behaviors that might put you 
at risk for getting HIV. If you were assigned female sex at birth, we will test you 
for pregnancy. 
We will review the test results with you. They may show that you are not eligible 
for the study, even if you want to join. 
Site: adapt the following section per the care available at your site 
7. If we find that you have a health problem during screening or during the 
study, we will tell you about the care that we can give here for free. 
For the care that we cannot give, we will help you get care elsewhere. For health 
problems unrelated to the study, we will not pay for care. 
8. If you were assigned female sex at birth and could become pregnant, you 
must use birth control to join this study. 
Site: If you want to include Appendix H, Approved birth control methods (for 
sample informed consent form), in this consent form, paste it below and delete 
paragraph below. 
It is important that you not become pregnant during the study because we do not 
know how the study vaccine could affect a developing baby. For this reason, you 
must agree to use effective birth control from 21 days before your first injection 
until 8 weeks after the time of your last study injection at 6 months (9 months 
total). We will talk to you about effective birth control methods. They are listed on 
a handout that we will give to you. 
Being in the study 
If you join the study, here is what will happen: 
9. You will come to the clinic for scheduled visits up to 12 times over 12 months. 
Site: Insert number of visits and range of visit lengths. (There is site-specific 
variation in screening protocols and in the number of possible follow-up visits 
between protocol-mandated visits.) 
Visits can last from [#] to [#] hours. 
You may have to come for more visits if you have a lab or health issue. We will 
also contact you 1 year after your last vaccination to check on your health. We 
may also contact you after the study ends (for example, to tell you about the study 
results). We may also contact you about other studies that you may want to join. 

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10. We will give you compensation for each study visit you complete. 
This amount is to cover the costs of [Site: Insert text]. There is also additional 
compensation for the leukapheresis procedure and lymph node cell collection 
procedure. We will tell you more about the procedures below. 
You will receive $xx.xx each time you have leukapheresis. This amount is to 
cover the costs of [Site: Insert text]. You will be paid ___________ [“following 
each completed visit”, “monthly”, “quarterly”, “at the end of your participation 
in the research study”, “following each completed visit or at the end of your 
participation in the research study, whichever you prefer”]. 
You will receive $xx.xx each time you have a lymph node cell collection. This 
amount is to cover the costs of [Site: Insert text]. You will be paid ___________ 
[“following each completed visit”, “monthly”, “quarterly”, “at the end of your 
participation in the research study”, “following each completed visit or at the 
end of your participation in the research study, whichever you prefer”]. 
The boxed text is for sites in the US 
You do not have to pay anything to be in this study. Site: Insert any costs to 
participants (eg, birth control costs for female participants who could become 
pregnant). 
11. We will give you the study vaccine on a schedule. 
We will give you injections of the study vaccines at 3 or 4 visits, according to the 
schedule in the table below. All injections will be given into the upper arm with a 
needle and syringe. The first 5 participants to enroll in the study will be assigned 
to Group 1. If you are not one of them, you have a small chance (10%) of being 
assigned to Group 1. Your chance of being assigned to either group 2 or group 3 
is slightly lower than 50%. Site: Modify the randomization metaphor in the next 
sentence as appropriate to your local culture. Your group assignment is 
completely random, like rolling dice or drawing straws.  
If you are in Group 1 or Group 2, you will only get the DNA vaccine. You will 
have 3 injection visits where you get 1 injection in each arm, followed by EP into 
the skin of each arm. 
If you are in Group 3, you will get the DNA vaccine at the first 2 injection visits. 
You will get 1 injection in each arm, followed by EP into your skin. At the third 
injection visit you will get the DNA vaccine injection in each arm, followed by 
EP into your skin, and you will also get a protein vaccine injection into the upper 
muscle of each arm. You will also have a fourth injection visit where you get just 
the protein vaccine into the upper muscle of each arm.  
Payments you receive for being in the study may be taxable. We may need to ask you 
for your Social Security number for tax reasons. 

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Because this is the first time the DNA vaccine is being given to people, only 1 
participant in  Group 1 will get the study vaccine each day until the first 5 
participants have gotten the DNA vaccine. After this, we will not enroll anyone 
else into the study until a safety review is completed. A team of experts who are 
not part of the study will look at the information about the first 5 people’s 
responses to the vaccine and decide if it is safe to continue the study. If it is, the 
rest of the participants in all groups will be enrolled. Since this is the first time the 
DNA and protein vaccines are given as a combination, at the Month 3 injection 
visit, only 1 participant in Group 3 will get the study vaccinations each day for the 
first five participants to reach this study visit, so that we can closely monitor their 
safety.  
Group 
# 
Number 
of 
People 
Products 
Dose 
Where 
How 
First 
injection 
visit 
1 
month 
later 
3 
months 
later 
6 
months 
later 
1 
9 
DNA 
vaccine  
Lower 
dose 
Skin of 
both upper 
arms 
Needle & 
syringe + 
EP 
 
 
 
 
2 
18 
DNA 
vaccine  
Higher 
dose 
Skin of 
both upper 
arms 
Needle & 
syringe + 
EP 
 
 
 
 
3 
18 
DNA 
vaccine  
Higher 
dose 
Skin of 
both upper 
arms 
Needle & 
syringe + 
EP 
 
 
 
 
Protein 
vaccine 
-- 
Muscle of 
both upper 
arms 
Needle & 
syringe 
 
 
 
  
For injections of the DNA vaccine between the layers of the skin (known as 
intradermal injections), we will give the injection and then press the EP device 
firmly against that same area. The device inserts 3 very short needles into your 
skin. We will activate the device and a very small amount of electricity will be 
sent in 4 short pulses from the needles into your arm. Each of these pulses will 
last less than 1 second. Your arm will move because of the electrical pulses. 
This injection procedure will take less than 1 minute. During the procedure and 
right after, you will feel some pain or discomfort. The intensity of that feeling 
lessens or may go away within a couple of minutes. After that, your arm may be 
sore for a day or two. 
People in Group 3 will also get injections of the protein vaccine and study 
adjuvant. These are given in the traditional way vaccines are given, using a needle 
and syringe to give the vaccine in the muscle of the upper arm. 
You will have to wait in the clinic for about 30 minutes to an hour after getting 
your injections to see if there are any problems.  
• If you are in Group 1 or Group 2, you will wait in the clinic for 60 minutes 
after your first injection and for 30 minutes after all other injections.  
• If you are in Group 3, you will wait for 60 minutes after your first, third 
and fourth injections and for 30 minutes after your second injection.  

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You will also use a secure electronic symptom log to keep track of how you are 
feeling. We call this symptom log an eDiary.  
• If you are in Group 1 or Group 2, after all injections you will use the 
eDiary to keep track of how you are feeling for that day and for 7 more 
days.  
• If you are in Group 3, after just the first 2 injections you will use the 
eDiary to keep track of how you are feeling for that day and for 7 more 
days. For the other injections, you will use the eDiary for that day and for 
2 more weeks.  
If you are unable or unwilling to use the eDiary, please talk with us about other 
options that may be available. Within 4 days after each injection visit, we will 
contact you to ask how you are doing. Contact us if you have any issues or 
concerns after getting an injection. 
As part of this research, you may need to use an app on your phone to access the 
eDiary. While using this app, information about you may be collected and shared 
with the researchers or people outside of the study who have been approved by 
the researchers. These data might include personal health information and 
information about your use of the app, such as the amount of time you spend on 
each screen. The eDiary does not collect personal information about your 
activities over time or from other websites or online services. It also does not 
allow third parties to collect that information. A complete description of the data 
collection and sharing can be found in the privacy policy associated with the app. 
If you would like to read these documents, the study doctor can tell you how to 
access this information. 
The privacy policy may include statements that limit your rights if you are 
harmed by using the app in this study. You do not release the study doctor, 
sponsor, this institution, or the research staff for responsibilities from mistakes. 
You also do not waive any of your rights as a research participant. 
We will ask you questions about any skin changes related to the EP procedure. 
We may ask to take photos of your arm where you got the injection or EP. This 
may help us learn more about reactions to the study vaccines or the EP procedure. 
We will not take photos of your face. Your name and other identifying 
information will not be included with the photos or in publications. The photos 
will only be used to research the safety of the vaccine and EP. 
12. We will do the procedures shown in the following table. 
 

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Time after first injection visit 
Procedure 
Screenin
g visit(s) 
First 
injection 
visit  
2 weeks 
1 month 
1½ months 
1¾ months 
3 months 
3½ months 
3¾ months 
6 months* 
6½ months 
12 months 
1 year + 6 
months 
Injection(s) 
 
√ 
 
√ 
 
 
√ 
 
 
√ 
 
 
 
Medical history 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
Complete 
physical 
√ 
 
 
 
 
 
 
 
 
 
 
√ 
 
Brief physical 
 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
 
 
Blood drawn 
√ 
 
√ 
 
√ 
 
√ 
√ 
 
 
√ 
√ 
 
Lymph node 
cell 
collection** 
 
 
 
 
 
√ 
 
 
√ 
 
 
 
 
Leukapheresis 
√ 
 
 
 
 
 
 
√ 
 
 
 
 
 
Pregnancy 
test*** 
√ 
√ 
 
√ 
 
√ 
√ 
√ 
√ 
√ 
 
 
 
HIV testing  
√ 
 
 
 
 
 
√ 
 
 
 
√ 
√ 
 
Risk reduction 
counseling 
√ 
√ 
√ 
√ 
√ 
 
√ 
√ 
 
√ 
√ 
√ 
 
Interview / 
questionnaire 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
Skin assessment 
 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
Health contact 
 
 
 
 
 
 
 
 
 
 
 
 
√ 
 
* The gray shading for the visit at 6 months is only for participants assigned to Group 3. 
** Lymph node cell collections are an optional procedure for people in Groups 2 and 3. 
*** Applies only to persons who were assigned female sex at birth and who are capable of becoming 
pregnant. Persons who have had a hysterectomy (removal of the uterus) or removal of both ovaries 
(verified by medical records) do not have to have pregnancy tests. 
When we take blood, the amount will depend on the lab tests we need to do. It 
will be some amount between 44 mL and 258 mL (about 3 tablespoons to about 1 
cup). Your body will make new blood to replace the blood we take out. We will 
review test results with you at your next visit, or sooner if necessary. We will tell 
you about any results that are important to your health. 
Site: Sites must obtain copies of the informed consent form and any other 
educational  materials that are available from the facilities where leukapheresis 
and FNA will be performed, so that they can be reviewed with potential 
participants during the study informed consent process. 
Leukapheresis procedure (for all participants): 
This procedure collects large amounts of white blood cells. Blood is made up of 
red cells that carry oxygen, white cells that fight infection, platelets that help form 
clots, and plasma, which is the fluid left over when all the cells are removed. 
During leukapheresis, white blood cells are removed from a vein in one arm and 
the rest of the blood is put back into a vein on the other arm.  

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The leukapheresis procedure will be done at [site: insert location]. Your 
eligibility to have this procedure will be assessed by the staff at our clinic and at 
the facility before you have the procedure. There will be another consent form for 
you to review and sign at the facility. It will provide additional details about the 
procedure and any risks involved. 
For the procedure, a clinician will insert a sterile needle into a vein in each of 
your arms. The needles are attached to tubes. Your blood will go out of your body 
through one tube and into a machine that separates the blood and takes out the 
white blood cells. After the white blood cells are taken out, the rest of the blood 
will go back into your body through the tube going into your other arm. 
Sometimes the fluid lost during the procedure is replaced by a sterile salt water 
solution, or a solution containing a protein called albumin. This protein is 
normally found in human blood. An anticoagulant may be added to your blood 
during the procedure. Anticoagulants prevent blood from clotting. 
It is normal to feel tired for up to 24 hours after having leukapheresis. 
The leukapheresis procedure will be done before your first injection visit and 
again about 2 weeks after your third injection visit. It will take about [Site: Insert 
timeframe] for the procedure. 
Risks of leukapheresis 
Generally, the risks of leukapheresis include pain, bruising, and rarely, infection. 
Rarely, albumin can cause an allergic reaction. If the leukapheresis procedure has 
to be stopped, it could result in the loss of up to 1 cup of blood. Your body makes 
new blood within 2 weeks. 
During leukapheresis, you may be given a blood thinner to prevent blood clots in 
the machine. This may cause you to have low calcium levels in your blood for a 
short time. Sometimes people with low calcium levels feel numbness and tingling 
in their body. Low calcium can be corrected by replacing what is lost with a 
calcium tablet during or shortly after the procedure. The leukapheresis centers 
know how to treat low calcium, so please let the staff know if you feel numbness 
or tingling. Very rarely low calcium levels can cause heart problems, but to our 
knowledge, this procedure should not cause you to have heart problems. 
If you notice any symptoms during leukapheresis, please let the nurse know 
immediately. Usually the symptoms can be reversed quickly by adding fluid or by 
slowing down the procedure. If there are any problems, the staff will use the 
appropriate medical procedures to treat you. 
Lymph node cell collection procedure (optional for Groups 2 and 3): 
People in Groups 2 and 3 will be asked to consider an additional procedure. The 
formal name for this procedure is fine needle aspiration (FNA). This procedure 
collects cells from lymph nodes. Lymph nodes are one of the key places where 
immune responses develop, and they are located in several places on the human 

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body. In this procedure, a clinician will use a very thin needle guided by 
ultrasound to collect cells from a lymph node on your arm or in your armpit. The 
researchers would like to have at least half of the people in Groups 2 and 3 agree 
to these cell collections. 
The lymph node cell collection procedure will be done at [site: insert location]. 
Your eligibility to have this procedure will be assessed by the staff at our clinic 
during screening and again at the facility before you have the procedure. There 
will be another consent form for you to review and sign at the facility. It will 
provide additional details about the procedure and any risks involved. We will 
also tell you about some medications you should avoid before having the 
procedure. 
To do the procedure, a clinician will find the appropriate lymph node by applying 
a cold gel to your skin and then pressing a handheld ultrasound device against 
your skin. If necessary, they may shave a small area near the lymph node. Once 
the node is found, they will apply a cleaning solution to the skin in that area. They 
will numb the area with an injection of a local anesthetic, such as Lidocaine; this 
is like Novocaine which is used for dental procedures. They will insert a very thin 
needle and use the images from the ultrasound to guide it into the lymph node. 
They may move the needle up and down to collect cells from the lymph node. 
Fluid may also be collected through the needle into a syringe or bottle. Each cell 
collection lasts about 10 to 15 seconds. It usually takes about 4 separate cell 
collections to get enough cells. 
After the procedure is done, the clinician will clean the area with warm water and 
give you a band-aid to cover the site where the needle was inserted. 
Lymph node cell collection will be done twice, about 3 weeks after your second 
and third injection visits. It will take about [Site: insert timeframe] for the 
procedure. 
Risks of lymph node cell collection 
The main risks of this procedure include pain, bruising, bleeding, and infection 
where the needle is inserted. The cleaning solution or shaving the area may cause 
skin irritation. Injecting the local anesthetic may sting or burn for a little while 
until the numbing takes effect. Rarely the local anesthetic can cause your arm to 
feel numb or weak. Examining the lymph node area may be uncomfortable.  
If discomfort, problems, or side effects happen during or after lymph node cell 
collection, the staff will use the appropriate medical procedures to treat you. 
13. We will counsel you about protecting yourself from HIV. 
We will ask you questions about your HIV risk factors such as sexual behavior, 
alcohol, and drug use. We will talk with you about ways to keep your risk of 
getting HIV low. 

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14. The HVTN will test your samples to see how your body, including your 
immune system, responds to the study vaccines. 
We will send your samples (without your name or other identifying information) 
to labs approved by the HVTN for this study. In rare cases, some of your samples 
may be sent to labs in other countries for research related to this study. 
Researchers may also do genetic testing on your samples. Your genes are passed 
to you from your birth parents. They affect how you look and how your body 
works. This genetic testing will involve only some of your genes, not all of your 
genes (your genome). It will involve genes related to the immune system and 
HIV. 
If you get HIV, the researchers may look at all of the genes of the virus found in 
your samples. The researchers will use this information to learn more about HIV 
and how the virus is impacted by the study vaccines. 
In some cases, researchers may take cells from your samples and grow more of 
them over time, so that they can continue to contribute to this study. 
These tests done on your samples are for research purposes, not to check your 
health. The labs will not give the results to you or this clinic because their tests 
are not approved for use in making health care decisions. These labs are only 
approved to do research tests. 
When your samples are no longer needed for this study, the HVTN will continue 
to store them. 
Site: Delete next section if using separate consent for use of samples and information 
in other studies 
15. When samples are no longer needed for this study, the HVTN may want to 
use them in other studies and share them with other researchers.  
The HVTN calls these samples “extra samples.” The HVTN will only allow your 
extra samples to be used in other studies if you agree to it. You will mark your 
decision at the end of this form. If you have any questions, please ask. 
Do I have to agree? No. You are free to say yes or no, or to change your mind 
after you sign this form. At your request, the HVTN will destroy all extra samples 
that it has. Your decision will not affect your being in this study or have any 
negative consequences here. 
Where are the samples stored? Extra samples are stored in a secure central place 
called a repository. [Site: choose one of the following two sentences. African sites 
should choose the sentence referencing the repository in South Africa. All other 
sites should choose the sentence referencing the repository in the United States.] 
Your samples will be stored in the HVTN repository in South Africa. Your 
samples will be stored in the HVTN repository in the United States. 

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How long will the samples be stored? There is no limit on how long your extra 
samples will be stored. [Site: Revise the previous sentence to insert limits if your 
regulatory authority imposes them.] 
Will I be paid for the use of my samples? No. Also, a researcher may make a new 
scientific discovery or product based on the use of your samples. If this happens, 
there is no plan to share any money with you. The researcher is not likely to ever 
know who you are. 
Will I benefit from allowing my samples to be used in other studies? Probably not. 
Results from these other studies are not given to you, this clinic, or your doctor. 
They are not part of your medical record. The studies are only being done for 
research purposes. 
Will the HVTN sell my samples and information? No, but the HVTN may share 
your samples with HVTN or other researchers. Once the HVTN shares your 
samples and information, it may not be able to get them back. 
How do other researchers get my samples and information? When a researcher 
wants to use your samples and information, their research plan must be approved 
by the HVTN. Also, the researcher’s Institutional Review Board (IRB) or Ethics 
Committee (EC) will review their plan. [Site: If review by your institution’s 
IRB/EC/RE is also required, insert a sentence stating this.] IRBs/ECs protect the 
rights and well-being of people in research. If the research plan is approved, the 
HVTN will send your samples to the researcher’s location. 
What information is shared with HVTN or other researchers? The samples and 
information will be labeled with a code number. The key to the code will stay at 
this clinic. It will not be shared with the HVTN, other researchers, or anyone else 
that does not need to know your name. Your name will not be part of the 
information. However, some information that the HVTN shares may be personal, 
such as your race, ethnicity, sex, health information from the study, and HIV 
status. The HVTN may share information about the study product you received 
and how your body responded to the study product. 
What kind of studies might be done with my extra samples and information? The 
studies will be related to HIV, vaccines, the immune system, and other diseases. 
Researchers may also do genetic testing on your samples. 
In some cases, researchers may take cells from your samples and grow more of 
them over time so that they can continue to do research with them. 
If you agree, your samples could also be used for genome-wide studies. In these 
studies, researchers will look at all of your genes (your genome). The researchers 
compare the genomes of many people, looking for common patterns of genes that 
could help them understand diseases. The researchers may put the information 
from the genome-wide studies into a protected database so that other researchers 
can access it, but your name and other personal information will not be included. 

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Usually, no one would be able to look at your genome and link it to you as a 
person. However, if another database exists that also has information on your 
genome and your name, someone might be able to compare the databases and 
identify you. If others found out, it could lead to discrimination or other problems. 
There may be other unknown risks. 
Who will have access to my information in studies using my extra samples? 
People who may see your information are: 
• Researchers who use your extra samples and information for other research 
• Government agencies that fund or monitor the research using your extra 
samples and information 
• The researcher’s IRB or EC 
• Any regulatory agency that reviews clinical trials 
• The people who work with the researcher 
All of these people will do their best to protect your information. The results of 
any new studies that use your extra samples and information may be published. 
No publication will use your name or identify you personally. 
16. We will do our best to protect your private information. 
US Site: Check HIPAA authorization for conflicts with this section. 
All of your samples and most of your study records will be labeled with a code 
number. Samples and study records are kept in secure locations. When you 
provide information in the eDiary after the injection visits, that information will 
only have your code number, not your name. Your data goes directly from the 
eDiary into your study record. 
Site: Any change to the following boxed text requires approval from HVTN 
Regulatory Affairs. You can remove the box around the text. 
We do need to share your name with the HVTN in case you need proof in the 
future that you participated in an HIV vaccine study. The HVTN will keep your 
name in a secure file with these items: 
• The name of your study 
• Your age or date of birth 
• Your study ID number 
• What study vaccines(s) you received 

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No HIV test results are kept in this file. The HVTN will not share any information 
that could identify you without your agreement. The HVTN will remove your 
name from the file if you do not want it there. 
Clinic staff can see your study records. Your records may also be reviewed by 
groups that watch over this study to see that we are protecting your rights, 
keeping you safe, and following the study plan. These groups include: 
• The NIH and its study monitors; 
• The US Food and Drug Administration (FDA); 
• Any regulatory agency that reviews clinical trials; 
• [Insert name of local IBC]; 
• US sites may, but are not required to include: [Insert name of local IRB/EC] ; 
• Advarra IRB; 
• [Insert name of local and/or national regulatory authority as appropriate]; 
• The Wistar Institute, Inovio Pharmaceuticals, AAHI and people who work for 
them; 
• The HVTN and people who work for them; 
• The HVTN Safety Monitoring Board (SMB); and 
• The US Office for Human Research Protections (OHRP). 
All reviewers will keep your records private. 
We cannot guarantee absolute privacy. If you have a medical condition that we 
are required to report by law, then some of your information may be shared. At 
this clinic, we have to report the following information: 
Site: Include any public health or legal reporting requirements. Bulleted 
examples should include all appropriate cases (reportable communicable disease, 
risk of harm to self or others, etc.) If your site does not have public health or legal 
reporting requirements, you may delete the last sentence in the paragraph above, 
along with the bullets below. 
• [Item 1] 
• [Item 2] 
• [Item 3] 

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US sites: Include the following boxed text. You can remove the box around the text. 
To help protect your privacy, we have a Certificate of Confidentiality from the US 
government. With the certificate, we do not have to release information about you to 
someone who is not connected to the study, such as the courts or police. However, we 
cannot withhold information from the US government because it funds this research. 
You can still give information about yourself and your study participation to others. 
The results of this study may be published. No publication will use your name or 
identify you personally. 
We may share information from the study with other researchers. We will not 
share your name or information that can identify you. 
A description of this clinical trial will be available on 
http://www.ClinicalTrials.gov. This Web site will not include information that can 
identify you. At most, the Web site will include a summary of the results. You can 
search this Web site at any time. 
17. We may stop your injections even if you want to stay in the study and even if 
you were scheduled for more injections. 
We will stop your injections if you become pregnant. We will encourage you to 
stay in the study and complete the other study procedures, but it will be your 
choice. If you leave the study while you are pregnant, we will contact you after 
your due date to ask some questions about your pregnancy and delivery if you 
agree. 
We will stop your injections if you get HIV. We will also take fewer samples and 
may not perform some of the procedures we told you about earlier. We will help 
you get care and support. We will counsel you about having HIV and about 
telling your partner(s). Site: Modify the following sentence as appropriate. We 
will not provide or pay for your HIV care. We will encourage you to stay in the 
study for up to 12 months after your last injection. It is up to you whether you 
choose to stay or leave. We will counsel you about having HIV and about telling 
your partner(s). Site: Modify the following sentence as appropriate. We will not 
provide or pay for your HIV care. 
We will stop your injections if you enroll in a different research study where you 
get another study product. 
18. We may take you out of the study at any time 
We may take you out of the study if: 
• You do not follow instructions, 
• We think that staying in the study might harm you, or 

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• The study is stopped for any reason. 
Other Risks 
19. There are other risks to being in this study. 
In addition to the risks of the study vaccine and the procedures that were 
described above, this section describes the other risks that we know about. There 
may be other risks not listed below, even serious ones. We will tell you if we 
learn anything new that may affect your decision to stay in the study. 
Risks of abnormal laboratory results: 
Minor changes in laboratory test results occasionally happen. This means that the 
test results can show something to be abnormal when it is not. If this happens, we 
will ask you to come back to the clinic to be retested. This may cause you to 
worry, and it may be inconvenient to come back to the clinic. If retesting confirms 
something to be abnormal, we will provide care or help you get the care you need. 
Risks of routine medical procedures: 
Routine medical procedures such as taking blood and giving injections can cause 
bruising, pain, fainting, soreness, redness, stinging, swelling, itching, a sore, 
bleeding, and rarely, infection where you got the injection. Taking blood can 
cause a low blood cell count (anemia), making you feel tired. 
Personal problems/discrimination/testing HIV antibody positive: 
Some people report personal problems or discrimination because they joined an 
HIV vaccine study. Family or friends may worry, get upset, or assume that you 
have HIV. Rarely, someone has lost a job because the study took too much time 
away from work, or because their employer thought they had HIV. 
Most vaccines cause the body to make antibodies to prevent infection. Your body 
may make antibodies to HIV because you received an HIV study vaccine. Those 
antibodies could cause you to test positive on some types of HIV tests, even if you 
do not have HIV. This is called vaccine-induced seropositivity (VISP). VISP 
means that after you get the study vaccine, a routine HIV test done outside this 
clinic is likely say you have HIV, even if you don’t. For this reason, you should 
get HIV tests only at this clinic. Our tests can tell the difference between true HIV 
infection and a positive result caused by the study vaccine. If you have VISP, we 
can arrange free HIV testing for as long as you need it.  
It is unlikely, but you could test antibody negative at the end of the study and then 
test positive sometime later, even though you don’t have HIV.  
Site: Modify the following paragraph if applicable. If someone believes you have 
HIV, you could face discrimination and other problems. In some countries, you 
could be denied medical or dental care, employment, insurance, a visa, or entry 

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into the military. If you have VISP, you will not be able to donate blood or 
organs. Your family and friends may treat you differently. We will give you a 
brochure that tells you more about VISP, and how you can avoid some of these 
problems. 
If you become pregnant during or after the study and have VISP, the antibodies 
might be passed to your baby. We know that this happens with some other 
vaccines. The antibodies are not a danger to the baby and they go away, usually in 
about 6 months. 
You should tell the delivery staff if you have VISP. However, you may still be 
tested for HIV using the antibody test when you deliver your baby. If your test is 
positive, your baby may be started on antiretroviral treatment when it is not 
needed. If this happens, we can arrange for you and the baby to have tests that can 
distinguish true HIV infection from VISP. If you or the baby continue to have 
VISP, we can arrange this testing for free for as long as it is needed. 
Embarrassment/anxiety: 
You may feel embarrassed when we ask questions about HIV risks, such as sex 
and using drugs. Also, waiting for HIV test results could make you feel anxious. 
You could feel worried if test results show that you have HIV. If you feel 
embarrassed or anxious, please tell us and we will try to help you. 
Risks of disclosure of your personal information: 
Although the risk is very low, it is possible for someone to see your personal 
information even if they should not be able to. If that happens, you could face 
discrimination, stress, and embarrassment. We can tell you more about how we 
will protect your personal information. 
Risks of genetic testing: 
It is possible that genetic tests could show that you may be at risk for certain 
diseases. If others found out, it could lead to discrimination or other problems. 
However, genetic test results are not part of your study record, so it is almost 
impossible for anyone to connect them to you personally. 
US Sites, include the following paragraph. 
Even if your genetic information somehow gets linked to your name, a federal law 
called the Genetic Information Nondiscrimination Act (GINA) helps protect you. 
GINA keeps health insurance companies and employers from seeing results of 
genetic testing when making decisions about giving you health insurance or 
offering you work. GINA does not help or protect you against discrimination by 
companies that sell life, disability, or long-term care insurance. 
Unknown risks: 

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We do not know if the study vaccines will increase, decrease, or not change your 
risk of getting HIV if exposed. If you get HIV, we do not know how the study 
vaccines might affect your HIV infection or how long it would take to develop 
AIDS. 
We do not know if getting these study vaccines will affect how you respond to 
any future approved HIV vaccine. Currently, no HIV vaccine has been approved 
for use. 
We do not know how the study vaccines will affect a pregnant participant or a 
developing baby. 
Benefits 
20. The study may not benefit you. 
We do not expect the study vaccine to benefit you in any way. However, being in 
the study might still help you in some ways. The counseling that you get as part of 
the study may help you avoid getting HIV. The lab tests and physical exams 
might detect health problems you don’t yet know about. 
When asked, most study participants say that participating in a study made them 
feel good about helping others, increased their knowledge about HIV, and 
improved their self-esteem. 
This study may help in the search for a vaccine to prevent HIV. However, if the 
study vaccine later becomes approved for sale, there are no plans to share any 
money with you. 
Your rights and responsibilities 
21. If you join the study, you have rights and responsibilities. 
We list these in the Bill of Rights and Responsibilities (BRR) for HIV Research. 
We will give you a copy of it. 
Leaving the study 
22. Tell us if you decide to leave the study. 
You are free to leave the study at any time and for any reason. This will not affect 
your care at this clinic and your legal rights. 
Previously collected information about you will remain in the study records and 
will be included in the analysis of results. Your information cannot be removed 
from the study records. 
Whether we take you out of the study or you leave on your own, we will ask you 
to come to the clinic one last time for a physical exam, a questionnaire, and a 

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pregnancy test, if indicated. We may ask to take some blood samples. Whether 
you come for this last visit is up to you. 
Injuries 
23. If you get sick or injured during the study, contact us immediately. 
Your health is important to us. (Sites: Adjust the following 2 sentences if 
applicable to the care available at your site) We will tell you about care that we 
can give here. For care that we cannot provide, we will explain how we will help 
you get care elsewhere. 
If you become sick or injured in this study, the HVTN has a process to decide if 
this is related to the study vaccine and/or procedures. If it is, we call it a “study-
related injury.” There are funds to pay for treatment of study-related injuries if 
certain conditions are met.Next paragraph for non-US sites. 
Sites: adjust the language in this paragraph so it is applicable to your site. Note 
that the ABPI guidelines apply to South Africa only. In this study, our clinic has 
insurance to cover your medical treatment in the case of a study-related injury. 
We will follow the Association of the British Pharmaceutical Industry guidelines 
for payment of study-related injury. We can give you a copy of these guidelines. 
In rare cases, the insurance funds may not be enough. 
In this study, Inovio Pharmaceuticals, Inc. will pay the cost of medical care that 
arises from injuries caused by the EP device and the DNA vaccine, INO-6172. 
AAHI will pay the cost of medical care from injuries caused by the 3M-052-AF 
adjuvant. For injuries caused by other study products or study procedures, the 
HVTN has limited funds to pay medical costs that it determines are reasonable. If 
the injury is not study related, then you and/or your health insurance will be 
responsible for treatment costs. 
Some injuries are not physical. For example, you might be harmed emotionally by 
being in an HIV vaccine study. Or you might lose wages because you cannot go 
to work. However, there are no funds to pay for these kinds of injuries, even if 
they are study related. 
You may disagree with the decision about whether your injury is study related. If 
you wish, the HVTN will ask independent experts to review the decision. You 
always have the right to use the court system if you are not satisfied. 
Questions  
24. If you have questions or problems at any time during your participation in 
this study, use the following important contacts. 
If you have questions about this study, contact 
[name or title and telephone number of the investigator or other study staff]. 

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If you have any symptoms that you think may be related to this study, contact 
[name or title and telephone number of the investigator or other study staff]. 
 
US sites:  
An Institutional Review Board (IRB) is an independent committee established to 
help protect the rights of research participants. If you have any questions about 
your rights as a research participant, and/or concerns or complaints regarding this 
research study, contact: 
By mail: 
Study Subject Adviser 
Advarra IRB 
6100 Merriweather Dr., Suite 600 
Columbia, MD 21044 
or call toll free:    877-992-4724 
or by email:          adviser@advarra.com 
Please reference the following number when contacting the Study 
Subject Adviser: Pro000xxx. 
Remainder of section for South African sites: The study has been structured in 
accordance with the Declaration of Helsinki (last updated October 2013), which 
deals with the recommendations guiding doctors in biomedical research involving 
human participants, the Ethics in Health Research: Principles, Structures and 
Processes Second Edition 2015 and Guidelines for Good Practice in the Conduct 
of Clinical Trials in Human Participants in South Africa. We can provide you 
with copies of these guidelines if you wish to review them. In addition, the recent 
Protection of Personal Information Act (POPIA) ensures that all South African 
institutions conduct themselves in a responsible manner when collecting, 
processing, storing, and sharing another entity’s personal information by holding 
them accountable should they abuse or compromise your personal information in 
any way. 
You may also contact the [name of local IRB/EC] at [insert contact information]. 
If you want to leave this study, contact 
[name or title and telephone number of the investigator or other study staff]. 
You can reach a study staff member 24 hours a day at [telephone number]. 
If you have questions about this trial, you should first discuss them with your 
doctor or the EC (contact details as provided on this form). After you have 
consulted your doctor or the EC, and if they have not provided you with answers 
to your satisfaction, you should write to the South African Healthcare Products 
Regulatory Authority (SAHPRA) at: 

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The Chief Executive Officer 
Dr Boitumelo Semete-Makokotlela 
South African Health Products Regulatory Authority Private Bag X828 
PRETORIA 
0001 
Tel: (012) 501 0410/13  
e-mail: Boitumelo.Semete@sahpra.org.za 
Your permissions and signature  
25. In Section 12 of this form, we told you about an optional sample collection 
procedure if you are in Groups 2 or 3. Please write your initials or make your 
mark in the box next to the procedure if you agree to have this done, or leave 
it blank if you do not want to provide these extra samples. You can change 
your mind after signing and dating this form. There will also be an 
additional consent form to sign and date at the time the procedure is done. 
 
 
I agree to lymph node cell collection. 
26. Above, we told you about possible other uses of your extra samples and 
information outside of this study. Please choose only one of the options below 
and write your initials or make your mark in the box next to it. Whatever 
you choose, the HVTN keeps track of your decision about how your samples 
and information can be used. You can change your mind after signing this 
form. 
 
 
I allow my extra samples and information to be used for other studies related to HIV, 
vaccines, the immune system, and other diseases. This may include genetic testing and 
keeping my cells growing over time.  
OR 
 
 
I agree to the option above and also to allow my extra samples and information to be 
used in genome wide studies. 
OR 
 
 
I do not allow my extra samples to be used in any other studies. This includes not 
allowing genetic testing, growing more of my cells, or genome wide studies.  
 
 

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27. If you agree to join this study, you will need to sign or make your mark 
below. Before you sign or make your mark on this consent form, make sure 
of the following: 
• You have read this consent form, or someone has read it to you. 
• You feel that you understand what the study is about and what will happen to 
you if you join. You understand what the possible risks and benefits are. 
• You have had your questions answered and know that you can ask more. 
• You agree to join this study. 
You will not be giving up any of your rights by signing this consent form. 
 
 
 
 
 
 
 
Participant’s name (print) 
 
Participant’s signature or mark 
 
Date 
 
Time 
 
 
 
 
 
 
 
Clinic staff conducting consent 
discussion (print) 
 
Clinic staff signature 
 
Date 
 
Time 
For participants who are unable to read or write, a witness should complete the 
signature block below: 
 
 
 
 
 
 
 
Witness’s name (print) 
 
Witness’s signature  
 
Date 
 
Time 
*Witness is impartial and was present for the entire discussion of this consent form. 
 
 

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Appendix E 
Sample consent form for use of samples and 
information in other studies 
Title: A phase 1 open-label clinical trial to evaluate the safety and 
immunogenicity of synthetic DNAs encoding NP-GT8 and IL-12, with or without 
aTLR-agonist-adjuvanted HIV Env Trimer 4571 boost, in adults without HIV 
HVTN protocol number: HVTN 305 
Site: [Insert site name] 
Key Information 
These are some of the things you should know about the use of your samples and 
information for other studies: 
• The extra samples will be labeled with a code number. They will not be 
labeled with your name. The extra samples are stored in a secure place. At 
your request, the HVTN will destroy all your extra samples. You can still join 
the main study even if you do not agree to use of your extra samples in other 
studies. 
• Researchers may do genetic testing on your samples, which could include 
genome wide studies. It is unlikely, but these tests could show you may be at 
risk for certain diseases. In the very unlikely event that others found out, this 
could lead to discrimination or other problems.  
• You will not be paid or otherwise benefit from allowing your extra samples to 
be used in other studies.  
The rest of this form gives more information about use of your extra samples for 
other studies. Please read it carefully. 
When samples are no longer needed for this study, the HVTN wants to use them 
in other studies and share them with other researchers. The HVTN calls these 
samples “extra samples.” The HVTN will only allow your extra samples to be 
used in other studies if you agree to this. You will mark your decision at the end 
of this form. If you have any questions, please ask. 
1. Do I have to agree? 
No. You are free to say yes or no, or to change your mind after you sign this form. 
At your request, we will destroy all extra samples that we have. Your decision 
will not affect your being in this study or have any negative consequences here. 
2. Where are the samples stored? 
Extra samples are stored in a secure central place called a repository. [Site: 
choose one of the following two sentences. African sites should choose the 
sentence referencing the repository in South Africa. All other sites should choose 

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the sentence referencing the repository in the United States.] Your samples will 
be stored in the HVTN repository in South Africa. Your samples will be stored in 
the HVTN repository in the United States. 
3. How long will the samples be stored? 
There is no limit on how long your extra samples will be stored. [Site: Revise the 
previous sentence to insert limits if your regulatory authority imposes them.] 
4. Will I be paid for the use of my samples? 
No. Also, a researcher may make a new scientific discovery or product based on 
the use of your samples. If this happens, there is no plan to share any money with 
you. The researcher is not likely to ever know who you are. 
5. Will I benefit from allowing my samples to be used in other studies? 
Probably not. Results from these other studies are not given to you, this clinic, or 
your doctor. They are not part of your medical record. The studies are only being 
done for research purposes. 
6. Will the HVTN sell my samples and information? 
No, but the HVTN may share your samples with HVTN or other researchers. 
Once we share your samples and information, we may not be able to get them 
back. 
7. How do other researchers get my samples and information? 
When a researcher wants to use your samples and information, their research plan 
must be approved by the HVTN. Also, the researcher’s Institutional Review 
Board (IRB) or Ethics Committee (EC) will review their plan. [Site: If review by 
your institution’s IRB/EC/RE is also required, insert a sentence stating this.] 
IRBs/ECs protect the rights and well-being of people in research. If the research 
plan is approved, the HVTN will send your samples to the researcher’s location. 
8. What information is shared with HVTN or other researchers? 
The samples and information will be labeled with a code number. The key to the 
code will stay at this clinic. It will not be shared with anyone who does not need 
to know your name. Your name will not be part of the information. However, 
some information that we share may be personal, such as your race, ethnicity, 
gender, health information from the study, and HIV status. We may share 
information about the study product you received and how your body responded 
to the study product. 

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9. What kind of studies might be done with my extra samples and information? 
The studies will be related to HIV, vaccines, the immune system and other 
diseases. 
Researchers may also do genetic testing on your samples. 
In some cases, researchers may take cells from your samples and grow more of 
them over time, so that they can continue to do research with them. 
If you agree, your samples could also be used for genome wide studies. In these 
studies, researchers will look at all of your genes (your genome). The researchers 
compare the genomes of many people, looking for common patterns of genes that 
could help them understand diseases. The researchers may put the information 
from the genome-wide studies into a protected database so that other researchers 
can access it, but your name and other personal information will not be included. 
Usually, no one would be able to look at your genome and link it to you as a 
person. However, if another database exists that also has information on your 
genome and your name, someone might be able to compare the databases and 
identify you. If others found out, it could lead to discrimination or other problems. 
The risk of this is very small. There may be other unknown risks. 
10. What are the risks of genetic testing? 
It is unlikely, but the genetic tests done on your samples could show you may be 
at risk for certain diseases. If others found out, it could lead to discrimination or 
other problems. However, it is almost impossible for you or others to know your 
test results from the genetic testing. The results are not part of your study records 
and are not given to you. 
11. Who will have access to my information in studies using my extra samples? 
People who may see your information are: 
• Researchers who use your extra samples and information for other research 
• Government agencies that fund or monitor the research using your extra 
samples and information 
• The researcher’s Institutional Review Board or Ethics Committee 
• Any regulatory agency that reviews clinical trials 
• The people who work with the researcher 
All of these people will do their best to protect your information. The results of 
any new studies that use your extra samples and information may be published. 
No publication will use your name or identify you personally. 
 

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Questions  
12. If you have questions or problems about allowing your samples and 
information to be used in other studies, use the following important contacts. 
If you have questions about the use of your samples or information or if you want 
to change your mind about their use, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you think you may have been harmed because of studies using your samples or 
information, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you have questions about your rights as a research participant, contact 
[name or title and telephone number of person on IRB/EC . 
The study has been structured in accordance with the Declaration of Helsinki (last 
updated October 2013) which deals with the recommendations guiding doctors in 
biomedical research involving human participants, the Ethics in Health Research: 
Principles, Structures and Processes Second Edition 2015, and Guidelines for 
Good Practice in the Conduct of Clinical Trials in Human Participants in South 
Africa. We can provide you with copies of these guidelines if you wish to review 
them. In addition, the recent Protection of Personal Information Act (POPIA) 
ensures that all South African institutions conducts themselves in a responsible 
manner when collecting, processing, storing and sharing another entity’s personal 
information by holding them accountable should they abuse or compromise your 
personal information in any way. 
You can reach a study staff member 24 hours a day at [telephone number]. 
If you have questions about this trial you should first discuss them with your 
doctor or the ethics committee (contact details as provided on this form). After 
you have consulted your doctor or the ethics committee and if they have not 
provided you with answers to your satisfaction, you should write to the South 
African Healthcare Products Regulatory Authority (SAHPRA) at: 
The Chief Executive Officer 
Dr Boitumelo Semete-Makokotlela 
South African Health Products Regulatory Authority Private Bag X828 
PRETORIA 
0001 
Tel: (012) 501 0410/13 
e-mail: Boitumelo.Semete@sahpra.org.za 
13. Please choose only one of the options below and write your initials or make 
your mark in the box next to it. Whatever you choose, the HVTN keeps track 

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of your choice about how your samples and information can be used. You 
can change your mind after signing this form. 
 
 
I allow my extra samples and information to be used for other studies related to 
HIV, vaccines, the immune system, and other diseases. This may include 
genetic testing and keeping my cells growing over time. 
OR 
 
 
I agree to the option above and also to allow my extra samples and information 
to be used in genome wide studies. 
 
OR 
 
 
I do not allow my extra samples to be used in any other studies. This includes 
not allowing genetic testing, growing more of my cells, or genome wide studies.  
 
 
 
 
 
 
 
 
 
 
Participant’s name (print) 
 
Participant’s signature or mark 
 
Date 
 
Time 
 
 
 
 
 
 
 
Clinic staff conducting consent 
discussion (print) 
 
Clinic staff signature 
 
Date 
 
Time 
For participants who are unable to read or write, a witness should complete the 
signature block below: 
 
 
 
 
 
 
 
Witness’s name (print) 
 
Witness’s signature  
 
Date 
 
Time 
*Witness is impartial and was present for the entire discussion of this consent form. 
 
 
 

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Appendix F 
Low-risk guidelines for US 
The following are intended as guidelines for the investigator to help identify potential 
vaccine trial participants at “low risk” for HIV infection. These guidelines are based 
on behaviors within the last 6 to 12 months prior to enrollment; however, it may be 
appropriate to consider a person’s behavior over a longer period of time than 
specified in order to assess the person’s likelihood of maintaining low-risk behavior. 
Some volunteers may not be appropriate for enrollment even if they meet these 
guidelines. These guidelines should be supplemented and interpreted with local 
epidemiologic information about HIV prevalence in your area and community 
networks. The investigator may review the risk level of any volunteer with the site PI 
and/or the Protocol Safety Review Team (PSRT). 
A volunteer may be appropriate for inclusion if he/she/they meets these guidelines: 
 
For US volunteers NOT on stable Pre-exposure prophylaxis 
(PrEP) 
1. SEXUAL BEHAVIORS 
In the last 12 months, did not: 
• Have oral, vaginal, or anal intercourse with an HIV-infected partner or a 
partner who uses injection drugs 
• Give or receive money, drugs, gifts, or services in exchange for oral, vaginal, 
or anal sex 
AND 
 
In the last 6 months, has abstained from penile/anal or penile/vaginal intercourse, 
OR 
In the last 6 months, had 4 or fewer partners of the opposite birth sex for vaginal 
and/or anal intercourse, OR 
Is a person born male with partner(s) born male (MSM) who, in the last 12 
months: 
• Had 2 or fewer MSM partners for anal intercourse and had no unprotected 
anal sex with MSM, OR 
• Had unprotected anal intercourse with only 1 MSM partner within a 
monogamous relationship lasting at least 12 months (during which neither 
partner had any other partners). If the monogamous relationship ended, the 

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volunteer may then have had protected anal intercourse with 1 other MSM 
partner (total 2 or fewer partners in the last 12 months), OR  
Is a transgender person, regardless of the point on the transition spectrum, having sex 
with men (born male) and/or other transgender persons, who in the last 12 months: 
• Had 2 or fewer partners for anal or vaginal intercourse, and had no 
unprotected anal or vaginal sex, OR  
• Had unprotected anal or vaginal intercourse sex with 1 partner only within 
a monogamous relationship lasting at least 12 months (during which 
neither partner had any other partners). If the monogamous relationship 
ended, may then have had protected anal or vaginal sex with 1 other 
partner (total 2 or fewer partners in the last 12 months). 
AND 
 
Uses or intends to use condoms in situations which may include penile/anal or 
penile/vaginal intercourse with new partners of unknown HIV status, occasional 
partners, partners outside a primary relationship, and/or partners known to have 
other partners. 
 
2. NONSEXUAL BEHAVIORS 
In the last 12 months, did not: 
• Inject drugs or other substances without a prescription 
• Use cocaine, methamphetamine, or excessive alcohol that, in the 
investigator’s judgment, rendered the participant at greater-than-low risk for 
acquiring HIV infection. 
The investigator’s judgment should consider local epidemiologic information 
about HIV prevalence in the area and community networks. 
A volunteer is NOT appropriate for inclusion if he/she: 
Acquired an STI (ie, new infection) in the last 12 months: 
• Syphilis 
• Gonorrhea 
• Nongonococcal urethritis 
• Herpes simplex virus type 2 (HSV2) 
• Chlamydia 

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• Pelvic inflammatory disease (PID) 
• Trichomonas 
• Mucopurulent cervicitis 
• Epididymitis 
• Proctitis 
• Lymphogranuloma venereum 
• Chancroid 
• Hepatitis B 
For US volunteers on Pre-exposure prophylaxis (PrEP) 
• PrEP ASSESSMENT 
• Reports 6 months (180 days) or longer of protective PrEP use 
• For daily oral PrEP use:  
o For persons AMAB who have sex with persons AMAB: Reports equal to 
or greater than 70% when asked the following: “Thinking about the past 4 
weeks, what percent of the time were you able to take all your PrEP 
medications?” 
o For people with a vagina having intravaginal intercourse: Reports equal to 
or greater than 90% when asked the following: “Thinking about the past 4 
weeks, what percent of the time were you able to take all your PrEP 
medications?” 
• For event-driven (on-demand or “2-1-1”) PrEP use1 in persons AMAB who 
have sex with persons AMAB: Reports use consistent with the guidance of the 
professional education organization, the International Antiviral Society–USA 
(IAS-USA): 
o For individuals with frequent use (> 15 pills per month): At least 80% 
of condomless sex acts are covered with on-demand PrEP at the 
recommended dose schedule 
o For individuals with less frequent use (≤ 15 pills per month): 
 A past history of high adherence (> 90%) 

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 Commitment to use on-demand PrEP for all condomless sex 
acts at the recommended dose schedule 
• For long-acting agents, such as injectable cabotegravir, please consult with the 
PSRT. 
• Commits to maintaining protective PrEP use throughout trial 
SEXUAL BEHAVIORS 
Persons stably taking PrEP as described above for 6 months or longer are considered 
low risk of HIV infection, regardless of any sexual behavior that might otherwise be 
associated with high risk of HIV exposure.  
NONSEXUAL BEHAVIORS 
In the last 12 months, did not: 
• Inject drugs or other substances without a prescription 
• Use cocaine, methamphetamine, or excessive alcohol that, in the 
investigator’s judgment, rendered the participant at greater-than-low risk for 
acquiring HIV infection. 
The investigator’s judgment should consider local epidemiologic information about 
HIV prevalence in the area and community networks. 

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Appendix G 
Low-risk guidelines for South Africa 
The following are intended as guidelines for the investigator to help identify potential 
vaccine trial participants at “low risk” for HIV infection. These guidelines are based 
on behaviors within the last 12 months prior to enrollment; however, it may be 
appropriate to consider a person’s behavior of maintaining low-risk behavior. Some 
volunteers may not be appropriate for enrollment even if they meet these guidelines. 
These guidelines should be supplemented and interpreted with local epidemiological 
information about the HIV prevalence in your area and community networks. The 
investigator may review the risk level of any volunteer with the site PI and/or the 
Protocol Safety Review Team (PSRT). 
Assessment of sexual behaviors 
Consider whether a volunteer would be appropriate for inclusion if, within 12 months 
prior to enrollment, the person: 
• Abstained from penile/vaginal and penile/anal intercourse, or 
• Was in a mutually monogamous relationship with a partner with a known 
HIV-uninfected status, or 
• Had 1 partner believed to be HIV-uninfected with whom he/she regularly used 
condoms for penile/vaginal or penile/anal intercourse. 
Exclude a volunteer if: 
Within 12 months prior to enrollment: a history of newly acquired syphilis; 
gonorrhea; chlamydia; trichomoniasis; active HSV lesions; chancroid; genital warts 
of the labia minora, vagina, or cervix; or any other symptomatic genital warts. 

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Appendix H 
Approved birth control methods 
Site: Any change to the following boxed text requires approval from HVTN 
Regulatory Affairs at vtn.core.reg@hvtn.org. You can remove the box around the text. 
It is important that you not become pregnant during the study because we do not 
know how the study vaccine could affect the developing baby. 
You must agree to use effective birth control from 21 days before your first 
injection until 8 weeks after your last study injection at 6 months (9 months total).  
Effective birth control means using any of the following methods every time you 
have sex: 
• Birth control drugs that prevent pregnancy—given by pills, shots, patches, 
vaginal rings, or inserts under the skin; 
• Male or female condoms, with or without a cream or gel that kills sperm; 
• Diaphragm or cervical cap with a cream or gel that kills sperm; 
• Intrauterine device (IUD); or 
• Any other contraceptive method approved by the researchers. 
You do not have to use birth control if: 
• You are only having sex with a partner or partners who have had a vasectomy. 
(We will ask you some questions to confirm that the vasectomy was 
successful.); 
• You have reached menopause, with no menstrual periods for one year; 
• You have had a hysterectomy (your uterus removed); 
• You have had your ovaries removed; 
• You have a tubal ligation (your “tubes tied”) or confirmed successful 
placement of a product that blocks the fallopian tubes; 
• You are having sex only with a partner(s) assigned female sex at birth; 
• You only have oral sex; or, 
• You are sexually abstinent (no sex at all). 

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Remember: If you are having sex, male and female condoms are the only birth 
control methods that also provide protection against HIV and other sexually 
transmitted infections. 
If you join the study, we will test you for pregnancy at some visits, including 
before each study injection. 

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Appendix I 
Adverse events of special interest (AESIs) 
AESIs for this protocol include, but are not limited to, potential immune-mediated 
diseases; representative examples of AESIs are listed below. Updates to AESIs will 
be provided as an appendix to the HVTN305 Study Specific Procedures (SSPs). 
Neuroinflammatory disorders 
Musculoskeletal disorders 
Skin disorders 
• 
Cranial nerve disorders, including 
paralyses/paresis (eg, Bell’s palsy)  
• 
Optic neuritis 
• 
Multiple sclerosis 
• 
Transverse myelitis 
• 
Guillain-Barré syndrome, including 
Miller Fisher syndrome and other 
variants 
• 
Acute disseminated 
encephalomyelitis, including site 
specific variants: eg, noninfectious 
encephalitis, encephalomyelitis, 
myelitis, myeloradiculoneuritis 
• 
Myasthenia gravis, including 
Lambert-Eaton myasthenic syndrome  
• 
Immune-mediated peripheral 
neuropathies and plexopathies, 
(including chronic inflammatory 
demyelinating polyneuropathy, 
multifocal motor neuropathy and 
polyneuropathies associated with 
monoclonal gammopathy). 
• 
Narcolepsy 
• 
Systemic lupus erythematosus and 
associated conditions 
• 
Systemic scleroderma (Systemic 
sclerosis), including diffuse systemic 
form and CREST syndrome 
• 
Idiopathic inflammatory myopathies, 
including dermatomyositis  
• 
Polymyositis  
• 
Antisynthetase syndrome 
• 
Rheumatoid arthritis, and associated 
conditions including juvenile chronic 
arthritis and Still’s disease 
• 
Polymyalgia rheumatica 
• 
Spondyloarthritis, including 
ankylosing spondylitis, reactive 
arthritis (Reiter's Syndrome) and 
undifferentiated spondyloarthritis  
• 
Psoriatic arthropathy 
• 
Relapsing polychondritis 
• 
Mixed connective tissue disorder 
• 
Psoriasis 
• 
Vitiligo 
• 
Erythema nodosum 
• 
Autoimmune bullous skin 
diseases (including pemphigus, 
pemphigoid and dermatitis 
herpetiformis) 
• 
Alopecia areata 
• 
Lichen planus 
• 
Sweet’s syndrome 
• 
Localized Scleroderma 
(Morphea)  
• 
Cutaneous lupus erythematosus 
 
Metabolic disorders 
• 
Addison’s disease 
• 
Autoimmune thyroiditis 
(including Hashimoto 
thyroiditis) 
• 
Diabetes mellitus type I 
• 
Grave's or Basedow’s disease 
Vasculitides 
Blood disorders 
Others 
• 
Large vessels vasculitis including: 
giant cell arteritis such as Takayasu’s 
arteritis and temporal arteritis.  
• 
Medium sized and/or small vessels 
vasculitis including: polyarteritis 
nodosa, Kawasaki's disease, 
microscopic polyangiitis, Wegener’s 
granulomatosis, Churg-Strauss 
syndrome (allergic granulomatous 
angiitis), Buerger’s disease 
(thromboangiitis obliterans), 
necrotizing vasculitis and 
antineutrophil cytoplasmic antibody 
(ANCA) positive vasculitis (type 
unspecified), Henoch-Schonlein 
purpura, Behcet’s syndrome, 
leukocytoclastic vasculitis. 
• 
Autoimmune hemolytic anemia 
• 
Autoimmune thrombocytopenia 
• 
Antiphospholipid syndrome 
• 
Pernicious anemia 
• 
Autoimmune aplastic anemia 
• 
Autoimmune neutropenia  
• 
Autoimmune pancytopenia 
 
Gastrointestinal disorders 
• 
Celiac disease 
• 
Crohn’s disease 
• 
Ulcerative colitis 
• 
Ulcerative proctitis 
 
Liver disorders 
• 
Autoimmune cholangitis 
• 
Autoimmune hepatitis 
• 
Primary biliary cirrhosis 
• 
Primary sclerosing cholangitis 
• 
Autoimmune 
glomerulonephritis (including 
IgA nephropathy, 
glomerulonephritis rapidly 
progressive, membranous 
glomerulonephritis, 
membranoproliferative 
glomerulonephritis, and 
mesangioproliferative 
glomerulonephritis) 
• 
Ocular autoimmune diseases 
(including autoimmune uveitis 
and autoimmune retinopathy) 
• 
Autoimmune 
myocarditis/cardiomyopathy 
• 
Sarcoidosis 
• 
Stevens-Johnson syndrome 
• 
Sjögren’s syndrome 
• 
Idiopathic pulmonary fibrosis 
• 
Goodpasture syndrome 
• 
Raynaud’s phenomenon 

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Appendix J 
Protocol team 
Protocol leadership 
Chair 
Sonya Heath 
University of Alabama, 
Birmingham 
205-996-2371 
slheath@uabmc.edu 
 
Statistician 
Ollivier Hyrien 
HVTN Statistics and Data 
Management Center (SDMC), 
Fred Hutch 
206-667-2809 
ohyrien@fredhutch.org 
Cochairs 
Amy Sherman 
Brigham and Women’s 
Hospital, Division of Infectious 
Diseases, Harvard Medical 
School  
617-525-3575 
acsherman@bwh.harvard.edu  
 
Nivashnee Naicker 
Centre for the AIDS 
Programme of Research In 
South Africa (CAPRISA), 
University of KwaZulu-Natal 
+27 (0)31-655-0618  
Nivashnee.naicker@caprisa.org  
 
DAIDS Medical 
officers 
Chris Duplessis 
DAIDS, NIAID 
240-292-0676 
chris.duplessis@nih.gov  
 
 
Protocol Team leaders 
Will Hahn 
HVTN LOC, Fred Hutch 
206-667-3431 
whahn@fredhutch.org  
 
Briana Furch 
HVTN LOC, Fred Hutch 
206-667-6607 
bfurch2@fredhutch.org 
 
 
Laboratory lead 
Guido Ferrari  
HVTN Laboratory Center 
919-684-2862 
guido.ferrari@duke.edu 
 
 
 
 
 
 
 
 
 

HVTN 305 Version 2.0 / Jan 06, 2023 
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Other contributors to the original protocol 
 
Core medical monitors 
Will Hahn 
HVTN LOC, Fred 
Hutch 
Briana Furch 
HVTN LOC, Fred 
Hutch 
Clinical safety specialists 
 
Regional Medical Liasons 
Nadia McCloskey 
Maija Anderson 
HVTN LOC, Fred Hutch 
Azwi Takalani 
Simone Hendricks 
HVTN LOC, Fred Hutch 
 
Study product developer 
representatives 
 
 
 
David Weiner 
The Wistar Institute 
Dan Kulp 
The Wistar Institute 
 
Clinical trials managers 
Lori Proulx-Burns 
HVTN LOC, Fred Hutch 
Shelly Ramirez 
HVTN LOC, Fred Hutch 
INOVIO representatives 
 
Laurent Humeau 
Linda Dickinson 
Clinical data managers 
Jennifer Schille 
Andrea Repetto 
HVTN SDMC, Fred Hutch 
VRC representative 
Lucio Gama 
VRC, NIAID 
SDMC Associate director of 
lab science 
April Randhawa 
HVTN SDMC, Fred Hutch 
Laboratory center representative 
Jessica Brown 
HVTN Laboratory 
Center, Fred Hutch 
DAIDS Product Lead 
Sujata Vijh 
DAIDS, NIAID 
Regulatory affairs associate 
Meg Brandon 
HVTN LOC, Fred 
Hutch 
Protocol development 
managers 
Smitha Sripathy 
HVTN LOC, Fred Hutch 
Meg Trahey 
HVTN LOC, Fred Hutch 
Clinic coordinators 
Bongi Zuma 
CAPRISA 
University of KwaZulu-
Natal, South Africa. 
 
Community engagement unit 
representatives 
Gail Broder  
HVTN LOC, Fred Hutch 
Kagisho Baepanye 
HVTN, Fred Hutch/HCRISA 
Matshidiso Malefo 
HVTN, Fred Hutch/HCRISA 
Community Advisory Board (CAB) 
members 
Sibusiso Mngadi 
CAPRISA CAB 
Rick Church 
New York City CAB 
Community 
educator/recruiter 
Andres Avila Paz 
Brigham and Women’s 
Hospital CRS 
DAIDS protocol pharmacists 
Justine Beck 
DAIDS, NIAID 
Rochelle Rogers 
DAIDS, NIAID 
 
Statistical research associate 
Randy Burnham 
HVTN SDMC, Fred Hutch 
 
 
 
Technical editor 
Sydney Strauss 
HVTN LOC, Fred Hutch 

HVTN 305 Version 2.0 / Jan 06, 2023 
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Appendix K 
Version history 
The Protocol Team may modify the original version of the protocol. Modifications 
are made to HVTN protocols via clarification memos, letters of amendment, or full 
protocol amendments. 
The version history of, and modifications to, Protocol HVTN 305 are described 
below. 
Protocol history and modifications 
Date: Jan 06, 2023 
Protocol version: Version 2.0 
Protocol modification: Full protocol Amendment 1 
  
Added in Section 5.1.1, Inclusion criteria; Appendix A, Schedule of Procedures: 
Participants undergoing FNA and Appendix B, Schedule of procedures: 
Participants not undergoing FNA : serum calcium 
  
Revised in Section 5.1.1, Inclusion criteria: criterion for blood pressure 
  
Removed in Section 9.1, AEs: grading Creatinine 
  
Added in Section 2.13.1, Potential Risks and in Appendix D, Sample informed 
consent form (SICF): risk of hypocalcemia 
  
Added in Appendix D, Sample informed consent form (SICF): study related injury 
compensation 
  
Updated in Section 5.1.2, Exclusion criteria and Section 5.2.1, Delaying 
vaccinations for a participant: nomenclature for Monkeypox 
  
Corrected in Section 7.1, Vaccine regimen: injection volume 
 
Updated in Appendix C, Visit windows: interval between last vaccination and 
AESI contact 
  
Corrected in Appendix C, Visit windows: Lower target window 
  Updated in Title page and Appendix K, Version History: IND number and 
contents of this amendment 
 
Corrected throughout the Protocol: minor errors in grammar, typography, 
formatting 

HVTN 305 Version 2.0 / Jan 06, 2023 
HVTN305_v2.0_Final_06Jan23 / Page 129 of 129 
Date: Sep 21, 2022 
Protocol version: v1.0 
Protocol modification: None