A Phase 1 Open-label Clinical Trial to Evaluate the Safety and Immunogenicity of Synthetic DNAs Encoding a Native-like HIV Env Trimer and Interleukin-12 (INO-6160), Alone or in a Prime-boost Regimen With 3M-052-AF + Alum Adjuvanted VRC HIV Env Trimer 4571 in Adult Participants Without HIV
Phase: PHASE1
Conditions: HIV-1-infection
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

PROTOCOL 
HVTN 304 
A phase 1 open-label clinical trial to evaluate the safety and 
immunogenicity of synthetic DNAs encoding a native-like 
HIV Env Trimer and Interleukin-12 (INO-6160), alone or in a 
prime-boost regimen with 3M-052-AF + Alum adjuvanted 
VRC HIV Env Trimer 4571 in adult participants without HIV 
DAIDS DOCUMENT ID 38861 
IND 029064 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 
Division of AIDS (DAIDS) 
Bethesda, Maryland, USA 
The Wistar Institute 
 Philadelphia, Pennsylvania, USA 
Access to Advanced Health Institute (AAHI) 
Seattle, Washington, USA 
The Dale and Betty Bumpers Vaccine Research Center (VRC) 
Bethesda, Maryland, USA 
Inovio Pharmaceuticals, Inc. 
Plymouth Meeting, Pennsylvania, USA 
January 11, 2023 
Final 
HVTN 304 
Version 2.0 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 2 of 107 
Protocol Signature Page 
 
A phase 1 open-label clinical trial to evaluate the safety and immunogenicity of 
synthetic DNAs encoding a native-like HIV Env Trimer and Interleukin-12 (INO-
6160), alone or in a prime-boost regimen with 3M-052-AF + Alum adjuvanted 
VRC HIV Env Trimer 4571 in adult participants 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 (U.S.) Health and Human Service regulations (45 
CFR 46); applicable U.S. Food and Drug Administration regulations; standards of 
the International Council 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, U.S. National Institutes of Health, Division of AIDS) and 
institutional policies 
 
 
 
 
 
 
 
 
 
Investigator of Record Name (print) 
 
Investigator of Record Signature 
 
Date 
 
 
 
DAIDS Protocol Number: HVTN 304 
DAIDS Protocol Version: Version 2.0 
Protocol Date: January 11, 2023 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 3 of 107 
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 
ALT 
 
alanine transaminase 
Alum 
 
aluminum hydroxide suspension 
ANCA 
 
anti-neutrophil cytoplasmic antibody 
AVEG 
 
AIDS Vaccine Evaluation Group 
AoU 
 
assessment of understanding 
BAMA 
 
binding antibody multiplex assay 
BCR 
 
B-cell receptor 
β-HCG 
 
beta human chorionic gonadotropin 
BMI 
 
body mass index 
bnAb 
 
broadly neutralizing antibody 
BRR 
 
Bill of Rights and Responsibilities 
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 
DHHS 
 
Department of Health and Human Services 
DM 
 
diabetes mellitus 
EAE 
 
expedited adverse event 
EC 
 
Ethics Committee 
ECLIA 
 
Electrochemiluminescence 
eGFR 
 
estimated glomerular filtration rate 
EIA 
 
enzyme immunoassay 
Env 
 
HIV envelope protein 
EP 
 
electroporation 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 4 of 107 
FDA 
 
US Food and Drug Administration 
GCP 
 
Good Clinical Practice 
GEE 
 
generalized estimating equation 
GINA 
 
Genetic Information Nondiscrimination Act 
GPP 
 
Good Participatory Practices 
HCV 
 
Hepatitis C antibody 
HVTN 
 
HIV Vaccine Trials Network 
IB 
 
Investigator’s Brochure 
IBC 
 
Institutional Biosafety Committee 
ICABA 
 
infected cell antibody-binding assay 
ID  
 
intradermal  
IFN-gamma 
interferon-gamma 
IM 
 
intramuscular 
IND 
 
investigational new drug application 
IRB 
 
Institutional Review Board 
IUD 
 
intrauterine device 
LABA 
 
long-acting beta agonist 
MAAE 
 
medically attended adverse event 
MAR 
 
missing at random 
MCAR 
 
missing completely at random 
MedDRA  
Medical Dictionary for Regulatory Activities 
MO 
 
Medical Officer 
MSD 
 
Meso Scale Discovery 
nAb 
 
neutralizing antibody 
NAT 
 
nucleic acid test 
NHP 
 
nonhuman primate 
NIAID 
 
National Institute of Allergy and Infectious Diseases 
NIH 
 
National Institutes of Health 
NLT 
 
native-like trimer 
NSAID 
 
non-steroidal anti-inflammatory drug 
OHRP 
 
Office for Human Research Protections 
PAB 
 
DAIDS Pharmaceutical Affairs Branch 
PBMC 
 
peripheral blood mononuclear cell 
PBS 
 
phosphate-buffered saline 
PI 
 
principal investigator 
PID 
 
pelvic inflammatory disease 
PrEP 
 
HIV pre-exposure prophylaxis 
PSRT 
 
Protocol Safety Review Team 
PV 
 
PENNVAX-B DNA vaccine 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 5 of 107 
RAB 
 
DAIDS Regulatory Affairs Branch 
RE 
 
Regulatory Entity 
RSC 
 
Regulatory Support Center 
SAE 
 
serious adverse event 
sD 
 
synthetic DNA 
SDMC 
 
Statistics and Data Management Center 
SHIV 
 
simian-human immunodeficiency virus 
SICF 
 
sample informed consent form 
SMB 
 
Safety Monitoring Board 
SPT 
 
DAIDS Safety and Pharmacovigilance Team 
SSP 
 
Study-specific Procedures 
SUSAR 
 
suspected unexpected serious adverse reaction 
Th1 
 
type 1 T helper 
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 
WFI 
 
water for injection 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 6 of 107 
Contents 
1 Executive summary ........................................................................................................8 
1.1 
Title ................................................................................................................8 
1.2 
Design .............................................................................................................8 
1.3 
Study products, diluents, and electroporation device .....................................8 
1.4 
Study participants ...........................................................................................9 
1.5 
Study plan and schema table ..........................................................................9 
1.6 
Duration per participant ...............................................................................10 
1.7 
Estimated total study duration ......................................................................10 
1.8 
Study sites ....................................................................................................10 
2 Introduction ..................................................................................................................11 
2.1 
Rationale for evaluation of sD-NLT-AB05 with IL-12 adjuvant and Trimer 
4571 adjuvanted with 3M-052-AF + Alum in a prime boost regimen.........11 
2.2 
Preclinical data with sD-NLT-AB05: mice and rabbits ...............................13 
2.3 
Clinical Experience with related DNA vaccines encoding HIV Env 
immunogens with and without IL-12 adjuvant ............................................16 
2.4 
Clinical experience with HIV Envelope proteins adjuvanted with 3M-052-
AF + Alum: safety data ................................................................................19 
2.5 
Rationale for Schedule .................................................................................21 
2.6 
Rationale for Dose of Trimer 4571 and 3M-052-AF + Alum adjuvant .......23 
2.7 
Rationale for Targeting Same Draining Lymph Node with both DNA and 
Protein ..........................................................................................................24 
2.8 
Risks and benefits .........................................................................................27 
3 Objectives and endpoints .............................................................................................29 
3.1 
Primary objectives and endpoints ................................................................29 
3.2 
Secondary objectives and endpoints ............................................................29 
3.3 
Exploratory objectives ..................................................................................30 
4 Laboratory strategy ......................................................................................................32 
4.1 
Assessment of immunogenicity endpoints ...................................................32 
5 Study design .................................................................................................................33 
5.1 
Study population ..........................................................................................33 
5.2 
Participant departure from vaccination schedule or withdrawal ..................38 
6 Statistical considerations ..............................................................................................42 
6.1 
Sample size justification and accrual ...........................................................42 
6.2 
Randomization .............................................................................................46 
6.3 
Blinding ........................................................................................................46 
6.4 
Statistical analyses ........................................................................................46 
7 Study vaccine preparation, storage, and administration ..............................................51 
7.1 
Vaccine Regimen .........................................................................................51 
7.2 
Study Product Formulation and Storage ......................................................51 
7.3 
Product Preparation ......................................................................................52 
7.4 
Acquisition of study products ......................................................................55 
7.5 
Study Vaccine Accountability ......................................................................56 
7.6 
Final Disposition of study product ...............................................................56 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 7 of 107 
8 Clinical procedures ......................................................................................................57 
8.1 
Screening ......................................................................................................57 
8.2 
Definition of Study Day and Study Visit .....................................................57 
8.3 
Reactogenicity Assessments ........................................................................57 
8.4 
EP-injection–site assessment ........................................................................58 
8.5 
Visit windows and missed visits ..................................................................59 
8.6 
Monitoring for HIV infection .......................................................................59 
8.7 
Early termination visit ..................................................................................60 
8.8 
AESI contact ................................................................................................60 
9 Safety and adverse events ............................................................................................61 
9.1 
Adverse events .............................................................................................61 
9.2 
Serious adverse events .................................................................................62 
9.3 
Safety monitoring .........................................................................................63 
9.4 
Total blood volume ......................................................................................64 
9.5 
Initial safety review ......................................................................................64 
9.6 
Safety pause and prompt PSRT AE review..................................................64 
9.7 
Study termination .........................................................................................66 
9.8 
Pregnancy .....................................................................................................66 
10 Protocol conduct and informed consent .......................................................................67 
10.1 Protocol conduct ...........................................................................................67 
10.2 Compliance with NIH guidelines for research involving products containing 
recombinant or synthetic nucleic acid molecules .........................................67 
10.3 Informed consent ..........................................................................................67 
11 Exploratory studies ......................................................................................................69 
11.1 Specimen storage and other use of specimens .............................................69 
12 Literature cited .............................................................................................................70 
Appendix A Schedule of procedures ................................................................................74 
Appendix B Visit windows ...............................................................................................76 
Appendix C Sample informed consent form ....................................................................77 
Appendix D Low-risk guidelines for US ..........................................................................99 
Appendix E Approved birth control methods ................................................................103 
Appendix F Adverse events of special interest ..............................................................105 
Appendix G Protocol team ..............................................................................................106 
Appendix H Version history ...........................................................................................107 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 8 of 107 
1 
Executive summary 
1.1 
Title  
A phase 1 open-label clinical trial to evaluate the safety and immunogenicity of 
synthetic DNAs encoding a native-like HIV Env Trimer and Interleukin-12 (INO-
6160), alone or in a prime-boost regimen with 3M-052-AF + Alum adjuvanted 
VRC HIV Env Trimer 4571 in adult participants without HIV. 
1.2 
Design 
This is a randomized open-label trial to examine the safety and immunogenicity 
of INO-6160 (synthetic DNAs encoding a native-like HIV Env Trimer and 
Interleukin-12), alone or in a prime-boost regimen with VRC HIV Env Trimer 
4571 adjuvanted with 3M-052-AF + Alum. The primary hypothesis is that the 
vaccine regimen will elicit HIV-1 envelope protein–specific binding antibody 
(Ab) and T-cell responses. 
1.3 
Study products, diluents, and electroporation device 
• INO-6160: sD-NLT-AB05 co-formulated with IL-12 DNA (pGX6001): sD-
NLT-AB05 consists of a single plasmid, pGX1060 (in pGX0001 vector 
backbone), encoding a soluble stabilized native-like trimer derived from clade 
A isolate BG505. pGX6001 (pGX0003 vector backbone), contains a dual 
promoter system for expression of both the human IL-12 p35 and p40 genes 
necessary for production of the active heterodimeric IL-12 protein. The 
plasmid ratio for the coformulated drug product is 4:1 (0.8 mg pGX1060/0.2 
mg pGX6001) per 0.1 mL/1 mg injection. The coformulation, INO-6160, in 
water-for-injection (WFI), is supplied at a concentration of 10 mg/mL and a 
volume of 0.4 mL in 2-mL glass vials. 
• Trimer 4571: HIV-1 Env Trimer 4571 (VRC-HIVRG096-00-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 adjuvant: This adjuvant is an aqueous formulation (AF) of the 
small molecule imidazoquinoline, which acts as a toll-like receptor (TLR) 7/8 
agonist. 3M-052-AF is supplied at a concentration of 50 mcg/mL and a fill 
volume of 0.4 mL in 2-mL glass vials. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 9 of 107 
• Aluminum Hydroxide Suspension, Adjuvant: Aluminum hydroxide 
suspension (Alum) is composed of Alhydrogel 2% (Brenntag Biosector, 
Frederikssund, Denmark) diluted with WFI to a concentration of 5 mg/mL. It 
is supplied as a sterile, pyrogen-free suspension filled into single-dose vials at 
a volume of 0.7 mL.  
• 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 and is provided by Inovio 
Pharmaceuticals.  
1.4 
Study participants 
20 healthy volunteers without HIV, 18 through 55 years of age. 
1.5 
Study plan and schema table 
Participants will be evaluated for safety and immune responses through blood 
collection at specified timepoints throughout the study. The study schema is 
below: 
Table 1-1 Schema 
 
 
 
 
Injection Schedule 
Group 
N 
Product/Dose 
Route 
Month 
0 
Month 
1 
Month 
3 
Month 
6 
1 
10 
INO-6160 / 2.0 mg 
ID EP 
X 
X 
X 
X 
2 
10 
INO-6160 / 2.0 mg 
ID EP 
X 
X 
X 
X 
Trimer-4571 / 100 mcg 
3M-052-AF (5 mcg) + 
Alum (500 mcg) 
IM 
-- 
-- 
X 
X 
Total 
     20* 
Table 1-1 Notes: The dose of INO-6160, 2 mg, will be administered as 2 separate 
intradermal (ID) injections because of the volume limitation of the device for 
intradermal injection. Each of 2 sites will receive 0.1 mL via ID injection (Mantoux 
injection) bilaterally, one on each upper arm. Following ID injections, EP will be 
performed with the Inovio CELLECTRA 2000 device. The dose of Trimer 4571, 100 
mcg, will be administered as 2 injections delivered intramuscularly (IM) via needle and 
syringe in both deltoid muscles. For all injections, if administration into the deltoid is 
contraindicated, the thigh muscle can be used (see Section 7.3.4). 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 10 of 107 
* Up to 5 additional participants may be enrolled (for a total of 25), if needed, with a 
goal of approximately 20 participants to contribute to the 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, or variations in the clinical care due to unpredictable events. Participants will not 
be replaced after visit 5, the visit 2 weeks post second vaccination, and replacement 
will require the assent of Protocol team leadership. 
Enrollment will be restricted to 1 participant per day for the first 5 participants (across 
both arms) and enrollment will pause after the first 5 participants are enrolled. The 
Protocol Safety Review Team (PSRT) will review cumulative safety information for all 
participants recorded through the visit scheduled 2 weeks post first vaccination for the 
first 5 participants and will determine whether it is safe to proceed with full enrollment. 
1.6 
Duration per participant 
12 months of scheduled clinic visits (main study) and an AESI health contact at 
month 18. 
1.7 
Estimated total study duration 
24 months (includes enrollment, planned safety holds, follow-up, and AESI health 
contact). 
1.8 
Study sites 
The HIV Vaccine Trials Network (HVTN) Clinical Research Sites (CRSs) will be 
located in the US and will be further specified in the Site Announcement Memo. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 11 of 107 
2 
Introduction 
This first-in-human clinical trial will examine the safety and immunogenicity of a 
vaccination regimen with a synthetic DNA–encoded stabilized HIV-1 Env 
Native-Like Trimer (sD-NLT-AB05 adjuvanted with IL-12 DNA), alone or in 
combination with HIV-1 Env Trimer 4571, adjuvanted with 3M-052-AF + Alum, 
as a boost. The DNA vectors will be delivered via ID EP. Vaccination with 
Native-Like Trimer (NLT) via a DNA platform (as opposed to recombinant 
protein) eliminates the need to express and purify NLTs in vitro, saving time and 
lowering costs.  
Advantages of the synthetic DNA vaccine platform with adjuvant IL-12 DNA 
(pGX6001), especially when administered in combination with NLT protein 
boosts, include: 
• The potential to elicit more robust immune responses than protein vaccine 
alone, 
• Potential dose sparing of protein vaccine required in the regimen, and 
• Unique induction of cellular responses to act synergistically with induced 
humoral responses. 
The clinical trial will inform future NLT and DNA clinical studies for both HIV 
vaccines and non-HIV vaccines, such as sD-NLT germline-targeting trimers to 
activate specific bnAb precursor lineages, sD-NLT heterologous boosting to 
broaden antibody recognition, and sD-NLT prime with protein NLT boosts with 
combinations of NLTs based on more diverse HIV isolates to generate broader 
responses. 
2.1 
Rationale for evaluation of sD-NLT-AB05 with IL-12 adjuvant and 
Trimer 4571 adjuvanted with 3M-052-AF + Alum in a prime boost 
regimen 
2.1.1 
sD-NLT-AB05 with IL-12 adjuvant 
Native-Like Env Trimers (NLTs): HIV envelope protein (Env) is a 
conformationally dynamic structure that is a key target for HIV vaccine 
development (1, 2). Preclinical studies demonstrate that recombinant NLTs, 
which resemble ‘native’ Env on the HIV virion surface, display surfaces 
recognized by all lineages of broadly neutralizing antibodies (bnAbs) capable of 
neutralizing HIV while simultaneously ensuring that many non-neutralizing 
epitopes remain hidden (3, 4). Stabilized NLTs can be engineered by 
incorporation of mutations to favor the prefusion state conformation. Recently, we 
optimized NLTs for an advanced synthetic DNA vaccine delivery platform (sD-

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 12 of 107 
NLT). In the sD-NLT formulation, NLT immunogens can be encoded in synthetic 
DNA cassettes as transgenes in a clinical plasmid (pVax).  
sD-NLT-AB05 encodes for an Env trimer derived from the HIV-1 clade A strain 
BG505(5). The DNA platform is thus referred to as “sD-NLT-AB05.” It is a 
gp140 format modified through a set of directed mutations to produce a stable 
NLT exposing epitopes targeted by bnAbs. It also occludes distracting epitopes 
targeted by non-neutralizing Abs (eg, on the V3 loop and inner domain surface) 
(5, 6). Similarly, the VRC-HIVRGP096-00-VP Env trimer (Trimer 4571) is a 
stabilized NLT, also derived from BG505 in a gp140 format, modified with a 
different set of mutations to prevent CD4-induced conformational triggering (7-
9). Specifically, the sD-NLT-AB05 encoded trimer and Trimer 4571 share the 
following amino acid sequence changes: SOS stabilizing mutations (501C, 605C), 
gp41 stabilizing mutation (559P), glycan introduction mutation (332N), and 
improved furin cleavage site mutations (RRRRRR between 507 and 512). They 
differ in 2 sets of changes: Trimer 4571 contains disulfide mutations (201C, 
433C) and sD-NLT-AB05 contains 11 changes to improve biophysical properties 
(T106E, M271I, F288L, N363Q, F519S, A561P, V570H, R585H, L568D, R304V 
and A319Y). The sD-NLT-AB05 and Trimer 4571 harbor the same bnAb 
epitopes and differ only by a small set of mutations, which are not anticipated to 
affect the B-cell or T-cell responses. 
IL-12 induces the production of interferon-gamma (IFN-gamma), favoring the 
differentiation of type 1 T helper (Th1) cells (10, 11). Biologically active IL-12 
has pleotropic effects but is generally considered to be a pro-inflammatory 
cytokine that biases the CD4+ T-cell response towards a Th1 phenotype. Previous 
studies have demonstrated that the inclusion of plasmid-encoded IL-12 
(GENEVAX IL-12-4532) with HIV PENNVAX-B DNA was dose sparing (12). 
The inclusion of IL-12 DNA did not affect the tolerability of an HIV DNA 
vaccine (13). 
2.1.2 
Trimer 4571 adjuvanted with 3M-052-AF + Alum 
In proof-of-concept preclinical studies in guinea pigs, Trimer 4571 adjuvanted 
with Alum-induced autologous neutralizing Abs to BG505.W6M.C2 after the 
second administration that increased after the third administration. Rhesus 
macaques immunized via IM injection with research-grade Trimer 4571 produced 
neutralizing Abs to BG505.W6M.C2.T332N (see the Investigator’s Brochure [IB] 
for more detail on these BG505 variants).  
3M-052-AF contains 3M-052 and an emulsifier in an AF. 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 (14-17).  
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 13 of 107 
adjuvanted with 3M-052-AF + Alum elicits neutralizing and binding Abs more 
quickly, to higher peak magnitudes, and with greater durability than any of the 
other protein-adjuvant combinations tested (see IB). 
In rhesus macaques, among 8 adjuvants combined with an Env gp140 
immunogen, the Alum/TLR7 agonist 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 3-10-fold compared to Alum 
alone (18). 
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 it 
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 (14). In NHPs 
immunized with SARS-CoV-2 RBD formulated with 3M-052 combined with 
Alum, several enhanced immune responses were observed, including 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 (19). 
For additional information, see the IB. 
2.2 
Preclinical data with sD-NLT-AB05: mice and rabbits 
In preclinical animal models, DNA-encoded NLTs are capable of inducing 
autologous neutralizing immune responses, and DNA-encoded NLTs act 
synergistically with recombinant protein NLTs in prime-boost regimens. NLTs 
formulated as synthetic DNA and delivered by electroporation have not been 
evaluated in humans.  
Preclinical studies in mice demonstrated that sD-NLT-AB05 can assemble in vivo 
with a native-like antigenic profile similar to the same recombinant NLT protein 
produced in vitro (see Figure 2-1, and reference (6)). 
This is the first time NLTs have been shown to assemble directly in vivo when 
launched from synthetic DNA by an advanced EP device. DNA-encoded NLTs 
folded properly in vivo and induced the appropriate humoral response, as 
demonstrated by the observation that sD-NLT-AB05 induces autologous tier-2 
neutralizing antibodies (nAbs) in mice. Comparison of protein versus DNA NLT 
vaccinations in mice demonstrated induction of trimer-binding antibodies by both 
vaccine platforms. High levels of CD4+ and CD8+ T-cell responses, as well as 
autologous tier-2 BG505.T332N neutralization was observed in mice immunized 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 14 of 107 
with sD-NLT-AB05 (Figure 2-2 A and B). However, recombinant protein failed 
to generate either CD8+ T-cell responses or any tier-2 responses in mice, as 
shown by us and others (20). Neutralizing clones have been isolated and Cryo-
EM employed to molecularly map the murine neutralizing epitope targeted by 
vaccination with sD-NLT-AB05, which is also a target of protective nAbs in 
primates (21, 22) (Figure 2-3 A and B). sD-NLT-AB05 has also been observed to 
generate autologous tier-2 neutralizing responses in rabbits (unpublished data, see 
the IB). In published macaque studies by other groups, DNA/Env protein co-
administration at the same site during each vaccination can improve induced 
binding and nAb responses (23), as well as challenge outcome (24). 
 
Figure 2-1 In vitro and in vivo characterization of sD-NLT-AB05. (A) Size-exclusion 
chromatography elution showing homogenous trimers are produced (B) Negative stain electron 
microscopy imaging of NLT-AB05 (C) Antigenic profile of NLT-AB05 (D) Muscle sections from mice 
treated intramuscularly with DNA-encoded BG505.gp120.foldon or sD-NLT-AB05 Stained with 
known monoclonal Abs recognizing either well folded trimers (PGT145) or poorly folded trimers 
(3074). Staining with non-nAb 3074 indicates poorly folded trimer while staining with bnAb PGT145 
indicates well-folded trimer. 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 15 of 107 
 
Figure 2-2 Immunogenicity of sD-NLT-AB05. (A) Comparison of BG505 Env-specific cellular 
responses in naïve mice or mice immunized with pVAX plasmid backbone, Sigma Adjuvant System 
(RIBI)-co-formulated protein BG505.MD39, or sD-NLT-AB05 without adjuvant by IFNγ ELIspot 
assay (left panel) or intracellular cytokine staining (right panel). Cells were stimulated with 
overlapping 15-mer peptide pools for wild-type BG505 gp140 divided into 3 pools from N-terminus 
to C-terminus. (B) Left panel: Binding antibodies induced by sD-NLT-AB05 comparable to RIBI-co-
formulated protein BG505.MD39 immunizations. Right panel: Unique induction of autologous tier-2 
neutralizing responses by sD-NLT-AB05 in contrast to recombinant protein. DNA (short) = 
immunization at weeks 0, 3, 6. DNA (long) = immunization at weeks 0, 3, 16. 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 16 of 107 
 
Figure 2-3 Immunogenicity of sD-NLT-AB05. (A) Neutralization ID50 titers of heat-inactivated 
mice sera for sD-NLT-AB05 immunized mice with strong neutralizing activity against listed 
pseudovirus isolates. Isolated nAbs using antigen-specific B-cell sorting of sD-NLT-AB05 
immunized mice. IC50 values of trimer-specific murine mAbs for BG505.T332N, 
BG505.T332N.T465N or MLV. (B) Structural model of nAb clone C24 with positions mutated from 
the germline sequences are displayed in red. Cryo-EM structure of C24 in complex with 
BG505.MD39. 
2.3 
Clinical Experience with related DNA vaccines encoding HIV Env 
immunogens with and without IL-12 adjuvant  
Prior clinical studies (HVTN 070, HVTN 080, HVTN 087, and HVTN 098) have 
demonstrated the safety of DNA-encoded HIV Env immunogens delivered via EP 
and demonstrated consistent induction of Ab and CD4+ T-cell and CD8+ T-cell 
responses. Key features of these trials and summaries of safety are shown in Table 
2-1. HVTN 098 tested optimized cocktail formulations administered via a device 
enhanced for ID EP delivery and resulted in an efficient platform, generating a 
potent, dose-sparing, and highly tolerable ID-based vaccine approach in humans. 
These studies established the importance of the optimized plasmid IL-12 for ID 
HIV DNA vaccine immune potency, response rates, and memory (25, 26).  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 17 of 107 
Table 2-1 Clinical trials with related DNA vaccines encoding HIV Env immunogens  
Study / 
ClinicalTrials.gov 
identifier 
Products 
Sample 
size 
(N) 
Route of 
administration 
Safety 
Reference 
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 
One discontinuation 
due to possibly 
related adverse 
event (cervical 
radiculopathy) 
Kalams, 
2013 (12) 
HVTN 080 / 
NCT00991354 
PENNVAX-B DNA 
vaccine 
IL-12 plasmid DNA: 
GENEVAX-IL-12-4532, 
formulated with 
bupivacaine  
 
48 
IM 
administration 
with 
electroporation 
using VGX 
CELLECTRA 
electroporation 
system 
No severe systemic 
reactogenicity 
No related severe 
adverse events 
Two 
discontinuations due 
to injection site pain  
Kalams, 
2013 (12) 
HVTN 087 / 
NCT01578889 
HIV multiantigen DNA: 
consists of 2 plasmid 
DNA expression vectors, 
HIV-1 gag/pol and HIV-1 
nef/tat/vif, env (clade B). 
IL-12 plasmid DNA: 
GENEVAX IL-12-4532, 
formulated with 
bupivacaine  
VSV-HIV Gag 
100 
IM 
administration 
with 
electroporation 
using Ichor 
Medical 
Systems 
TriGrid 
Delivery 
System (TDS) 
EP device 
No severe systemic 
reactogenicity.  
No severe related 
adverse events. 
Three 
discontinuations due 
to injection site 
pain. 
Elizaga, 
2018 (27) 
(28) 
HVTN 098 / 
NCT02431767 
PENNVAX-GP DNA 
Vaccine: admixture of 2 
plasmids encoding 
consensus clade A and C 
HIV-1 envelope protein 
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 
electroporation 
using 
CELLECTRA 
3P EP for ID 
delivery and 
CELLECTRA 
5P EP for IM 
delivery. 
No severe systemic 
reactogenicity.  
No severe related 
adverse events. 
One participant 
discontinued due to 
pain, anxiety and 
presyncopal episode 
Edupuganti, 
2020 (26) 
De Rosa 
2020 (25) 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 18 of 107 
2.3.1 
Safety 
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 
300 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 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 in HVTN 098 (26). 
Systemic reactogenicity symptoms with IM injection of DNA vaccines and with 
ID injection of DNA vaccines did not differ (26). 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. 
When the injection and EP are given in the skin, the needles 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 (26). 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 using 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 IB. 
2.3.2 
Immunogenicity  
EP improves immunogenicity of DNA vaccines. 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 (HVTN 070) IM EP (12). 
Comparing the comparable 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 have not been induced well by DNA. The responses were 
primarily directed to Gag and Pol, and not to Env. Because of this, in HVTN 098, 
the Env expressing DNA vaccine was modified, including two env plasmids 
encoding clade A and C Env immunogens, and including a substitution of an 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 19 of 107 
optimized IgE leader as well as the deletion of their cytoplasmic tail to improve 
surface expression (25). 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 EP and intradermal EP, 
with the ID EP administration at 1/5th the dose of the IM EP. 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 highest 
responses for the ID + IL-12 group. Neutralizing antibodies were mainly detected 
only to a tier 1A viral isolate. Only a few participants developed detectable 
antibody-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 the 
ID route was dose-sparing.  
Inclusion of IL-12 plasmid DNA . Regarding the benefit of IL-12, in HVTN 
080, the sample size for the group without IL-12 was small, likely reducing the 
ability to detect significant differences. Although not statistically significant, the 
response rates including IL-12 were higher than 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 
without and with IL-12 after the fourth dose. The binding Ab responses to Env 
were similar for the ID groups with and without IL-12 after the fourth vaccination 
but tended to be lower after the third vaccination without IL-12, suggesting that 
inclusion of IL-12 accelerated the response, achieving near maximal after the 
third rather than the fourth. The effect of IL-12 was examined in another HVTN 
study, HVTN 087, that tested increasing doses of IL-12 included with DNA 
administered IM with EP for 3 doses followed by vesicular stomatitis virus (VSV) 
boost (28). 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, 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 
Clinical experience with HIV Envelope proteins adjuvanted with 3M-
052-AF + Alum: safety data 
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 20 of 107 
mcg (Group 2), both combined with 500 mcg Alum. The study is ongoing and 
remains blinded to within-group treatment assignment. As of April 27, 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) 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 post-vaccination. 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 post-injection. 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 post-vaccination 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. 
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), adverse events of special interest 
(AESIs), or deaths in either group after this third dose. Both local and systemic 
reactogenicity was similar between all 3 doses in participants who received 3 
doses. 
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 27, 2022, Part B is 
fully enrolled but remains blinded. 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 one of the HVTN 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 21 of 107 
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 a 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 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. 
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 these 5 participants received a subsequent 
vaccination without any Grade 3 reactogenicity, one (1) experienced an additional 
Grade 3 systemic reactogenicity symptom, one (1) has not yet received a 
subsequent vaccination, and two (2) declined to receive additional vaccinations 
after their first Grade 3 systemic reactogenicity event. 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 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 (12, 27). Specifically for ID 
delivery, both Abs and T-cell responses were induced (25, 26). In HVTN 098, the 
fourth vaccination boosted the magnitude of total Ab response for all treatment 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 22 of 107 
groups against gp140 antigens from clade A and B as compared to the magnitude 
of total Ab response after the third dose (25).  
 
Figure 2-4 IgG binding antibody responses as measured by binding antibody multiplex 
assay (BAMA) against consensus Env gp140 antigens for clades A, B and C. Assays were 
performed 2 weeks after the third and fourth vaccinations and 6 months after the fourth vaccination. 
Positive responses are shown in filled circles in color, negative responses are shown in open gray 
triangles. Box plots represent the distribution for the positive responders only. Bar plots show 
response rates. Numbers below the bars indicate numbers of positive responders and total 
participants. Positive response rates were compared using the Fisher’s exact test for unpaired data 
(between treatment groups) and the McNemar test for paired data (between visits). Response 
magnitudes among positive responders were compared using the Wilcoxon rank sum test for 
unpaired data and the Wilcoxon signed rank test for paired data. All p-values are two-sided. False-
discovery rate adjusted q values were calculated to account for multiple antigens, multiple 
timepoints, or treatment groups (Figure from supplemental data (25)). 
By including 2 boosts of Trimer 4571 at months 3 and 6 in Group 2, we can 
assess whether additional doses of protein and DNA lead to enhanced 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 23 of 107 
immunogenicity compared to DNA alone in this study (Group 1) and previous 
studies. By maintaining a standard schedule, data will be directly comparable to 
previous DNA vaccine studies. 
2.6 
Rationale for Dose of Trimer 4571 and 3M-052-AF + Alum adjuvant 
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 is 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 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. 
Table 2-2 Clinical trials with Trimer 4571 
Study 
ClinicalTrials.go
v NCT # / Study 
Status 
Participant 
HIV status 
Number 
receiving 
Trimer 
4571 + 
Alum 
Route /  
Dose 
Schedule 
Safety data 
(ClinicalTrials.gov) 
VRC 018 
NCT03783130 / 
Completed 
Negative 
16 
IM or SC 
at 100 or 
500 mcg 
Trimer and 
500 mcg 
Alum  
0, 2, 5 
months 
As of August 2021, final 
study data: no related 
SAEs, related grade 4 
AEs, or related MAAEs 
were reported or other 
unexpected reactions, 
and no study pause 
criteria were met at any 
time. 
NIAID 19-I-
0069 
NCT03878121 / 
Enrolling  
Negative 
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. 
Therapeutic 
immunization 
study (NETI) 
PI: M 
Choudhary, 
Univ of 
Pittsburg 
NCT04985760 
Enrolling 
Positive 
24 
IM at 100 
or 500 mcg 
Trimer and 
500 mcg 
Alum 
0, 2, 5 
months 
As of Jan 1, 2022, 5 
participants have 
received Trimer 4571. 
No related SAEs, no 
SUSARs. 
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 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 24 of 107 
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. 
2.7 
Rationale for Targeting Same Draining Lymph Node with both DNA 
and Protein 
Direct targeting of the draining lymph node with a multipronged immune stimulus 
consisting of both a DNA vaccine and an adjuvanted protein vaccine leads to 
improved outcomes in a variety of several preclinical models (23, 29-31). The 
most directly relevant study is that of Felber and colleagues in rhesus macaques 
(31). It involved a comparison of vaccination with monomeric HIV envelope 
administered via both DNA and an adjuvanted protein delivered either into the 
same limb or into contralateral limbs, allowing 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 lead to an improvement in a variety of 
immune outcomes, including functional Ab responses (both increased ADCC 
activity and improved FcRIIIa binding) and cellular responses (both 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 simian-human immunodeficiency virus (SHIV) 
challenge (see Figure 2-5). 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 25 of 107 
 
Figure 2-5 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 two vaccine groups (n 
= 18 and 17, respectively) and the control group (n = 20). The RMs were exposed to 15 weekly 
intravaginal challenges. Infection was defined by 2 consecutive positive plasma VL measurements. 
No RMs were censored. P-value, exact log-rank test. Figure from Felber et al (31). 
In keeping with the well-tolerated safety profile of simultaneous delivery of HIV 
immunogens via both DNA+ envelope protein administration strategies (32, 33), 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 26 of 107 
no AEs were noted with either strategy (separate versus same draining lymph 
node), including assessments via clinical chemistry, complete 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 (34) and the DNA will be delivered intradermally whereas the protein 
will be delivered intramuscularly. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 27 of 107 
2.8 
Risks and benefits 
2.8.1 
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 antibody test result 
• 
Visible lesion(s) at the injection site, such as erythematous papules with eschar, 
hypopigmentation, hyperpigmentation, or scar related to the EP injection 
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 by EP 
• 
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 infection/disease if the participant is infected with HIV 
• 
Effects on the fetus and on pregnancy 
Blood drawing may cause pain, bruising, fainting, and, rarely, infection at the site 
where the blood is taken.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 28 of 107 
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 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 the 
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 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. 
2.8.2 
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 infection. 
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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 29 of 107 
3 
Objectives and endpoints 
3.1 
Primary objectives and endpoints 
Objectives 
Endpoints 
1. To evaluate the safety and tolerability of 2 doses of sD-
NLT-AB05 + IL-12 DNA adjuvant followed by 2 doses of 
sD-NLT-AB05 + IL-12 DNA adjuvant alone or in 
combination with Trimer 4571 adjuvanted with 3M-052-
AF + Alum 
a) Local and systemic reactogenicity signs and 
symptoms will be collected for a minimum of 2 
weeks following receipt of any study vaccine 
b) Serious adverse events (SAEs), medically 
attended adverse events (MAAEs), adverse events of 
special interest (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 product. 
Additionally, all adverse events will be collected for 
30 days after any receipt of study vaccination. 
 
2. To evaluate the immunogenicity of 2 doses of sD-NLT-
AB05 + IL-12 DNA adjuvant followed by 2 doses of sD-
NLT-AB05 + IL-12 DNA adjuvant alone or in combination 
with Trimer 4571 adjuvanted with 3M-052-AF + Alum 
 
 a) Response rate and magnitude of vaccine-matched 
IgG binding Ab responses as assessed by multiplex 
assay 2 weeks following the fourth vaccination  
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 fourth vaccination  
3.2 
Secondary objectives and endpoints 
Objectives 
Endpoints 
1. To further evaluate and compare the ability of 2 doses of 
sD-NLT-AB05 + IL-12 DNA adjuvant followed by 1 or 2 
doses of sD-NLT-AB05 + IL-12 DNA adjuvant alone or in 
combination with Trimer 4571 adjuvanted with 3M-052-
AF + Alum to elicit humoral immune responses  
a) Neutralizing Ab magnitude and breadth against 
autologous and tier 1a HIV-1 isolates as assessed by 
TZM-bl neutralization assay following the third and 
fourth vaccinations 
b) Response rate, magnitude, and epitope specificity 
of HIV-1 specific IgG binding Ab responses as 
assessed by multiplex assay 2 weeks following third 
vaccination  
2. To further evaluate and compare the ability of 2 doses of 
sD-NLT-AB05 + IL-12 DNA adjuvant alone or in 
combination with Trimer 4571 adjuvanted with 3M-052-
AF + Alum to elicit cellular immune responses  
a) 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 
third vaccination 
 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 30 of 107 
Objectives 
Endpoints 
3. To evaluate the durability of cellular and humoral 
immune responses elicited by 2 doses of sD-NLT-AB05 + 
IL-12 DNA adjuvant followed by 2 doses of sD-NLT-
AB05 + IL-12 DNA adjuvant alone or in combination with 
Trimer 4571 adjuvanted with 3M-052-AF + Alum  
a) Magnitude and response rate of CD4 + and 
CD8+ T-cell responses measured by flow cytometry, 
to HIV-1-specific Env peptide pools 6 months post 
last vaccination 
b) Response rate and magnitude of HIV-1 specific 
IgG binding Ab responses as assessed by multiplex 
assay 6 months post last vaccination 
c) Neutralizing Ab magnitude and breadth against 
autologous tier 2 HIV-1 isolates as assessed by 
TZM-bl neutralization assay 6 months post last 
vaccination 
 
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. 
2. To evaluate the response rate and magnitude of HIV-1 specific IgG binding Ab 
responses as assessed by multiplex assay 2 weeks following second vaccination. 
3. To evaluate the 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 second vaccination. 
4. To evaluate the frequency of Env-specific B cells measured by flow cytometry 2 
weeks following the second, third, and fourth vaccinations and 6 months post last 
vaccination. 
5. To evaluate serum Ab specificities and elicitation of trimer-degrading Abs using 
polyclonal epitope mapping as assessed by Electron Microscopy 2 weeks post 
third and fourth vaccinations. 
6. To evaluate the neutralizing Ab magnitude and breadth against heterologous and 
tier 2 HIV-1 isolates as assessed by TZM-bl neutralization assay following the 
third, and fourth vaccinations. 
7. To evaluate of HIV-1 specific IgG binding Ab responses to the trimer base 2 
weeks following second, third, and fourth vaccinations. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 31 of 107 
8. To evaluate Ab avidity and Fc Receptor functions such as FcR binding, ADCC, 
antibody-dependent cellular phagocytosis (ADCP), and infected cell antibody-
binding assay (ICABA) after the second, third, and fourth vaccinations. 
9. To evaluate B-cell receptor (BCR) repertoires and sequences (including analysis 
of rare B-cell lineages associated with bnAb precursors), including cellular 
phenotyping and mutational frequency analysis suggestive of somatic 
hypermutation and affinity maturation following immunization. 
10. To characterize monoclonal Abs derived from BCR sequences from Env-specific 
B cells. 
11.  To conduct analyses related to furthering the understanding of HIV, 
immunology, vaccines, and clinical trial conduct. 
12.  To further evaluate immunogenicity of each vaccine regimen, additional 
immunogenicity assays may be performed in a subset of participants, including on 
samples from other timepoints, based on the HVTN Laboratory Assay Portfolio. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 32 of 107 
4 
Laboratory strategy  
4.1 
Assessment of immunogenicity endpoints 
The primary goal of HVTN 304 is to determine whether the sD-NLT-AB05/ IL-
12 DNA alone or in a prime-boost regimen with VRC Env Trimer 4571 
adjuvanted with 3M-052-AF + Alum will elicit HIV-1 envelope protein specific 
binding antibody (Ab) and T-cell responses. To this end, HVTN immunogenicity 
assays will be used to measure both Env-binding Abs and both CD4+ and CD8+ 
Env-specific T cells.  
For the primary endpoints, vaccine matched HIV-specific binding Ab responses 
will be assessed by multiplex assay, and response rate and magnitude of CD4 + 
and CD8+ T-cell responses will be assessed by flow cytometry. These data will 
also be used to identify and prioritize samples for in-depth immunogenicity 
analyses (secondary and exploratory endpoints, see Sections 3.2 and 3.3). 
Additional assays may be performed on selected responders and at additional 
timepoints at the discretion of Protocol Team leadership based on its evaluation of 
primary results following the fourth vaccination (see Section 3.1).  
The 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 protocol webpage. 
Descriptions of the standard HVTN laboratory assays can be found online at 
https://www.hvtn.org/content/dam/hvtn/scientific-programs/hvtn-laboratory-
assay-descriptions.pdf. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 33 of 107 
5 
Study design 
This is a randomized trial to examine the safety and immunogenicity of INO-6160 
(sD-NLT-AB05/ IL-12 DNA) alone or in combination with Trimer 4571 
adjuvanted with 3M-052-AF + Alum in healthy adults. The primary hypothesis is 
that INO-6160 alone or in combination with Trimer 4571 adjuvanted with 3M-
052-AF + Alum vaccinations can induce seroconversion binding antibody 
responses and induce antigen specific T-cell responses (both CD4+ and CD8+).  
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 in 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: volunteer demonstrates understanding of this 
study; completes a questionnaire prior to first vaccination with verbal 
demonstration of understanding of questionnaire items that were answered 
incorrectly. 
2. 18-55 years old, inclusive, on day of enrollment. 
3. Available for clinic follow-up through the last clinic visit and willing to be 
contacted 12 months after the last vaccine administration. 
4. Agrees not to enroll in another study of an investigational agent during 
participation in the trial. 
5. In good general health according to the clinical judgment of the site investigator. 
6. 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. 
7. Assessed as low risk for HIV acquisition per low risk guidelines (see Appendix 
D), agrees to discuss HIV infection risks, agrees to risk reduction counseling, and 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 34 of 107 
agrees to avoid behavior associated with high risk of HIV exposure through the 
final study visit. Low risk may include persons stably taking PrEP as prescribed 
for 6 months or longer. 
8. 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 men who have been on hormone therapy for more than 6 
consecutive months 
• ≥ 12.0 g/dL for transgender women 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 
their sex assigned at birth  
9. 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). 
10. Platelets = 125,000-550,000/mm3 
11. Alanine aminotransferase (ALT) < 2.5 x upper limit of normal (ULN) based on 
the institutional normal range 
12. Serum creatinine ≤ 1.1 x ULN based on the institutional normal range 
13. Blood pressure in the range of 90 to < 140 mmHg systolic and 50 to < 90 mmHg 
diastolic. 
14. Negative results for HIV infection by a Food and Drug Administration (FDA)-
approved enzyme immunoassay (EIA) or chemiluminescent microparticle 
immunoassay (CMIA). 
15. Negative for anti-Hepatitis C antibodies (anti-HCV) or negative HCV nucleic acid 
test (NAT) if anti-HCV antibodies are detected. 
16. Negative for Hepatitis B surface antigen. 
17. 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 35 of 107 
days prior to enrollment through 8 weeks after their last scheduled fifth 
vaccination timepoint (see Appendix E). 
• Has negative β-HCG (human chorionic gonadotropin) pregnancy test (urine or 
serum) on day of enrollment. 
• Volunteers who were assigned female sex at birth must also agree not to seek 
pregnancy through alternative methods, such as oocyte retrieval, artificial 
insemination or in vitro fertilization through 8 weeks after their last scheduled 
vaccination timepoint. 
5.1.2 
Exclusion criteria 
1. Volunteer who is breast-feeding or pregnant. 
2. Body mass index (BMI). Enrollment of individuals with BMI ≥ 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 diet alone or 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. Congenital or acquired immunodeficiency, including systemic medication use 
likely to impair immune response to vaccine in the opinion of the site investigator 
such as glucocorticoid use equal to or greater than prednisone 10 mg/day within 3 
months prior to enrollment. 
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. 
8. ACAM2000 vaccine for Monkeypox received within 30 days prior to enrollment 
or receipt of study product, or if ACAM2000 received greater than 30 days prior 
to enrollment, or prior to receipt of study product, vaccination scab still present; 
or planned administration within 30 days after enrollment or receipt of study 
product. 
9. 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 monkeypox disease are not considered to be live vaccines.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 36 of 107 
10. Receipt of non-HIV experimental vaccine(s) received within the last 1 year. 
Exceptions may be made by the PSRT for vaccines that have subsequently 
undergone licensure or Emergency Use Authorization by the FDA or, if outside 
the United States, by the national regulatory authority or World Health 
Organization. For volunteers who have received control/placebo in an 
experimental vaccine trial, the PSRT will determine eligibility on a case-by-case 
basis.  
11. 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. 
12. 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. 
13. History of serious reaction (eg, hypersensitivity, anaphylaxis) to any related 
vaccine or component of the study vaccine regimen. 
14. Hereditary angioedema, acquired angioedema, or idiopathic forms of angioedema. 
15. Idiopathic urticaria within the past year. 
16. Bleeding disorder diagnosed by a doctor (eg, factor deficiency, coagulopathy, or 
platelet disorder requiring special precautions). 
17. Seizure disorder; febrile seizures as a child or seizures secondary to alcohol 
withdrawal more than 5 years ago are not exclusionary.  
18. Asplenia or functional asplenia. 
19. Active duty and reserve US military personnel. 
20. 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). 
21. 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 37 of 107 
• 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 
urgent/emergent care solely for asthma medication refills or co-existing 
conditions unrelated to asthma); OR 
• Uses a short-acting rescue inhaler more than 2 days/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 one medication for maintenance therapy daily. Inclusion of 
anyone on a stable dose of more than one medication for maintenance therapy 
daily for greater than two years requires PSRT approval. 
22. A participant with a history of an immune-mediated disease, either active or 
remote. Specific examples are listed in Appendix F (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 
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 38 of 107 
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 pre-vaccination 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 antibodies or other 
immunomodulators (other than nonsteroidal anti-inflammatory drugs 
[NSAIDs]), with the exception that study injection may continue per principal 
investigator (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 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 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 2 weeks 
prior to study vaccine administration. Replication incompetent vaccines such 
as the Jynneos vaccine for the prevention of monkeypox disease are not 
considered to be live vaccines. 
Vaccinations should not be administered outside the visit window period specified 
in Appendix B without PSRT approval.  
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 infection 
• 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 39 of 107 
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, as per the Second National Institute of Allergy and 
Infectious Disease/Food Allergy and Anaphylaxis Network symposium 
criteria for anaphylaxis (35), is provided below:  
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 PEF, hypoxemia)  
b. Reduced 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 40 of 107 
• Newly disclosed AESI (see Appendix F) 
• 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 304 PSRT) 
Participants discontinuing study vaccine for reasons other than HIV infection 
should be encouraged to participate in follow-up visits and procedures per the 
protocol for at least 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 becomes HIV-infected during the course of the study, no 
additional study vaccine will be administered. Participants will be encouraged to 
continue scheduled study visits for at least 12 months following their last study 
vaccine administration. At post-infection 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 still eligible to receive 
future vaccinations.  
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) 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, CBC with differential, serum chemistry, physical examination, and if 
indicated, a pregnancy test (see Appendix A). For participants with HIV infection, 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 41 of 107 
please see Section 8.6. If the site PI has questions regarding a termination visit, 
they should consult with the PSRT.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 42 of 107 
6 
Statistical considerations 
6.1 
Sample size justification and accrual 
Recruitment will target enrolling 10 healthy, adult participants without HIV per 
group for the 2 study groups receiving either 2 doses of INO-6160 followed by 2 
doses of INO-6160 alone or in combination with Trimer adjuvanted with 3M-052-
AF + Alum (for study schema see Table 1-1). Up to 5 additional participants, or a 
total of 25 participants, may be enrolled to ensure sufficient samples for 
immunogenicity analyses, with a goal of at least 20 contributing to the final 
analyses. Examples of reasons that might necessitate enrollment of additional 
participants are provided in Section 1.5.  
Since enrollment is concurrent with receiving the first vaccination, all participants 
will provide some safety data. It is possible, however, for immunogenicity data to 
be missing; previous HVTN and AIDS Vaccine Evaluation Group (AVEG) 
studies suggest 10% is a reasonable estimate for the rate of missing data. For this 
reason, the sample size calculations account for 10% of enrolled participants 
having missing data for the primary immunogenicity endpoints. This 10% 
missingness accounts for both post-processing variables (eg, problems with 
samples) and is mitigated by the potential for replacing participants who do not 
continue with the study. 
6.1.1 
Power calculations for immunogenicity 
The main goal for the analysis is to evaluate the difference of the frequency and 
magnitude of HIV-1 specific binding Ab responses, and the rate of CD4+ and 
CD8+ T-cell responses between the 2 study groups, following the third and fourth 
vaccination. The precision with which the true response rate can be estimated 
from the observed data depends on the true underlying response rate and the 
sample size. Two-sided 95% confidence intervals (CIs) for the response rate 
based on observing a particular rate of responses in the study groups is shown in 
Table 6-1. The table indicates n = 9 per group, assuming a 10% loss of data.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 43 of 107 
Table 6-1 Two-sided 95% CIs for the true response rate based on observing a particular rate 
of responses in the study groups, n = 9 (total 10 with 10% missing) 
N of 
responses 
Observed 
response rate (%) 
95% CI (%) 
0/9 
0 
0 
37.1 
1/9 
11.1 
0.6 
49.3 
2/9 
22.2 
3.9 
59.8 
3/9 
33.3 
9 
69.1 
4/9 
44.4 
15.3 
77.3 
5/9 
55.6 
22.7 
84.7 
6/9 
66.7 
30.9 
91 
7/9 
77.8 
40.2 
96.1 
8/9 
88.9 
50.7 
99.4 
9/9 
100 
62.9 
100 
The estimated differences in response rate, as well as their 95% CIs are shown in 
Table 6-2. Due to the relatively small sample sizes, it is recommended (36) to 
construct the two-sided response rate difference CIs with Agresti-Caffo interval 
(37) that is less conservative. The group with higher response rate is assumed to 
have positive response rate of 50%, 70%, and 90%, and the assumed range of 
response rate differences between 2 groups is 0% to 50%, 70%, and 90%, 
respectively. With sample size of 10 for each group, the statistically significant 
difference in response rates of approximately 50% (44.4%-55.6%) can be detected 
while controlling Type-I error rate at 5%.  
Table 6-2 Observed rate difference between study groups and their two-sided 95% 
confidence intervals from 10 total study participants per group. Assuming 10% missing rate 
for immunogenicity endpoints, n = 9 (total 10 with 10% missing). 
Group 1 
positive 
response 
Group 2 
positive 
response 
Observed rate 
difference (%) 
Rate difference 
95% CI (%) 
(Agresti-Caffo) 
5 
5 
0 
-41.6 
41.6 
5 
4 
11.1 
-32.5 
50.7 
5 
3 
22.2 
-22.7 
59.1 
5 
2 
33.3 
-12.2 
66.8 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 44 of 107 
Group 1 
positive 
response 
Group 2 
positive 
response 
Observed rate 
difference (%) 
Rate difference 
95% CI (%) 
(Agresti-Caffo) 
5 
1 
44.4 
-0.9 
73.6 
5 
0 
55.6 
11.5 
79.4 
7 
7 
0 
-37.2 
37.2 
7 
6 
11.1 
-29.6 
47.8 
7 
5 
22.2 
-21.3 
57.7 
7 
4 
33.3 
-12.2 
66.8 
7 
3 
44.4 
-2.4 
75.1 
7 
2 
55.6 
8.2 
82.7 
7 
1 
66.7 
19.7 
89.4 
7 
0 
77.8 
32.3 
95 
9 
9 
0 
-24 
24 
9 
8 
11.1 
-19.3 
37.5 
9 
7 
22.2 
-13.1 
49.5 
9 
6 
33.3 
-5.8 
60.4 
9 
5 
44.4 
2.4 
70.3 
9 
4 
55.6 
11.5 
79.4 
9 
3 
66.7 
21.4 
87.7 
9 
2 
77.8 
32.3 
95 
 
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 
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. The SAEs will be monitored following each dose. For the first 2 doses, 
study participants in both groups will receive the same sD-NLT-AB05 + IL-12 
DNA adjuvant; thus, the safety sample size can be doubled. Specifically, for this 
two-arm study with sample size n = 10 per group (n = 20* for safety after the first 
2 doses), there is at least a 90% chance of observing at least 1 event if the true rate 
of such an event is 20.6% (10.9%*) or more and there is at least a 90% chance of 
observing no events if the true rate is 1.05% (0.53%*) or less. Safety data will be 
evaluated using historical controls. As a reference, in HVTN vaccine trials 
conducted in the US from April 2008 through March 2018, about 1% of 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 45 of 107 
participants who received placebos experienced an SAE. Binomial probabilities of 
observing 0 events, 1 or more events, and 2 or more events among 10 (20*) 
participants receiving study vaccine are presented in Table 6-3 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. 
Table 6-3 Probability of observing 0 events, 1 or more events, and 2 or more events among a 
group of 10 (20* for the first two doses) study participants for different true event rates. 
True 
event rate 
(%) 
Group size 
0 events 
1+ events 
2+ events 
1 
10 
0.9 
0.1 
0 
4 
10 
0.66 
0.34 
0.06 
10 
10 
0.35 
0.65 
0.26 
20 
10 
0.11 
0.89 
0.62 
30 
10 
0.03 
0.97 
0.85 
True 
event rate 
(%) 
Safety size for 
the first two 
doses* 
0 events 
1+ events 
2+ events 
1 
20* 
0.82* 
0.18* 
0.02* 
4 
20* 
0.44* 
0.56* 
0.19* 
10 
20* 
0.12* 
0.88* 
0.61* 
20 
20* 
0.01* 
0.99* 
0.93* 
30 
20* 
0* 
1* 
0.99* 
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 
6-4 shows the 2-sided 95% confidence intervals for the probability of an event 
based on a particular observed rate. If none of the 10 (20* for safety after first 2 
doses) participants receiving the study vaccine experience a safety event, the 95% 
2-sided upper confidence bound for the true rate of such events in the total 
vaccinated population is 32.6% (19.4%*). 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 46 of 107 
Table 6-4 Two-sided 95% confidence intervals for the probability of observing a safety event 
based on observing a particular rate of safety endpoints in a group of 10 study participants 
(n*=20 for safety after first two doses). 
Observed 
event 
n 
95% CI (%) 
0 
10 
0 
32.6 
1 
10 
0 
42.9 
2 
10 
4.9 
52.2 
3 
10 
10.6 
60.8 
Observed 
event 
n* 
95% CI (%) 
0 
20* 
0* 
19.4* 
1 
20* 
0* 
25.7* 
2 
20* 
1.8* 
31.6* 
3 
20* 
4.6* 
37.1* 
6.2 
Randomization 
The randomization sequence will be obtained by computer-generated random 
numbers and provided to each HVTN CRS through the Statistics and Data 
Management Center’s (SDMC) Web-based randomization system. The 
randomization will be done in blocks to ensure balance across study groups. At 
each institution, the pharmacist with primary responsibility for dispensing study 
products is charged with maintaining security of the treatment assignments. 
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. 
6.4 
Statistical analyses 
This section describes the final study analysis. All data from enrolled participants 
will be analyzed according to the initial randomization assignment regardless of 
how many vaccinations they received. The analysis is a modified intent-to-treat 
analysis in that individuals who are randomized but not enrolled do not contribute 
data and hence are excluded. Because of the brief length of time between 
randomization and enrollment—typically no more than 4 working days—very 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 47 of 107 
few such individuals are expected. All analyses will be performed using SAS and 
R.  
No formal multiple comparison adjustments will be employed for multiple safety 
endpoints, multiple primary immunogenicity endpoints, or secondary endpoints. 
However, multiplicity adjustments will be made for certain immunogenicity 
assays, as discussed below, when the assay endpoint is viewed as a collection of 
hypotheses (eg, testing multiple peptide pools to determine a positive response). 
6.4.1 
Analysis variables 
The analysis variables consist of baseline participant characteristic, safety, and 
immunogenicity for primary- and secondary-objective analyses. 
6.4.2 
Baseline demographics 
Participants’ baseline characteristics will be summarized using descriptive 
statistics.  
6.4.3 
Safety/tolerability analysis 
Since enrollment is concurrent with receiving the first vaccination, all participants 
will have received at least 1 vaccination and therefore will provide some safety 
data. 
Reactogenicity: The number and percentage of participants experiencing each 
type of reactogenicity sign or symptom will be tabulated by severity and 
treatment. For a given sign or symptom, each participant’s reactogenicity will be 
counted once under the maximum severity for all injection visits. Wilcoxon rank 
sum tests will be used to test for differences in severity between groups. 
AEs and SAEs: AEs will be coded into Medical Dictionary for Regulatory 
Activities (MedDRA)–preferred terms. The number and percentage of 
participants 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. 
6.4.4 
Immunogenicity analyses 
6.4.4.1 
Statistical Analysis to address primary objectives 
The first component of the analysis will entail comparing the frequency of HIV-
1–specific binding Ab responses 2 weeks following the fourth vaccination. 
Comparison between the 2 vaccine groups will use Fisher’s exact test, while 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 48 of 107 
McNemar’s test will be used for paired data (between visits), with a significant 
difference declared if the 2-sided p-value is ≤ 0.05. 
To compare the magnitude of HIV-1–specific binding Ab responses 2 weeks 
following the fourth vaccination. Between the 2 vaccine groups, a two-sided 
Wilcoxon rank sum test with 5% type-I error rate will be performed. Wilcoxon 
signed rank test with 5% type-I error rate will be performed for paired data. 
With the response rates of CD4+ and CD8+ T cell to HIV-1 specific Env peptide 
pools 2 weeks following the fourth vaccination, the 2 vaccine groups will be 
compared using Fisher’s exact test, and McNemar’s test will be used for paired 
data), with a significant difference declared if the 2-sided p-value is ≤ 0.05. For 
each T-cell subset, the positivity call for each peptide pool will include a multiple-
comparison adjustment for the number of peptide pools used in the assay using 
the discrete Bonferroni adjustment. The magnitude of response will be analyzed 
with two-sided Wilcoxon rank sum test for independent data and Wilcoxon signed 
rank test for paired data, with 5% type-I error rate. Graphs will be used to display 
the background-subtracted magnitudes for each participant by protein, treatment 
arm, and timepoint, with a box plot of data from positive responders 
superimposed on the individual data values. Statistical testing comparing the 
magnitudes will be based on positive responders only. 
6.4.4.2 
General approach for secondary and exploratory 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 they received. Additional analyses may be 
performed, limited to participants who received all scheduled injections per 
protocol. Assay results that are unreliable, from specimens collected outside of 
the visit window, or from HIV-infected participants are excluded. Since the exact 
date of HIV infection is unknown, any assay data from blood draws 4 weeks, or 
less, prior to an infected participant’s last seronegative sample and thereafter may 
be excluded. If an HIV-infected participant does not have a seronegative sample 
post enrollment, then all data from that participant may be excluded from the 
analysis.  
Discrete categorical assay endpoints (eg, response rates) will be analyzed by 
tabulating the frequency of positive response for each assay by antigen and 
treatment arm at each timepoint for which an assessment is performed. Crude 
response rates will be presented with their corresponding 95% CI estimates 
calculated using the score test method (38). Fisher’s exact tests will be used to 
compare the response rates of between 2 vaccine groups, with a significant 
difference declared if the 2-sided p-value is ≤ 0.05. 
For quantitative assay data (eg, magnitude of HIV-1 Env-specific binding Ab 
responses), graphical and tabular summaries of the distributions by antigen, 
treatment arm, and timepoint will be made. The difference between arms at a 
specific timepoint will be tested with a nonparametric Wilcoxon rank sum test if 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 49 of 107 
the data are not normally distributed and with a 2-sample t-test if the data appear 
to be normally distributed. An appropriate data transformation (eg, log10 
transformation) may be applied to better satisfy assumptions of symmetry and 
homoscedasticity (constant variance). More sophisticated analyses employing 
repeated measures methodology (for example, repeated measures analysis of 
variance or generalized estimating equations) may be utilized to incorporate 
immune responses over several timepoints and to test for differences over time. 
However, inference from such analyses would be limited by the small sample size 
of this study. All statistical tests will be 2-sided and will be considered 
statistically significant if p  0.05.  
6.4.4.3 
Missing Data 
Based upon previous AIDS Vaccine Evaluation Group and HVTN trials, missing 
10% of immunogenicity results for a specific assay is common due to study 
participants terminating from the study early, problems in shipping specimens, or 
low cell viability of processed peripheral blood mononuclear cells (PBMCs). To 
achieve unbiased statistical estimation and inferences with nonparametric tests 
and generalized linear models fit by generalized estimating equation (GEE) 
methods, missing data need to be missing completely at random (MCAR). MCAR 
assumes that the probability of an observation being missing does not depend 
upon the observed responses or upon any unobserved covariates but may depend 
upon covariates included in the model (eg, missing more among whites than 
nonwhites). When missing data are minimal (specifically if no more than 20% of 
participants are missing any values), then nonparametric tests and GEE methods 
will be used, because violations of the MCAR assumption will have little impact 
on the estimates and hypothesis tests. These models will include as covariates all 
available baseline predictors of the missing outcomes.  
If a substantial amount of immunogenicity data is missing (at least 1 value 
missing from more than 20% of participants), then using the methods that require 
the MCAR assumption may give misleading results. In this situation, analyses of 
the immunogenicity endpoints at a specific timepoint will be performed using 
parametric generalized linear models fit by maximum likelihood. These methods 
provide unbiased estimation and inferences under the parametric modeling 
assumptions and the assumption that the missing data are missing at random 
(MAR). MAR assumes that the probability of an observation being missing may 
depend upon the observed responses and upon observed covariates, but not upon 
any unobserved factors. Generalized linear models for response rates will use a 
binomial error distribution and for quantitative endpoints, a normal error 
distribution. For assessing repeated immunogenicity measurement, linear mixed 
effects models will be used. If the immunological outcomes are left- and/or right-
censored, then the linear mixed effects models of Hughes (39) will be used 
because they accommodate the censoring. In addition, secondary analyses of 
repeated immunogenicity measurements may be done using weighted GEE (40) 
methods, which are valid under MAR. All of the models described above will 
include as covariates all available baseline predictors of the missing outcomes. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 50 of 107 
6.4.5 
Analyses prior to end of scheduled follow-up visits 
Any analyses conducted prior to the end of the scheduled follow-up visits should 
not compromise the integrity of the trial in terms of participant retention or safety 
or immunogenicity endpoint assessments. In particular, early analyses by 
treatment assignment require careful consideration and should be made available 
on a need-to-know basis only. 
6.4.5.1 
Safety 
During the course of the trial, analyses of safety data will be prepared 
approximately every 4 months for review by the Safety Monitoring Board (SMB). 
Ad hoc safety reports may also be prepared for SMB review at the request of the 
HVTN 304 PSRT. The HVTN leadership must approve any other requests for 
safety data prior to the end of the scheduled follow-up visits. 
6.4.5.2 
Immunogenicity 
A statistical analysis by treatment assignment of a primary immunogenicity 
endpoint may be performed. The HVTN Laboratory Program will review the 
analysis report prior to distribution to the protocol chairs, Division of AIDS 
(DAIDS), vaccine developer, and other key HVTN members and investigators. 
Distribution of reports will be limited to those with a need to know for the 
purpose of informing future trial-related decisions. The HVTN leadership must 
approve any other requests for HVTN immunogenicity analyses prior to the end 
of the scheduled follow-up visits. Any analyses conducted prior to the end of the 
study should not compromise the integrity of the trial in terms of participant 
retention or safety or immunogenicity endpoint assessments. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 51 of 107 
7 
Study vaccine preparation, storage, and 
administration 
7.1 
Vaccine Regimen 
The schedule of vaccination is shown in Section 1 and additional information is 
given below. 
Group 1  
Treatment 1 (T1):  
INO-6160, 2 mg dose, to be administered as 2 separate intradermal (ID) injections 
(0.1 mL each) followed by electroporation (EP) using the CELLECTRA 2000 EP 
device at months 0, 1, 3, and 6. 
Group 2  
Treatment 2 (T2):  
INO-6160, 2 mg dose, 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, 3 
and 6. 
Trimer 4571, 100 mcg, admixed with 3M-052-AF, 5 mcg, and Alum, 500 mcg, to 
be administered as 2 separate intramuscular (IM) injections (0.2 mL each) at 
months 3 and 6. 
7.2 
Study Product Formulation and Storage 
7.2.1 
INO-6160 
INO-6160 Drug Product is sD-NLT-AB05 coformulated with IL-12 DNA at a 
plasmid ratio of 4:1 (0.8 mg sD-NLT-AB05 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 size glass vials. Vials may 
contain a volume greater than the labeled volume. 
Store INO-6160 frozen at -25°C to -15°C. The study product is described in 
further detail in the IB. 
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 52 of 107 
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 
a preservative. 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 WFI 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 technique and sterile preparation 
procedures/guidance as outlined in USP <797>, utilizing a pharmacy biosafety 
cabinet/isolator or better. Local regulations and site institutional policies and 
procedures for 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 portion of entered vials, or unused prepared study product 
should be discarded in a biohazard container and disposed of in accordance with 
institutional or pharmacy policy. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 53 of 107 
7.3.1 
INO-6160, 2 mg (Group 1 and Group 2) 
1. Remove 1 vial of INO-6160, 10 mg/mL from the freezer and record this as the 
study product preparation start time. 
2. Once contents of vial are thawed, insert the sterile BD 0.5 mL Tuberculin 
Syringe with permanently attached needle, 27G x ½ inch (BD Reference 
Number 305620), into the sterile glass vial containing INO-6160, 10 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-6160, 10 mg/mL, 0.1mL 
each. 
3. Label the syringes according to Section 7.3.3 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 syringes may be stored at room temperature and used within 4 hours 
of study product preparation start time.  
7.3.2 
Trimer 4571, 100 mcg, admixed with 3M-052-AF, 5 mcg, and Alum, 
500 mcg (Group 2) 
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 the 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 54 of 107 
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 7.3.3 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.   
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.  
7.3.3 
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  
7.3.4 
Study Vaccine Administration 
7.3.4.1 
INO-6160 
INO-6160, 2 mg, will be administered as 2 separate 0.1 mL injections 
intradermally, bilaterally, one on each arm at months 0, 1, 3 and 6. Following ID 
injections of INO-6160, EP will be performed using the Inovio CELLECTRA 
2000 EP Device.  
The ID injections will be administered in 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 55 of 107 
contents will be injected to form a small bleb. One injection should be completed 
(including EP) prior to administering the second injection. 
If an injection cannot be given in the upper arm due to a medical contraindication, 
it should be administered in the thigh (not in the contralateral upper arm). If both 
upper arms are unsuitable for injection, the injections should be administered in 
the thigh. The appropriate study staff should document this clearly. 
7.3.4.2 
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 2 participants at months 3 and 6, following 
administration of INO-6160, as 2 separate 0.2 mL injections (1 in each arm) 
intramuscularly into the deltoid muscles by needle and syringe.  Immediately 
prior to administration, gently invert each syringe 10 times.  
If an injection cannot be given in a deltoid muscle due to a medical 
contraindication, it should be administered in the thigh (not in the contralateral 
deltoid). If both deltoids are unsuitable for injection, the injections should be 
administered in the thigh muscle. The appropriate study staff should document 
this clearly.  
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. 
7.4 
Acquisition of study products 
sD-NLT-AB05 and IL-12 DNA are manufactured under quality controls and 
production protocols developed and tech transferred by Inovio Pharmaceuticals. 
INO-6160, the co-formulation containing sD-NLT-AB05 and 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 are manufactured by Leidos Biomed (Vaccine Research 
Center, Frederick, MD, USA). Trimer 4571 will be provided by Dale and Betty 
Bumpers Vaccine Research Center (VRC), and Alum will be provided by 
DAIDS, the National Institute of Allergy and Infectious Diseases (NIAID), the 
US National Institutes of Health (NIH) (Rockville, MD, USA). 
3M-052-AF is manufactured and provided by Access to Advanced Health 
Institute (AAHI) (Seattle, Washington, USA). 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 56 of 107 
Once an HVTN clinical research site (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. 
7.5 
Study Vaccine Accountability 
The HVTN CRS pharmacist is required to maintain complete records of all study 
products.  
7.6 
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. 
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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 57 of 107 
8 
Clinical procedures 
All required clinic and laboratory procedures for each study visit are summarized 
in Appendix A, Schedule of 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) 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 Assessment of Understanding (AoU) will be completed prior to enrollment. 
Records will be kept documenting the reason(s) that screened participants did not 
enroll. 
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 B), 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 14 full days 
following vaccine administration. Clinicians will follow and collect resolution 
information for any reactogenicity signs and symptoms that have not resolved 
within the reactogenicity assessment period. 
Pre-vaccine Administration: Intercurrent illness assessment and evaluations 
(including vital signs and planned injection-site evaluation) are performed prior to 
each vaccine administration. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 58 of 107 
Guidance for participants scheduling licensed vaccines or allergy 
immunotherapy: In order to prevent interference with reactogenicity assessment, 
participants who plan to receive licensed vaccines or allergy immunotherapy 
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). 
Post-vaccine administration in-clinic evaluation: Following the enrollment 
vaccine administration, participants will be observed for a minimum of 60 
minutes post study product administration. For all subsequent administrations, 
participants will be observed for a minimum of 30 minutes with the following 
exception: for Group 2, participants will be observed for 60 minutes following the 
first administration of Trimer 4571 + 3M-052 AF + Alum scheduled for Month 3. 
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.  
Post-vaccine 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 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. 
Symptom diaries: Participants are asked to record symptoms on a daily basis 
using an electronic participant diary (eDiary). A paper alternative is available. All 
participants will be given a thermometer for oral temperature measurement, a 
ruler, and access to the eDiary (paper alternative available). All participants will 
be provided training on diary completion, proper thermometer usage, and the 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 the day of vaccination and 14 full days 
following each vaccine administration. 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. 
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 
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 59 of 107 
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 reactogenicity. An abbreviated assessment will take 
place during the AESI contact (Section 8.8). Please see HVTN 304 Study-specific 
Procedures (SSP) for further details. 
8.5 
Visit windows and missed visits 
The schedule of visits and evaluations performed at each visit is shown in 
Appendix A. Visit windows are shown in Appendix B. The procedures for 
documenting missed visits and out of window visits are described in the HVTN 
304 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. 
8.6 
Monitoring for HIV infection 
Study participants will be tested for HIV infection periodically throughout the 
study per the HVTN HIV testing algorithm as noted in Appendix A. 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 infection in accordance with the most recent Centers for 
Disease Control and Prevention HIV counseling guidelines. 
Although the study vaccine will not cause HIV infection, it may induce Abs 
detectable by standard HIV infection 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. 
• 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 the 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 60 of 107 
potential for VISP) and to qualify former participants for post-study HIV 
testing to distinguish between VISP and HIV infection. Information in the 
VISP registry is not used for research.  
8.7 
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, physical examination, and if 
indicated, a pregnancy test (see Appendix A). If a participant acquires HIV, 
please see Section 5.2.2. If the site PI has questions regarding a termination visit, 
they should consult with the PSRT. 
8.8 
AESI contact 
CRS staff will contact study participants 12 months after the last vaccination to 
collect the information listed below and will perform an abbreviated EP-
injection–site assessment 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 AESI assessment. AESIs are described further in Appendix 
F. 
• 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; 
▪ AESIs (a sample list of AESIs is provided in Appendix F). AESIs are 
reported regardless of relationship to study product(s); 
▪ Medically attended adverse events (MAAEs), defined as any AEs leading 
to an unscheduled visit to a healthcare professional, which are reported 
regardless of relationship to study product(s); 
▪ New diagnosis of HIV infection; 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.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 61 of 107 
9 
Safety and adverse events 
9.1 
Adverse events 
Unsolicited 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 Appendix F. 
In addition, a limited set of AEs will be collected and reported for 12 months 
following the last vaccine administration the participant receives: 
• SAEs/EAEs, 
• AESIs, 
• MAAEs,  
• 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 
304 SSP). 
• 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 
▪ Grade 4 is: Potentially life-threatening consequences (eg, abscess, 
exfoliative dermatitis, necrosis involving dermis or deeper tissue) 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 62 of 107 
9.2 
Serious adverse events 
The term “Serious Adverse Event” (SAE) is defined in 21 CFR 312.32 as follows:  
“An adverse event or suspected adverse reaction is considered serious if, in the 
view of either the investigator or the sponsor, it results in any of the following 
outcomes: 
• Death,  
• A life-threatening adverse event, inpatient hospitalization or prolongation of 
existing hospitalization,  
• A persistent or significant incapacity or substantial disruption of the ability to 
conduct normal life functions, 
• 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 
Regulatory Support Center (RSC) website at https://rsc.niaid.nih.gov/clinical-
research-sites/manual-expedited-reporting-adverse-events-daids. 
The DAIDS Adverse Experience Reporting System (DAERS), an internet-based 
reporting system, must be used for expedited AE (EAE) reporting to DAIDS. In 
the event of system outages or technical difficulties, EAEs may be submitted 
using the DAIDS EAE Form. This form is available on the DAIDS RSC website 
at https://rsc.niaid.nih.gov/clinical-research-sites/paper-eae-reporting. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 63 of 107 
For questions about DAERS, please contact NIAID CRMS Support at 
CRMSSupport@niaid.nih.gov. Please note that site queries may also be sent from 
within the DAERS application itself.  
For questions about expedited 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 and investigational device for which expedited reporting are 
required are: 
• INO-6160, 
• [Trimer 4571 + 3M-052-AF + Alum], and 
• EP Device. 
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 
Protocol Safety Review Team (PSRT) 
The PSRT comprises: DAIDS Medical Officer, Protocol Chair, Protocol Cochair, 
Protocol Team Leader, and Clinical Safety Specialist (CSS). Other members of 
the Protocol Team (Clinic Coordinator, Clinical Data Manager, Clinical Trial 
Manager, vaccine developer representative(s), and others) may also be included in 
HVTN 304 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 (SMB) 
The 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 64 of 107 
months. The SMB conducts additional special reviews at the request of the HVTN 
304 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). 
9.4 
Total blood volume 
Required blood volumes per visit are shown in Appendix A. 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 
Enrollment will be restricted to 1 participant per day for the first 5 participants 
and then enrollment will pause. Participants who have been enrolled will continue 
to receive vaccinations during the pause. The PSRT will review cumulative safety 
information recorded through the visit scheduled 2 weeks post first vaccination 
for the first 5 participants and will determine whether it is safe to proceed with 
full enrollment. 
9.6 
Safety pause and prompt PSRT AE review 
The PSRT (see Section 9.3.1) 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 an immediate safety pause or prompt HVTN 304 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 304 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 65 of 107 
Table 9-1 Pause Rules  
Event and 
relationship to 
study vaccine 
Severity 
Grade 
HVTN Site Actions 
HVTN Core 
Action 
SAE, related 
Any 
Phone 24/7 Safety Phone immediately 
Email vtn.clin.safety.spec@hvtn.org 
Submit CRFs immediately 
Immediate pause 
and PSRT 
review 
AE, related  
(see Grade 3 
exceptions below) 
4 or 3 
Email CSS 
Submit CRFs immediately 
Prompt PSRT 
AE review to 
consider a pause 
*Once a CRF is submitted to the database that meets a Pause Rule, the CSS receives an immediate 
email alert in order to follow up with the site and inform the PSRT.  
Exceptions to the 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 office phone upon learning 
of any unrelated participant deaths. The site will also email the CSS and 
immediately submit CRFs. The CSS will then promptly notify the PSRT. 
If you need to contact the CSS, refer to phone numbers and email addresses found 
on the Protocol home page on the HVTN Members’ site 
(https://members.hvtn.org/protocols/hvtn304). 
9.6.1 
Plan for review of pause rules  
For all safety pauses, HVTN Core notifies the HVTN 304 PSRT, HVTN 
Regulatory Affairs, DAIDS Pharmaceutical Affairs Branch (PAB), DAIDS 
Regulatory Affairs Branch (RAB), DAIDS Safety and Pharmacovigilance Team 
(SPT), and participating HVTN CRSs. When an immediate safety pause is 
triggered, HVTN Core notifies the SMB. 
Once a trial is paused, the HVTN 304 PSRT reviews safety data and decides 
whether the pause can be lifted or 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 66 of 107 
study vaccinations. Based on the HVTN 304 PSRT assessment, DAIDS RAB 
notifies the FDA as needed. 
If an immediate HVTN 304 PSRT notification or prompt HVTN 304 PSRT AE 
review is triggered, HVTN Core notifies the HVTN 304 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 304 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 304 PSRT, 
the NIH, the US Department of Health and Human Services (DHHS) 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 the 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 
304 SSP. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 67 of 107 
10 
Protocol conduct and informed consent 
10.1 
Protocol conduct 
This research study will be conducted in compliance with the protocol, Good 
Clinical Practice (GCP) (ICH E6 (R2)), HVTN and DAIDS policies and 
procedures as specified in the HVTN Manual of Operations and the 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 304 SSP.  
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 (CAB) are required by DAIDS and supported at all 
HVTN research sites to ensure community input in accordance with Good 
Participatory Practices (GPP) and all local and national guidelines. 
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, 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. 
10.3 
Informed consent 
The sample informed consent form (SICF) in Appendix C 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 68 of 107 
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 level of understandability. 
If any new information is learned that might affect the participants’ decisions to 
stay in the trial, this information will be shared with trial participants. 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 69 of 107 
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 accord with the language in the SICF (Appendix C) 
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 C). 
This research may relate to HIV, vaccines, the immune system, and 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 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 a Web-based tool that documents their 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 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 70 of 107 
12 
Literature cited 
1. Burton DR, Hangartner L. Broadly Neutralizing Antibodies to HIV and Their 
Role in Vaccine Design. Annu Rev Immunol. 2016;34:635-59. 
2. Medina-Ramirez M, Garces F, Escolano A, Skog P, de Taeye SW, Del 
Moral-Sanchez I, et al. Design and crystal structure of a native-like HIV-1 
envelope trimer that engages multiple broadly neutralizing antibody 
precursors in vivo. J Exp Med. 2017;214(9):2573-90. 
3. Kulp DW, Steichen JM, Pauthner M, Hu X, Schiffner T, Liguori A, et al. 
Structure-based design of native-like HIV-1 envelope trimers to silence non-
neutralizing epitopes and eliminate CD4 binding. Nat Commun. 
2017;8(1):1655. 
4. Sanders RW, Moore JP. Native-like Env trimers as a platform for HIV-1 
vaccine design. Immunol Rev. 2017;275(1):161-82. 
5. Steichen JM, Kulp DW, Tokatlian T, Escolano A, Dosenovic P, Stanfield RL, 
et al. HIV Vaccine Design to Target Germline Precursors of Glycan-
Dependent Broadly Neutralizing Antibodies. Immunity. 2016;45(3):483-96. 
6. Xu Z, Walker S, Wise MC, Chokkalingam N, Purwar M, Moore A, et al. 
Induction of tier-2 neutralizing antibodies in mice with a DNA-encoded HIV 
envelope native like trimer. Nat Commun. 2022;13(1):695. 
7. Chuang GY, Geng H, Pancera M, Xu K, Cheng C, Acharya P, et al. 
Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer 
with Reduced CD4 Affinity and Improved Immunogenicity. J Virol. 
2017;91(10). 
8. Kwon YD, Pancera M, Acharya P, Georgiev IS, Crooks ET, Gorman J, et al. 
Crystal structure, conformational fixation and entry-related interactions of 
mature ligand-free HIV-1 Env. Nat Struct Mol Biol. 2015;22(7):522-31. 
9. Sanders RW, Derking R, Cupo A, Julien JP, Yasmeen A, de Val N, et al. A 
next-generation cleaved, soluble HIV-1 Env trimer, BG505 SOSIP.664 
gp140, expresses multiple epitopes for broadly neutralizing but not non-
neutralizing antibodies. PLoS Pathog. 2013;9(9):e1003618. 
10. Trinchieri G. Interleukin-12: a proinflammatory cytokine with 
immunoregulatory functions that bridge innate resistance and antigen-specific 
adaptive immunity. Annu Rev Immunol. 1995;13:251-76. 
11. Trinchieri G. Interleukin-12 and the regulation of innate resistance and 
adaptive immunity. Nat Rev Immunol. 2003;3(2):133-46. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 71 of 107 
12. Kalams SA, Parker SD, Elizaga M, Metch B, Edupuganti S, Hural J, et al. 
Safety and comparative immunogenicity of an HIV-1 DNA vaccine in 
combination with plasmid interleukin 12 and impact of intramuscular 
electroporation for delivery. J Infect Dis. 2013;208(5):818-29. 
13. Kalams SA, Parker S, Jin X, Elizaga M, Metch B, Wang M, et al. Safety and 
immunogenicity of an HIV-1 gag DNA vaccine with or without IL-12 and/or 
IL-15 plasmid cytokine adjuvant in healthy, HIV-1 uninfected adults. PLoS 
ONE. 2012;7(1):e29231. 
14. Fox CB, Orr MT, Van Hoeven N, Parker SC, Mikasa TJ, Phan T, et al. 
Adsorption of a synthetic TLR7/8 ligand to aluminum oxyhydroxide for 
enhanced vaccine adjuvant activity: A formulation approach. J Control 
Release. 2016;244(Pt A):98-107. 
15. Smirnov D, Schmidt JJ, Capecchi JT, Wightman PD. Vaccine adjuvant 
activity of 3M-052: an imidazoquinoline designed for local activity without 
systemic cytokine induction. Vaccine. 2011;29(33):5434-42. 
16. Van Hoeven N, Fox CB, Granger B, Evers T, Joshi SW, Nana GI, et al. A 
Formulated TLR7/8 Agonist is a Flexible, Highly Potent and Effective 
Adjuvant for Pandemic Influenza Vaccines. Sci Rep. 2017;7:46426. 
17. Gupta SG-O, J; Hong, D; Marabelle, A; Munster, P; Aggarwal, R; Aspeslagh, 
S; G. Dixon, R; Patel, M; Subbiah, V; Morehouse, C; Wu, Y; Zha, J; Tseng, 
L; Cooper, Z; Morris, S; Brody, J. Abstract CT091: Safety and 
pharamcodynammic activity of MEDI9197, a TLR 7/8 agonist, administered 
intratumorally in subjects with solid tumors (Proceedings: AACR Annual 
Meeting 2017; April 1-5, 2017; Washington, DC). Cancer Research 
[Internet]. 2017; 77 
(13):[http://cancerres.aacrjournals.org/content/77/13_Supplement/CT091 p.]. 
Available from: 
http://cancerres.aacrjournals.org/content/77/13_Supplement/CT091. 
18. 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. 
19. Pino M, Abid T, Pereira Ribeiro S, Edara VV, Floyd K, Smith JC, et al. A 
yeast expressed RBD-based SARS-CoV-2 vaccine formulated with 3M-052-
alum adjuvant promotes protective efficacy in non-human primates. Sci 
Immunol. 2021;6(61). 
20. Hu JK, Crampton JC, Cupo A, Ketas T, van Gils MJ, Sliepen K, et al. Murine 
Antibody Responses to Cleaved Soluble HIV-1 Envelope Trimers Are Highly 
Restricted in Specificity. J Virol. 2015;89(20):10383-98. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 72 of 107 
21. Dingens AS, Pratap P, Malone K, Hilton SK, Ketas T, Cottrell CA, et al. 
High-resolution mapping of the neutralizing and binding specificities of 
polyclonal sera post-HIV Env trimer vaccination. Elife. 2021;10. 
22. Zhao F, Joyce C, Burns A, Nogal B, Cottrell CA, Ramos A, et al. Mapping 
Neutralizing Antibody Epitope Specificities to an HIV Env Trimer in 
Immunized and in Infected Rhesus Macaques. Cell Rep. 
2020;32(10):108122. 
23. Jalah R, Kulkarni V, Patel V, Rosati M, Alicea C, Bear J, et al. DNA and 
protein co-immunization improves the magnitude and longevity of humoral 
immune responses in macaques. PLoS ONE. 2014;9(3):e91550. 
24. Singh S, Ramirez-Salazar EG, Doueiri R, Valentin A, Rosati M, Hu X, et al. 
Control of Heterologous Simian Immunodeficiency Virus SIVsmE660 
Infection by DNA and Protein Coimmunization Regimens Combined with 
Different Toll-Like-Receptor-4-Based Adjuvants in Macaques. J Virol. 
2018;92(15). 
25. De Rosa SC, Edupuganti S, Huang Y, Han X, Elizaga M, Swann E, et al. 
Robust antibody and cellular responses induced by DNA-only vaccination for 
HIV. JCI Insight. 2020;5(13). 
26. Edupuganti S, De Rosa SC, Elizaga M, Lu Y, Han X, Huang Y, et al. 
Intramuscular and Intradermal Electroporation of HIV-1 PENNVAX-
GP((R)) DNA Vaccine and IL-12 Is Safe, Tolerable, Acceptable in Healthy 
Adults. Vaccines (Basel). 2020;8(4). 
27. Elizaga ML, Li SS, Kochar NK, Wilson GJ, Allen MA, Tieu HVN, et al. 
Safety and tolerability of HIV-1 multiantigen pDNA vaccine given with IL-
12 plasmid DNA via electroporation, boosted with a recombinant vesicular 
stomatitis virus HIV Gag vaccine in healthy volunteers in a randomized, 
controlled clinical trial. PLoS One. 2018;13(9):e0202753. 
28. Li SS, Kochar NK, Elizaga M, Hay CM, Wilson GJ, Cohen KW, et al. DNA 
priming increases frequency of T-cell responses to a VSV HIV vaccine with 
specific enhancement of CD8+ T-cell responses by IL-12 pDNA. Clin 
Vaccine Immunol. 2017. 
29. Li J, Valentin A, Kulkarni V, Rosati M, Beach RK, Alicea C, et al. HIV/SIV 
DNA vaccine combined with protein in a co-immunization protocol elicits 
highest humoral responses to envelope in mice and macaques. Vaccine. 
2013;31(36):3747-55. 
30. Patel V, Jalah R, Kulkarni V, Valentin A, Rosati M, Alicea C, et al. DNA and 
virus particle vaccination protects against acquisition and confers control of 
viremia upon heterologous simian immunodeficiency virus challenge. Proc 
Natl Acad Sci USA. 2013;110(8):2975-80. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 73 of 107 
31. Felber BK, Lu Z, Hu X, Valentin A, Rosati M, Remmel CAL, et al. Co-
immunization of DNA and Protein in the Same Anatomical Sites Induces 
Superior Protective Immune Responses against SHIV Challenge. Cell Rep. 
2020;31(6):107624. 
32. Hosseinipour MC, Innes C, Naidoo S, Mann P, Hutter J, Ramjee G, et al. 
Phase 1 Human Immunodeficiency Virus (HIV) Vaccine Trial to Evaluate 
the Safety and Immunogenicity of HIV Subtype C DNA and MF59-
Adjuvanted Subtype C Envelope Protein. Clin Infect Dis. 2021;72(1):50-60. 
33. Rouphael NG, Morgan C, Li SS, Jensen R, Sanchez B, Karuna S, et al. DNA 
priming and gp120 boosting induces HIV-specific antibodies in a randomized 
clinical trial. J Clin Invest. 2019;129(11):4769-85. 
34. Sardesai NY, Weiner DB. Electroporation delivery of DNA vaccines: 
prospects for success. Curr Opin Immunol. 2011;23(3):421-9. 
35. Sampson HA, Munoz-Furlong A, Campbell RL, Adkinson NF, Jr., Bock SA, 
Branum A, et al. Second symposium on the definition and management of 
anaphylaxis: summary report--Second National Institute of Allergy and 
Infectious Disease/Food Allergy and Anaphylaxis Network symposium. J 
Allergy Clin Immunol. 2006;117(2):391-7. 
36. Fagerland MW, Lydersen S, Laake P. Recommended confidence intervals for 
two independent binomial proportions. Stat Methods Med Res. 
2015;24(2):224-54. 
37. Agresti A, Caffo B. Simple and effective confidence intervals for proportions 
and differences of proportions result from adding two success and two 
failures. American Statistician. 2000;54(4):280-8. 
38. Agresti A, Coull BA. Approximate is better than "exact" for interval 
estimation of binomial proportions. Am Stat. 1998;52(2):119-26. 
39. Hughes JP. Mixed effects models with censored data with application to HIV 
RNA levels. Biometrics. 1999;55(2):625-9. 
40. Rotnitzky A, Robins J. Analysis of semi-parametric regression models with 
non-ignorable non-response. Stat Med. 1997;16(1-3):81-102. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 74 of 107 
Appendix A 
Schedule of procedures 
Visit Number 
01 
02 
03 
04 
05 
06 
07 
08 
09 
10 
AESI13 
Study Week 
 
0 
2 
4 
6 
12 
14 
24 
26 
52 
78 
Study Month 
 
0 
0.5 
1 
1.5 
3 
3.5 
6 
6.5 
12 
18 
Study Day 
-56 to 1 
1 
15 
29 
43 
85 
99 
169 
183 
365 
547 
Procedure 
Screen1 
Vac 1 
 
Vac 2 
 
Vac 3 
 
Vac 4 
 
 
 
Study procedures 
 
 
 
 
 
 
 
 
 
 
 
 
Assessment of Understanding 
(AoU) 
 
√ 
 
 
 
 
 
 
 
 
 
 
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 
 
 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
Clinical labs 
Tube 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy test (urine or serum) 9, 10 
 
√ 
√ 
 
√ 
 
√ 
 
√ 
 
 
 
HBsAg/anti-HCV 10 
SST 
5 
 
 
 
 
 
 
 
 
 
 
HIV screening test 10, 11 
SST 
5 
 
 
 
 
 
 
 
 
 
 
CBC/Differential 10 
EDTA 
5 
 
5 
 
5 
 
5 
 
5 
 
 
ALT & Creatinine 10 
SST 
5 
 
5 
 
5 
 
5 
 
5 
 
 
HIV diagnostic test 
EDTA 
 
 
 
 
 
10 
 
10 
 
20 
 
Research samples12 
 
 
 
 
 
 
 
 
 
 
 
 
PBMC for assays & storage 
ACD 
136 
 
 
 
136 
51 
136 
 
136 
178.5 
 
Serum for assays and storage 
SST 
34 
 
 
 
34 
34 
34 
 
34 
34 
 
Daily volume (mL) 
190 
0 
10 
0 
180 
95 
180 
10 
180 
232.5 
 
56-day total volume (mL) 
190 
190 
200 
200 
380 
275 
455 
10 
190 
232.5 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 75 of 107 
Blue shading: Vaccination Visit 
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, vital signs, and clinical assessments of head, ears, 
eyes, nose, and throat; neck, 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 report 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. 
5 Risk reduction counseling per CRS Standard Operating Procedures. 
6 Concomitant medications, including prescription and non-prescription 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 each vaccination and prior to clinic discharge, participants will have vital signs taken, the 
injection site will be assessed, and systemic symptoms will be assessed.  
8 Reactogenicity: Reactogenicity assessments performed daily from the day of vaccination through 14 full days following vaccine administration. (see 
Section 8.3) 
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 AESI contact: CRS staff will contact study participants 12 months after the last vaccination received to collect the information listed in Section 8.8. The 
EP-injection–site assessment will be an abbreviated form that can be conducted over the phone.  
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 76 of 107 
Appendix B 
Visit windows 
 
Visit 
Number 
Visit Type 
Lower 
Allowable 
Window 
(-) 
Lower 
Target 
Window 
(-) 
Target 
Day* 
Upper 
Target 
Window 
(+) 
Upper 
Allowable 
Window 
(+) 
01.0 
Screening 
-56 
- 
- 
- 
- 
02.0 
Enrollment / 
Vaccination 1 
- 
- 
1 
- 
- 
03.0 
2 weeks post-
vaccination 1 
 
-4 
15 
+4 
+7 
 
04.0 
Vaccination 2 
 
- 
29 
+9 
+28 
05.0 
2 weeks post-
vaccination 2 
 
-4 
43 
+4 
+7 
06.0 
Vaccination 3 
 
- 
85 
+14 
+28 
07.0 
2 weeks post-
vaccination 3 
 
- 
-4 
99 
+4 
+7 
08.0 
Vaccination 4 
- 
-7 
169 
+14 
+28 
09.0 
2 weeks post-
vaccination 4 
- 
-4 
183 
+4 
+7 
10.0 
6 months post 
vaccination 4 
-28 
-14 
365 
+14 
+28 
AESI* 
Final Visit 
-21 
-14 
547 
 
+14 
+28 
All target dates are relative to Day 1, with the exception of the post-vaccination visits, visits 3, 5, 
7, and 9 which are relative to the vaccination immediately preceding the visit. 
*This contact must be at least 12 months post last vaccination. 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 77 of 107 
Appendix C 
Sample informed consent form 
Title: A phase 1 open-label clinical trial to evaluate the safety and 
immunogenicity of synthetic DNAs encoding a native-like HIV Env Trimer and 
Interleukin-12 (INO-6160), alone or in a prime-boost regimen with 3M-052-AF + 
Alum adjuvanted VRC HIV Env Trimer 4571 in adult participants without HIV 
HVTN protocol number: HVTN 304 
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. 
Key information 
• This is the first study in which one of the study vaccines will be given 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 age 18-55, do 
not have HIV, and are generally healthy. 
• The purpose of this study is to see if the study vaccines are safe and to see 
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 1 year after your last vaccination to check on your health. 
• Procedures will include blood draws and injections of study vaccines. 
• 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. We will tell you more about this procedure and 
its risks later in this consent form.  
• There are risks from participating. We will tell you more about all of them 
later in this consent form. The most common risks are as follows: 
▪ Taking blood and giving injections can cause bruising, pain, fainting, 
soreness, redness, swelling, itching, a sore and bleeding. 
▪ 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 12 months or longer. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 78 of 107 
▪ 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 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 25 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 three 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 vaccines? (Your 
immune system protects you from infections and disease.) 
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. 
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 and a study adjuvant.  
One is called INO-6160. We will call it the DNA vaccine. It is made up of 2 
different pieces of DNA, sD-NLT-AB05 + interleukin-12 or IL-12. This DNA is 
very much like natural DNA, but it was 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 study vaccine is called Trimer 4571. We will call it the protein 
vaccine. It is a small, lab-made piece of protein that looks like a part of a protein 
found on the surface of the HIV virus. In this study, it is being mixed with an 
adjuvant called 3M-052-AF and another adjuvant called Alum. We will call the 
combination of these 2 adjuvants the study adjuvant. Adjuvants are products that 
help alert the immune system to have a stronger response. 3M-052 was originally 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 79 of 107 
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. This study is using the improved 3M-052-AF 
adjuvant. It is dissolved in water and 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. 
Everyone in this study will get the DNA vaccine. It will tell your body to make 
small amounts of 2 different proteins. One of them is a protein that is found in 
HIV, and the second protein is called IL-12. IL-12 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 
make antibodies and T cells more efficiently. Antibodies and T cells are parts of 
an immune response that defends the body against diseases. 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 and the study adjuvant. 
The immune system might be able to see this piece of protein and learn how to 
recognize HIV if you are ever exposed to it in the future.  
Where do the study products come from? 
The DNA vaccine was developed by the Wistar Institute (Philadelphia, 
Pennsylvania) and formulated by Inovio Pharmaceuticals. It is provided by the 
Wistar Institute through a grant from the NIH.  
The protein vaccine was developed and is provided by the Dale and Betty 
Bumpers Vaccine Research Center (VRC), which is part of the NIH.  
The 3M-052-AF adjuvant was developed, manufactured and is being provided by 
the Access to Advanced Health Institute (AAHI) in Seattle, Washington. Alum is 
being provided by the Division of AIDS (DAIDS) in Bethesda, Maryland, which 
is part of the NIH. 
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 4 other studies. In general, people who got DNA vaccines in those studies were 
not too uncomfortable and did not have any serious health problems related to the 
vaccine. Six 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 as part of 2 
different ongoing studies. In one study, VRC018, 16 people got the same protein 
vaccine mixed with the Alum adjuvant that we will use in this study. Each person 
got 3 injections of the vaccine and adjuvant at the same dose or higher than the 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 80 of 107 
dose we will use in this study. The injections did not cause any serious health 
problems. Most people in that study had at least one of the side effects listed in 
the Risks of vaccines described below. They said the 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 the same 
protein vaccine combined with Alum at a higher dose than the one we will use in 
this study. These injections did not cause any serious health problems. This study 
is still ongoing and plans to enroll up to 100 people.  
Another protein vaccine and the same 3M-052-AF + Alum adjuvant being used in 
this study are being tested for the first time in people in 2 other studies called 
HVTN 137 and HVTN 300. As of Spring 2022, 48 people in these 2 studies have 
had at least 1 study injection and at least 45 people have had 2 injections. These 
injections have not caused any serious health problems. 
HVTN 300 enrolled 13 people. All people in the study had at least some injection 
site reactions during the trial, mostly mild to moderate. One (1) 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. These severe side effects went away within 2 days. Of 
the 5 people who had severe side effects, 2 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 
one (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. 
One person enrolled in HVTN 137 had some redness in the area where they got 
the injection that lasted for about 3 days. They also had significant swelling on 
their arm for about 4 days. It took about 6 weeks for the swelling to completely go 
away. They also had mild pain, which did not prevent them from going to work, 
and the pain went away on its own. This person decided to stop getting any more 
injections. There were 4 people who experienced stronger side effects. These 
included chills, headache, muscle aches, and generally feeling unwell. Because 
the study is ongoing and is blinded, we do not know if these people got the study 
vaccine with the adjuvant or a placebo (sterile salt water with no vaccine in it).  
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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 81 of 107 
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. 
Risks of vaccines: 
All vaccines can cause fever, chills, rash, aches and pains, nausea, dizziness, 
feeling tired and generally feeling unwell. Vaccines can also cause pain, 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 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 the “Risks of vaccines” section above, but no serious health 
concerns have been reported so far. 
These are the side effects 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 82 of 107 
4. The DNA vaccine is given using electroporation. 
Besides adjuvants, another way to improve immune responses to DNA vaccines is 
to use EP. EP uses an electrical pulse to briefly open tiny pores in the cells. The 
DNA can enter the cells through these pores. EP has been used for many years in 
the laboratory to get DNA or other substances into cells. One study also showed 
increased immune responses to another experimental DNA vaccine. 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. 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, together with IL-12 adjuvant, were given into 
the skin with EP to 55 people. EP has not caused 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 
9,300 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 and scabs. Later the marks may heal but might 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 83 of 107 
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 
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/friends if it felt the same as their study experience, and 96% of these 
participants said that the appearance at injection sites was acceptable.  
We do not know if EP will change the risks for the DNA vaccine. 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 not with this DNA vaccine.  
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, that will not affect your other care at this clinic 
and 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. 
During the study, you should not donate blood or tissue. 
Site: Remove item 8 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-
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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 84 of 107 
We will review the test results with you. They may show you are not eligible for 
the study, even if you want to.  
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 E, 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 (for a total of 
9 months). We will talk to you about effective birth control methods. They are 
listed on a handout that we will give to you. 
You also should not begin the process to have your eggs collected until 8 weeks 
after your last scheduled study injection. If this is something you are considering, 
please discuss it with your study doctor and your fertility specialist. 
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 10 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 
contact you 12 months 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).  
10. We will pay you for each study visit you complete. 
This payment is to cover the costs of [Site: insert text].  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 85 of 107 
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. 
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 vaccines on a schedule. 
We will give participants injections of the study vaccines at 4 visits, according to 
the schedule in the table below. All participants will get 2 injections of the DNA 
vaccine with a needle and syringe into the skin of both upper arms followed by 
electroporation at all 4 vaccination visits. People in Group 2 will also get 2 
injections of the protein vaccine plus the study adjuvant with a needle and syringe 
into the upper arm muscle of both arms at the last 2 vaccination visits. If for some 
reason we cannot give an injection in your arm, we may give it in the thigh 
instead.  
You have an equal chance of being assigned to Group 1 or Group 2. Site: Modify 
the randomization metaphor in the next sentence as appropriate to your local 
culture. Whether you are assigned to Group 1 or 2 is completely random, like 
rolling dice or drawing straws. 
Because this is the first time the DNA vaccine is being given to people, only 1 
participant will get the study vaccine each day, until a total of 5 participants 
across both groups have been vaccinated. After this, we will pause the study for a 
safety review by a team of experts who are not part of the study. They will look at 
information about participant responses to the study vaccines and decide if it is 
safe to continue the study. If it is, the rest of the participants will be enrolled. 
The injections of the DNA vaccine will be given between the layers of the skin 
(known as intradermal injections with a needle and syringe. Then we will press 
the EP device firmly against that same area. The device will insert 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 
Group 
Vaccine 
type 
Where 
How 
First 
injection 
visit 
1 
month 
later 
3 
months 
later 
6 
months 
later 
1 
DNA 
vaccine 
Skin of both 
upper arms 
Needle & 
syringe with 
EP 
✓ 
✓ 
✓ 
✓ 
2 
DNA 
vaccine 
Skin of both 
upper arms 
Needle & 
syringe with 
EP 
✓ 
✓ 
✓ 
✓ 
Protein 
vaccine 
plus 
adjuvant 
Muscle of both 
upper arms 
Needle & 
syringe 
 
 
✓ 
✓ 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 86 of 107 
these pulses will last less than 1 second. Your arm will move because the 
electrical pulses.  
The injection procedure with EP 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 and may go away in a couple of minutes. After that, your arm 
may be sore for a day or two.  
You will have to wait in the clinic for about an hour after your first study injection 
of the DNA vaccine and your first study injection of the protein vaccine plus 
adjuvant to see if there are any problems. For the other injections, you will have 
to wait for about 30 minutes. Then for that night and for 14 more days, you will 
use a secure electronic symptom log to keep track of how you are feeling. We call 
this symptom log an eDiary. 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, we 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 changes on your skin related to the EP 
procedure. We may ask to take photos of your arm where you got an 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 study vaccines and EP.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 87 of 107 
12. We will do the procedures shown in this table. 
 
 
Time after first injection visit 
Procedure 
Screening 
visit(s) 
First 
injection 
visit  
2 weeks 
1 month 
1½ months 
3 months 
3½ months  
6 months  
6½ months 
12 months 
1½ years 
Injection(s) 
 
√ 
 
√ 
 
√ 
 
√ 
 
 
 
Medical 
history 
√ 
 
 
 
 
 
 
 
 
 
 
Complete 
physical 
√ 
 
 
 
 
 
 
 
 
√ 
 
Brief physical 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
 
Blood drawn 
√ 
 
√ 
 
√ 
√ 
√ 
√ 
√ 
√ 
 
Pregnancy 
test* 
√ 
√ 
 
√ 
 
√ 
 
√ 
 
 
 
HIV testing  
√ 
 
 
 
 
√ 
 
√ 
 
√ 
 
Risk reduction 
counseling 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
Interview / 
questionnaire 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
Skin 
assessment 
 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
Health contact 
 
 
 
 
 
 
 
 
 
 
√ 
* For 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 10 mL and 243 mL (2 teaspoons to about 1 cup). 
These samples are below the maximum of 500 mL that is allowed when people 
donate blood. Your body will make new blood to replace the blood we take out 
within 4 to 8 weeks. 
Site: You may want to add a sentence to the end of the previous paragraph 
contextualizing the blood volumes described (eg, “To compare, people who 
donate blood in the US can give a total of about 500 mL in an 8-week period.”) 
Modify the example for cultural relevance and alter blood volumes as necessary. 
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. 
13. We will counsel you about protecting yourself from HIV. 
We will ask you questions about your risk factors for being exposed to HIV, such 
as sexual behavior, alcohol, and drug use. We will talk with you about ways to 
keep your risk of exposure to HIV low. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 88 of 107 
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. 
15. 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. 
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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 89 of 107 
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 with anyone 
else who 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 90 of 107 
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. 
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. 
16. We will do our best to protect your private information. 
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 online symptom log after the injection visits, that 
information only has your code number. 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 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 91 of 107 
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; 
• US sites may, but are not required to include: [Insert name of local IRB/EC] ; 
• Advarra IRB; 
• Site: [Insert name of local IBC] ; 
• [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; and 
• The US Office for Human Research Protections. 
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] 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 92 of 107 
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, but it will be your choice. You may complete study procedures 
unless there is a medical reason not to or applicable regulations require you leave 
the study. 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. We will 
also contact you to check on your health 12 months after your last injection. 
We will stop your injections if you get HIV. We will also take fewer samples, and 
we will help you get care and support. We will encourage you to stay in the study 
for up to 12 months if you choose. We will also contact you to check on your 
health 12 months after your last injection. 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. 
If we stop your injections, we may ask you to stay in the study to complete other 
study procedures for up to 12 months after your last injection, if you agree to do 
so. 
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, 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 93 of 107 
• We think that staying in the study might harm you, or 
• 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 we know about. There may 
be other risks, 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.  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 94 of 107 
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 
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 that someone who should not may see 
your personal information. 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 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. 
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 deciding 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: 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 95 of 107 
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 takes 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 vaccines 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 vaccines later become 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. 
We will ask you to come to the clinic one last time for a physical exam, and a 
pregnancy test, if indicated. We may ask to take some blood samples. Whether 
you come for this last visit is up to you. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 96 of 107 
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. 
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-6160. 
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]. 
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]. 
If you have any questions about your rights as a research participant, or problems 
with or concerns about how you are being treated in this study, contact: 
• 
By mail: 
Study Subject Adviser 
Advarra IRB 
6100 Merriweather Dr., Suite 600 
Columbia, MD 21044 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 97 of 107 
• 
or call toll free:    877-992-4724 
• 
or by email:          adviser@advarra.com 
Please reference the following number when contacting the Study Subject 
Adviser: [Advarra to provide]. 
US sites may include if applicable: 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]. 
Your permissions and signature  
25. In Section 15 of this form, we told you about possible other uses of your extra 
samples and information, outside 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.  
 
 
26. 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 98 of 107 
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. 
 
 
 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 99 of 107 
Appendix D 
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 6-12 months prior to enrollment; 
however, it may be appropriate to consider a person’s behavior over a longer 
period of time than specified 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. 
 
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 an MSM (person born male with partner(s) born male) 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 
volunteer may then have had protected anal intercourse with 1 other MSM 
partner (total 2 or fewer partners in the last 12 months).  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 100 of 107 
Is a transgender person, regardless of the point on the transition spectrum, having 
sex with men (assigned male sex at birth) 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. NON-SEXUAL BEHAVIORS 
In the last 12 months, did not: 
• Inject drugs or other substances without a prescription 
• Use cocaine, methamphetamine, or excessive alcohol, which 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/they: 
Acquired an STI (ie, new infection) in the last 12 months: 
• Syphilis 
• Gonorrhea 
• Non-gonococcal urethritis 
• Herpes Simplex Virus type 2 (HSV2) 
• Chlamydia 
• Pelvic inflammatory disease (PID)  
• Trichomonas  
• Mucopurulent cervicitis  
• Epididymitis 
• Proctitis  
• Lymphogranuloma venereum  

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 101 of 107 
• Chancroid  
• Hepatitis B 
 
For US volunteers on PrEP 
1. PrEP ASSESSMENT 
• Reports six months (180 days) or more 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%) 
▪ 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 
 
1 See Study-specific Procedures (SSP) for additional guidance on on-demand PrEP. 
 
2. SEXUAL BEHAVIORS 
Persons stably taking PrEP as prescribed 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. 
 
3. NON-SEXUAL BEHAVIORS 
In the last 12 months, did not: 
• Inject drugs or other substances without a prescription 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 102 of 107 
• Use cocaine, methamphetamine, or excessive alcohol, which 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 103 of 107 
Appendix E 
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. 
You should 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 the time of your last study injection at 6 months (for a 
total of 9 months).  
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; 
• External or internal 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). 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 104 of 107 
Remember: If you are having sex, external and internal 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. 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 105 of 107 
Appendix F 
Adverse events of special interest 
Adverse events of special interest (AESI) for this protocol include but are not limited 
to potential immune-mediated diseases; representative examples of AESI are listed 
below. Updates to AESI will be provided as an appendix to the HVTN 304 Study-
specific Procedures (SSP). 
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, non-infectious 
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 anti-
neutrophil 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 
• 
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 
Gastrointestinal disorders 
• 
Celiac disease 
• 
Crohn’s disease 
• 
Ulcerative colitis 
• 
Ulcerative proctitis 
Liver disorders 
• 
Autoimmune cholangitis 
• 
Autoimmune hepatitis 
• 
Primary biliary cirrhosis 
• 
Primary sclerosing cholangitis 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 106 of 107 
Appendix G 
Protocol team 
Protocol leadership 
Chair 
Steve De Rosa 
HVTN Laboratory 
Center 
206-667-1681 
sderosa@fredhutch.org 
Statistician 
Yifan Zhu 
HVTN SDMC, Fred Hutch 
206-667-4526 
yzhu2@fredhutch.org 
Cochair 
Ronnie Matt Gravett 
University of Alabama at 
Birmingham 
205-934-9792 
rgravett@uabmc.edu 
DAIDS Medical 
Officer 
Laura Polakowski 
DAIDS, NIAID 
240-627-3040 
laura.polakowski@nih.gov 
Protocol Team Leaders 
Will Hahn 
HVTN LOC, Fred Hutch 
206-667-3431 
whahn@fredhutch.org 
Manuel Villaran 
HVTN LOC, Fred Hutch 
206-491-6206 
mvillar2@fredhutch.org 
Laboratory Lead 
Steve De Rosa 
HVTN Laboratory Center 
206-667-1681 
sderosa@fredhutch.com 
Other contributors to the original protocol 
Core Medical Monitors 
Will Hahn 
Manuel Villaran 
HVTN LOC, Fred Hutch 
Clinical Safety 
Specialist 
Megan Jones 
HVTN LOC, Fred Hutch 
Study Product Developer 
Representatives 
David Weiner 
Dan Kulp 
Mansi Purwar 
The Wistar Institute 
Laurent Humeau 
Inovio Pharmaceuticals 
Zachary Sagawa 
AAHI  
Clinical Trials 
Managers 
Carrie Sopher 
HVTN LOC, Fred Hutch 
VRC Representative 
Lucio Gama 
VRC, NIAID  
Regulatory Affairs 
Associate 
Liz Briesemeister 
HVTN LOC, Fred Hutch 
Laboratory Center Representative 
Jen Hanke 
HVTN Laboratory 
Center, Fred Hutch 
Statistical Research 
Associate 
Lorel Schmitzberger 
HVTN SDMC, Fred Hutch 
DAIDS Laboratory Lead 
Amy Palin 
DAIDS, NIAID 
Clinical Data Manager 
Sophie Arogbonlo 
Brian Ingersoll 
HVTN SDMC, Fred Hutch 
Clinic Coordinator 
Heather Logan 
UAB CRS 
SDMC Associate 
Director of Lab Science 
April Randhawa 
HVTN SDMC, Fred Hutch 
Community Advisory Board (CAB) 
Members 
Shane Hebel 
Fenway CAB 
DAIDS Product Lead 
Sujata Vijh 
DAIDS, NIAID 
Community Engagement Unit 
Representatives 
Gail Broder 
Rafael Gonzalez  
HVTN LOC, Fred Hutch 
Protocol Development 
Manager 
Meg Trahey 
Smitha Sripathy 
HVTN LOC, Fred Hutch 
Community Educator/Recruiter 
Jorge Benitez 
Columbia Research Unit 
DAIDS Protocol 
Pharmacist 
Rochelle Rogers 
240-651-6818 
Kelly Colsh 
240-669-5721 
DAIDS, NIAID 
Technical Editor 
Sydney Strauss 
HVTN LOC, Fred Hutch 
Project Coordinator 
Anders McConachie 
HVTN LOC, Fred Hutch 

HVTN 304 Version 2.0 / January 11, 2023 
HVTN304_v2.0_Final / Page 107 of 107 
Appendix H 
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 304 are described 
below. 
Protocol history and modifications 
Date: January 11, 2023 
Protocol version: Version 2.0 
Protocol modification: Full protocol amendment 1 
Item 1 Revised in Section 5.1.1, Inclusion criteria, inclusion criterion #13 :blood 
pressure range 
Item 2 Deleted in Section 9.1, Adverse events: creatinine as an exception to the 
DAIDS Table for Grading the Severity of Adult and Pediatric Adverse 
Events 
Item 3 Revised in Section 9.6, Safety pause and prompt PSRT AE review, Table 9-
1: pause rules for related SAEs 
Item 4 Revised in Appendix C, Sample informed consent form, item 23: study-
related injury language and role of Inovio Pharmaceuticals, Inc. and Access 
to Advanced Health Institute (AAHI) 
Item 5 Revised in Appendix B, Visit windows: lower allowable window for AESI 
contact 
Item 6 Minor formatting changes throughout 
Item 7 Added to Appendix H, Version history: contents of this amendment 
Date: November 3, 2022 
Protocol version: 1.0 
Original protocol