A Phase 1 Clinical Trial to Evaluate the Safety and Immunogenicity of the HIV-1 CH505 Transmitted/Founder gp120 Adjuvanted With GLA-SE in Healthy, HIV-exposed Uninfected Infants
Phase: PHASE1
Conditions: HIV Infections
Sponsor: HIV Vaccine Trials Network

 
 
PROTOCOL 
HVTN 135 
A phase 1 clinical trial to evaluate the safety and 
immunogenicity of the HIV-1 CH505 transmitted/founder 
gp120 adjuvanted with GLA-SE in healthy, HIV-exposed 
uninfected infants  
DAIDS DOCUMENT ID 38606 
A non-IND study 
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 PRODUCT(S) PROVIDED BY 
DAIDS, NIAID, NIH, DHHS 
Bethesda, Maryland, USA 
December 18, 2020  
HVTN 135 
Final 
Version 3.0 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 2 of 151 
Contents 
1 Ethical considerations ....................................................................................................5 
2 IRB/EC review considerations .......................................................................................7 
2.1 
Minimized risks to participants ......................................................................7 
2.2 
Reasonable risk/benefit balance .....................................................................7 
2.3 
Equitable participant selection .......................................................................8 
2.4 
Appropriate informed consent ........................................................................8 
2.5 
Adequate safety monitoring ...........................................................................9 
2.6 
Protect privacy/confidentiality .....................................................................10 
3 Overview ......................................................................................................................11 
3.1 
Protocol Team ..............................................................................................15 
4 Background ..................................................................................................................17 
4.1 
Rationale for trial concept ............................................................................17 
4.2 
CH505TF gp120 ...........................................................................................22 
4.3 
GLA-SE adjuvant .........................................................................................22 
4.4 
Trial design rationale ....................................................................................23 
4.5 
Plans for future product development and testing ........................................28 
4.6 
Preclinical safety studies ..............................................................................29 
4.7 
Preclinical immunogenicity studies .............................................................30 
4.8 
Clinical studies .............................................................................................34 
4.9 
Potential risks of study products and administration ...................................40 
5 Objectives and endpoints .............................................................................................42 
5.1 
Primary objectives and endpoints ................................................................42 
5.2 
Secondary objectives and endpoints ............................................................42 
5.3 
Exploratory objectives ..................................................................................43 
6 Statistical considerations ..............................................................................................45 
6.1 
Accrual and sample size calculations ...........................................................45 
6.2 
Randomization .............................................................................................50 
6.3 
Blinding ........................................................................................................50 
6.4 
Statistical analyses ........................................................................................50 
7 Selection and withdrawal of participants .....................................................................57 
7.1 
Inclusion criteria: Mother .............................................................................57 
7.2 
Exclusion criteria: Mother ............................................................................58 
7.3 
Inclusion criteria: Infant ...............................................................................59 
7.4 
Exclusion criteria: Infant ..............................................................................59 
7.5 
Participant departure from vaccination schedule or withdrawal ..................60 
8 Study product preparation and administration .............................................................63 
8.1 
Vaccine regimen ...........................................................................................63 
8.2 
Study product formulation ...........................................................................64 
8.3 
Preparation of study products .......................................................................65 
8.4 
Administration ..............................................................................................69 
8.5 
Acquisition of study products ......................................................................69 
8.6 
Pharmacy records .........................................................................................70 
8.7 
Final disposition of study products ..............................................................70 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 3 of 151 
9 Clinical procedures ......................................................................................................71 
9.1 
Informed consent ..........................................................................................71 
9.2 
Pre-enrollment procedures ...........................................................................73 
9.3 
Enrollment and vaccination visits, Infant .....................................................74 
9.4 
Follow-up visits, Infants ...............................................................................75 
9.5 
Enrollment and Follow-up Visits, Mother ...................................................76 
9.6 
HIV counseling and testing ..........................................................................77 
9.7 
Assessments of reactogenicity .....................................................................79 
9.8 
Visit windows and missed visits ..................................................................80 
9.9 
Early termination visit ..................................................................................81 
9.10 Procedures for Infants Identified as HIV infected during the study ............81 
10 Laboratory ....................................................................................................................82 
10.1 HVTN CRS laboratory procedures ..............................................................82 
10.2 Total blood volume ......................................................................................82 
10.3 Primary immunogenicity timepoints ............................................................82 
10.4 Endpoint assays: cellular ..............................................................................83 
10.5 Endpoint assays: humoral .............................................................................83 
10.6 Microbiome analysis ....................................................................................85 
10.7 Systems Biology ...........................................................................................86 
10.8 Lab assay algorithm .....................................................................................86 
10.9 Exploratory studies .......................................................................................86 
10.10 Specimen storage and other use of specimens .............................................86 
10.11 Biohazard containment .................................................................................87 
11 Safety monitoring and safety review ...........................................................................88 
11.1 Safety monitoring and oversight ..................................................................88 
11.2 Safety reporting ............................................................................................89 
11.3 Safety reviews ..............................................................................................92 
11.4 Safety pause and prompt PSRT AE review..................................................93 
11.5 Review of cumulative safety data ................................................................95 
11.6 Study termination .........................................................................................95 
12 Protocol conduct ..........................................................................................................96 
12.1 Social impacts ..............................................................................................97 
12.2 Emergency communication with study participants ....................................97 
13 Version history .............................................................................................................98 
14 Document references (other than literature citations) ................................................101 
15 Acronyms and abbreviations......................................................................................104 
16 Literature cited ...........................................................................................................108 
Appendix A Sample informed consent form ..................................................................117 
Appendix B Sample consent form for use of samples and information in other studies 137 
Appendix C Table of procedures (for sample informed consent form) ..........................142 
Appendix D Laboratory procedures: Infant ....................................................................143 
Appendix E Laboratory procedures: Mother ..................................................................145 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 4 of 151 
Appendix F Procedures at HVTN CRS: Infant ..............................................................146 
Appendix G Procedures at HVTN CRS: Mother ............................................................147 
Appendix H Adverse events of special interest ..............................................................148 
Appendix I Infant visit windows ...................................................................................149 
Appendix J Mother visit windows .................................................................................150 
Appendix K Protocol Signature Page .............................................................................151 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 5 of 151 
1 
Ethical considerations 
Multiple candidate HIV vaccines will need to be studied simultaneously in 
different populations around the world before a successful HIV preventive 
vaccine is found. It is critical that universally accepted ethical guidelines are 
followed at all sites involved in the conduct of these clinical trials. The HIV 
Vaccine Trials Network (HVTN) has addressed ethical concerns in the following 
ways: 
• HVTN trials are designed and conducted to enhance the knowledge base 
necessary to find a preventive vaccine, using methods that are scientifically 
rigorous and valid, and in accordance with International Council for 
Harmonisation of Technical Requirements for Pharmaceuticals for Human 
Use (ICH) and/or other Good Clinical Practice (GCP) guidelines. 
• HVTN scientists and operational staff incorporate the philosophies underlying 
major codes (1-3), declarations, and other guidance documents relevant to 
human subjects research into the design and conduct of HIV vaccine clinical 
trials. 
• HVTN scientists and operational staff are committed to substantive 
community input—into the planning, conduct, and follow-up of its research—
to help ensure 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. 
• HVTN clinical trial staff counsel study participant mothers routinely on how 
to reduce HIV transmission risk to their infants.  
• Infants enrolled at birth who become HIV infected during the trial will be 
referred to accredited antiretroviral therapy (ART) treatment sites for their 
care. If clinical trials are available at the site, infants may be referred for 
screening. In the event of ART stockouts (supply shortages), ART will be 
made available until supply issue is resolved. ART will be supplied to the site 
through a mechanism established by the South African Medical Research 
Council.  
• The HVTN provides training so that the participating site protects the privacy 
of research participants, and obtains meaningful informed consent. During the 
study, participants will have their wellbeing monitored, and to the fullest 
extent possible, their privacy protected. Participants, and/or their 
parent(s)/guardian(s) may withdraw from the study at any time. 
• Prior to implementation, HVTN trials are rigorously reviewed by scientists 
who are not involved in the conduct of the trials under consideration. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 6 of 151 
• HVTN trials are reviewed by local and national regulatory bodies and are 
conducted in compliance with all applicable national and local regulations. 
• The HVTN designs its research to minimize risk and maximize benefit to both 
study participants and their local communities. For example, HVTN protocols 
provide enhancement of participants’ knowledge of HIV and HIV prevention, 
as well as counseling, guidance, and assistance with any social impacts that 
may result from research participation. HVTN protocols also include careful 
medical review of each research participant’s health conditions and reactions 
to study products while in the study. 
• HVTN research aims to benefit local communities by directly addressing the 
health and HIV prevention needs of those communities and by strengthening 
the capacity of the communities through training, support, shared knowledge, 
and equipment. Researchers involved in HVTN trials are able to conduct other 
critical research in their local research settings. 
• The HVTN values the role of in-country Institutional Review Boards (IRBs), 
Ethics Committees (ECs), and other Regulatory Entities (REs) as custodians 
responsible for ensuring the ethical conduct of research in each setting. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 7 of 151 
2 
IRB/EC review considerations 
US federal regulations require IRBs/ECs/REs to ensure that certain requirements 
are satisfied on initial and continuing review of research (Title 45, Code of 
Federal Regulations (CFR), Part 46.111(a) 1-7; 21 CFR 56.111(a) 1-7). The 
following section highlights how this protocol addresses each of these research 
requirements. Each HVTN Investigator welcomes IRB/EC/RE questions or 
concerns regarding these research requirements. 
This trial is being conducted in South Africa, with funding from the US NIH 
among others. Due to this, the trial is subject to both US and local regulations and 
guidelines on the protection of human research subjects and ethical research 
conduct. Where there is a conflict in regulations or guidelines, the HVTN strives 
towards maximum protection of human research participants. 
In compliance with international and local (as appropriate) ICH and/or other GCP 
guidelines, each research location has a locally-based Principal Investigator (PI) 
who is qualified to conduct (and supervise the conduct of) the research. The 
investigators take responsibility for the conduct of the study and the control of the 
study products, including obtaining all appropriate regulatory and ethical reviews 
of the research. 
2.1 
Minimized risks to participants 
45 CFR 46.111 (a) 1 and 21 CFR 56.111 (a) 1: Risks to subjects are 
minimized. 
This protocol minimizes risks to participants by (a) correctly and promptly 
informing participants and/or their parent(s)/guardian(s) about risks so that they 
can join in partnership with the researcher in recognizing and reporting harms; (b) 
respecting local/national blood draw limits; (c) performing direct observation of 
participants postvaccination and collecting information regarding side effects for 
several days postvaccination; (d) having staff properly trained in administering 
study procedures that may cause physical harm or psychological distress, such as 
blood draws, vaccinations, HIV testing and counseling; (e) providing HIV risk 
reduction counseling for the infant; and (e) providing safety monitoring. 
2.2 
Reasonable risk/benefit balance 
45 CFR 46.111(a) 2 and 21 CFR 56.111(a) 2: Risks to subjects are 
reasonable in relation to anticipated benefits, if any, to subjects, and 
the importance of the knowledge that may reasonably be expected to 
result. 
In all public health research, the risk-benefit ratio may be difficult to assess 
because the benefits to a healthy participant are not as apparent as they would be 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 8 of 151 
in treatment protocols, where a study participant may be ill and may have 
exhausted all conventional treatment options. However, this protocol is designed 
to minimize the risks to participants while maximizing the potential value of the 
knowledge it is designed to generate. 
2.3 
Equitable participant selection  
45 CFR 46.111 (a) 3 and 21 CFR 56.111 (a) 3: Subject selection is 
equitable 
This protocol has specific inclusion and exclusion criteria for investigators to 
follow in admitting participants into the protocol. Participants are selected 
because of these criteria and not because of positions of vulnerability or privilege. 
Investigators are required to maintain screening and enrollment logs to document 
volunteers who screened into and out of the protocol and for what reasons. 
2.4 
Appropriate informed consent 
45 CFR 46.111 (a) 4 and 5 and 21 CFR 56.111 (a) 4 and 5: Informed 
consent is sought from each prospective adult subject and the infant 
subject’s parent(s) or guardian(s) as required by 45 CFR 46.116 and 
21 CFR Part 50; informed consent is appropriately documented as 
required by 45 CFR 46.117 and 21 CFR 50.27 
The study site must comply with the requirements of the DAIDS policy on 
Enrolling Children (including Adolescents) in Clinical Research, which is 
available at: https://www.niaid.nih.gov/research/daids-clinical-research-protocol-
informed-consent 
HIV-infected pregnant women who are 18 years of age and older will provide 
written informed consent for their own and their infant’s study participation 
before any study-specific procedures are performed. Further, informed consent is 
assessed throughout the trial (see Section 9.1). The site is provided training in 
informed consent by the HVTN as part of its entering the HVTN. The HVTN 
requires a signed consent document for documentation, in addition to chart notes 
or a consent checklist. 
As indicated above, it is generally expected that mothers will provide informed 
consent for their own and their infant’s participation in this study. However, 
parental consenting requirements at each site will depend on the IRB/EC risk 
determination described above and all IRB/EC requirements will be followed. 
Should the consenting mother of an enrolled infant die or no longer be available 
for any reason, no further study-specific visits or procedures may be performed 
until informed consent for continued study participation is obtained from a 
guardian. The guardian will not replace the mother as a participant but will be 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 9 of 151 
asked to continue bringing the infant for scheduled study visits. In accordance 
with the DAIDS policy on Enrolling Children (including Adolescents) in Clinical 
Research (available at the website referenced above), the study site must establish 
and maintain written procedures describing the standards that will be followed to 
identify who may serve as guardian for an enrolled infant, reflective of applicable 
IRB/EC guidance for conduct of human subjects research within the context of 
available local law, regulation, or government policy.  
Children enrolled in this study are considered vulnerable participants per 45 CFR 
46 Subpart D. Site IRBs/ECs must consider the potential benefits, risks, and 
discomforts of the study to children and assess the justification for their inclusion 
in the study. As part of this assessment, IRBs/ECs must determine the level of risk 
to children in the categories specified in 45 CFR 46.404-407. Documentation of 
this determination is required to complete the DAIDS protocol registration 
process described in Section 13.2 in 45 CFR 46.404-407.  
The risk category assigned by the IRB/EC determines the parental informed 
consent requirements for the study at each site. Per 45 CFR 46.408 (b), the 
IRB/EC may find that the permission of one parent is sufficient for research to be 
conducted under 46.404 or 46.405. Where research is covered by 46.406 or 
46.407, both parents must give their permission unless one parent is deceased, 
unknown, incompetent, or not reasonably available, or when only one parent has 
legal responsibility for the care and custody of the child (as determined locally). 
IRBs/ECs must document their risk determination and the study site should adapt 
the signature pages of their site-specific informed consent forms (ICFs) as needed 
to accommodate the parental consent requirements associated with the IRB/EC 
determination.  
2.5 
Adequate safety monitoring 
45 CFR 46.111 (a) 6 and 21 CFR 56.111 (a) 6: There is adequate 
provision for monitoring the data collected to ensure the safety of 
subjects. 
This protocol has extensive safety monitoring in place (see Section 11). Safety is 
monitored daily by HVTN Core and routinely by the HVTN 135 Protocol Safety 
Review Team (PSRT). In addition, the HVTN Safety Monitoring Board (SMB) 
periodically reviews study data. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 10 of 151 
2.6 
Protect privacy/confidentiality 
45 CFR 46.111 (a) 7 and 21 CFR 56.111 (a) 7: There are adequate 
provisions to protect the privacy of subjects and maintain the 
confidentiality of data. 
Privacy refers to an individual’s right to be free from unauthorized or 
unreasonable intrusion into his/her private life and the right to control access to 
individually identifiable information about him/her. The term “privacy” concerns 
research participants or potential research participants as individuals whereas the 
term “confidentiality” is used to refer to the treatment of information about those 
individuals. This protocol respects the privacy of participants by informing them 
about who will have access to their personal information and study data (see 
Appendix A). The privacy of participants is protected by assigning unique 
identifiers in place of the participant’s name on study data and specimens. In 
addition, each staff member at each study site in this protocol signs an Agreement 
on Confidentiality and Use of Data and Specimens with the HVTN. In some 
cases, a comparable confidentiality agreement process may be acceptable. The 
study site participating in the protocol is required to have a standard operating 
procedure on how the staff members will protect the confidentiality of study 
participants. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 11 of 151 
3 
Overview 
Title  
A phase 1 clinical trial to evaluate the safety and immunogenicity of the HIV-1 
CH505 transmitted/founder gp120 adjuvanted with GLA-SE in healthy, HIV-
exposed uninfected infants 
Primary objectives 
Primary objective 1 
To evaluate the safety and tolerability of HIV-1 CH505 transmitted/founder virus 
Env gp120 immunogen (CH505TF gp120) adjuvanted with Glucopyranosyl Lipid 
A - stable emulsion (GLA-SE) in healthy HIV-1 exposed uninfected (HEU) 
infants 
Primary objective 2 
To determine whether vaccination with CH505TF gp120 adjuvanted with GLA-
SE initiates B-cell lineages potentially capable of generating a broadly 
neutralizing antibody response 
Study products and routes of administration 
• CH505TF gp120: CH505TF gp120 with the adjuvant GLA-SE. GLA-SE is 
an oil-in-water stable emulsion (SE) containing the immunological adjuvant 
Glucopyranosyl Lipid A (GLA). The vaccine will be administered by 
intramuscular (IM) injection in the thigh at 2 doses of gp120 (20 or 5 mcg) 
and 2 doses of adjuvant (2.5 mcg or 5 mcg).  
• Placebo: Sodium Chloride for Injection, 0.9%, administered IM in the thigh at 
volumes to match the active product. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 12 of 151 
Table 3-1 Schema 
Group 
 
N 
Injection schedule in weeks 
Week 0 
Week 8 
Week 16 
Week 32 
Week 54 
Part A 
Initial Safety  
1 
5 
20 mcg 
CH505TF 
gp120 
+2.5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+2.5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+2.5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+2.5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+2.5 mcg 
GLA-SE 
2 
2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Part B 
Safety Ramp-Up  
3 
 
2 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
4 
 
2 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Part C 
Immunogenicity of CH505TF Recombinant Protein 
5 
 
16 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
20 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
6 
 
3 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
7 
 
5 
5 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
5 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
5 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
5 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
5 mcg 
CH505TF 
gp120 
+5 mcg 
GLA-SE 
8 
 
3 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
Total 
 
38 
28 vaccinees, 10 placebos 
Notes: To ensure safety, enrollment will proceed in stages, see Section 11.3.1.  
Groups 3 and 5 (20 mcg CH505TF and 5 mcg GLA-SE) and Groups 2, 4, 6, and 8 (placebo) will be 
pooled in the ultimate analyses.  
Participants 
Healthy HIV-1–uninfected infants born to HIV-1–infected mothers through 
caesarian delivery in South Africa. To quantify maternal HIV antibody response, 
mothers will also be enrolled in the study but will not receive study product. 
Sample size 
38 mother-infant pairs 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 13 of 151 
Design 
Single center, double-blind, randomized, controlled trial 
Duration per participant 
24.5 months of scheduled clinic visits  
Estimated total study duration 
32 months (includes enrollment, planned safety holds, and follow-up) 
Clinical trial sponsor 
DAIDS, NIAID, NIH, DHHS (Bethesda, Maryland, USA) 
Study product providers 
• CH505TF gp120: DAIDS, NIAID, NIH, DHHS (Bethesda, Maryland, USA) 
• GLA-SE adjuvant: DAIDS, NIAID, NIH, DHHS (Bethesda, Maryland, USA) 
Core operations 
HVTN Vaccine Leadership Group/Core Operations Center, Fred Hutchinson 
Cancer Research Center (Fred Hutch) (Seattle, Washington, USA) 
Statistical and data management center (SDMC) 
Statistical Center for HIV/AIDS Research and Prevention (SCHARP), Fred Hutch 
(Seattle, Washington, USA) 
HIV diagnostic laboratory 
HIV Sero-Molecular Laboratory, National Institute for Communicable Diseases 
(HSML-NICD) (Johannesburg, South Africa) 
Endpoint assay laboratories 
• Fred Hutchinson Cancer Research Center (Seattle, Washington, USA) 
• Duke University Medical Center (Durham, North Carolina, USA) 
• Duke Human Vaccine Institute (Durham, North Carolina, USA) 
• Precision Vaccines Program, Boston Children’s Hospital/Harvard Medical 
Center (Boston, Massachusetts, USA) 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 14 of 151 
• University of British Columbia (Vancouver, Canada) 
• South Africa Immunology Laboratory and National Institute for 
Communicable Diseases (SAIL-NICD) (Johannesburg, South Africa) 
Study site 
Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital (Johannesburg, 
South Africa) 
Safety monitoring 
Protocol Safety Review Team (PSRT); HVTN Safety Monitoring Board (SMB) 
 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 15 of 151 
 
3.1 
Protocol Team 
Protocol leadership 
Chairs 
Avy Violari 
Perinatal HIV Research Unit 
Chris Hani Baragwanath Hospital 
+27 11 989 9700 
violari@mweb.co.za 
Co-chair 
Glenda Gray 
Perinatal HIV Research Unit 
Chris Hani Baragwanath Hospital 
President, South African Medical 
Research Council 
+27 11 989 9702 
glenda.gray@mrc.ac.za 
 
Georgia Tomaras 
Duke University, HVTN Laboratory 
Program 
919 681-5598 
gdt@duke.edu 
Laboratory 
lead 
Nono Mkhize 
Centre for HIV and STIs, National 
Institute for Communicable Diseases 
+27 11 386 6541  
nonom@nicd.ac.za 
Protocol Team 
leader 
Will Hahn 
HVTN Core, Fred Hutch 
206-667-3431 
whahn@fredhutch.org 
Statisticians 
Holly Janes 
SCHARP, Fred Hutch 
206-667-6353  
hjanes@fredhutch.org 
Medical officer 
Maggie Brewinski Isaacs 
DAIDS, NIAID 
301-761-7009 
maggie.brewinskiisaacs@nih.gov 
 
Kennedy Otwombe 
Perinatal HIV Research Unit 
Chris Hani Baragwanath Hospital 
+27 11 989 9807 
otwombek@phru.co.za 
 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 16 of 151 
Other contributors to the original protocol 
Core medical 
monitor 
Will Hahn 
HVTN Core, Fred Hutch 
Clinical safety 
specialist 
Sophie Hasan 
HVTN Core, Fred Hutch 
Study product 
developer 
representatives 
Barton Haynes 
M. Anthony Moody  
Duke Human Vaccine Institute 
Duke University Medical 
Center 
Regional medical 
liaison 
Simba Takuva 
HVTN Core, Fred Hutch 
 
Zachary Sagawa 
Infectious Disease Research 
Institute (IDRI) 
Clinical trials 
manager 
Shelly Ramirez 
HVTN Core, Fred Hutch 
Consultants 
Sallie Permar 
Genevieve Fouda 
Coleen Cunningham 
Department of Pediatrics 
Duke University Medical 
Center 
Clinic coordinator 
Nastassja Ramsagar 
Perinatal HIV Research Unit 
Chris Hani Baragwanath Hospital 
 
Leo Stamatatos 
VIDD, Fred Hutch  
Clinical data 
manager 
Vicky Kim 
SCHARP, Fred Hutch 
 
Ofer Levy 
Precision Vaccines Program 
Boston Children’s Hospital 
Statistical 
research associate 
Brittany Prigmore 
SCHARP, Fred Hutch 
 
Guido Ferrari 
Duke University School of 
Medicine 
Regulatory affairs 
associate 
Megan Brandon 
HVTN Core, Fred Hutch 
HVTN Chief-of - 
Staff 
Troy Martin 
HVTN Core, Fred Hutch 
SDMC Associate 
director of lab 
science 
April Randhawa 
SCHARP, Fred Hutch 
SDMC Associate 
Director of 
Operations 
Jessica Andriesen 
VIDD, Fred Hutch 
Protocol 
development 
manager 
Meg Trahey 
HVTN Core, Fred Hutch 
DAIDS protocol 
pharmacists 
Cynthia Parker 
DAIDS, NIAID 
301-761-7199 
Community 
engagement unit 
representatives 
Kagisho Baepanye 
HVTN Core, Fred Hutch 
 
Katie Shin 
DAIDS, NIAID 
240-627-3047 
Community 
educator/recruiter 
Valerie Khemese 
PHRU 
Laboratory center 
representative 
Jen Hanke  
HVTN Laboratory Program, 
Fred Hutch 
Community 
Advisory Board 
(CAB) members 
Susan Moloto 
Winnie Frazenburg 
PHRU 
DAIDS Product 
Lead 
 
Sujata Vijh 
DAIDS, NIAID 
Technical editors 
Richa Chaturvedi 
Frankie Rentas 
HVTN Core, Fred Hutch 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 17 of 151 
4 
Background 
4.1 
Rationale for trial concept 
Despite the success of antiretroviral (ARV) prophylaxis to prevent vertical HIV-1 
transmission, more than 180,000 infants acquired HIV-1 in 2017 (4). These infant 
infections are partly due to suboptimal HIV-1 testing and ARV coverage in areas 
with high HIV-1 incidence. Yet, even if 90% ARV coverage is achieved for 
pregnant women known to be infected with HIV-1 globally, it is estimated that 
more than 100,000 infants will still become infected with HIV-1 annually (5). 
This high burden is due to several settings in which suppressive antiretroviral 
therapy may not be achieved or diagnosis of maternal infection may not be timely, 
including: 1) acute infection of the mother during pregnancy or breastfeeding, 2) 
late presentation to prenatal care, 3) maternal non-adherence, which can be 
particularly challenging for the full duration of breastfeeding, 4) other 
psychosocial factors that are barriers to care, and 5) the development of drug-
resistant variants. Thus, there remains a critical need to develop easy-to-
implement immune-based prophylaxis strategies to prevent infant HIV-1 
acquisition, such as infant HIV-1 immunization.  
Maternal ARV-based strategies cannot achieve universal protection of infants 
against HIV-1 
Young women living in areas of high HIV-1 incidence comprise a highly 
vulnerable population. In fact 30% of new HIV-1 infections in sub-Saharan Africa 
affect young women aged 15-24 years (6). This population includes young 
women who are or may become pregnant. Acute infection late in pregnancy and 
during the postpartum period puts the infant at high risk of HIV acquisition. In 
Mozambique for example, the rate of postpartum acute HIV-1 acquisition was 
3.2/100-person years and up to 4.9/100-person years in 18-19-year-old women 
(7). Most of these women are not aware of their HIV status if acquired 
postpartum. Safe and effective, universal infant vaccination is necessary for 
eliminating postpartum HIV acquisition and for achieving an HIV-1-free 
generation in areas of high HIV-1 prevalence. 
Late ARV initiation results in high rates of mother to child transmission (MTCT) 
of HIV-1. A South African study reported that women who received < 4 weeks of 
ARV had a transmission rate of 9.3%, which declined to 5.5% in those with 4-16 
weeks of ARV, and further to 0.5% in women who received 16-32 weeks of ARV 
(8). Up to one-third of new mothers have disengaged from care and discontinued 
their own ARV therapy by six months postpartum (9). A universal infant HIV-1 
vaccine that can be incorporated into the World Health Organization Expanded 
Program on Immunization (EPI) vaccine schedule is a rational approach towards 
elimination of the pediatric HIV-1 epidemic. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 18 of 151 
Induction of broadly neutralizing antibodies is a highly desirable feature for an 
HIV vaccine 
Passive immunization studies in nonhuman primates have provided proof of 
principle that broadly neutralizing antibodies (bnAbs) can protect from virus 
exposure (10, 11). However, inducing bnAbs by vaccination has proven 
challenging. The difficulty of inducing bnAbs constitutes a major obstacle to the 
development of an efficacious preventive HIV vaccine. 
There are several explanations for the difficulties of inducing bnAbs, including 
the conformational structure of the viral envelope, the antigenic similarity 
between envelope (Env) conserved epitopes and host antigens, and the 
requirement for complex B cell maturation pathways (12). About 10-25% of HIV-
1–infected individuals develop bnAbs after several years of infection and these 
antibodies generally exhibit specific characteristics including extensive somatic 
hypermutation (generally associated with affinity maturation), long heavy chain 
complementarity determining regions, and/or autoreactivity with host antigens. It 
has been hypothesized that bnAbs can be induced following priming with a 
transmitted/founder (TF) Env followed by sequential immunization with different 
Env immunogens over time to mimic bnAb development in chronically HIV-1–
infected individuals (13).  
B cell immunogen lineage design is a process to circumvent host 
immunoregulatory mechanisms to give the bnAb lineages that are typically 
subdominant in infected patients a survival advantage. This design consists of 
selecting sequential Env immunogens with high affinity to the B cell receptors 
(BCRs) of the unmutated common ancestor (UCA) antibody and intermediate 
ancestor (IA) antibodies that are functional maturation steps leading to the mature 
bnAb of interest (11). Analysis of the co-evolution of mutations in both the 
infecting HIV-1 viruses and the resulting BCRs of discrete B cell clones from the 
time of seroconversion to the development of plasma bnAb induction allows for 
mapping the pathways that lead to generation of bnAbs. For example, early 
detection of acute HIV-1 infection in one adult donor (individual CH505) allowed 
evolutionary development mapping of a bnAb directed against the CD4 binding 
site (CD4bs) of gp120 (bnAb CH103) (12). Follow up studies in this individual 
revealed that the same transmitted founder virus also induced the development of 
a second CD4bs bnAb lineage (bnAb CH235). While CH103 and CH235 evolved 
though distinct evolutionary paths, CH235 cooperatively selected for resistant 
viruses that drove the evolutionary development of the CH103 bnAb lineage (14, 
15). Such cooperative immunological pressure may be important for inducing rare 
viral variants that stimulate the maturation of lineages such as CH103. 
Importantly, despite the diversity of the human antibody repertoire, for many 
bnAbs there are only a few B cell lineages that can lead to bnAb development and 
these lineages are the same across individuals (13, 14). Therefore, immunization 
with an Env lineage that drives bnAb development in one individual is likely 
generalizable to others. A key question in such designs is whether each 
immunogen stimulates the early members of these B cell lineages in vaccinees 
and how much expansion of those populations can be achieved.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 19 of 151 
This study will address this key question in infants and will be directly compared 
to similar data obtained in the adult participants in HVTN 115. Preliminary 
immunogenicity data collected after the third (of five total) immunizations with 
CH505TF in HVTN115 Part A (see schema Table 4-6) has been analyzed. 
Remarkably, a high percentage of CD4bs specific antibodies to the vaccine 
envelope were elicited, as detected by serum binding and B cell assays. Although 
there were no CD4bs or V2 glycan precursors detected by the specialized bnAb 
precursor neutralization assays, these data suggest that the right specificities were 
elicited early on and may be capable of further maturation. If, as hypothesized, the 
neonatal immune system is more permissive for the rapid development of bnAbs, 
it may be possible to elicit bnAbs in infants without sequential immunizations 
with lineage-directed immunogens. 
Sequential immunization is a potential approach to achieve neutralization 
breadth 
The Duke CHAVI-ID team described the co-evolution of HIV-1 and a CD4bs 
bnAb clonal lineage (CH103) in an African HIV-infected individual (CH505) 
(14). From this analysis, four evolutionarily related Envs were selected as vaccine 
immunogens (Figure 4-1). The safety and immunogenicity of this 4-valent 
sequential gp120 Env immunogen (EnvSeq-1), adjuvanted with GLA-SE, will be 
tested in a phase 1 adult trial (HVTN 115, Part B). Results from preclinical 
studies have provided support for sequential immunization strategies. Notably, 
CH505TF gp120 engaged a CD4bs bnAb germline B cell receptor on naïve B 
cells in CH103 bnAb knock-in mice (15). Moreover, sequential immunization of 
infant rhesus macaques with CH505 Env immunogens induced plasma 
neutralizing activity against their respective autologous viruses and recombinant 
memory B cell-derived monoclonal antibodies capable of neutralizing tier 2 HIV 
strains (Han, V and Haynes, BF unpublished). As noted above, the critical 
question is how efficiently infant bnAb precursor B cells are expanded compared 
with adults. Overall, human and NHP data suggest that in infants, bnAb evolution 
may be achieved in a shorter time frame than in adults. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 20 of 151 
 
Figure 4-1 CH505 transmitted-founder (TF) and Env variants generated during viral evolution 
drove affinity maturation of CH103 bnAb lineage. Figure adapted from (14). 
Development of broadly neutralizing antibodies in children 
Two independent research groups have reported differences in bnAb responses 
between adults and children. Goo, Overbaugh et al. (16) demonstrated that two 
years after infection, the neutralization breadth in some infants is comparable to 
the breadth observed in the top 1% of adult neutralizers several years after 
infection. They also isolated a bnAb from an infected infant that had developed 
with lower levels of somatic hypermutation and variable gene usage than typically 
seen in isolated adult bnAbs (17). Similarly, Goulder and colleagues measured 
neutralizing antibody responses in HIV-1–infected children and found that 70% of 
slow progressor children, but only 15% of HIV-1 clade C chronically-infected 
adults, were able to neutralize at least 50% of viruses tested (18). Children also 
had higher neutralization titers than adults (18). Together, these data suggested 
that it may be easier to induce bnAbs in children than in adults.  
The rationale for assessing HIV vaccines within the first weeks of life is based 
upon (a) the desire to test whether the unique immune status of neonates is more 
conducive to HIV vaccination strategies designed to produce broadly neutralizing 
antibodies and to unequivocally establish the safety of multiple immunizations 
with a vaccine-adjuvant combination; (b) birth being a reliable point of healthcare 
contact worldwide such that neonatal immunization is practical and achieves high 
population penetration in resource poor settings; and (c) the substantial 
preadolescent window provides a unique opportunity to develop mature immune 
responses over years (19). If, as hypothesized, infants are capable of more rapidly 
generating a wider repertoire of high affinity, somatically hypermutated B cell 
clones when compared to adults given the same complex immunogen, this would 
substantially increase the prospects for a vaccine to drive a broadly neutralizing 
antibody response. Furthermore, it would strongly support further testing of 
lineage-directed vaccines in a neonatal population. The results of the proposed 
study have important implications for HIV vaccinology that merit careful 
exploration in this vulnerable population.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 21 of 151 
Development of broadly neutralizing serum in children infected with HIV is 
associated with a more diverse antibody repertoire capable of recognizing distinct 
epitopes when compared to antibodies isolated from mothers infected with the 
same virus family (20). In addition to increases in the breadth of response, infants 
vaccinated with similar doses of gp120 vaccines develop higher titer antibodies to 
the specific V1V2 loop (a response associated with protection in RV144) when 
compared to adults vaccinated with the same vaccine (21). Thus, the magnitude, 
breadth, and rate of affinity maturation may be improved in infants as compared 
to adults.  
The mechanisms underpinning the differences between the infant immune and 
adult immune system are complex and under active investigation, but specific 
differences in the underlying B cell repertoire have important implications for the 
development of broadly neutralizing antibodies. It has been known for >50 years 
that the neonatal immune system is an environment with more permissive 
peripheral tolerance checkpoints and infant B cells are more polyreactive than 
adults (22). Over the first four weeks of life, there is a profound reduction in the 
percentage of transitional B cells, a B cell subset enriched for autoreactivity (23, 
24). The number of potentially self-reactive B cells present at birth is important in 
the context of HIV as many of the bnAbs identified to date cross-react with self-
antigens that are recognized by B cells thought to be eliminated by tolerance 
checkpoints in adults (25, 26). Vaccinating as early as possible maximizes the 
probability of determining whether these differences in the neonatal immune 
system are important in the context of HIV vaccination. In addition, breaching 
peripheral tolerance checkpoints is associated with improved development of 
HIV-neutralizing titers in experimental systems (27). Our hypothesis is that 
neonates have a wider repertoire of B cells capable of generating broadly 
neutralizing antibodies in response to vaccination as compared to adults (28). Our 
incorporation of powerful systems biology approaches (eg. high resolution flow 
cytometry, RNASeq, proteomics, metabolomics, multiplex cytokines, etc.), 
recently optimized for use in small volumes (<1 mL) of newborn peripheral 
blood, as tertiary study goals will provide fresh insight into dynamic changes in 
the infant immune system over the first months of life in relation to vaccine 
immunogenicity (29). Moreover, use of some of the same systems biology assay 
platforms (eg, plasma proteomics and metabolomics) in this infant protocol and 
the adult HVTN 115 study will enable meaningful comparisons to understand the 
impact of ontogeny on the response to the same adjuvanted HIV vaccine. 
In this proof of concept study, we propose to test the safety, tolerability and 
immunogenicity of the CH505TF gp120 adjuvanted with GLA-SE in healthy 
HEU newborn infants. We will also test the ability of the vaccine regimen to 
initiate both CD4bs and V1V2 lineage specific antibodies that have the potential 
to develop neutralization capacity (ie, bnAb lineage) in an infant population. 
Although the proposed immunogen (CH505TF gp120) was designed to elicit 
antibodies against the CD4bs, it contains other regions of the gp120 protein and 
also elicits bnAbs against the V1V2 region in preclinical models (15). The 
overarching hypothesis guiding development of HIV vaccines for an infant 
population is that establishment of both an early and durable broad neutralizing 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 22 of 151 
antibody response could protect against vertical transmission during breastfeeding 
and, perhaps, horizontal HIV-1 transmission at sexual debut. 
4.2 
CH505TF gp120 
The CH505TF gp120 to be used in this study was derived from the clade C 
transmitted/founder virus which was isolated from a single acutely HIV-1–
infected donor from Malawi (CH505) (14, 30, 31). CH505TF was chosen based 
on its binding affinity to the CH103 bnAb lineage as shown in Table 4-1. 
CH505TF binds with high affinity to the unmutated common ancestor (UCA) of 
the CH103 lineage (30).  
The CH505TF gp120 immunogen has an N-terminal deletion to facilitate 
production by decreasing protein dimer formation and increasing production yield 
(30). CH505TF is currently being evaluated in combination with GLA-SE in adult 
participants in HVTN 115. 
Table 4-1 Binding of CH103 lineage antibodies to autologous CH505 Envs. Choice of 
transmitted/founder CH505 Env by binding EC50 levels to recombinant sequential CH505 Envs. 
Data are in EC50 binding titers in ELISA. (Red shading highlights the antigens being assessed in 
HVTN 115 and the antibodies they bind to.) 
 
4.3 
GLA-SE adjuvant 
To compare immunogenicity results between infants immunized with CH505TF 
gp120 and adults in HVTN 115, this study will use the same adjuvant system. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 23 of 151 
GLA-SE is a synthetic TLR4 agonist formulated in a stable nano-emulsion of 
squalene oil and promotes a robust Th1-type immune response to vaccine 
antigens (32, 33). GLA-SE has been tested in over 2500 individuals in phase 1, 2 
and 3 trials with a variety of antigens. No significant safety concerns were 
reported. Previous clinical trial experience with GLA-SE and other antigens in 
adults is described in Section 4.8.2. Two trials are underway in children 
evaluating GLA-SE with other vaccines: a protein subunit vacine for 
schistosomiasis (NCT03799510), and a tuberculosis vaccine (NCT03806699). 
Our study will further compliment these studies in elucidating the safety and 
tolerability of this adjuvant in combination with other vaccines in infants. 
Importantly, the Th1-type immune response elicited is of interest in this 
population because of potential benefit towards more rapid cellular responses and 
more rapid bnAb development.  
A study in infant rhesus macaques testing HIV-1 Env bivalent clade C gp120 
C1086 and gp120 TV1 evaluated various adjuvants including GLA-SE at the dose 
as proposed in this study. This adjuvant demonstrated good immunogenicity in 
these infants, which weigh on average 500 g, indicating the immunogenicity of 
this dose in low-weight NHPs. GLA-SE adjuvanted vaccines has been previously 
studied in more than 2500 human adults and these GLA-SE adjuvanted vaccines 
have been demonstrated to be safe (see Table 4-7). Moreover, no major safety 
signal has been observed in adults who received a GLA-SE adjuvated gp120 
vaccine in the Part A of HVTN 115. 
4.4 
Trial design rationale 
An important goal for the vaccine field is to evaluate the concept of inducing 
broadly neutralizing antibodies against HIV-1. This study complements the work 
done in adults that evaluates a series of immunogens that, in an NHP model, were 
found to be capable of recreating the natural evolution of Env that promotes 
CD4bs specific antibody evolution. We will provide key insight into whether the 
infant immune milieu is more supportive of the development of several classes of 
affinity matured, somatically hypermutated bnAbs than the adult immune system, 
with important biological implications for the development of a generally 
effective HIV vaccine. In addition, we will also assess whether the promising 
adjuvant GLA-SE is as safe and tolerable in an infant population as it has been in 
adult populations.  
It is critical to evaluate whether repeated administrations of GLA-SE with a single 
protein are safe in a neonatal population and do not interfere with required EPI 
vaccinations (secondary endpoint 1). This study will lay the groundwork for 
subsequent testing with sequential immunogens (eg, more directed lineage 
shaping approach, such as in HVTN 115 Part B), which will likely require a larger 
number of participants. All currently envisioned testing regimens require five (or 
more) doses of vaccine product. By including five doses with a placebo group 
powered for a robust consideration of safety, the study will allow for assessment 
of whether five doses of a GLA-SE adjuvanted vaccine is safe in infants. In 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 24 of 151 
addition, the measurement of EPI vaccine responses after one year of vaccination 
will provide additional insight on whether GLA-SE causes interference with other 
important vaccines, as “bystander” interference has been observed with prior 
neonatal vaccination approaches (34). 
The use of a placebo arm does not compromise standard of care for either the 
mother or the infant as both groups will receive the local standard of care for 
prevention of HIV transmission through breastfeeding. 
4.4.1 
Study Population 
HIV exposed infants represent the population that may potentially benefit from 
this vaccine approach. Our overall objective is to evaluate interventions that 
substantially reduce the risk of HIV-1 transmission through breastmilk, hence the 
use of this population of HIV-exposed, uninfected infants.  
4.4.2 
Cord Blood 
To minimize blood draws and to optimize our understanding of cellular responses, 
we will be using cord blood to evaluate baseline immunogenicity. Without the use 
of cord blood, some of the baseline immunologic studies we are proposing would 
require volumes of blood that would be unsafe to draw directly from neonates. 
For logistical reasons, cord blood can only reliably be obtained during planned 
Caesarian sections and planned Caesarian sections occur more frequently in this 
population. We will only enroll women who will require a planned Caesarian 
section, as determined by their obstetrical care provider. 
4.4.3 
Immunization number 
Several interlinked considerations have informed the design of our study, utilizing 
five immunizations of CH505TF gp120 starting at birth, with the primary 
immunogenicity analysis at one year. These include: 1) to begin vaccination at 
birth to engage the unique qualities of the neonatal immune system, 2) the need to 
test for infant antibody responses at a timepoint where maternal antibodies have 
waned, and 3) to compare the infant immune response to the adult immune 
response in HVTN 115. 
4.4.4 
Dose (amount and number) 
The intention of this study is to evaluate the safety and immunogenicity of 
CH505TF gp120 adjuvanted with GLA-SE in infants and to compare these data to 
the findings of the adult HVTN 115 study using the same adjuvanted vaccine 
formulation. In this study, HEU infants will be enrolled within 5 days of birth to 
receive vaccination with the CH505TF gp120 immunogen adjuvanted with GLA-
SE or placebo. The highest GLA-SE dose (5 mcg) chosen for this study is 
supported by data coming from a nonhuman primate study that demonstrated the 
immunogenicity of GLA-SE in infant rhesus macaques (35). It is important to 
note that this is half the dose of GLA-SE administered to adults in the HVTN 115 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 25 of 151 
study. This is consistent with the approach for vaccines licensed for use in 
pediatric populations such as Hepatitis A and Hepatitis B, both of which use half 
the adult dose in children. We aim to do a dose escalation for the adjuvant, 
starting with a lower 2.5 mcg GLA-SE dose in a small number of infants, use a 
planned enrollment hold to review safety data from these infants, and then 
increase to the 5 mcg GLA-SE dose with an additional enrollment hold for PSRT 
review of safety data.  
There are HIV-specific concerns regarding somatic hypermutation, a necessary 
but not sufficient element for driving a bnAb response, that support the 5 mcg 
dose. In preclinical studies of other adjuvants (cholera toxin A1-DD and delta-
inulin), increasing the dose of adjuvant is clearly associated with increasing 
somatic hypermutation (36, 37). Insufficient TLR4 stimulation (the target of 
GLA-SE) has been demonstrated to mediate the poor affinity maturation and lack 
of germinal center responses observed with formalin-inactivated RSV vaccines 
(38). In addition, pre-clinical data would suggest that there is likely to be less 
reactogenicity in infants to the GLA-SE adjuvant. In vitro studies have 
demonstrated that both neonatal and infant whole blood stimulated with GLA 
produces reduced amounts of the pyrogens TNF, IL-12 (p40), and IFN- when 
compared to adult blood (39). These inflammatory cytokines are likely mediators 
of reactogenicity (40). 
The immunogen dose is the lowest dose (20 mcg) of CH505TF gp120 evaluated 
in HVTN 115 Part A. In HVTN 115 Part A, there were no statistically significant 
differences in magnitude of elicited antibodies or in response rates between the 20 
mcg, 100 mcg, and 400 mcg dosing groups. In the adult cohort, no suppressive 
effect was seen with higher protein doses in HVTN 115 Part A; however, 
consistent with other vaccines used in both adult and infant populations, we have 
elected to use the lower 20 mcg dose to reduce the risk of high-dose suppression. 
It is possible that we may observe interference from maternal polyclonal 
antibodies vertically transferred to the infant; given the proven efficacy of EPI 
vaccines in this population where similar maternal antibody interference could 
occur, we believe the dose of 20 mcg offers the best balance between safety and 
immunogenicity. 
In addition to the 20 mcg dose of CH505TF gp120, we propose to include a 5 
mcg arm to further explore immunogenicity of CH505TF gp120 recombinant 
protein. This choice is supported both by preclinical studies of CH505TF gp120 
and by the results of HVTN 115 Part A. As described below in Section 4.7.3, we 
performed a dose ranging study in rhesus macaques. After the second and third 
immunizations, neutralization titers were similar in all dose groups (5 to 600 
mcg/dose; Figure 4-2). 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 26 of 151 
 
Figure 4-2 Neutralization activity elicited by CH505TF gp120 immunization against three 
isolates in the TZM-bl assay. There are no statistically significant differences between groups. 
In HVTN 115 Part A, the frequency of antigen-specific B cells in peripheral blood 
after the fifth dose of vaccine was similar in all groups, with a slightly higher 
frequency and response rate observed in the higher dose groups compared with 
the 20 mcg/dose group. In this pediatric study, the selection of lower doses is 
consistent with vaccine strategies for other pathogens where lower doses are often 
used. While this study will not be powered for a formal comparison between these 
arms, previous pediatric vaccine studies have demonstrated that immune 
responses induced by a lower dose of protein were comparable to or greater than 
those seen with the higher dose. In the malaria RTS,S vaccine regimen, a delayed 
fractional immunogen dose equal to 20 mcg induced higher antibody avidity and 
antibody somatic hypermutation as compared to the higher dose (41). The 
inclusion of the 5 mcg dose in this study will permit a comparison of antibody 
avidity with the 20 mcg dose in infants and with all doses (20, 100 and 400 mcg) 
in adult immune responses from the HVTN 115 study. 
The inclusion of five doses of a single product with a placebo arm will allow for 
directly assessing safety and testing whether the neonatal immune system has 
characteristics favorable to lineage-directed vaccination strategies.  
4.4.5 
Schedule 
In this study, the CH505TF gp120 vaccine will be given at 0, 8, 16, 32, and 54 
weeks. The first dose will be given within the first five days of life and, to avoid 
potential confounding of reactogenicity assessments, subsequent boosts will be 
given at least 7 days after scheduled EPI vaccinations (see Table 4-2). If an infant 
is found to have delayed EPI vaccinations, those vaccines will be administered by 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 27 of 151 
the site as soon as possible, followed by the next scheduled study product 
administration no sooner than 7 days later. The boost at week 54 is proposed to 
guide affinity maturation and the potential development of bnAb B cell lineages. 
This schedule is informed by the results of the PACTG 230 study (see Section 
4.8.3). The PACTG 230 study immunized infants at 0, 4, 12, 20 weeks (extended 
schedule) and at 0, 2, 8 and 20 weeks (accelerated schedule) –both with high titers 
of antibodies that were similar at weeks 24 and 52 (42). A recent infant NHP 
immunization study comparing three accelerated (0, 3, and 6 weeks) dosing Env 
arms (protein only, MVA/protein prime boost, combination MVA/protein) vs. a 
fourth extended dosing (0, 6, 12 and 32 weeks) Env regimen (combination 
MVA/protein) demonstrated highest avidity antibody in the extended dosing arm 
(43). In vaccine studies with adult macaques, higher avidity anti-Env antibodies 
have been correlated with protection against rectal and vaginal simian-human 
immunodeficiency virus (SHIV) or simian immunodeficiency virus (SIV) 
challenge (44-47).  
Table 4-2 Relationship of HVTN 135 Study Product Administration Schedule to National EPI 
schedule in South Africa  
Infant 
Age 
Vaccine 
Proposed 
Schedule in 
HVTN 135 
Birth 
Oral Polio (OPV) 
Tuberculosis (BCG) 
 
Day 5 
 
Dose 1 
Week 6 
Oral Polio (OPV) 
Rotavirus (RV) 
Diphtheria-tetanus-acellular pertussis-injectable polio- Haemophilus 
influenza B Hepatitis B (DTaP-IPV-Hib-Hep B) 
Pneumococcal Conjugate Vaccine (PCV) 
 
Week 8 
 
Dose 2 
Week 10 
Diphtheria-tetanus-acellular pertussis-injectable polio- Haemophilus 
influenza B Hepatitis B (DTaP-IPV-Hib-Hep B) 
 
Week 14 
Rotavirus (RTV) 
Diphtheria-tetanus-acellular pertussis-injectable polio- Haemophilus 
influenza B Hepatitis B (DTaP-IPV-Hib-Hep B)  
Pneumococcal Conjugate (PCV) 
 
Week 16 
 
Dose 3 
Week 24 
Measles Vaccine 
 
Week 32 
 
Dose 4 
Week 36 
Pneumococcal Conjugate (PCV) 
 
Week 52 
Measles Vaccine 
 
Week 54 
 
Dose 5 
In addition, there are several important scientific questions favoring keeping the 
schedule the same as the adult schedule in HVTN 115 Part A. As described 
above, a high degree of somatic hypermutation is thought to be required for the 
development of most, but not all (17), bnAbs. While there have been no detailed 
kinetic studies (to our knowledge) of how repeated vaccinations in neonates 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 28 of 151 
affects the rate of somatic hypermutation, a higher degree of somatic 
hypermutation with five vaccinations as compared to two vaccinations was 
demonstrated in a nonhuman primate model (48). In humans there are even fewer 
data. Although direct interpretation is difficult due to the long lag between 
immunizations, data from RV144/305 strongly suggest that a fifth vaccination 
leads to further somatic hypermutation (49). Data from this study will be 
compared to data derived from the adult population (HVTN 115) to assess if the 
quality and kinetics of vaccine-induced somatic hypermutation differ in neonatal 
vs adult B cells. 
4.4.6 
Choice of control 
Sodium Chloride for Injection, 0.9% will be used as placebo.  
Inclusion of placebo groups in this study is deemed necessary to maintain 
blinding when evaluating clinical events and their relationship to study vaccine. In 
addition, comparisons of safety and immunogenicity data between vaccine and 
placebo recipients could provide useful descriptive results despite the small 
sample size.  
4.5 
Plans for future product development and testing 
A goal of iterative human clinical trials to learn how to induce broadly 
neutralizing antibodies. This study will allow us to compare infant and adult 
immune responses to CH505TF gp120 and initiate the first steps in the CH103 
bnAb lineage. Considerable data have been reported that bnAb precursors are rare 
and subdominant and, in some cases, are made so by tolerance controls of bnAb 
precursors in bone marrow (12, 50, 51). Thus, this trial represents the first in a 
potential series of iterative phase 1 clinical studies to begin learning how to 
induce bnAbs in infant vaccinees. Importantly, the match of the immunogens and 
adjuvants with Part A of the adult trial (HVTN 115) will address the question of 
whether infants respond differently than adults. Next steps following this trial will 
be informed by the results of the lineage based design strategy being tested in 
adults through HVTN 115 as well as several upcoming adult trials testing next 
generation immunogens which may stimulate bnAb lineages. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 29 of 151 
4.6 
Preclinical safety studies 
Table 4-3 Summary of preclinical safety studies  
Study 
number 
Product 
Type of 
study 
Animal 
N per 
group 
Dose groups 
Route 
Schedule 
1726-031 
Stable 
CH505TF 
Toxicity 
New 
Zealand 
White 
Rabbits 
10 M, 
10 F  
Group 1: saline control 
Group 2: CH505TF + GLA-SE 
Group 3: DNA + CH505TF + 
GLA-SE 
Group 4: GLA-SE 
IM 
Days 1, 15, 29, 
43, 57, 71, 85 
 
4.6.1 
Toxicology study of CH505TF gp120 adjuvanted with GLA-SE in 
rabbits 
A toxicity study with Stable CH505TF gp120 in New Zealand White (NZW) 
rabbits was conducted in compliance with Good Laboratory Practices (GLP). 
Seven biweekly IM injections of 400 mcg CH505TF gp120 with 20 mcg GLA-SE 
adjuvant or 20 mcg GLA-SE alone, to NZW rabbits for 13 weeks were well 
tolerated. Test article-related changes in dermal scores, clinical pathology 
parameters, and microscopic injection site findings were resolved/partially 
resolved and/or trending toward recovery by the recovery necropsy, and none of 
the test article-related changes were deemed adverse. A more detailed description 
of the toxicity study can be found in the Investigator’s Brochure (IB). 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 30 of 151 
4.7 
Preclinical immunogenicity studies 
Table 4-4 Summary of preclinical immunogenicity studies  
Study 
number 
Product 
Animal 
N per 
group 
Regimen groups 
Route 
Schedule 
(weeks) 
Assay 
501 
CH505TF 
gp120 Env 
with GLA-
SE 
Guinea pigs 
4 
1 mcg CH505TF only 
1 mcg CH505TF, 5 mcg GLA-SE 
1 mcg CH505TF, 10 mcg GLA-SE 
1 mcg CH505TF, 25 mcg GLA-SE 
5 mcg CH505TF only 
5 mcg CH505TF, 5 mcg GLA-SE 
5 mcg CH505TF, 10 mcg GLA-SE 
5 mcg CH505TF, 25 mcg GLA-SE 
50 mcg CH505TF only 
50 mcg CH505TF, 5 mcg GLA-SE 
50 mcg CH505TF, 10 mcg GLA-SE 
50 mcg CH505TF, 25 mcg GLA-SE 
IM 
0, 3, 6, 9, 
12 
Ab binding 
NHP 79 
EnvSeq-1 
gp120 Env 
with GLA-
SE 
Rhesus 
Macaques 
4 
100 mcg CH505TF only 
100 mcg CH505 sequential Envs 
100 mcg CH505 additive Envs 
IM 
0, 6, 12, 
19, 24, 57 
nAb, binding 
Ab, blocking 
Ab 
NHP 
106 
CH505TF 
gp120 Env 
with GLA-
SE 
Rhesus 
Macaques 
4 
5 mcg CH505TF Env 
20 mcg CH505TF Env 
100 mcg CH505TF Env 
300 mcg CH505TF Env 
600 mcg CH505TF Env 
 
IM 
0, 4, 12, 
29, 37 
nAb, binding 
Ab, 
differential 
Ab, blocking 
Ab 
The CH505TF gp120 used in the initial animal studies (guinea pig 501 and NHP 
79) was generated in transiently-transfected 293 cells. The 293-produced 
CH505TF gp120 has a different glycosylation profile than that of the CHO-
produced protein used for the rabbit toxicology study, mouse immunogenicity, the 
nonhuman primate (NHP) dose response study (NHP 106). 
4.7.1 
Immunogenicity of CH505-derived gp120 envelopes with GLA-SE in 
guinea pigs, study # 501 
In a guinea pig study, the GLA-SE adjuvant increased binding antibody 
magnitude to gp120 at all doses over the no adjuvant alone group (Barton Haynes, 
personal communication; see Figure 4-3).  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 31 of 151 
 
Figure 4-3 GLA-SE enhanced immunogenicity of CH505TF gp120 protein in guinea pigs. 
Binding antibodies from guinea pigs immunized with varying doses of CH505TF gp120 and varying 
amounts of GLA-SE were measured by enzyme-linked immunosorbent assay (ELISA). The data 
were used to calculate the log-transformed area under the curve (AUC) as shown on the y-axis. 
CH505TF gp120 was immunogenic at all doses. Groups 1, 5 and 9 received no GLA-SE and had 
significantly less anti-gp120 antibodies than the groups with 5 mcg (Groups 2, 6 and 10), 10 mcg 
(Groups 3, 7, and 11) or 25 mcg (Groups 4, 8, and 12) of GLA-SE. Moreover, there was no 
difference between the three GLA-SE doses (statistical analysis not shown). 
4.7.2 
Immunogenicity of CH505-derived gp120 Envelopes, study NHP 79 
The EnvSeq-1 immunogens consist of four gp120 proteins derived from CH505: 
CH505TF, CH505w53, CH505w78, and CH505w100. These proteins were 
administered to 3 groups of four rhesus macaques (NHP 79) with the GLA-SE 
adjuvant: 1) CH505TF Env gp120 alone; 2) the Envs given in a sequential 
regimen (CH505TF, CH505w53, CH505w78, and CH505w100); and 3) an 
additive-immunization regimen consisting of CH505TF gp120 in combination 
with the evolved Env variants (CH505TF, then CH505TF + CH505w53, then 
CH505TF + CH505w53 + CH505w78, then CH505TF + CH505w53 + 
CH505w78 + CH505w100). These Envs were research grade products made by 
transient transfection of 293 cells and have different glycosylation profiles than 
the CHO-produced CH505TF gp120s to be used in this clinical trial. This study 
demonstrates the proof of concept of using sequential Env variants in NHPs; the 
first step of this process using CH505TF is what will be tested in this clinical 
protocol. A follow on study employing the EnvSeq-1 immunogens administered 
in additive or sequential combinations may be a next step and will be informed by 
results from the HVTN 115 Part B trial which will test this approach in adults. 
The CH505 gp120 Env derivatives (including CH505TF) were found to be 
immunogenic. An important readout in this study was the development of 
antibodies capable of binding a specific epitope on the CD4bs of the envelope 
protein. A defining feature of the CH103-like bnAbs is the capacity to bind wild 
type CH505 Env gp120 but not to CH505 Env with a deletion of isoleucine at aa 
position 371 (CH505 Env IΔ371 gp120). The data from the NHP 79 study suggest 
that the UCAs of bnAb lineages could be expanded by vaccination, but that they 
L o g  A U C
0
5
1 0
1 5
2 0
Group #:
CH505TF (mcg):
GLA-SE (mcg):
1
1
0
6
5
5
7
5
10
8
5
25
9
50
0
10
50
5
11
50
10
12
50
25
2
1
5
3
1
10
4
1
25
5
5
0

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 32 of 151 
did not evolve into bnAbs. In this study of 12 monkeys, 10/12 monkeys had 
differential binding antibodies isolated but overall the Env differential binding 
antibodies were subdominant and comprised only about 13% of the total activity 
of the antibody response, which is consistent with the low rate of bnAb 
development in HIV-infected adults (52) (Table 4-5). 
Table 4-5 Frequency of antibodies isolated from CH505 envelope-vaccinated macaques. 
CH505 differential-binding antibodies (CH505 TF gp120+, CH505 TF gp120 Δ371I+/− (greater than 
or equal to threefold vs. Wild Type)) were referred to as CH505 differentials. Adapted from Williams 
et al. (52). 
 
4.7.3 
Immunogenicity of CH505TF gp120, study NHP 106 
A dose ranging study was performed in rhesus macaques using 5 to 600 mcg of 
Env CH505TF gp120 (produced by stably transfected CHO-DG44 cells) along 
with 25 mcg GLA-SE with each dose administered 5 times (Barton Haynes, 
personal communication). Immunogenicity was demonstrated at all dose levels 
and no adverse reactions were observed. Env binding antibody (Figure 4-4) and 
CD4bs antibody (Figure 4-5) levels were determined and the CH505 TF gp120 
was found to be immunogenic after 2 immunizations. All dose levels gave similar 
binding responses. When we assessed the frequency of memory B cells that bind 
antigen probes in a CD4bs differential manner (called differential binding 
memory B cells), all animals in the 20, 100, 300, and 600 mcg protein dose 
groups demonstrated differential binding after 3 injections (Figure 4-6). In this 
group of adult macaques, the frequency of differential binding B cells in the 5 
mcg dose group was lower than that seen in the other groups. The inclusion of the 
5 mcg dose in this study will permit evaluation of both antibody levels and B cell 
frequencies in human infants, enabling us to determine if the permissive 
environment of the infant immune system results in a higher frequency of 
differential binding B cells in a dose sparing manner. 
 
Total Abs
CH505 Env +
CH505 Env +
Vaccine Groups
Animal ID
Count
% of total
Count
% of total
5346
15
13
4
27%
13
11
2
15%
5356
2
1
0
0%
2
1
0
0%
5360
11
9
0
0%
10
8
0
0%
5361
78
74
9
12%
68
64
7
10%
5362
2
2
1
50%
2
2
1
50%
5363
14
14
1
7%
14
14
1
7%
5551
11
10
1
9%
11
10
1
9%
5553
35
24
3
9%
34
23
3
9%
5554
23
19
3
13%
21
17
3
14%
5556
49
39
9
18%
43
33
7
16%
5558
28
23
4
14%
25
21
4
16%
5560
10
9
1
10%
10
9
1
10%
Summary
278
237
36
13%
253
213
30
12%
Unique clonal lineages
CH505 differentials
Unique 
lineages
Group 1: CH505TF 
alone
Group 4: EnvSeq1 
given sequentially
Group 5: EnvSeq1 
given additively
CH505 differentials
All antibody lineages isolated

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 33 of 151 
 
Figure 4-4 Binding of Immunized NHP 106 Plasma to CH505TF gp120. Each data point 
represents a single animal. Animals were immunized with the indicated dose at weeks 0, 4 and 12. 
X-axis shows bleed time. Bleed 0 was performed before immunization and subsequent bleeds were 
performed 2 weeks after each respective immunization. No animals had binding antibodies prior to 
immunization; binding was detected in all animals after the second immunization. After the third 
immunization, binding antibodies were similar among all groups. While all doses gave a similar 
binding response, the higher doses of 300 and 600 mcg gave earlier responses and trended to 
higher responses compared to the lower doses. No additional rise in antibody levels was observed 
after the fourth or fifth immunizations (data not shown). 
 
 
 
 
 
Figure 4-5 Inhibition of CD4 binding by NHP 106 plasma after immunization with 5 different 
doses of CH505TF gp120. Each data point represents a single animal. Animals were immunized 
with the indicated dose at weeks 0, 4 and 12. Note that the x-axis shows bleed time. Bleed 0 was 
performed before immunization and subsequent bleeds were performed 2 weeks after each 
respective immunization. The ability of plasma antibody to block the binding of CD4bs mAb CH106 
to heterologous B.63521 gp120 was quantified by ELISA. Blocking antibodies were detectable in 
the high dose groups after the second immunization and increased in Group 3 after the third 
immunization. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 34 of 151 
 
Figure 4-6 Percentage of differential binding memory B cells. Peripheral blood mononuclear 
cells (PBMC) from immunized macaques were assayed to determine the frequency of memory B 
cells that bind to CH505TF gp120 but not to CH505TF gp120 ΔI371 that disrupts the CD4 BS. 
Each data point represents a single animal. Animals were immunized at the indicated dose at 
weeks 0, 4 and 12. Note that the x-axis shows bleed time. Bleed 0 was performed before 
immunization and subsequent bleeds were performed 2 weeks after each respective immunization. 
A pattern of increased differential binding memory B cell induction with increasing vaccine dose is 
noted starting after two immunizations and increasing after a third immunization. 
4.8 
Clinical studies 
4.8.1 
Clinical studies of CH505TF gp120 with GLA-SE adjuvant in adults, 
HVTN 115 
CH505TF gp120 is being evaluated in HVTN 115, the first clinical trial designed 
to use a B-cell lineage approach to elicit CD4bs bnAbs. In Part A, the dose 
ranging portion of HVTN 115, the CH505TF gp120 was administered at months 
0, 2, 4, 8, and 12 at doses of 20, 100, and 400 mcg with 10 mcg of GLA-SE 
(Table 4-6).  
Table 4-6 Schema for HVTN 115 Part A 
Study 
group 
N 
Month 0 
 (Day 0) 
Month 2 
(Day 56) 
Month 4 
(Day 112) 
Month 8 
(Day 224) 
Month 12 
(Day 364) 
1 
12 
20 mcg 
CH505TF 
20 mcg 
CH505TF 
20 mcg 
CH505TF 
20 mcg 
CH505TF 
20 mcg 
CH505TF 
 
 
 
 
 
 
 
2 
12 
100 mcg 
CH505TF 
100 mcg 
CH505TF 
100 mcg 
CH505TF 
100 mcg 
CH505TF 
100 mcg 
CH505TF 
 
 
 
 
 
 
 
3 
12 
400 mcg 
CH505TF 
400 mcg 
CH505TF 
400 mcg 
CH505TF 
400 mcg 
CH505TF 
400 mcg 
CH505TF 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
4 
6 
Placebo 
placebo 
placebo 
Placebo 
placebo 
 
 
 
 
 
 
 
Total 
Part A 
42 
(36/6) 
 
 
 
 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 35 of 151 
Based on safety and immunogenicity data from Part A, HVTN 115 Part B will 
administer Env gp120 immunogens that induced a CD4bs bnAb clonal lineage in 
an HIV-infected individual in both sequential and additive strategies at a dose of 
400mcg. The primary immune signal is differential binding antibodies between 
CH505TF Env gp120 (wild type) and CH505 Env IΔ371 gp120 (mutant): a 
differential binding ratio of wild-type/mutant ≥ 2.5 is characteristic of a CD4bs 
specificity.  
HVTN 115 Part A reached full enrollment with 42 participants in May 2018. The 
last participant received their final vaccination on May 7, 2019. As of  August 20, 
2020, all scheduled study visits have been completed and the study remains 
blinded from a safety perspective. Most participants reported none or mild local 
signs and symptoms after vaccine administrations. There have been 2 cases of 
severe (grade 3) erythema/induration, which resolved within days. One of those 
participants experienced moderate injection site cellulitis, which resolved with 
oral antibiotics. Most participants reported none or mild systemic reactogenicity 
signs or symptoms after vaccinations. One case of severe (grade 3) pain at the 
injection site and two cases of severe (grade 3) headache were reported during the 
reactogenicity period. Beyond the reactogenicity period, there have been no 
severe adverse events related to study product.  
Preliminary analysis of the immunogenicity data from Part A suggests that there 
were no statistically significant differences between dose groups for magnitude 
and response rates for antibodies to CH505, CD4 binding site, or B-cell responses 
between the treatment groups. For this reason, the lower doses have been chosen 
for this study to maximize safety and immunogenicity in human infants. 
4.8.2 
Clinical studies of the GLA-SE adjuvant and protein combinations in 
adults 
GLA-SE is a synthetic TLR4 agonist formulated in a stable nano-emulsion of 
squalene oil and promotes a strong Th1-type immune response to vaccine 
antigens (32, 33). Although the CH505TF gp120 protein in combination with 
GLA-SE has not been evaluated in humans to date, as summarized in Section 
4.8.1, the Stable CH505TF gp120 is under evaluation in HVTN 115. In addition, 
there is considerable clinical trial experience with the GLA-SE adjuvant in 
combination with other antigens. These data are summarized below and in Table 
4-7 and Table 4-8. Over 2500 individuals have received at least one dose of the 
GLA-SE adjuvant at doses ranging from 0.5 to 20 mcg with no significant safety 
concerns identified to date. Please see the IB for additional details of the clinical 
trials performed with GLA-SE.  
Five studies have included the 10 mcg dose of the GLA-SE adjuvant, a higher 
dose than proposed for this study. The first was an open-label phase 1 clinical trial 
conducted in Brazil with a Schistosoma mansoni antigen (Sm14) (53). Twenty 
healthy males received 3 IM doses of 50 mcg Sm14 + 10 mcg GLA-SE at one-
month intervals. The vaccine was safe and generally well tolerated with no 
serious adverse events (SAEs) or Grade 4 adverse events (AEs) (53). Injection 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 36 of 151 
site pain was commonly reported (80%, 50%, and 41% after the first, second, and 
third dose, respectively), but generally mild and self-limited. There were no 
abnormalities in physical exams, serum chemistries, and hematology values 
related to study vaccine. Humoral and CD4+ T cell responses to Sm14 were 
reported (53). 
The second clinical trial using the 10 mcg dose of GLA-SE is a recently 
completed phase 1, open-label evaluation of the safety, tolerability, and 
immunogenicity of a Leishmania vaccine (LEISH-F3) in combination with SLA-
SE (second generation glucopyranosyl lipid A in stable oil-in-water emulsion) 
adjuvant compared to LEISH-F3 with GLA-SE in healthy adults (NCT02071758; 
Protocol IDRI-LVVPX-117). The SLA-SE adjuvant is a next generation TLR4 
adjuvant formulation. Thirty-nine participants were randomized to 4 arms: high 
dose LEISH-F3 (20 mcg) and low dose SLA-SE (5 mcg); high dose LEISH-F3 
and high dose GLA-SE adjuvant (10 mcg); low dose LEISH-F3 (5 mcg) and high 
dose GLA-SE adjuvant (10 mcg); and high dose LEISH-F3 and high dose SLA-
SE adjuvant (10 mcg). Participants received 3 injections at one-month intervals. 
This study is complete. The LEISH-F3 + GLA-SE and LEISH-F3 + SLA-SE 
vaccines were safe and well tolerated in adult subjects. No deaths, no dose-
limiting toxicities (DLTs), no Grade 3 or 4 AEs, no SAEs, and no AE of special 
interest (AESIs) occurred during the study. All study injection reactions were 
Grade 1 or Grade 2. The most frequently reported reactions were injection site 
tenderness/pain and fatigue. 
Three other clinical trials are evaluating GLA-SE in oncology. Two ongoing trials 
are investigating GLA-SE alone as an immunotherapy and one trial tested GLA-
SE in combination with a prostate cancer antigen. No data are currently available 
from these trials. 
Clinical experience with GLA-SE as an adjuvant for other vaccines at doses 
ranging from 1 mcg to 5 mcg suggests that it is safe and well-tolerated. Treanor et 
al. tested an avian influenza hemagglutinin (H5) subunit vaccine at a range of 
doses with and without a fixed dose of GLA-SE (1 mcg) and reported mild to 
moderate injection site pain and/or tenderness in 50–70% of H5 + GLA-SE 
recipients, with myalgias and headaches reported by a minority (25-27%) of 
vaccinees. No other safety findings were noted, and the GLA-SE adjuvant 
substantially increased the immunogenicity of the vaccine (54). 
A recent phase 1 clinical trial of a respiratory syncytial virus (RSV) vaccine in 
older adults (≥ 60 years of age) found that a combination of the RSV fusion (F) 
protein at three different doses with 2.5 mcg of GLA-SE was safe, well-tolerated, 
and immunogenic. Compared with the unadjuvanted vaccine, GLA-SE increased 
local reactogenicity, with 40-65% of subjects reporting mild-to-moderate, self-
limited injection site pain and/or tenderness (55). No other safety concerns were 
reported. Immune responses were F protein dose-dependent, and the adjuvant 
enhanced both humoral and cellular immune responses (55). A follow-up study 
using a higher dose of F protein and three doses of GLA-SE has been completed 
(NCT02289820). The vaccine was safe, tolerable, and immunogenic and the data 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 37 of 151 
supported the selection of 120 mcg F protein /5 mcg of GLA-SE for further 
evaluation (56). 
GLA-SE was also found to be safe and well-tolerated at doses of 2 mcg and 5 
mcg in a phase 1 trial of LEISH-F3 given three times at one month intervals (57). 
Injection site pain and/or tenderness was quite common (90-100%) with fatigue 
noted by 40-60% of vaccinees receiving the adjuvanted product versus 33% of 
subjects who received unadjuvanted vaccine (57). The unadjuvanted LEISH-F3 
was essentially non-immunogenic while antibody and cytokine responses were 
noted in vaccinees in both dose groups of GLA-SE (57). 
GLA-SE at a 5 mcg dose has also been tested together with protein antigens in 
several other clinical trials which are included in Table 4-7 and Table 4-8. 
Table 4-7 Completed clinical trials using IDRI GLA-SE adjuvant formulations in combination 
with other vaccines (as of March 20, 2018) 
Sponsor/Partner 
Disease Area/Antigen 
GLA-SE 
Dose 
# Receiving 
GLA-SE 
Total 
# in 
Study 
Oswaldo Cruz Foundation / IDRI 
NCT01154049  
Schistosomiasis (Sm14)  
10 mcg  
20 
20 
IDRI / Rockefeller University 
NCT01397604 and NCT01864876  
Adjuvant only 
2 mcg  
5 mcg  
10 
7 
49 
IDRI  
NCT01484548  
Leishmaniasis (LEISH-F3)  
2 mcg  
5 mcg  
12 
12 
36 
NIAID / IDRI  
NCT01751048  
Leishmaniasis (LEISH-F3)  
5 mcg  
16 
 
48 
IDRI 
NCT02071758  
Leishmaniasis (LEISH-F3)  
10 mcg 
18 
39 
WRAIR / IDRI 
NCT01540474  
Malaria (CelTOS)  
2 mcg  
5 mcg  
10 
20 
30 
European Vaccine Initiative / IDRI 
NCT01949909  
Malaria (p27A)  
2.5 mcg 
5 mcg  
24 
8 
56 
European Vaccine Initiative / IDRI 
NCT02014727  
Malaria (AMA-1 DiCo)  
2.5 mcg  
33 
66 
IDRI / Aeras 
NCT01599897  
TB (ID93)  
2 mcg  
5 mcg  
24 
24 
60 
IDRI / Aeras 
NCT01927159 
TB (ID93) 
2 mcg 
5 mcg 
39 
15 
66 
IDRI  
NCT02465216 
TB (ID93) 
2 mcg 
5 mcg 
20 
28 
60 
CONFIDENTIAL  
Seasonal influenza  
0.5 mcg  
1 mcg  
2.5 mcg  
5 mcg  
6 
12 
36 
4 
96 
Protein Sciences / Immune Design  
NCT01147068  
Pandemic influenza 
(recombinant protein)  
1 mcg  
220 
392 
Novavax / Immune Design 
NCT01596725  
Pandemic influenza (H5-VLP)  
2.5 mcg  
169 
333 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 38 of 151 
Sponsor/Partner 
Disease Area/Antigen 
GLA-SE 
Dose 
# Receiving 
GLA-SE 
Total 
# in 
Study 
Medicago / Immune Design 
NCT01991561  
Pandemic influenza (H5-VLP)  
5 mcg  
130 
390 
Immune Design 
NCT02015416 
Cancer (NY-ESO-1) 
2 mcg 
5 mcg 
10 mcg 
3 
3 
6 
12 
Medimmune / Immune Design 
NCT02115815  
Respiratory syncytial virus (sF) 
2.5 mcg 
60 
144 
Medimmune / Immune Design 
NCT02289820  
Respiratory syncytial virus (sF) 
1 mcg 
2.5 mcg 
5 mcg 
39 
99 
79 
261 
Medimmune / Immune Design 
NCT02508194 
Respiratory syncytial virus (sF) 
5 mcg 
946 
1894 
 
Table 4-8 Ongoing clinical trials using IDRI GLA-SE adjuvant formulations in combination 
with other vaccines (as of March 20, 2018) 
Sponsor/Partner 
Disease Area/Antigen 
GLA-SE 
Dose 
# Receiving 
GLA-SE 
Total # in 
Study 
IDRI  
NCT03302897 
Leprosy (LEP-F1)  
5 mcg  
24 
24 
NIH/NIAID/DMID 
NCT02508376 
TB (ID93) 
5 mcg 
 
20 
 
70 
University Hospital Tuebingen 
NCT02647489 
Malaria (PAMVAC) 
5 mcg 
 
21 
 
63 
Institut National de la Santé Et 
de la Recherche Médicale 
NCT02658253 
Malaria (PRIMVAC) 
2.5 mcg 
29 
68 
Immune Design 
NCT02035657 
Merkel cell carcinoma 
5 mcg 
9 
9 
Immune Design  
NCT02501473 
Non-Hodgkin's lymphoma  
5 mcg 
10 mcg 
20 mcg 
3 
3 
4 
10 
Immune Design  
NCT02180698 
Sarcoma  
5 mcg 
10 mcg 
20 mcg 
4 
4 
4 
12 
Immune Design  
NCT02387125 
Cancer (CMB305, G305) 
Not 
Available 
 
 
Immune Design  
NCT02609984 
Cancer (CMB305) 
Not 
Available 
 
 
Immune Design  
NCT02320305 
Melanoma (MART1) 
Not 
Available 
 
 
HVTN 115 
NCT03220724 
HIV Vaccine 
10 mcg 
116 
132 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 39 of 151 
4.8.3 
Clinical studies with related HIV-1 Env gp120 immunogens in infants 
(PACTG 230 and PACTG 326) 
The adult RV144 pox prime-rgp120 protein boost trial was the first HIV-1 
vaccine study to show some efficacy against HIV-1 acquisition. A vaccine 
efficacy of 31% was achieved in a heterosexual adult Thai population at three 
years post vaccination (58) and up to 60% protection was observed one-year post 
vaccination (58, 59). The subsequent immune correlate analysis showed that an 
IgG response, IgG3 in particular, against the variable loops 1 and 2 (V1V2) 
predicted decreased HIV-1 acquisition risk (60-63) whereas an Env-specific IgA 
response to specific HIV epitopes was associated with a lack of vaccine efficacy 
(64). Moreover, an IgG response against the variable loop 3 (V3) region was also 
associated with protection in the setting of low levels of vaccine-elicited 
responses (62). 
Samples from two pediatric trials (PACTG 230 and PACTG 326) were then 
evaluated to determine immune responses in infants (see Table 4-9). In PACTG 
230, infants were vaccinated between 0-20 weeks of age with four doses of 
Chiron rgp120 (SF2 strain) with MF59 (n=74), VaxGen rgp120 (MN strain) with 
aluminum hydroxide (alum, n=80), or adjuvant only placebo (MF59 or alum) 
(n=30) (65). In PACTG 326, infants received four doses of ALVAC-HIV alone 
(n= 12), ALVAC-HIV with AIDSVAX B/B with the adjuvant alum (n=10) or 
placebo (n=12; n=8 saline & n=3 alum/saline) between 0-12 weeks of age (66). 
Infants developed robust Env-specific antibody responses to these first-generation 
HIV-1 Env vaccines. At one year of age, most maternally-acquired antibodies had 
waned, and vaccine Envelope-specific IgG responses were significantly higher in 
vaccinees compared to placebo recipients (21). Importantly, infants receiving the 
Chiron vaccine had higher and more durable IgG responses than adult vaccinees 
in the RV144 vaccine trial, with vaccine-elicited IgG responses still detectable in 
56% of infants at two years of age. Remarkably, at peak immunogenicity, the 
concentration of anti-V1V2 IgG, a response associated with reduced risk of HIV 
acquisition in the RV144 adult vaccine trial, was 22-fold higher in infants who 
received the Chiron rgp120/MF59 vaccine/adjuvant combination than in RV144 
vaccinees. V1V2-specific IgG3 responses, the subclass of the IgG response that 
best associated with protection in the RV144 study, were detected  in 43% of the 
vaccinated infants at 24 weeks after vaccination. Finally, vaccine-elicited IgA 
responses rarely detected in the vaccinated infants. To determine if the difference 
in infant and RV144 adult responses was due to distinct vaccine regimens, the 
infant vaccine-elicited antibody responses were then compared to that of adults 
immunized with the same vaccine regimens (67). At peak immunogenicity, no 
difference was observed in the magnitude of gp120 or V1V2-specific IgG 
between adults and infants immunized with the Alum adjuvanted VaxGen 
vaccines; however, infants who received the MF59 adjuvanted Chiron vaccine 
had more robust responses than adults immunized with the same vaccine. Overall, 
these studies have multiple implications for the proposed clinical trial: 1) they 
demonstrated that infant vaccination can elicit high levels and durable Env-
specific antibodies that have been associated with protection in vaccinated adults; 
2) they indicate possible differences in how adjuvants modulate immune 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 40 of 151 
responses in adults and infants (68-71); 3) they also indicate that presence of 
maternal anti-HIV-1 responses are not likely impact the immunogenicity of the 
vaccine. Ongoing studies will determine if the higher levels of antigen-specific 
antibodies generated in infant immunization have antiviral activity comparable to 
those elicited in adults. 
Table 4-9 Previous pediatric HIV vaccine trials including gp120 immunogen 
Study 
Name 
Vaccine (dose)/Adjuvant 
Schedule 
Population 
Related 
Serious 
Adverse 
Events 
 
Reference 
PACTG 
218 
 
Chiron rgp120 (15 or 50 
mcg)) 
Microgene sys rgp160 (40 or 
320 mcg) 
Genentech rgp120 (75 or 300 
mcg) 
Alum/MF59 
0, 1, 2, 3, 4, 6 
months 
HIV-infected 
children enrolled 
between ages 1 
month to 18 
years/n=79/USA 
None 
(72) 
PACTG 
230 
VaxGen rgp120/Alum 
Chiron rgp120/MF59 
0, 4, 12, 20 
weeks OR 
0, 2, 8, 20 
weeks 
HIV-exposed 
children enrolled 
at birth 
/n=183/USA 
None 
(65) 
PACTG 
326 
Part 1:ALVAC vCP205  
 
Part 2: ALVAC vCP1452 +/- 
AIDSVAX B/B (200 mcg 
total)/Alum 
0, 4, 8, 12 
weeks 
 
HIV-exposed 
children enrolled 
at 
birth/n=28/USA 
HIV-exposed 
children enrolled 
at 
birth/n=30/USA 
None 
(66, 73) 
4.9 
Potential risks of study products and administration 
Table 4-10 includes general risks of vaccine administration along with risks 
known from prior clinical studies of CH505TF gp120 products and similar 
envelope proteins with GLA-SE adjuvant. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 41 of 151 
Table 4-10 Summary of potential risks of study products and administration 
Common  
• Mild to moderate injection site pain, tenderness, erythema, or 
swelling/induration/edema 
• A vaccine-induced positive HIV antibody test result 
Less common 
• Severe injection site pain or tenderness 
• Fever, chills, flu-like syndrome, arthralgia, rash, decreased appetite  
• 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 
• Muscle damage at the injection site 
Theoretical risks 
• Autoimmune disease  
• 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 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 42 of 151 
5 
Objectives and endpoints 
5.1 
Primary objectives and endpoints 
Primary objective 1: 
To evaluate the safety and tolerability of HIV-1 CH505 transmitted/founder virus 
Env gp120 immunogen (CH505TF gp120) adjuvanted with Glucopyranosyl Lipid 
A - stable emulsion (GLA-SE) in healthy HIV-1 exposed uninfected (HEU) 
infants 
Primary endpoint 1: 
Local and systemic reactogenicity signs and symptoms, laboratory measures of 
safety, weight gain, and AEs and SAEs 
Primary objective 2: 
To determine whether vaccination with CH505TF gp120 adjuvanted with GLA-
SE initiates B-cell lineages potentially capable of generating a broadly 
neutralizing antibody response 
Primary endpoints 2: 
Magnitude of HIV-1 Env gp120, CD4 binding site and V1V2-specific serum IgG 
binding antibodies, as assessed by BAMA two weeks after the 5th vaccination  
Quantification and phenotypic characterization of peripheral B cells capable of 
binding HIV-1 Env gp120, the CD4 binding site (including differential binding to 
the IΔ371 mutant) and the V1V2 binding site, as assessed by flow cytometry two 
weeks after the 3rd and 5th vaccinations 
5.2 
Secondary objectives and endpoints 
Secondary objective 1: 
To evaluate the effect of vaccination with CH505TF gp120 adjuvanted with 
GLA-SE on antibody responses to the South African Expanded Program on 
Immunization (EPI) schedule vaccinations 
Secondary endpoint 1: 
EPI vaccine-specific antibody responses, as assessed by Pediatric Vaccine 
Multiplex Assay (PVMA) 2 weeks after the 5th vaccination  
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 43 of 151 
Secondary objective 2: 
To evaluate the ability of the vaccine regimen to elicit HIV-specific nAbs. 
Secondary endpoint 2: 
Magnitude and breadth of serum neutralization of vaccine-matched viral isolates, 
and viruses engineered to detect precursors of CD4 binding site and V1V2 
antibodies 2 weeks after the 5th vaccination.  
Secondary objective 3: 
To measure Fc-mediated antibody effector functions 
Secondary endpoints 3: 
Response rate and magnitude of vaccine-elicited serum binding antibodies to FcR 
proteins, as assessed by BAMA 2 weeks after the 5th vaccination 
Response rate and magnitude of serum Antibody-dependent cellular cytotoxicity 
(ADCC), as assessed by flow cytometry and/or luciferase assays 2 weeks after the 
5th vaccination 
Response rate and magnitude of serum Antibody-dependent cellular phagocytosis 
(ADCP), as assessed by flow cytometry 2 weeks after the 5th vaccination 
5.3 
Exploratory objectives 
Exploratory objective 1: 
To compare antibody and B cell responses generated in infants in response to 
CH505TF gp120 adjuvanted with GLA-SE with adults vaccinated with the same 
immunogen and same adjuvant on the same schedule (HVTN 115) 
Exploratory objective 2:  
To characterize B cell lineages and evaluate the amount of B-cell receptor somatic 
hypermutation  
Exploratory objective 3: 
To evaluate effects of maternal antibody in cord blood, serum/plasma, and breast 
milk on the humoral response of infants 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 44 of 151 
Exploratory objective 4:  
To evaluate additional vaccine-induced antibody effector functions such as 
Antibody-dependent neutrophil phagocytosis (ADNP) and/or Antibody dependent 
complement deposition (ADCD) 
Exploratory objective 5: 
To measure the kinetics and maturation of the polyclonal antibody response with 
biophysical measurements (eg, BioLayer Interferometry, Surface Plasmon 
Resonance) 
Exploratory objective 6: 
Evaluation of breast milk and fecal microbiome at birth and over time to explore 
the effect of the microbiome on the development of the immune system and on 
vaccine immunogenicity 
Exploratory objective 7: 
To use systems biology approaches (eg, high resolution flow cytometry. 
transcriptomics, proteomics, metabolomics and/or plasma cytokine/chemokine 
assessment) to determine whether the pre- and/or postvaccination systemic 
immune milieu is associated with vaccine immunogenicity. 
Exploratory objective 8: 
To further evaluate immunogenicity, additional assays may be performed based 
on the HVTN Lab Assay Algorithm 
Exploratory objective 9: 
To conduct analyses related to furthering the understanding of HIV, immunology, 
vaccines, and clinical trial conduct 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 45 of 151 
6 
Statistical considerations 
6.1 
Accrual and sample size calculations 
Recruitment will target enrolling 38 healthy, HIV-uninfected infant participants 
born by Caesarian section to HIV-infected mothers (HIV exposed, uninfected 
infants). Although this study is proposed to be a direct comparison with 
HVTN 115, there are important differences in the populations that justify the 
inclusion of a larger placebo group. Specifically, we anticipate a higher rate of 
background severe adverse events in this population, as previous vaccine studies 
have determined a rate of 18% SAE in the placebo arm (74). This is markedly 
different from the background rate of severe adverse events in typical HVTN 
adult studies (~1-2%), supported by emerging safety data from the HVTN 115 
study in adults (see Section 4.8.1). Therefore, we propose including a larger 
placebo group to ensure capacity for determining a safety signal in this vulnerable 
population. 
There are both technical and logistical considerations that favor enrolling infants 
delivered via Caesarian section. From a technical standpoint, assessment of the 
evolution of antibodies requires an understanding of the baseline nature of B cells 
prior to vaccination. Standard assessment of antigen-specific B cell lineages 
requires several milliliters of blood, a volume that is unsafe in neonates. We 
intend to protect the participants of the study by performing baseline cellular 
assays on cord blood that would otherwise be discarded. For logistical reasons 
related to the study site, cord blood can only be collected during Caesarian 
sections. 
Since enrollment of mother-infant pairs is concurrent with infants receiving the 
first study vaccination, all participants will be included in the safety analysis. 
However, for immunogenicity analyses, it is possible that data may be missing for 
various reasons, such as participants terminating from the study early, problems in 
shipping specimens, low cell viability of processed peripheral blood mononuclear 
cells (PBMCs), or high assay background. While the site staff report a historical 
rate of 10% missingness in infant studies conducted at the site, immunogenicity 
data from previous phase 1 and 1 phase 2a HVTN trials indicate that 25% is a 
reasonable estimate for the rate of missing data for the later timepoint (1 year). 
For this reason, the sample size calculations below account for 25% of enrolled 
participants having missing data for the primary immunogenicity endpoint. 
6.1.1 
Sample size calculations for safety 
The goal of the safety evaluation for this study is to identify safety concerns 
associated with product administration. The ability of the study to identify SAEs 
can be expressed by the true event rate above which at least 1 event would likely 
be observed and the true event rate below which no events would likely be 
observed. Specifically, in the pooled Groups 3 and 5 (20 mcg CH505TF gp120 + 
5 mcg GLA-SE vaccine arms) of the study (n = 18), there is 90% chance of 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 46 of 151 
observing at least one event, if the true rate is 12% or more, and there is a 90% 
chance of observing no events, if the true rate is 0.58% or less. In the pooled 
placebo group of the study (n = 10), there is 90% chance of observing at least one 
event, if the true rate is 20% or more. There is a 90% chance of observing no 
events, if the true rate is 1% or less.  
As a reference, in HVTN vaccine trials in Southern Africa from April 2008 
through March 2018, about 1.7% of adult participants who received placebos 
experienced an SAE. Based on recent infant clinical trial studies including infants 
at the PHRU site, approximately 18% of infant participants who received 
placebos experienced an SAE (54). 
Binomial probabilities of observing 0, 1 or more, and 2 or more events among 
arms of size 18 and 10 are presented in Table 6-1 for a range of possible true 
adverse event rates. These calculations provide a more complete picture of the 
sensitivity of this study design to identify potential safety problems with the 
vaccine. 
Table 6-1 Probability (Pr) of observing 0 events, 1 or more events, and 2 or more events, 
among arms of size 18 and 10, for different true event rates 
True event rate (%) 
Pr(0/18) 
Pr(1+/18) 
Pr(2+/18) 
Pr(0/10) 
Pr(1+/10) 
Pr(2+/10) 
1 
83.5 
16.5 
1.4 
90.4 
9.6 
0.4 
5 
39.7 
60.3 
22.6 
59.9 
40.1 
8.6 
10 
15 
85 
55 
34.9 
65.1 
26.4 
20 
1.8 
98.2 
90.1 
10.7 
89.3 
62.4 
30 
0.2 
99.8 
98.6 
2.8 
97.2 
85.1 
40 
0 
100 
99.9 
0.6 
99.4 
95.4 
50 
0 
100 
100 
0.1 
99.9 
98.9 
60 
0 
100 
100 
0 
100 
99.8 
70 
0 
100 
100 
0 
100 
100 
80 
0 
100 
100 
0 
100 
100 
90 
0 
100 
100 
0 
100 
100 
An alternative way of describing the statistical properties of the study design is in 
terms of the 95% confidence interval for the true rate of an adverse event based 
on the observed data. Table 6-2 shows the 2-sided 95% confidence intervals for 
the probability of an event based on an observed rate. Calculations are done using 
the score test method (75). If none of the 18 participants in Groups 3 and 5 
experience a safety event, the 95% 2-sided upper confidence bound for the true 
rate of such events is 18%. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 47 of 151 
Table 6-2 Two-sided 95% confidence intervals based on observing a particular rate of safety 
endpoints for arms of size 18 and 10 
Observed event rate 
95% 
Confidence interval (%) 
0/18 
[0, 17.6] 
1/18 
[1, 25.8] 
2/18 
[3.1, 32.8] 
3/18 
[5.8, 39.2] 
4/18 
[9, 45.2] 
5/18 
[12.5, 50.9] 
6/18 
[16.3, 56.3] 
7/18 
[20.3, 61.4] 
8/18 
[24.6, 66.3] 
9/18 
[29, 71] 
10/18 
[33.7, 75.4] 
11/18 
[38.6, 79.7] 
12/18 
[43.7, 83.7] 
 
 
0/10 
[0, 27.8] 
1/10 
[1.8, 40.4] 
2/10 
[5.7, 51] 
3/10 
[10.8, 60.3] 
4/10 
[16.8, 68.7] 
5/10 
[23.7, 76.3] 
6/10 
[31.3, 83.2] 
6.1.2 
Sample size calculations for immunogenicity 
To address binding antibody endpoints, the analysis will descriptively summarize 
binding response positivity call rates to test superiority of IgG binding Ab 
responses for CH103-like CD4bs and V1V2 for comparisons versus zero using 
binomial exact tests with 5% type-I error rate each. Note that if in HIV-infected 
women, maternal antibodies that differentially bind to the CD4bs occur at a rate 
of 51% [estimated from n = 14 CAPRISA 002 women infected 2-3 years (76) and 
are transmitted to the infants, comparisons at early timepoints may be confounded 
by background positivity. Therefore, we have chosen the 2 weeks post 5th 
vaccination timepoint (~ one year) for the power calculation. By this time, both 
transplacental and breastmilk-associated antibodies (breast-feeding typically 
ceases at 6 months of age in the South Africa setting) are expected to have waned 
to below detectable levels, obviating the requirement to increase the placebo 
recipient sample size for a comparison with this high background rate. The 
following calculations assume a 25% rate of missing data to accommodate 
missingness at the 1-year timepoint. 
The main goals of this trial regarding immunogenicity outcomes involve a 
preliminary estimation of rates of “response” (defined for Primary Endpoint 2 as 
initiation of either the CD4bs or the V1V2 B cell lineage) based on data from 
immune assays among vaccinees. No adjustment for multiple comparisons will be 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 48 of 151 
made for the use of multiple assays. 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 for the 
response rate based on observing a rate of responses in the vaccinees is shown in 
Table 6-3. Calculations are done using the score test method (75). The n = 13 
assumes a 25% rate of missing immunogenicity data.  
Table 6-3 Two-sided 95% confidence intervals for the true response rate based on observing 
a particular rate of responses in the vaccinees (n = 13) 
No. of responses 
Observed response rate (%) 
95% Confidence interval 
1/13 
7.7 
[1.4, 33.3] 
2/13 
15.4 
[4.3, 42.2] 
3/13 
23.1 
[8.2, 50.3] 
4/13 
30.8 
[12.7, 57.6] 
5/13 
38.5 
[17.7, 64.5] 
6/13 
46.2 
[23.2, 70.9] 
7/13 
53.8 
[29.1, 76.8] 
8/13 
61.5 
[35.5, 82.3] 
9/13 
69.2 
[42.4, 87.3] 
10/13 
76.9 
[49.7, 91.8] 
11/13 
84.6 
[57.8, 95.7] 
12/13 
92.3 
[66.7, 98.6] 
13/13 
100 
[77.2, 100] 
As a reference, the range of the lineage initiate response rates seen in adult 
participants in HVTN 115, Part A was 30%-40%, varying by dose. The sample 
size of n = 13 yields greater than 90% power to detect a response if the true rate is 
30% or greater. 
Group 7 is not powered for a formal immunogenicity analysis but is instead 
intended to provide data for a cursory investigation of immunogenicity of a 
variant regimen with a lower protein dose: (5 mcg instead of 20 mcg of protein). 
After an expected loss of 25% of immunogenicity data, the sample sizes of n = 4 
for this group is sufficient to detect an extreme loss of immunogenicity due to the 
lower dose. If 0 of 4 participants in this group is a responder, the upper end of the 
95% confidence interval for the true response rate for that regimen is 49%. 
As shown in Table 6-4, there is low power for a formal comparison of 
immunogenicity response rates between the Group 3 and 5 pooled vaccine arms 
of size n = 13 versus a pooled placebo arm of size n = 8 after accounting for 
missingness. The sizes of differences that the trial is powered to detect are around 
70%. These calculations use a Fisher’s exact 2-sided test with a Type I error rate 
of 0.05. There is insufficient power for a formal comparison across the vaccine 
regimens or between Group 7 and the pooled placebo group. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 49 of 151 
Table 6-4 Power for comparison of response rates between 2 pooled arms (n1 = 8, n2 = 13) 
 
Minimum true response rate in Arm 2 in order to detect a 
difference 
True response rate Arm 1 
(%) 
80% power 
90% power 
0 
64 
73 
10 
79 
87 
20 
89 
95 
30 
96 
- 
6.1.3 
Considerations for comparison of immunogenicity to adults in 
HVTN 115 
To address Exploratory Objective 1 (comparison to adults), the analysis will 
descriptively summarize binding response positivity call rates, and test superiority 
of the frequency of differential memory B cell binding (to wild type vs mutant 
IΔ371 CH505) and of the magnitude and breadth of the IgG binding Ab response 
to a panel of gp120 proteins using a two-sided Wilcoxon rank sum test with 5% 
type-I error rate per comparison. This exploratory objective involves a 
comparison across populations (infants versus adults, in different demographic 
settings). Since this factor is necessarily non-randomizable, these comparisons are 
of the “observational” category, in the sense that they do not share the statistical 
benefits of the randomized controlled clinical trial design. Specifically, while the 
“study” factor differentiates adults from infants, these cohorts differ in many other 
respects. Therefore, the interpretation of any differences observed will require 
more careful consideration of the potential confounding effects of measured and 
unmeasured differences across the populations besides the age of the vaccine 
recipient. 
Immunogenicity in the HVTN 115 Part B vaccine recipients (n = 20 per group, n 
= 60 total), pooled with the corresponding dose group from Part A (n = 12) will 
be compared to immunogenicity in the HVTN 135 vaccine recipients. After an 
expected 15% of immunogenicity data missingness in HVTN 115 at the 
immunogenicity timepoint post 5th vaccination (at one year), sample sizes for 
comparisons across these study populations will be approximately n = 13 infants 
vs n = 27 (one HVTN 115 Part B arm, and one HVTN 115 Part A arm) or n = 61 
(all 3 Part B vaccine arms, and 1 Part A arm). Comparisons are well-powered 
under this scenario for both binary responses and for comparisons of continuous 
responses (eg measuring antibody binding magnitudes). If the true response rate 
in adults is 30%, there is 80% power to detect a difference vs infant response rates 
if the true rate in infants is 82% or greater when comparing to n = 27 adults, and 
there is 80% power to detect a difference vs n = 61 adults when the true rate in 
infants exceeds 77%. 
For continuous endpoints, there is 80% power to detect a difference across adult 
vs infant groups of 1.16 standard deviations (SDs) when considering n = 27 
adults, and 1.04 SDs when considering n = 61 adults. As a point of reference, 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 50 of 151 
using the mean and SD values from the HVTN 088 trial, the difference of 1.04 
SD translates to a difference of 0.38 units of log10 binding magnitude; if one of 
the comparison groups has the same mean as was seen among non-negative 
values among vaccine recipients in the HVTN 088 trial (4.05 units), then after 
incorporating the expected 6% zero-valued (negative responder) measures, this 
corresponds to observing an increase or decrease in the overall mean from 3.78 in 
group 1 to 4.16 in group 2. These calculations assume a 15% loss-to-follow-up 
rate in HVTN 115 and a 25% rate in HVTN 135 and the (94%) response rate 
observed in the HVTN 088 vaccine recipients. The same approach will be used to 
test superiority of the magnitude of the IgG binding Ab response to each 
individual gp120 antigen in the panel. 
6.2 
Randomization  
A participant’s randomization assignment will be computer generated and 
provided to the HVTN clinical research site (CRS) pharmacist through a Web-
based randomization system. At the institution, the pharmacist with primary 
responsibility for dispensing study products is charged with maintaining security 
of the treatment assignments (except in emergency situations as specified in the 
HVTN Manual of operations [MOP]). 
6.3 
Blinding 
Participants and site staff (except for site pharmacists) will be blinded as to 
participant treatment arm assignments (eg, vaccine or placebo). Study product 
assignments are accessible to those HVTN CRS pharmacists, DAIDS protocol 
pharmacists and contract monitors, and SDMC staff who are required to know 
this information to ensure proper trial conduct. Any discussion of study product 
assignment between pharmacy staff and any other HVTN CRS staff is prohibited. 
The HVTN SMB members also are unblinded to treatment assignment to conduct 
review of trial safety. 
When a participant leaves the trial prior to study completion, the participant will 
be told he or she must wait until all participants are unblinded to learn his or her 
treatment assignment. 
In some cases, the CRS, PSRT, or study sponsor may believe unblinding of the 
site PI and participant would be appropriate to facilitate the clinical management 
of an AE or SAE. The HVTN Unblinding MOP specifies procedures for 
emergency unblinding, and for early unblinding for medical reasons. 
6.4 
Statistical analyses 
This section describes the final study analyses, unblinded as to treatment arm 
assignment. All data from enrolled participants will be analyzed according to the 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 51 of 151 
initial randomization assignment regardless of how many vaccinations they 
received. In the rare instance that a participant receives the wrong treatment at a 
specific vaccination time, the Statistical Analysis Plan (SAP) will address how to 
analyze the participant’s safety data. Analyses are modified intent-to-treat in that 
individuals who are randomized but not enrolled do not contribute data and hence 
are excluded. Because of blinding and the brief length of time between 
randomization and enrollment—typically no more than 4 working days—very 
few such individuals are expected. 
Analyses for primary endpoints will be performed using SAS and R. All other 
descriptive and inferential statistical analyses will be performed using SAS, 
StatXact, or R statistical software. 
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 characteristics, safety, and 
immunogenicity for primary- and secondary-objective analyses. 
6.4.2 
Baseline comparability 
Treatment arms will be compared for baseline participant characteristics using 
descriptive statistics. 
6.4.3 
Safety/tolerability analysis 
Since enrollment of mother-infant pairs is concurrent with the infant receiving the 
first vaccination, all infants will have received at least 1 vaccination and therefore 
will provide some safety data. 
6.4.3.1 
Reactogenicity 
The number and percentage of participants experiencing each type of 
reactogenicity sign or symptom will be tabulated by severity and treatment arm 
and the percentages displayed graphically by arm. For a given sign or symptom, 
each participant’s reactogenicity will be counted once under the maximum 
severity for all injection visits. In addition, to the individual types of events, the 
maximum severity of local pain or tenderness, induration or erythema, and of 
systemic symptoms will be calculated. Kruskal-Wallis and Wilcoxon rank sum 
tests will be used to test for differences in severity between arms. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 52 of 151 
6.4.3.2 
AEs and SAEs 
AEs will be summarized using MedDRA System Organ Class and preferred 
terms. Tables will show by treatment arm the number and percentage of 
participants experiencing an AE within a System Organ Class or within preferred 
term category by severity or by relationship to study product. For the calculations 
in these tables, a participant with multiple AEs within a category will be counted 
once under the maximum severity or the strongest recorded causal relationship to 
study product. Formal statistical testing comparing arms is not planned since 
interpretation of differences must rely heavily upon clinical judgment. 
A listing of SAEs reported to the DAIDS Regulatory Support Center (RSC) 
Safety Office will provide details of the events including severity, relationship to 
study product, time between onset and last vaccination, and number of 
vaccinations received. 
6.4.3.3 
Local laboratory values 
Box plots of local laboratory values will be generated for baseline values and for 
values measured during the study by treatment arm and visit. Each box plot will 
show the first quartile, the median, and the third quartile. Outliers (values outside 
the box plot) will also be plotted. If appropriate, horizontal lines representing 
boundaries for abnormal values will be plotted. 
For each local laboratory measure, summary statistics will be presented by 
treatment arm and timepoint, as well as changes from baseline for postenrollment 
values. In addition, the number (percentage) of participants with local laboratory 
values recorded as meeting Grade 1 AE criteria or above as specified in the 
Division of AIDS Table for Grading the Severity of Adult and Pediatric Adverse 
Events (see Section 9.7) will be tabulated by treatment arm for each 
postvaccination timepoint. Reportable clinical laboratory abnormalities without an 
associated clinical diagnosis will also be included in the tabulation of AEs 
described above. 
6.4.3.4 
Reasons for vaccination discontinuation and early study termination 
The number and percentage of participants who discontinue vaccination and who 
terminate the study early will be tabulated by reason and treatment arm. 
6.4.4 
Immunogenicity analysis 
6.4.4.1 
General approach 
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 postinfection are excluded. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 53 of 151 
Since the exact date of HIV infection is unknown, any assay data from blood 
draws 4 weeks 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 postenrollment, 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% confidence interval 
estimates calculated using the score test method (75). Because of the small 
numbers of control participants in each group, no adjustment will be made to the 
vaccine arm estimates for the false positive rates in the control arms. Barnard or 
Fisher’s exact tests, as specified in the SAP, will be used to compare the response 
rates across treatment groups, with a significant difference declared if the 2-sided 
p-value is ≤ 0.05; however the primary analysis will prioritize the one-sample 
comparison of those response rates to zero due to the power considerations 
discussed above. In general Barnard’s is preferred since under most circumstances 
it is more powerful than Fisher’s (77). 
In addition to response rate estimates for each timepoint, the probability of 
observing at least 1 positive response by a given timepoint and the probability of 
observing more than 1 positive response by a given timepoint will be estimated, 
with corresponding confidence intervals, for each vaccine arm using maximum 
likelihood-based methods (78). 
For quantitative assay data (eg, magnitudes and magnitude-breadth AUCs from 
the neutralizing antibody multiplex assay), graphical and tabular summaries of the 
distributions by antigen, treatment arm, and timepoint will be made. For all 
primary and secondary immunogenicity endpoints, box plots and plots of 
estimated reverse cumulative distribution curves will be used for graphical display 
of all of the study arms. Typically, the results will be shown for each vaccine arm 
and for the set of control arms pooled into one group.  
Some immunologic assays have underlying continuous or count-type readout that 
are dichotomized into responder/nonresponder categories (eg, nAb multiplex 
assay positivity). If treatment arm differences for these assays are best 
summarized by a mixture model, then Lachenbruch’s test statistic (79) will be 
used to evaluate the composite null hypothesis of equal response rates in the 2 
arms and equal response distributions among responders in the 2 such arms. This 
test statistic equals the square of a binomial Z-statistic for comparing the response 
rates plus the square of a Wilcoxon statistic for comparing the response 
distributions in the subgroup of responders. A permutation procedure is used to 
obtain a 2-sided p-value. For estimation, differences in response rates between 
arms will be estimated using the methods described above, and in the subgroup of 
positive responders, differences in location parameters between arms will be 
estimated using the methods described above.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 54 of 151 
Based upon previous HVTN trials as well as experience conducting infant clinical 
trials in the study population, missing up to 25% of immunogenicity results for a 
specific assay is anticipated 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 standard methods applied in a complete-case 
manner (only including participants with observed data in the analysis), missing 
data need to be missing completely at random (MCAR). Following the most 
commonly used definition, MCAR assumes that the probability of an observation 
being missing does not depend on any participant characteristics (observed or 
unobserved). When missing data are minimal (specifically if no more than 20% of 
participants are missing any values), then standard complete-case methods will be 
used, because violations of the MCAR assumption will have little impact on the 
estimates and hypothesis tests. 
If a substantial amount of immunogenicity data is missing for an endpoint (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 (80) will be used, 
because they accommodate the censoring. In addition, secondary analyses of 
repeated immunogenicity measurements may be done using weighted GEE (81) 
methods, which are valid under MAR. All the models described above in this 
paragraph will include as covariates all available baseline predictors of the 
missing outcomes. 
Some “resource-intensive” immunogenicity endpoints are only measured in 
subset of participants, eg, immunogenicity endpoints based on mucosal samples. 
For such endpoints, exploratory analyses will be conducted to assess the 
correlation of participant characteristics measured in (nearly) all participants with 
the resource-intensive endpoints. For example, if the same assay is performed on 
blood and mucosal samples, then a scatterplot and Spearman rank correlation 
coefficient (r) will be used to assess the correlation of responses. If at least 
moderate correlations exist (eg, r ≥ 0.3), then the semiparametric efficient analysis 
method of Rotnitzky and Robins (82) will be used (described in Gilbert, Sato et 
al. for application to vaccine studies (83)) to estimate the mean of the resource-
intensive endpoint for each group and to compare means between groups. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 55 of 151 
6.4.4.2 
Multivariate display of immunogenicity endpoints 
Data visualization techniques may be used to explore the relationship among 
immunogenicity readouts. The set of readouts may be based on one of the primary 
endpoints on the set of primary endpoints, or on immunogenicity endpoints that 
also include secondary or exploratory endpoints. To understand the relationship 
between pairs of readouts, scatter plots may be used when the number of readouts 
is small or for a larger number of readouts, a heatmap showing the degree of 
correlation between any two pairs. Principal component analysis (PCA) and 
associated ‘biplots’ of the scores and loadings are particularly useful to 
understand associations between readouts, especially when readouts are correlated 
(84). PCA is a method to reduce the dimensionality of the number of readouts to a 
smaller set of values (principal components) that are normalized linear 
combinations of the readouts in such a way that the first principal component 
accounts for the most variability in the data and subsequent components, while 
maximizing variability, are uncorrelated with each other. A ‘biplot’ displays the 
first and second principal component scores and principal component loadings. 
The x-axis is the value from the first principal component and the y-axis is the 
second principal component, where each axis label includes the percentage of 
variation in the total set of readouts captured by the principal component. The top 
axis is the first principal component loadings and the right axis is the second 
principal component loadings. An arrow is drawn for each immunogenicity 
readout (eg, antibody binding magnitudes) from the origin to the point defined by 
its first two principal component loadings. The length of the arrow represents the 
amount of total variation of the set of readouts captured by the given readout. The 
direction of an arrow conveys the extent to which the variation of a readout is in 
the direction of the first or second principal component. The angle between two 
arrows conveys information about the correlation of the two readouts, with a zero-
degree angle denoting perfect correlation and a 90-degree angle denoting no 
correlation. Each arrow on the biplot is labeled by the immunogenicity readout it 
represents. A biplot is annotated with key meta-information such as the treatment 
arm (most common application) or a demographic category. Depending on the 
application, K-means clustering and hierarchical clustering may also be applied 
for multivariate graphical display of immunogenicity readouts. 
6.4.4.3 
Analysis of multiplexed immunoassay data 
When a small panel of analytes (eg, ≤ 5) is being assessed in a multiplexed 
immunoassay, the analysis of response rates and response magnitudes will be 
evaluated and compared as described under the general approach. Details for 
calculating a positive response and response magnitude will be provided in the 
SAP. When a larger panel is being assessed, two approaches may be considered to 
evaluate the magnitude and breadth of these responses. First, Magnitude–Breadth 
(M-B) curves maybe employed to display individual- and group-level response 
breadth as a function of magnitude. Response breadth of neutralizing antibodies is 
defined as the average of the log10 nAb titer over the panel of isolates, where 
titers that are below the limit-of-detection are set to half of that limit. Response 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 56 of 151 
breadth of binding antibodies is similarly defined as the average of the response 
over the panel of antigens. Details of the approach will be described in the SAP. 
6.4.5 
Analyses and data sharing 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 unblinded analyses 
by treatment assignment require careful consideration and should be made 
available on a need to know basis in accordance with Sections 6.4.5.1 and 6.4.5.2. 
Interim blinded safety and immunogenicity data should not be shared outside of 
the SMB, PSRT, the protocol team leadership, the HVTN Executive Management 
Team, the study product developer, and the study sponsor and/or its designee(s) 
for their regulatory reporting unless approved by the protocol leadership and the 
HVTN leadership. 
6.4.5.1 
Safety analyses 
During the course of the trial, unblinded analyses of safety data will be prepared 
approximately every 4 months for review by the SMB. Ad hoc safety reports may 
also be prepared for SMB review at the request of the PSRT. Refer to the process 
described in the HVTN Unblinding MOP for any requests for unblinded safety 
data prior to the end of the scheduled follow-up visits. 
6.4.5.2 
Immunogenicity analyses 
An unblinded statistical analysis by treatment assignment of a primary 
immunogenicity endpoint may be performed when all participants within any 
study Part (A, B, or C) have completed the corresponding primary 
immunogenicity visits and data are available for analysis from at least 80% of 
these participants. Similarly, an unblinded statistical analysis by treatment 
assignment of a secondary or exploratory immunogenicity endpoint may be 
performed when all participants within any study Part have completed the 
corresponding immunogenicity visit and data are available for analysis from at 
least 80% of these participants. The Laboratory Program will review the analysis 
report prior to distribution to the protocol chairs, DAIDS, study product 
developer, and other key HVTN members and investigators. Reports for 
distribution or presentation should use PubIDs and not PTIDs for individual 
responses. Distribution of reports will be limited to those with a need to know for 
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. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 57 of 151 
7 
Selection and withdrawal of participants 
This study will be conducted among 38 healthy infants born to HIV-infected 
mothers in South Africa. Infants will be enrolled in pairs with their mothers, 
although only infants will receive study product. Mothers will be HIV-infected 
(seropositive) adults on cART (combination anti-retroviral therapy) who are 
clinically stable and who comprehend the purpose of the study and have provided 
written informed consent for their and their infant’s participation in the study. 
Mother-infant pairs will be selected for the study according to the criteria in 
Sections 7.1, 7.2, 7.3, and 7.4. Final eligibility determination will depend on 
information available at the time of enrollment, including results of screening 
laboratory tests, medical history, physical examinations, and answers to self-
administered and/or interview questions. 
Investigators should always use good clinical judgment in considering a mother-
infant pair’s overall fitness for trial participation. Some mother-infant pairs 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 mother-infant 
pairs may be poor candidates for retention. 
Due to the logistical requirements that require identification of mothers during 
pregnancy but an inability to fully assess inclusion criteria for infants until birth 
the screening periods will be different for the infants and their mothers. All 
inclusion and exclusion criteria for the infant must be within 5 days prior to 
enrollment. To allow for identification of women during pregnancy, the screening 
time-frame will be different. All inclusion and exclusion criteria for the mother 
must be assessed within 273 days (9 months) of enrollment, unless otherwise 
specified in the eligibility criteria below.  
7.1 
Inclusion criteria: Mother 
General and Demographic Criteria 
1. Mother’s age is at least 18 years, and willing and able to provide written 
informed consent for her and her infant’s participation in this study. 
2. Mother is in the second or third trimester of singleton pregnancy, as 
determined by a clinical exam, or sonography and reported menstrual history. 
3. Mother agrees to donate umbilical cord blood. 
4. Mother has a planned Caesarian Section at Chris Hani Baragwanath Academic 
Hospital, Soweto and plans to remain in the area after delivery. 
5. Mother is determined by the site investigator to be in good overall health at 
the time of delivery based on medical history, and physical exam. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 58 of 151 
6. Mother has a documented CD4 count > 350 cells/microliter during her 
pregnancy. 
7. Mother has documented SARS CoV-2 negative PCR test within 2 days before 
delivery to 5 days after delivery  
8. Mother has access to the participating HVTN CRS and willingness to be 
followed for the planned duration of the study. 
9. Assessment of understanding: Mother demonstrates understanding of this study; 
completes a questionnaire prior to delivery with verbal demonstration of 
understanding of all questionnaire items answered incorrectly. 
10. Mother agrees not to enroll either herself or her infant in another research 
study for the duration of the trial without prior approval of the HVTN 135 PSRT. 
11. Mother has confirmed HIV-1 infection documented by medical records at any 
time during or prior to screening, and confirmed by the HVTN CRS by serology. 
12. Mother has been on cART for at least sixteen weeks prior to delivery and 
intends to continue with cART for the duration of breastfeeding. 
13.  Mother has a viral load of less than 400 copies/mL between two weeks before 
and 5 days after delivery. 
7.2 
Exclusion criteria: Mother 
1. Any WHO Grade IV illness within one year prior to study enrollment as 
determined by the history and physical examination and review of the medical 
record (if available). These include HIV wasting syndrome, PJP Pneumonia, 
Cerebral Toxoplasmosis, extrapulmonary Cryptococcosis, Progressive Multifocal 
Leukoencephalopathy, any disseminated endemic mycosis (histoplasmosis), 
candidiasis of the esophagus, trachea, bronchi or lung, disseminated atypical 
mycobacteria, non-typhoid Salmonella septicemia, extrapulmonary tuberculosis, 
lymphoma, Kaposi’s sarcoma. 
2. Prior participation in any HIV-1 vaccine or anti-HIV antibody-mediated 
prevention trial. 
3. Receipt of any investigational agent during this pregnancy. 
4. Receipt of blood products, immunoglobulin, or immunomodulating therapy 
within 45 days prior to delivery of the placenta. 
5. Any medical, psychiatric, occupational, or other condition that, in the 
judgment of the investigator, would interfere with, or serve as a contraindication 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 59 of 151 
to, protocol adherence, assessment of safety or reactogenicity, or a volunteer’s 
ability to give informed consent. 
6. Any condition that places the newborn at higher risk of early-onset sepsis, 
such as concern for active maternal infection at delivery as determined by local 
site investigators (eg, fever). 
7. Detectable Hepatitis B surface antigen. 
7.3 
Inclusion criteria: Infant 
1. Born via Caesarean section to an HIV-1–infected woman who meets all 
maternal inclusion/exclusion criteria listed above. 
2. Estimated gestational age at birth is at least 37 weeks. 
Note: If gestational age at birth is not documented in the infant’s available birth 
records, study staff may assess gestational age at the earliest possible opportunity 
during the screening period and use this assessment for purposes of eligibility 
determination. 
3. Weight at birth is at least 2.5 kg. 
4. Has initiated antiretroviral prophylaxis consistent with current site-specific 
standard of care. 
5. Hemoglobin >14.0 g/dL. 
6. White Blood Cell Count ≥ 7000 cells/mm3 
7. Platelets > 100,000 cells/mm3 
8. Alanine aminotransferase (ALT) <1.25 times upper limit of age adjusted 
normal. 
9. Creatinine < 1.1 times upper limit of age adjusted normal. 
10. Negative HIV-1 nucleic acid test on specimen drawn within 72 hours of birth.  
11. Written informed consent provided by mother. 
12. Age is equal to or less than five days. 
7.4 
Exclusion criteria: Infant 
1. Any clinically significant congenital anomaly/birth defect. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 60 of 151 
2. Documented or suspected serious medical illness, infection, clinically 
significant finding from physical examination or immediate life-threatening 
condition, including requirement for ongoing supplemental oxygen, as judged by 
the examining clinician. 
3. Receipt of or anticipated need for blood products, immunoglobulin, or 
immunosuppressive therapy. This includes infants who require Hepatitis B 
Immunoglobulin (HBIG) but does not require exclusion of infants who receive 
Hepatitis B vaccine in the newborn period. 
4. Receipt of any other investigational product. 
7.5 
Participant departure from vaccination schedule or withdrawal 
This section concerns an individual participant’s departure from the study product 
administration schedule. Pause rules for the trial are described in Section 11.4. 
7.5.1 
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: 
• Within 45 days prior to any study injection 
▪ Receipt of blood products or immunoglobulin 
• Prevaccination abnormal vital signs or clinical symptoms that may mask 
assessment of vaccine reaction 
• Child becomes moderately or severely underweight as defined by the DAIDS 
AE Grading Table for Underweight < 2 years of age. 
• If an infant has delayed EPI immunizations at the time of a scheduled study 
vaccine visit, the EPI immunizations will be provided to the infant at least 7 
days prior to administration of the study vaccination. The only exception is 
the initial vaccination. 
Vaccinations should not be administered outside the visit window, as specified in 
Appendix I. 
7.5.2 
Participant departure from vaccination schedule 
Every effort should be made to follow the vaccination schedule per the protocol. 
If an infant misses a vaccination and the visit window period for the vaccination 
has passed, that vaccination cannot be given. The mother-infant pair should be 
asked to continue study visits. The infant should resume the vaccination schedule 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 61 of 151 
with the next vaccination unless there are circumstances that require further delay 
or permanent discontinuation of vaccination (see Sections 7.5.3 and 7.5.4). 
7.5.3 
Discontinuing vaccination for a participant 
Under certain circumstances, an individual infant’s vaccinations will be 
permanently discontinued. Specific events that will result in stopping an 
individual’s vaccination schedule include: 
• Co-enrollment 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 PSRT) 
• Clinically significant condition (a condition that affects the immune system or 
for which continued vaccinations and/or blood draws may pose additional 
risk), including but not limited to the following: 
▪ HIV infection 
▪ Any grade 4 local or systemic reactogenicity symptom, lab abnormality, or 
AE that is subsequently considered to be related to vaccination 
▪ Any grade 3 lab abnormality that is subsequently considered to be related 
to vaccination 
▪ Other grade 3 clinical AE that is subsequently considered to be related to 
vaccination except for fever (temporal artery temperature), vomiting, and 
subjective local and systemic symptoms. For grade 3 injection site 
erythema and/or induration, upon review, the PSRT may allow 
continuation of vaccination 
▪ SAE that is subsequently considered to be related to vaccination 
▪ Any type 1 hypersensitivity reaction considered to be related to study 
vaccination.  
• Investigator determination in consultation with Protocol Team leadership (eg, 
for repeated nonadherence to study staff instructions). 
The mothers of infants discontinuing study product for reasons other than HIV 
infection should be counseled on the importance of continuing with the study and 
strongly encouraged to participate in follow-up visits and protocol-related 
procedures per the protocol for the remainder of the trial, unless medically 
contraindicated (see HVTN 135 Study Specific procedures [SSP]). 
Infants diagnosed with HIV infection during the study should be encouraged to 
participate in follow-up visits for sole purposes of safety monitoring as indicated 
in Section 9.10. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 62 of 151 
7.5.4 
Participant termination from the study 
Under certain circumstances, an individual mother-infant pair may be terminated 
from participation in this study. Specific events that will result in early 
termination include: 
• Infant’s mother/guardian voluntarily withdraws consent to participate, 
• Mother-infant pair relocates and remote follow-up is not possible, 
• HVTN CRS determines that the mother-infant pair is lost to follow-up, or 
• Investigator decides, in consultation with Protocol Team leadership, to 
terminate participation (eg, if infant’s mother exhibits inappropriate behavior 
toward clinic staff). 
• Any condition where termination from the study is required by applicable 
regulations. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 63 of 151 
8 
Study product preparation and administration 
CRS pharmacists should consult the Pharmacy Guidelines and Instructions for 
DAIDS Clinical Trials Networks for standard pharmacy operations. The protocol 
schema is shown in Table 3-1. See the Investigator’s Brochures (IBs) for further 
information about study products. 
8.1 
Vaccine regimen 
The schedule of vaccination is shown in Section 3 and additional information is 
given below. 
Part A 
Group 1 
Treatment 1 (T1): 20 mcg Stable CH505TF gp120 admixed with 2.5 mcg GLA-
SE, to be administered as a 0.25 mL intramuscular (IM) injection into either thigh 
at Weeks 0, 8, 16, 32, and 54. 
Group 2 
Control 2 (C2): Placebo for 20 mcg Stable CH505TF gp120 + 2.5 mcg GLA-SE 
(Sodium Chloride for Injection, 0.9%) to be administered as a 0.25 mL IM 
injection, into either thigh at Weeks 0, 8, 16, 32, and 54. 
Part B 
Group 3  
Treatment 3 (T3): 20 mcg Stable CH505TF gp120 admixed with 5 mcg GLA-
SE, to be administered as a 0.5 mL IM injection into either thigh at Weeks 0, 8, 
16, 32, and 54. 
Group 4  
Control 4 (C4): Placebo for 20 mcg Stable CH505TF gp120 + 5 mcg GLA-SE 
(Sodium Chloride for Injection, 0.9%) to be administered as a 0.5 mL IM 
injection, into either thigh at Weeks 0, 8, 16, 32, and 54. 
Part C 
Group 5  
Treatment 5 (T5): 20 mcg Stable CH505TF gp120 admixed with 5 mcg GLA-
SE, to be administered as a 0.5 mL IM injection into either thigh at Weeks 0, 8, 
16, 32, and 54. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 64 of 151 
Group 6  
Control 6 (C6): Placebo for 20 mcg Stable CH505TFgp120 + 5 mcg GLA-SE 
(Sodium Chloride for Injection, 0.9%) to be administered as a 0.5 mL IM 
injection, into either thigh at Weeks 0, 8, 16, 32, and 54. 
Group 7  
Treatment 7 (T7): 5 mcg Stable CH505TF gp120 admixed with 5 mcg GLA-SE, 
to be administered as a 0.5 mL IM injection into either thigh at Weeks 0, 8, 16, 
32, and 54. 
Group 8  
Control 8 (C8): Placebo for 5 mcg Stable CH505TF gp120 + 5 mcg GLA-SE 
(Sodium Chloride for Injection, 0.9%) to be administered as a 0.5 mL IM 
injection, into either thigh at Weeks 0, 8, 16, 32, and 54. 
8.2 
Study product formulation 
8.2.1 
Stable CH505TF gp120; labeled as CH505TF gp120 
The Stable CH505TF gp120 will be provided at a concentration of 0.8 mg/mL of 
protein in phosphate-buffered saline per vial. Each sterile, single use vial contains 
0.75 mL of study product. The study product should be stored frozen at ≤ -65°C. 
When thawed, the Stable CH505TF gp120 will be clear, and colorless to slightly 
yellow, liquid. The study product is described in further detail in the IB. 
8.2.2 
GLA-SE (Glucopyranosyl Lipid A-Stable Emulsion); labeled as AP 10-
201 
The GLA-SE adjuvant will be provided at a concentration of 20 mcg/mL GLA in 
a 4% oil-in-water emulsion in a vial. Each sterile, single use vial contains 0.6 mL 
of this mixture, which appears as a milky-white liquid. GLA-SE must be stored at 
2-8°C and must not be frozen. The study product is described in further detail in 
the IB. 
8.2.3 
Placebo for Stable CH505TF gp120 + GLA-SE 
Sodium Chloride for Injection, 0.9% will be used as the placebo. It must be stored 
in accordance with manufacturer’s recommendation. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 65 of 151 
8.3 
Preparation of study products 
8.3.1 
Group 1 (T1): 20 mcg Stable CH505TF gp120 + 2.5 mcg GLA-SE 
One vial of Stable CH505TF gp120 (0.8 mg/mL) and one vial of GLA-SE (20 
mcg/mL) will be needed to prepare the dose.  
Prior to admixture, the pharmacist will remove one vial of Stable CH505TF 
gp120 from the freezer and allow to thaw completely at room temperature. Once 
thawed completely, invert the vial 10 times to ensure a homogeneous product. 
Remove one vial of GLA-SE from the refrigerator and allow to equilibrate to 
room temperature.  
Using aseptic technique, the pharmacist will add 0.2 mL of Stable CH505TF 
gp120 and 0.8 mL of Sodium Chloride for injection, 0.9% to an empty sterile vial. 
Mix the contents of this vial thoroughly using a vortex machine at high speed for 
3 seconds. The final concentration of Stable CH505TF gp120 is 160 mcg/mL.  
Next, using aseptic technique, withdraw 0.4 mL of the diluted 160 mcg/mL Stable 
CH505TF gp120 admixture and 0.4 mL of GLA-SE (20 mcg/mL) and inject it 
into an empty sterile vial. Mix thoroughly by gently inverting 10 times, yielding a 
concentration of 80 mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE.  
Finally, using aseptic technique, withdraw 0.25 mL from the vial containing 80 
mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE, using a 1 mL size 
syringe. Remove the needle for preparation and attach the needle for 
administration to the prepared study product in syringe before dispensing.  
The prepared syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into the thigh, with an expiration date and 
time of 8 hours following the last mixing procedure. 
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.2 
Group 2 (C2): Placebo for 20 mcg Stable CH505TF gp120 + 2.5 mcg 
GLA-SE 
The pharmacist using aseptic technique will withdraw 0.25 mL of Sodium 
Chloride for Injection, 0.9% into a 1 mL size syringe, remove the needle for 
preparation and attach the needle for administration to the prepared study product 
in syringe before dispensing.  
The final syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into thigh, with an expiration date and time 
of 8 hours following the last mixing procedure.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 66 of 151 
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.3 
Group 3 (T3): 20 mcg Stable CH505TF gp120 + 5 mcg GLA-SE 
One vial of Stable CH505TF gp120 (0.8 mg/mL) and one vial of GLA-SE (20 
mcg/mL) will be needed to prepare the dose.  
Prior to admixture, the pharmacist will remove one vial of Stable CH505TF 
gp120 from the freezer and allow to thaw completely at room temperature. Once 
thawed completely, invert the vial 10 times to ensure a homogeneous product. 
Remove one vial of GLA-SE from the refrigerator and allow to equilibrate to 
room temperature.  
Using aseptic technique, the pharmacist will add 0.1 mL of Stable CH505TF 
gp120 and 0.9 mL of Sodium Chloride for Injection, 0.9% to an empty sterile vial. 
Mix the contents of this vial thoroughly using a vortex machine at high speed for 
3 seconds. The final concentration of Stable CH505TF gp120 is 80 mcg/mL.  
Next, using aseptic technique, withdraw 0.4 mL of the diluted 80 mcg/ml Stable 
CH505TF gp120 admixture  and 0.4 mL of GLA-SE (20 mcg/mL) and inject it 
into an empty sterile vial.  Mix thoroughly by gently inverting 10 times, yielding a 
concentration of 40 mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE.  
Finally, using aseptic technique, withdraw 0.5 mL from the vial containing 40 
mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE, using a 1 mL size 
syringe. Remove the needle for preparation and attach the needle for 
administration to the prepared study product in syringe before dispensing.  
The prepared syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into the thigh, with an expiration date and 
time of 8 hours following the last mixing procedure. 
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.4 
Group 4 (C4): Placebo for 20 mcg Stable CH505TF gp120 + 5 mcg 
GLA-SE 
The pharmacist using aseptic technique will withdraw 0.5 mL of Sodium Chloride 
for Injection, 0.9%  into a 1 mL size syringe, remove the needle for preparation 
and attach the needle for administration to the prepared study product in syringe 
before dispensing.  
The final syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 67 of 151 
also be labeled for IM administration into the thigh, with an expiration date and 
time of 8 hours following the last mixing procedure.  
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.5 
Group 5 (T5): 20 mcg Stable CH505TF gp120 + 5 mcg GLA-SE 
One vial of Stable CH505TF gp120 (0.8 mg/mL) and one vial of GLA-SE (20 
mcg/mL) will be needed to prepare the dose.  
Prior to admixture, the pharmacist will remove a vial of Stable CH505TF gp120 
from the freezer and allow to thaw completely at room temperature. Once thawed 
completely, invert the vial 10 times to ensure a homogeneous product. Remove 
one vial of GLA-SE from the refrigerator and allow to equilibrate to room 
temperature.  
Using aseptic technique, the pharmacist will add 0.1 mL of Stable CH505TF 
gp120 and 0.9 mL of Sodium Chloride for Injection, 0.9% to an empty sterile vial. 
Mix the contents of this vial thoroughly using a vortex machine at high speed for 
3 seconds. The final concentration of Stable CH505TF gp120 is 80 mcg/mL.  
Next, using aseptic technique, withdraw 0.4 mL of the diluted 80 mcg/mL Stable 
CH505TF gp120 admixture and 0.4 mL of GLA-SE and inject it into an empty 
sterile vial. Mix thoroughly by gently inverting 10 times, yielding a concentration 
of 40 mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE.  
Finally, using aseptic technique, withdraw 0.5 mL from the vial containing 40 
mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE, using a 1 mL size 
syringe. Remove the needle for preparation and attach the needle for 
administration to the prepared study product in syringe before dispensing.  
The prepared syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into the thigh, with an expiration date and 
time of 8 hours following the last mixing procedure. 
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.6 
Group 6 (C6): Placebo for 20 mcg Stable CH505TF gp120 + 5 mcg 
GLA-SE 
The pharmacist using aseptic technique will withdraw 0.5 mL of Sodium Chloride 
for Injection, 0.9% into a 1 mL size syringe, remove the needle for preparation 
and attach the needle for administration to the prepared study product in syringe 
before dispensing.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 68 of 151 
The final syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into thigh, with an expiration date and time 
of 8 hours following the last mixing procedure.  
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.7 
Group 7 (T7): 5 mcg Stable CH505TF gp120 + 5 mcg GLA-SE 
One vial of Stable CH505TF gp120 (0.8 mg/mL) and one vial of GLA-SE (20 
mcg/mL) will be needed to prepare the dose.  
Prior to admixture, the pharmacist will remove one vial of Stable CH505TF 
gp120 from the freezer and allow to thaw completely at room temperature. Once 
thawed completely, invert the vial 10 times to ensure a homogeneous product. 
Remove one vial of GLA-SE from the refrigerator and allow to equilibrate to 
room temperature.  
Using aseptic technique, the pharmacist will add 0.1 mL of Stable CH505TF 
gp120 and 3.9 mL of Sodium Chloride for Injection, 0.9% to an empty sterile vial. 
Mix the contents of this vial thoroughly using a vortex machine at high speed for 
3 seconds. The final concentration of Stable CH505TF gp120 is 20 mcg/mL.  
Next, using aseptic technique, withdraw 0.4 mL of the diluted 20 mcg/mL Stable 
CH505TF gp120 admixture and 0.4 mL of GLA-SE (20 mcg/mL) and inject it 
into an empty sterile vial.  Mix thoroughly by gently inverting 10 times, yielding a 
concentration of 10 mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE.  
Finally, using aseptic technique, withdraw 0.5 mL from the vial containing 10 
mcg/mL Stable CH505TF gp120 and 10 mcg/mL GLA-SE, using a 1 mL size 
syringe. Remove the needle for preparation and attach the needle for 
administration to the prepared study product in syringe before dispensing.  
The prepared syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into the thigh, with an expiration date and 
time of 8 hours following the last mixing procedure. 
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.3.8 
Group 8 (C8): Placebo for 5 mcg Stable CH505TF gp120 + 5 mcg 
GLA-SE 
The pharmacist using aseptic technique will withdraw 0.5 mL of Sodium Chloride 
for Injection, 0.9%  into a 1 mL size syringe, remove the needle for preparation 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 69 of 151 
and attach the needle for administration to the prepared study product in syringe 
before dispensing.  
The final syringe for administration must be covered with an overlay and then 
labeled as “Stable CH505TF gp120 + GLA-SE or Placebo”. The syringe must 
also be labeled for IM administration into thigh, with an expiration date and time 
of 8 hours following the last mixing procedure.  
Any unused portion of vials or expired prefilled syringes should be disposed of in 
accordance with institutional or pharmacy policy. 
8.4 
Administration 
All injections should be administered IM in the thigh of either leg as listed in 
Section 8.1.  
The prepared study product in the syringe should be rolled gently prior to 
administration.  
The prepared study product in the syringe is to be stored between 4o C and 25o C 
and must be administered as soon as possible and before the 8-hour expiration.  
Any administrator of study product will be blinded to the individual participant's 
treatment assignment.  
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. 
8.5 
Acquisition of study products 
Stable CH505TF gp120 is manufactured by Berkshire Sterile Manufacturing and 
GLA-SE is manufactured by IDRI. Stable CH505TF gp120 and GLA-SE will be 
provided by DAIDS. Sodium Chloride for Injection, 0.9%  will not be provided 
through the protocol and must be obtained by the site.  
Once an HVTN CRS is protocol registered, the pharmacist can obtain study 
products from the NIAID Clinical Research Products Management Center 
(CRPMC) by following the ordering procedures outlined in Pharmacy Guidelines 
and Instructions for DAIDS Clinical Trials Networks. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 70 of 151 
8.6 
Pharmacy records 
The HVTN CRS pharmacist is required to maintain complete records of all study 
products. The pharmacist must also record the lot # and manufacturer of the 
Sodium Chloride for Injection, 0.9%  used to prepare the participant’s study 
product injection. The pharmacist of record is responsible for maintaining 
randomization codes and randomization confirmation notices for each participant 
in a secure manner. 
8.7 
Final disposition of study products 
For non-US clinical research sites, 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 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 71 of 151 
9 
Clinical procedures 
The schedule of clinical procedures is shown in Appendix F and Appendix G. 
9.1 
Informed consent 
Informed consent is the process of working with mothers so that they fully 
understand what will and may happen to them and their infant while participating 
in this research study. The HVTN informed consent form documents that the 
mother (1) has been informed about the potential risks, benefits, and alternatives 
to participation, and (2) is willing for her infant and herself to participate in this 
study. Informed consent encompasses all written or verbal study information 
HVTN CRS staff provide, before and during the trial. HVTN CRS staff will 
obtain informed consent according to HVTN policies and procedures. 
The informed consent process continues throughout the study. Key study concepts 
should be reviewed periodically with the participant and the review should be 
documented. At each study visit, HVTN CRS staff should consider reviewing the 
procedures and requirements for that visit and for the remaining visits. 
Additionally, if any new information is learned that might affect the 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 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. 
Note: As defined in the DAIDS Protocol Registration Manual, an RE is “Any 
group other than the local IRB/EC responsible for reviewing and/or approving a 
clinical research protocol and site-specific ICFs [informed consent forms] prior to 
implementation at a site.” CRSs are responsible for knowing the requirements of 
their applicable REs. 
Mothers will provide written informed consent for their own and their infant’s 
study participation before any study-specific procedures are performed. The 
informed consent process will be conducted during pregnancy. The study will be 
discussed again with the mother after delivery and her prior consent decision will 
be confirmed at that time. If the mother changes her mind and withdraws her 
consent at that time, the mother-infant pair will not be enrolled in the study and no 
further study-specific procedures will be performed. 
Due to the need to draw samples prior to enrollment (with the first vaccination), 
the informed consent will specifically address the need to collect biospecimens 
including peripheral and cord blood prior to full enrollment. The protocol-specific 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 72 of 151 
consent process will specify that if the mother-infant pair is never enrolled in the 
study, any biospecimens will be destroyed.  
Should the consenting mother of an enrolled infant die or no longer be available 
for any reason, no further study-specific visits or procedures may be performed 
until informed consent for continued study participation is obtained from a 
guardian. In accordance with the DAIDS policy on Enrolling Children (including 
Adolescents) in Clinical Research (available at the website referenced in Section 
14), the CRS must establish and maintain written procedures describing the 
standards that will be followed to identify who may serve as guardian for an 
enrolled infant, reflective of applicable IRB/EC guidance for conduct of human 
subjects research within the context of available local law, regulation, or 
government policy. 
The consent forms describe the study products to be used and all aspects of 
protocol participation, including screening and enrollment procedures. A sample 
consent form for the main study is located in Appendix A. A separate sample 
consent form for other uses of specimens is located in Appendix B. 
The HVTN CRS is responsible for developing a protocol-specific consent form(s) 
for local use, based on the sample protocol-specific consent forms in Appendix A 
and Appendix B. The consent form(s) must be developed in accordance with 
requirements of the following: 
• CRS’s IRB/EC and any applicable REs, 
• CRS’s institution, and 
• Elements of informed consent as described in Title 45, CFR Part 46 and Title 
21 CFR, Part 50, and in ICH E6, Good Clinical Practice: Consolidated 
Guidance 4.8. 
The study site is strongly encouraged to have their local CABs review their site-
specific consent forms. This review should include, but should not be limited to, 
issues of cultural competence, local language considerations, and the level of 
understandability. 
The sample informed consent form(s) include instructions for developing specific 
content. Regarding protocol registration, the site should follow procedures 
outlined in the current version of the DAIDS Protocol Registration Manual. 
9.1.1 
Assessment of Understanding 
Study staff are responsible for ensuring that the mother fully understands the 
study requirements for both her and her infant before enrollment. This process 
involves reviewing the informed consent form, allowing time to reflect on the 
procedures and issues presented, and answering all questions completely. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 73 of 151 
An Assessment of Understanding is used to document the mother’s understanding 
of key concepts in this HIV vaccine trial. The mother must complete the 
Assessment of Understanding before enrollment. Staff may provide assistance in 
reading and understanding the questions and responses, if necessary. The mother 
must verbalize understanding of all questions answered incorrectly. This process 
and the participant’s understanding of the key concepts should be recorded in 
source documentation at the site. 
IRB/EC and any applicable RE may require that a consent document be signed 
prior to administering the Assessment of Understanding. The consent process 
(including the use of the Assessment of Understanding) should be explained 
thoroughly to the IRB/EC and any applicable RE, whose recommendations should 
be followed.  
9.2 
Pre-enrollment procedures 
Screening may occur over the course of several contacts/visits, up to and 
including before vaccination on day 0. All inclusion and exclusion criteria for the 
infant must be assessed within 5 days prior to enrollment. To allow for 
identification of women during pregnancy, the screening time-frame will be 
different for the mothers. All inclusion and exclusion criteria for the mother 
must be assessed within 273 days (9 months) of enrollment, unless otherwise 
specified in the eligibility criteria (or below in this section). Enrollment for the 
mother-infant pair is simultaneous with the infant’s first vaccination 
9.2.1 
Pre-enrollment procedures, Mother 
After the appropriate informed consent has been obtained and before enrollment, 
the following procedures are performed: 
• Medical history, documented in the case history record 
• Complete physical examination, including 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 
• Assessment of concomitant medications the mother is taking, including 
antiretroviral regimen, prescription and nonprescription drugs, vitamins, 
topical products, alternative/complementary medicines (eg, herbal and health 
food supplements), recreational drugs, vaccinations, and allergy shots 
• Risk reduction counseling 
• Assessment of Understanding 
• Specimen collection 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 74 of 151 
• Laboratory tests, including: 
▪ CD4 count, if not available from medical records 
▪ SARS-CoV-2 PCR test, if not available from medical records 
▪ Hepatitis B surface antigen (HBsAg), if not available from medical 
records during pregnancy 
▪ HIV enzyme immunoassay (EIA) test 
▪ HIV polymerase chain reaction (PCR) Viral Load 
• Obtaining of volunteer demographics in compliance with the NIH Policy on 
Reporting Race and Ethnicity Data: Subjects in Clinical Research, Aug. 8, 
2001 (available at http://grants.nih.gov/grants/guide/notice-files/NOT-OD-01-
053.html) 
9.2.2 
Pre-enrollment procedures, Infant 
After the appropriate informed consent has been obtained and before enrollment, 
the following procedures are performed: 
• Medical history  
• Complete physical examination, see HVTN 135 SSP for details 
• Assessment of concomitant medications the infant is taking, including 
prescription and nonprescription drugs, antiretroviral prophylaxis regimen, 
vitamins, topical products, alternative/complementary medicines (eg, herbal 
and health food supplements), vaccinations 
• Specimen collection 
• Laboratory tests, including: 
▪ Complete blood count (CBC)  
▪ Chemistry panel (ALT, Creatinine) 
▪ Screening HIV Test 
• Obtaining of volunteer demographics in compliance with the NIH Policy on 
Reporting Race and Ethnicity Data: Subjects in Clinical Research, Aug. 8, 
2001 (available at http://grants.nih.gov/grants/guide/notice-files/NOT-OD-01-
053.html) 
9.3 
Enrollment and vaccination visits, Infant 
Once the mother has provided written informed consent for their own and their 
infant’s study participation and the mother-infant pair has been found to meet all 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 75 of 151 
eligibility criteria (see Sections 7.1, 7.2, 7.3, and 7.4), the HVTN CRS requests 
the randomization assignment via a Web-based randomization system. Enrollment 
for the mother-infant pair is simultaneous with the infant’s first vaccination and 
must be done within five days of life. In general, the time interval between 
randomization and enrollment should not exceed 4 working days.  
At all vaccination visits, the following procedures are performed on the infant 
before vaccination: 
• Abbreviated physical examination, including weight, vital signs, and a 
symptom-directed evaluation by history and/or appropriate physical exam 
based on the mother’s reported symptoms for her infant (see HVTN 135 SSP); 
• Feeding History; 
• Assessment of baseline reactogenicity parameters; 
• Assessment of concomitant medications (as described in Section 9.2); 
• Assessment of any new or unresolved AEs/intercurrent illnesses; and 
Following completion of all procedures in the preceding list, and if results 
indicate that vaccination may proceed, vaccination is prepared and administered 
(see Sections 8.3 and 8.4). 
Immediately following vaccination, the participant remains in the clinic for 
observation and ongoing monitoring for a minimum of 60 minutes. An initial 
reactogenicity assessment is made at a target of 30 minutes after injection, with an 
acceptable range of 25-60 minutes. Before leaving the clinic, the mother is given 
the Participant Diary and is instructed on how to complete it. The site will make 
arrangements to be in contact with the mother during the reactogenicity period (as 
described in Section 9.7). 
The following procedures will be performed as specified in Appendix D:  
• Blood collection performed prior to vaccination 
• Stool specimen collection, optional, performed prior to or following 
vaccination 
9.4 
Follow-up visits, Infants 
The following procedures are performed at all scheduled follow-up visits: 
• Assessment of new or continuing concomitant medications (as described in 
Section 9.2) 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 76 of 151 
• Assessment of new or unresolved AEs/intercurrent illnesses. 
• Blood collection  
• Feeding History 
Additional procedures will be performed at scheduled follow-up visits as 
specified in Appendix D and Appendix F: 
• Laboratory tests, including: 
▪ Complete blood count (CBC)  
▪ Chemistry panel (ALT, Creatinine) 
• HIV infection assessment including pretest counseling and HIV testing. A 
subsequent follow-up contact is conducted to provide post-test counseling and 
to report results to participant 
• Abbreviated physical examination including weight, vital signs, and a 
symptom-directed evaluation by history and/or appropriate physical exam 
based on the mother’s reported symptoms for her infant 
• Complete physical examination, see HVTN 135 SSP for details  
• Stool specimen collection, optional 
9.5 
Enrollment and Follow-up Visits, Mother 
The following procedures will be performed at every visit: 
• Risk reduction counseling 
• Social Impact Assessment 
•  Assessment of new or continuing concomitant medications (as described 
in Section 9.2.1) 
The following procedures will be performed at scheduled follow-up visits as 
specified in Appendix E and Appendix G: 
• Blood collection 
• Breast milk collection, optional 
• Stool collection, optional 
• Outside testing of baby questionnaire 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 77 of 151 
• Belief questionnaire  
9.6 
HIV counseling and testing 
HIV counseling will be performed in compliance with the CDC’s guidelines or 
other local guidelines for HIV counseling and referral. 
The current RSA standard of care requires close monitoring for the development 
of HIV infection in infants born to HIV-infected women. To minimize 
phlebotomy in this vulnerable population, infants will receive HIV testing through 
well baby clinics according to the local standard of care. Results from well-baby 
clinic testing will be obtained by CRS staff. 
Since the RSA standard of care requires antibody testing at month 18 for all 
infants regardless of risk, all babies will have HIV testing performed using the 
HVTN HIV testing algorithm at month 17 so the results can be provided to the 
babies’ pediatrician at the 18-month timepoint. Any testing required after month 
18 should be conducted through the CRS using the HVTN HIV testing algorithm 
until there is no evidence of Vaccine Induced Seropositivity (VISP), see Section 
9.6.1.  
Additionally, to further enhance surveillance, infants of mothers who have a viral 
load of ≥ 400 copies/mL after initial enrollment will receive additional 
monitoring, including HIV testing (see HVTN 135 SSP). Mothers who have a 
viral load ≥ 400 copies/mL will also have a reflex testing performed to look for 
drug resistant viral variants and offered counseling and options for alternative 
ARV therapy if necessary.  
Mothers will be counseled routinely during the trial on the prevention of HIV 
infection in their baby and on the potential negative social impacts of testing 
antibody positive due to VISP (see HVTN 135 SSP). They will also be counseled 
on the risks of HIV antibody testing outside of the HVTN CRSs after 18 months 
of age and will be discouraged from doing so during study participation and/or 
during any period of VISP.  
Study staff will take particular care to inform mothers of the potential for routine 
or emergency HIV testing being offered or performed outside the study CRS at 
emergency rooms, clinics, and medical offices after 18 months of age. CRS staff 
should also inform mothers of the need to maintain study blinding by getting HIV 
testing only at the study CRS. CRS staff should provide mothers with CRS 
contact information and should encourage mothers to ask medical providers to 
contact the CRS. The CRS can verify that the infant is in an HIV vaccine clinical 
trial and should only be tested at the study CRS. 
Infants identified as being HIV-infected during screening are not enrolled. 
Potential and enrolled infants identified as being HIV-infected will be urgently 
referred for medical treatment, counseling, and management of the HIV infection 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 78 of 151 
as per local standard of care. Infants who are found to be HIV-infected after 
enrollment will not receive any additional study product. The mothers will be 
encouraged to maintain their infants’ participation in follow-up visits for safety 
assessments. These individuals may also be referred to appropriate ongoing 
clinical trials or observational studies. 
9.6.1 
Distinguishing intercurrent HIV infection from vaccine-induced 
positive serology 
The study product may elicit an antibody response to HIV proteins. Therefore, 
vaccine-induced positive serology, also known as vaccine-induced seropositivity 
(VISP), may occur in this study. Several precautionary measures will be taken to 
distinguish intercurrent HIV infection from vaccine-induced positive serology. 
These precautionary measures include: 
• Infants will have physical examinations at visits specified in Appendix F. 
Signs or symptoms of an acute HIV infection syndrome, an intercurrent illness 
consistent with HIV-1 infection, or probable HIV exposure would prompt a 
diagnostic workup per the HVTN algorithm for Recent Exposure/Acute 
Infection Testing to determine HIV infection. 
• The current RSA standard of care for infants (less than 18 months of age) born 
to HIV-infected women is capable of distinguishing between infection and 
antibody positivity as it relies on PCR-based testing to confirm HIV infection. 
• Beginning at age 17 months, or earlier if indicated as above, HIV testing will 
be performed via the CRS according to the HVTN HIV testing algorithm (see 
HVTN 135 SSP), which is able to distinguish vaccine-induced antibody 
responses from actual HIV infections. 
• All infants can receive HIV-1 diagnostic testing from the site following their 
last scheduled visit until they are told that they did not receive an HIV vaccine 
or that they do not have vaccine-induced seropositivity. 
• All infants who received vaccine product and who have vaccine-induced 
positive or indeterminate HIV-1 serology (as measured by the standard anti-
HIV antibody screening tests) at or after the study is unblinded will be offered 
poststudy HIV-1 diagnostic testing (per the HVTN poststudy HIV-1 testing 
algorithm) periodically and free of charge as medically/socially indicated 
(approximately every 6 months) unless or until HIV Ab testing is no longer 
the standard test in clinical settings. 
9.6.2 
VISP registry  
Experimental HIV vaccines may induce antibody production to HIV antigens, 
producing reactive results on commercially available HIV test kits. This is called 
“vaccine-induced seropositivity” (VISP). In order to provide poststudy HIV 
testing to distinguish between VISP and HIV infection, and to mitigate potential 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 79 of 151 
social harms resulting from VISP in HIV vaccine recipients who are not infected 
with HIV, the HVTN has created a VISP registry. Following study unblinding, 
the registry will allow trained staff to verify that an individual has received an 
HIV vaccine, and therefore has the potential for VISP. Information in the VISP 
registry will not be used for research. Rather, the registry exists to support 
provision of poststudy testing and counseling services to HIV vaccine recipients. 
The registry contains the names of all study participants, unless the mother 
requests that her baby’s name be removed. 
9.7 
Assessments of reactogenicity 
For all infant participants, reactogenicity assessments are performed before and 
after each vaccination. All reactogenicity symptoms are followed until resolution 
and graded according to the Division of AIDS Table for Grading the Severity of 
Adult and Pediatric Adverse Events, Corrected Version 2.1, July 2017, except as 
noted in Section 11.2.2. 
The reactogenicity assessment period is 7 full days following each vaccination per 
the assessment schedule shown in Table 9-1. Mothers (or caregivers) are 
instructed to record their infant’s symptoms using a Participant Diary. Contacts 
between the mother and the site staff should take place at least once between 1-3 
days postvaccination. In general, a mother who reports any infant postvaccination 
reaction greater than mild should have their infant seen by a CRS clinician, or a 
clinician at a local health facility within 24 hours after onset, unless the reaction is 
improving and/or has completely resolved. Clinic staff will follow new or 
unresolved reactogenicity symptoms present at day 7 to resolution. 
Reactogenicity events are reported using CRFs that correspond to the time of 
assessment in Table 9-1. Reactogenicity assessments include assessments of 
systemic and local symptoms, and vaccine-related lesions. Events not listed on a 
CRF, or with an onset after the reactogenicity assessment period (day of 
vaccination and 7 full days after), or those meeting SAE/adverse events requiring 
expedited reporting according to DAIDS criteria, are recorded on an adverse 
experience log form. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 80 of 151 
Table 9-1 Schedule of reactogenicity assessments 
Day 
Time 
Performed by 
0a 
Baseline: before vaccination 
HVTN CRS clinician 
 
Early: 25-60 minutes after 
vaccination 
HVTN CRS clinician 
 
Between early assessment and 
11:59pm day 0 
HVTN CRS clinician or 
mother/caregiver 
1-7b 
Between 12:00am and 11:59pm on 
the respective day 
HVTN CRS clinician or 
mother/caregiver 
a Day of vaccination 
b New or unresolved reactogenicity symptoms present on day 7 are followed until resolution 
9.7.1 
Assessment of systemic and local symptoms 
Systemic symptoms include fever, sleepiness/lethargy, rash, vomiting, anorexia, 
seizure. Local symptoms include pain and/or tenderness, erythema/redness, and 
induration/swelling at the injection site. The daily maximum severity reached for 
each symptom during the assessment period is reported.  
Body temperature is measured by temporal artery thermometry (TAT). This 
includes temperatures taken in the clinic, as well as temperatures taken by 
mothers (or caregivers) during the reactogenicity period. Refer to the HVTN 135 
SSP for details. 
Temperature is reported in degrees Celsius. A measurement is taken once daily 
during the assessment period and should be repeated if an infant appears feverish. 
9.7.2 
Assessment of injection site 
Typical injection site reactions are erythema/redness and induration/swelling. The 
maximum diameter for all injection site reactions is recorded. 
All injection site reactions are monitored until resolution. Reactions with 
diameters greater than 2.5 cm are followed daily; otherwise, the frequency of 
follow-up is based on clinician judgment. See HVTN 135 SSP for detail. 
9.8 
Visit windows and missed visits 
Visit windows are included in Appendix I and Appendix J. The procedures for 
documenting missed visits and out of window visits are described in HVTN 135 
SSP. If the missed visit is one that required safety assessments or local safety labs, 
HVTN CRS staff should attempt to bring the mother-infant pair in for an interim 
visit as soon as possible. 
Procedures performed at an interim visit are usually toxicity/safety assessments 
(including local safety labs) and HIV testing. With the exception of HIV testing, 
these procedures are performed only if they were required at the missed visit or if 
clinically indicated. HIV testing may be performed as deemed appropriate by the 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 81 of 151 
study staff. Blood samples for immunogenicity assays are not typically collected 
at interim visits. 
If a missed visit required vaccination, please refer to Sections 7.5.2 and 7.5.3 for 
resolution. 
9.9 
Early termination visit 
In the event of early participant termination, site staff should consider if the 
following assessments are appropriate: a final physical examination, clinical 
laboratory tests (including CBC and chemistry panel), and social impact 
assessment. If the infant is older than 18 months, an HIV test should also be 
performed. Prior to 18 months of age, the mother’s antibodies may impact the 
result of VISP testing so VISP testing should be deferred until after 18 months. 
For infants who have a confirmed diagnosis of HIV infection, referrals for care 
should be provided.  
9.10 
Procedures for Infants Identified as HIV infected during the study 
If HIV infection is confirmed, no additional study product will be administered. 
All infants identified with HIV infection will be actively referred to non-study 
sources of HIV care and treatment. Mothers will be encouraged to continue with 
all scheduled study visits. Follow-up duration for infants diagnosed with HIV 
infection may be adjusted in consultation with the CRS investigator and the 
HVTN 135 PSRT (eg, to avoid interference with infant’s initiation of HIV 
treatment). At postinfection follow-up visits, only specimens required for 
protocol-specified safety laboratory tests, will be collected (see Appendix D). 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 82 of 151 
10 
Laboratory 
10.1 
HVTN CRS laboratory procedures 
The Site Lab Instructions and HVTN 135 SSP provide further guidelines for 
operational issues concerning the clinical and processing laboratories. These 
documents include guidelines for general specimen collection, special 
considerations for phlebotomy, specimen labeling, whole blood processing, HIV 
screening/diagnostic testing, and general screening and safety testing. 
Tube types for blood collection are specified in Appendix D and Appendix E. For 
tests performed locally, the local lab may assign appropriate tube types. 
In specific situations, the blood collection tubes may be redirected to another 
laboratory or may require study-specific processing techniques. In these cases, 
laboratory special instructions will be posted on the protocol-specific section of 
the HVTN website. 
Of note, all assays described below are performed as research assays to evaluate 
the ability of the vaccine to induce immune responses in the context of the 
participants’ genetic background and are not approved for use in medical care. 
Results from these assays are not made available to participants or medical 
professionals to guide treatment decisions. 
10.2 
Total blood volume 
Required blood volumes per visit are shown in Appendix D and Appendix E. Not 
shown is any additional blood volume that would be required if a safety lab needs 
to be repeated. The additional blood volume would likely be minimal. The 
cumulative blood draw volume will not exceed 5% of total blood volume in any 
30-day period for each infant participant and will not exceed 500mL in any 56-
day (8-week) period for each adult participant. Additionally, the maximum daily 
blood draw volume will not exceed 2.5% of total blood volume for each infant 
participant. The blood volume limits are conservative, designed to minimize 
participant harm, and are within the general consensus of the pediatric research 
community both in Africa and the US (85).  
10.3 
Primary immunogenicity timepoints 
The primary immunogenicity timepoint in this study occurs 2 weeks after third 
and fifth vaccinations for cellular assays, and 2 weeks after the fifth vaccination 
for humoral assays. Endpoint assays are performed on specimens collected from 
participants at the primary immunogenicity timepoint and may be performed on 
samples collected from participants at other timepoints; the schedule is shown in 
Appendix D. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 83 of 151 
10.4 
Endpoint assays: cellular 
10.4.1 
Env-specific B cell phenotyping  
HIV-1 Env-specific B cells and plasmablasts induced by vaccination will be 
identified and characterized using fluorescently-labeled recombinant Env proteins 
(including CH505) in combination with a flow cytometry antibody panel. In 
particular, Env-reactive B cells and plasmablasts will be enumerated and may be 
further characterized for expression of memory, activation, inhibitory or other 
markers of interest.  
10.4.2 
B-cell lineage and repertoire analysis 
Single memory B cells or plasmablasts isolated from PBMCs may be single-cell 
or bulk sorted. For memory B cells, Env reactive memory B cells will be sorted 
using CH505 Env hooks, and then single-cell sorted into individual wells of 96 
well plates. VH and VL genes will be amplified and cloned into an IgG1 
backbone and tested for Env binding and HIV neutralization. In some cases, 
memory B cells will be cultured in limiting dilution cultures and screened for 
binding and neutralization before PCR and VH and VL rescue. For plasmablasts, 
VH and VL genes will be amplified and cloned into an IgG1 backbone for 
expression and determination of binding and neutralization. Finally, to evaluate 
levels of BCR somatic hypermutation next generation sequencing for all VH and 
VL families may be performed on memory B cells or plasmablasts for VH and 
VL genes for either single-cell or bulk sorted Env-specific B cells. 
10.5 
Endpoint assays: humoral 
10.5.1 
Binding antibody multiplex assay (BAMA) 
HIV-1 Env binding IgG antibodies to the vaccine immunogens (EnvSeq-1 gp120 
TF) will be assessed on serum/plasma samples from study participants taken at 
the primary immunogenicity timepoint and, due to the contributions of maternal 
antibody at birth, will be compared to matched samples from placebo controls. 
Conformational epitopes (eg, CD4BS specificities using differential binding to 
EnvSeq-1 gp120 TF compared to CH505 I371 protein and sCD4 blocking) will 
be determined. Specimens from other timepoints as well as other HIV antigens 
(ie, other conformational epitopes and linear epitopes), IgA and antibody 
subclasses (IgG1, IgG2, IgG3, IgG4) may also be assayed based on the results of 
the initial assay. Cord blood, serum and/or breast milk samples may be tested to 
evaluate the effects of maternal antibodies. 
10.5.2 
Pediatric Vaccine Multiplex Assay (PVMA) 
The PVMA will assess serum samples for interaction (ie, additivity, antagonism 
or synergy) of the candidate vaccine with standard infant vaccines. The PVMA 
simultaneously measures antibodies against hepatitis B, Haemophilus influenzae 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 84 of 151 
type B, diphtheria, tetanus, pertussis, rubella and respiratory syncytial virus. Polio 
antigens will also be assessed. The assay developed by Itell, Permar and Fouda et 
al. is a reliable, high-throughput technique that requires minimal sample volume 
and measures multiple antibody concentrations (86). Serum samples will be tested 
at the primary immunological timepoint after the 5th vaccination. 
10.5.3 
Neutralizing antibody assay 
HIV-1–specific nAb assays will be performed on serum samples from study 
participants taken at the primary immunogenicity timepoint(s). Specimens from 
other timepoints may also be analyzed at the discretion of the HVTN Laboratory 
Program, which may be contingent on the results of the primary immunogenicity 
timepoints. The TZM-bl assay will test neutralization of the vaccine strain 
(CH505TF) and a single highly neutralization-sensitive autologous Tier 1 virus as 
a positive control (CH505.w4.3). The global panel and/or clade-specific panels 
may be used to assess heterologous Tier 2 neutralization. (87, 88). Additional 
assays will be performed to detect early precursors of CH103, CH235 and CH01 
bnAb lineages. 
10.5.4 
Antibody avidity 
Antibody avidity may be measured utilizing the binding antibody multiplex assay 
with the addition of a dissociation step to calculate the antibody avidity index 
(BAMA-AI) and will be assessed on serum samples from study participants taken 
at the primary immunogenicity timepoints and due to the contributions of 
maternal antibody at birth, will be compared to age-matched samples from 
placebo controls. Biolayer Interferometry (BLI) and/or Surface Plasmon 
Resonance (SPR) technologies may also be utilized to define antibody avidity. 
10.5.5 
FcγR binding assay 
The Fc-FcγR complex plays a critical role in eliciting cytotoxicity by mediating 
antibody Fc effector functions (such as ADCC and ADCP). Vaccine elicited HIV 
specific antibody binding to FcγR proteins will be assessed by the FCR BAMA. 
The FCR BAMA is a modification of the binding assay where fluorescently 
labeled FcR proteins are utilized as the detection reagent for serum antibodies 
bound to HIV proteins on microspheres. Serum samples from study participants 
will be taken at the primary immunogenicity timepoint. Specimens from other 
timepoints may also be assayed based on the results of the initial assay. 
10.5.6 
Antibody-dependent cellular cytotoxicity (ADCC) 
ADCC activity may be assessed using serum samples from study participants 
taken at a primary immunogenicity timepoint. Specimens from other timepoints 
may also be analyzed at the discretion of the HVTN Laboratory Program, which 
may be contingent on the results of the primary immunogenicity timepoint. For 
the Granzyme B flow-based cytotoxicity assay (89), participant sera are incubated 
with effector cells and gp120-coated target cells. ADCC is quantified as net 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 85 of 151 
percent granzyme B activity which is the percent of target cells positive for 
GranToxiLux (GTL), an indicator of granzyme B uptake, minus the percent of 
target cells positive for GTL when incubated with effector cells but without sera. 
For the Luciferase-based cytotoxicity assay (90), participant sera are incubated 
with IMC-infected cells and percent killing is measured using ViviRen 
luminescence.  
10.5.7 
Antibody-dependent cellular phagocytosis (ADCP) 
To assess the ability of vaccine-elicited antibodies to engage cellular FcR for 
potential antiviral function, ADCP may be measured using serum samples from 
vaccine recipients at a primary immunogenicity timepoint and compared to age-
matched samples from placebo controls. ADCP is measured by assessing the 
ability of vaccine elicited antibodies to mediate monocyte phagocytosis of HIV-1 
antigen coated fluorescent beads by flow cytometry (91, 92). An array of antigens 
or viruses in addition to specimens from other timepoints may also be analyzed at 
the discretion of the HVTN Laboratory Program, which may be contingent on the 
results of the primary immunogenicity timepoint. 
10.5.8 
Antibody-dependent neutrophil phagocytosis (ADNP) 
To assess the ability of vaccine-elicited antibodies to engage cellular FcR for 
potential antiviral function, ADNP may be measured using serum samples from 
study participants at a primary immunogenicity timepoint and compared to age-
matched samples from placebo controls. ADNP is measured by assessing the 
ability of vaccine elicited antibodies to mediate neutrophil cell line phagocytosis 
of HIV-1 antigen coated fluorescent beads by flow cytometry (93). An array of 
antigens or viruses in addition to specimens from other timepoints may also be 
analyzed at the discretion of the HVTN Laboratory Program, which may be 
contingent on the results of the primary immunogenicity timepoint. 
10.5.9 
Antibody dependent complement deposition (ADCD) 
To assess the ability of vaccine-elicited antibodies to bind complement, 
complement activation may be measured using serum samples from vaccine 
recipients and compared with placebo controls at a primary immunogenicity 
timepoint (94, 95). Additional specimens from other timepoints may also be 
analyzed at the discretion of the HVTN Laboratory Program, which may be 
contingent on the results of the primary immunogenicity timepoint. 
10.6 
Microbiome analysis 
Aliquots of breast milk and swabs of stool and/or bulk stool samples will be 
shipped to a central laboratory. Samples will be subjected to culture to obtain 
microbial samples. These samples will be cryopreserved and/or processed to yield 
a homogeneous flora lysate of each culture type to study their impact on 
immunity. These lysates are then available to use to assess antibody binding by 
Western blot, ELISA, or other methods. In addition, DNA will be extracted from 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 86 of 151 
samples and subjected to quality control qPCR assays to measure total microbial 
load and assess for PCR inhibitors. Broad-range 16S rRNA PCR and/or 
metagenomics will be performed. Microbial community composition will be 
assessed using taxonomic assignment of reads to a custom reference set of gene 
sequences from the human gut. Finally, bulk stool samples may also have 
antibodies extracted for analysis of the presence of mucosal antibodies elicited by 
the vaccine. 
10.7 
Systems Biology 
In vitro (cord blood) and in vivo (infant peripheral blood) samples will be 
assessed for correlates of vaccine safety and immunogenicity. In vitro studies will 
include stimulation of cryopreserved cord blood mononuclear cells (CBMC) and 
infant cord blood under 4 conditions: control, adjuvant (GLA-SE), antigen 
(gp120) and adjuvant/antigen combined. Samples will be fractionated and 
cryopreserved for downstream analysis of analytes. In vitro and in vivo samples 
will be used for plasma- and cell-based OMIC assays which may include 
RNASeq, high resolution flow cytometry, leukocyte epigenetics, plasma 
cytokines/chemokines, proteomics, and metabolomics in relation to vaccine 
immunogenicity. To identify molecular signatures that correlate with the 
immunogenicity of adjuvanted HIV vaccine, in silico analyses will employ 
programs such as NetworkAnalyst, DIABLO and related bioinformatic tools. 
10.8 
Lab assay algorithm 
The Lab Assay Algorithm lists assays to characterize cellular, humoral, and innate 
immune responses as well as host genetics that may be conducted to determine 
endpoints in HVTN vaccine trials. The type of assay(s) employed will be 
dependent on the response obtained by the primary immunogenicity assays at 
relevant timepoints. Please note that the Lab Assay Algorithm will be updated 
periodically to include new assays. 
10.9 
Exploratory studies 
Maternal and infant 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. 
10.10 
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 required by IRB/EC, or RE. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 87 of 151 
Other use of specimens is defined as studies not covered by the protocol or the 
informed consent form for the main study (see Appendix A and Appendix B). 
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 CRS. 
As part of consenting for the study, participants document their initial decision to 
allow or not allow their or their infant’s 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 or their infant’s 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. 
10.11 
Biohazard containment 
As the transmission of HIV and other blood-borne pathogens can occur through 
contact with contaminated needles, blood, and blood products, appropriate 
precautions will be employed by all personnel in the drawing of blood and 
shipping and handling of all specimens for this study, as currently recommended 
by the CDC and the NIH or other applicable agencies. 
All dangerous goods materials, including Biological Substances, Category A or 
Category B, must be transported according to instructions detailed in the 
International Air Transport Association Dangerous Goods Regulations. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 88 of 151 
11 
Safety monitoring and safety review 
11.1 
Safety monitoring and oversight 
11.1.1 
HVTN 135 PSRT 
The HVTN 135 PSRT is composed of the following members: 
• DAIDS medical officer representative 
• Protocol chairs and cochairs 
• Protocol Team leader 
• Core medical monitor 
• Clinical safety specialist 
• Regional medical liaison 
The clinician members of HVTN 135 PSRT are responsible for decisions related 
to participant safety. 
The Protocol Team clinic coordinator, clinical data manager, vaccine developer 
representative, clinical trial manager, and others may also be included in HVTN 
135 PSRT meetings. 
11.1.2 
HVTN 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, unblinded as to treatment arm, approximately 
every 4 months. The reviews consist of evaluation of cumulative reactogenicity 
events, AE, laboratory safety data, and individual reports of adverse events 
requiring expedited reporting to DAIDS. The SMB conducts additional special 
reviews at the request of the HVTN 135 PSRT. 
The study site will receive SMB summary minutes and are responsible for 
forwarding them to their IRB/EC and any applicable RE. 
11.1.3 
SDMC roles and responsibilities in safety monitoring  
The roles and responsibilities of the SDMC in relation to safety monitoring 
include: 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 89 of 151 
• Maintaining a central database management system for HVTN clinical data; 
• Providing reports of clinical data to appropriate groups such as the HVTN 135 
PSRT and HVTN SMB (see Sections 11.1.1 and 11.1.2); 
11.1.4 
HVTN Core roles and responsibilities in safety monitoring 
• Daily monitoring of clinical data for events that meet the safety pause and 
HVTN 135 PSRT AE review criteria (see Section 11.4); 
• Notifying HVTN CRSs and other groups when safety pauses or planned holds 
are instituted and lifted (see Section 11.4); 
• Querying HVTN CRSs for additional information regarding reported clinical 
data; and 
• Providing support to the HVTN 135 PSRT. 
11.2 
Safety reporting 
11.2.1 
Submission of safety forms to SDMC 
Site staff must submit all safety forms (eg, reactogenicity, adverse experience, 
local lab results, and concomitant medications) before the end of the next business 
day, excluding federal or bank holidays. The forms should not be held in 
anticipation of additional information at a later date. If additional information is 
received at a later date, the forms should be updated and resubmitted before the 
end of the next business day after receiving the new information. For the case of a 
longer site holiday closure, site staff must submit the data by the end of the 5th 
day (local time) after receiving the information even if this day is a holiday. 
For example: If the site becomes aware of an AE on Thursday (Day 0), the site 
must submit the data by the end of the next business day, on Friday. If there is a 
longer site holiday closure, then this AE must be reported no later than the end of 
the fifth day, Monday (Day 4). If Monday is a holiday as well, all safety forms 
still need to be submitted by the end of Monday (Day 4). 
11.2.2 
AE reporting 
An AE is any untoward medical occurrence in a clinical investigation participant 
administered a study product/procedure(s) and which does not necessarily have a 
causal relationship with this treatment. An AE can therefore be any unfavorable 
and unintended sign (including an abnormal laboratory finding), symptom, or 
disease temporally associated with the use of an investigational study 
product/procedure(s), whether or not related to the investigational study 
product/procedure(s). AEs will be collected for infants only. All AEs are graded 
according to the Division of AIDS (DAIDS) Table for Grading the Severity of 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 90 of 151 
Adult and Pediatric Adverse Events, Corrected Version 2.1, July 2017, available 
on the RSC website at https://rsc.niaid.nih.gov/clinical-research-sites/daids-
adverse-event-grading-tables, except as shown below in Table 11-1, rows for 
pain/tenderness at the injection site and sleepiness/lethargy were extracted from 
IMPAACT 2004. The grading of creatinine is a required modification to the 
DAIDS table dictated by the inappropriateness of using a “baseline” creatinine in 
an infant as the reference range as this analyte fluctuates greatly throughout the 
first year of life. 
Table 11-1 Study-Specific Severity Grading Table 
 
Grade 1 
Mild 
Grade 2 
Moderate 
Grade 3 
Severe 
Grade 4 
Potentially Life-
Threatening 
Pain/tenderness at 
injection site 
Mild reaction to 
light touch with no 
or minimal 
limitation of 
movement of limb 
Persistent crying 
(up to one hour) 
with no or light 
touch or 
significant 
limitation of 
movement of limb 
Persistent crying 
for more than one 
hour or 
interference with 
infant’s ability to 
sleep or eat 
Inconsolable 
crying for more 
than two hours 
Sleepiness/lethargy 
Transiently 
lethargic but 
otherwise normal 
routine 
More sleeping 
than usual, not on 
normal routine 
without 
alternative 
explanation 
Refuses to 
play/smile with 
parent/caregiver 
or somnolent, 
needs to be 
stimulated to take 
feedings 
Non-responsive, 
unable to rouse 
Creatinine, High 
1.1 to 1.3 x ULN 
> 1.3 to 1.8 x 
ULN 
> 1.8 to < 3.5 x 
ULN 
≥ 3.5 x ULN 
Platelets, 
Decreased 
(cells/mm3) 
≤ 5 days of age 
 75,000 to < 
100,000 
50,000 to < 
75,000 
25,000 to < 
50,000 
<25,000 
 
Platelets, 
Decreased 
(cells/mm3) 
> 5 days of age 
100,000 to < 
125,000 
50,000 to 
<100,000 
25,000 to < 
50,000 
<25,000 
 
Additionally, to report weight loss in an infant as an AE, the “Underweight” row 
on the DAIDS Grading Table should be used. Weight loss should not be graded 
using the “Unintentional Weight Loss” row. 
All AEs are reported to the SDMC on the appropriate CRF. Clinic staff should 
evaluate every AE to determine if (1) the AE meets the requirements for 
expedited reporting (Section 11.2.3), (2) if the AE meets the criteria for a safety 
pause/prompt AE review (Section 11.3), and (3) if the AE is a potential immune-
mediated disease that may be listed as an AE of special interest (AESI). A sample 
list of AESI is provided in Appendix H. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 91 of 151 
The study site is expected to notify HVTN clinical safety staff of any serious 
safety concern requiring their attention (Table 11-3). Telephone numbers and 
email addresses are found on the protocol home page on the HVTN Members’ 
site (https://members.hvtn.org/protocols/hvtn135). Concerns requiring immediate 
attention should be communicated by calling the clinical safety phone. 
In the case of email notification, clinical safety staff will reply within one 
business day. Serious events that meet pause rule criteria will be addressed 
immediately (as outlined in Table 11-3). If email service is not available, the CRS 
should notify clinical safety staff of the event by telephone, and then submit 
CRFs.  
In addition, site investigators are required to submit AE information in accordance 
with IRB/EC and any applicable RE requirements. 
11.2.3 
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 RSC 
website at https://rsc.niaid.nih.gov/clinical-research-sites/manual-expedited-
reporting-adverse-events-daids. The SAE Reporting Category will be used for this 
study. 
The internet-based DAIDS Adverse Experience Reporting System (DAERS) must 
be used for expedited AE (EAE) reporting to DAIDS. In the event of system 
outages or technical difficulties, expedited AE reports may be submitted via 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. 
For questions about DAERS, please contact CRMSsupport@niaid.nih.gov or 
from within the DAERS application itself. 
For questions about EAE reporting, please contact the DAIDS RSC Safety Office 
at (DAIDSRSCSafetyOffice@tech-res.com). 
The study products for which expedited reporting are required are:  
• CH505TF gp120 with GLA-SE or Placebo 
The NIAID/DAIDS will report all unexpected SAEs related to the study products 
observed in this clinical trial to the South African Healthcare Products Regulatory 
Authority (SAHPRA). However, because safety is a primary study endpoint, the 
study sponsor Medical Officer will not routinely be unblinded to study treatment 
assignment when there is an assessment of relatedness of the SAE with the study 
product(s); and the safety report will be sent to the SAHPRA based on the blinded 
attribution assessment. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 92 of 151 
In some cases, the PSRT or CRS may believe unblinding of the site PI and 
participant would be appropriate to facilitate the clinical management of an AE or 
SAE. The HVTN MOP specifies procedures for emergency unblinding, and for 
early unblinding for medical reasons. 
11.3 
Safety reviews 
11.3.1 
Safety Evaluations for moving from Part A to Part B and Part B to 
Part C  
To ensure safety, enrollment will proceed in stages. In addition to monitoring 
participant safety throughout the study period, the HVTN 135 PSRT will review 
safety data as shown in Table 11-2.  
There are two planned enrollment holds and one planned safety review. An 
enrollment hold prevents advancement of the study to the next part but does not 
prevent participants from receiving further vaccinations. Part B cannot enroll until 
the safety data following the first vaccination in all seven infants in Part A has 
been reviewed (Enrollment Hold #1). Part C cannot begin enrollment until all 
safety data following the third vaccination in Part B has been reviewed 
(enrollment hold #2). 
There is also a planned safety review to occur after the final infant in part B (the 
fourth infant) has receives the first vaccination. During this safety review, no 
infants in Part B will receive further vaccinations until the PSRT has reviewed all 
cumulative safety information. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 93 of 151 
Table 11-2 Summary of planned enrollment holds and safety reviews 
Planned Holds 
and Reviews 
Timepoint/Data Reviewed 
Actions 
Planned Safety 
Hold #1 
 
Enrollment in 
Parts B and C 
are on hold. 
Begins after the 7th participant receives 
first vaccination in Part A. 
 
Review of all cumulative safety data 
available for the seven participants in 
Part A (Groups 1 and 2) up to and 
including the 2-week visit after the 
first vaccination. 
The PSRT will make a decision based on 
this safety data regarding the 
appropriateness of beginning enrollment 
in Part B from a safety perspective. 
  
Planned Safety 
Review in Part B 
 
All vaccinations 
in Part B are on 
hold.  
Enrollment in 
Part C is on hold. 
Begins after the 4th participant in Part 
B (Groups 3 and 4) receives their first 
vaccination.  
 
Review of all cumulative safety data 
available for: 
• 
all participants in Part A. 
• 
all four participants in Part B 
(Groups 3 and 4) up to and 
including the 2-week visit 
after the first vaccination. 
The PRST will make a decision based on 
this safety data regarding the 
appropriateness of continuing Part B 
vaccinations from a safety perspective. 
Planned Safety 
Hold #2 
 
Enrollment in 
Part C is on hold. 
Begins after the 4th participant in Part 
B (Groups 3 and 4) receives their third 
vaccination. 
 
Review of all cumulative safety data 
available for: 
• 
all participants in Part A. 
• 
the four participants in Part B 
(Groups 3 and 4) up to and 
including the 2-week visit 
after the third vaccination. 
The PSRT will make a decision based on 
this safety data regarding the 
appropriateness of beginning enrollment 
in Part C from a safety perspective. 
 
11.4 
Safety pause and prompt PSRT AE review 
When a trial is placed on safety pause, all enrollment and vaccination with the 
product related to the event that triggered the pause will be held until further 
notice. The AEs that will lead to a safety pause or prompt HVTN 135 PSRT AE 
review are summarized in Table 11-3. 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 135 PSRT, participant safety may be threatened. 
Criteria for an individual participant’s departure from the schedule of vaccinations 
are listed in Section 7.5. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 94 of 151 
Table 11-3 AE notification and safety pause/AE review rules 
Event and relationship to study products 
Severity 
HVTN CRS action 
a 
HVTN Core action 
SAE, related 
Any 
Grade 
Phone 
immediately, email 
and submit forms 
immediately 
Immediate pause 
SAE, not related 
Grade 5 
Phone 
immediately, email 
and submit forms 
immediately 
Immediate PSRT 
notification 
AE, related 
Grade 4 
or 3 
Email and submit 
forms immediately 
Immediate PSRT 
notification and 
prompt PSRT AE 
review to consider 
pause 
25% or more infants (four or more infants 
if fewer than 20 have been enrolled) with 
a grade 3 or higher reactogenicity signs 
and symptoms within the first full 3 days 
after study product administration 
Grade 4 
or 3 
Phone 
immediately, email 
and submit forms 
immediately 
Immediate PSRT 
notification and 
prompt PSRT AE 
review to consider 
pause 
a Phone numbers and email addresses are found on the Protocol home page on the HVTN Members’ site 
(https://members.hvtn.org/protocols/hvtn135) 
For all immediate safety pauses, HVTN Core notifies the HVTN 135 PSRT, 
HVTN Regulatory Affairs, DAIDS Pharmaceutical Affairs Branch (PAB), 
DAIDS Regulatory Affairs Branch (RAB), DAIDS Safety and Pharmacovigilance 
Team (SPT), and the participating HVTN CRS. When an immediate safety pause 
is triggered, HVTN Core notifies the SMB, and the SMB will be convened as 
soon as possible to review the event. 
Once an immediate safety pause is triggered and the trial is paused, the HVTN 
135 PSRT and the SMB review safety data and decide whether the pause can be 
lifted or permanent discontinuation of vaccination is appropriate. HVTN Core 
notifies the participating HVTN CRS, HVTN Regulatory Affairs, DAIDS PAB, 
DAIDS RAB, and DAIDS SPT of the decision regarding resumption or 
discontinuation of study vaccinations. Based on the HVTN 135 PSRT assessment, 
DAIDS RAB notifies the SAHPRA as needed. 
If an immediate HVTN 135 PSRT notification or prompt HVTN 135 PSRT AE 
review is triggered, HVTN Core notifies the HVTN 135 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 135 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. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 95 of 151 
In addition, all other AEs are reviewed routinely by the HVTN 135 PSRT (see 
Section 11.5.2). 
11.5 
Review of cumulative safety data 
Routine safety review occurs at the start of enrollment and then throughout the 
study. 
Reviews proceed from a standardized set of protocol-specific safety data reports. 
These reports are produced by the SDMC and include queries to the HVTN CRSs. 
Events are tracked by internal reports until resolution. 
11.5.1 
Daily review 
Blinded daily safety reviews are routinely conducted by HVTN Core for events 
requiring expedited reporting to DAIDS, and events that meet safety pause criteria 
or prompt HVTN 135 PSRT AE review criteria. 
11.5.2 
Weekly review 
During the injection phase of the trial, the HVTN 135 PSRT reviews clinical 
safety reports on a weekly basis and conducts calls to review the data as 
appropriate. After the injections and the final 2-week safety visits are completed, 
less frequent reporting and safety reviews may be conducted at the discretion of 
the HVTN 135 PSRT. HVTN Core reviews reports of clinical and laboratory 
AEs. Events identified during the review that are considered questionable, 
inconsistent, or unexplained are referred to the HVTN CRS clinic coordinator for 
verification. 
11.6 
Study termination 
This study may be terminated early by the determination of the HVTN 135 PSRT, 
a pertinent national regulatory authority, SAHPRA, NIH, Office for Human 
Research Protections (OHRP), or study product developer(s). In addition, the 
conduct of this study at the HVTN CRS may be terminated by the determination 
of the IRB/EC and any applicable RE. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 96 of 151 
12 
Protocol conduct 
This protocol and all actions and activities connected with it will be conducted in 
compliance with the principles of GCP (ICHe6), and according to DAIDS and 
HVTN policies and procedures as specified in the HVTN Manual of Operations, 
DAIDS Clinical Research Policies and Standard Procedures Documents including 
procedures for the following: 
• Protocol registration, activation, and implementation; 
• Informed consent, screening, and enrollment; 
• Study participant reimbursement; 
• Clinical and safety assessments; 
• Safety monitoring and reporting; 
• Data collection, documentation, transfer, and storage; 
• Participant confidentiality; 
• Study follow-up and close-out; 
• Unblinding of staff and participants; 
• Quality control; 
• Protocol monitoring and compliance; 
• Advocacy and assistance to participants regarding negative social impacts 
associated with the vaccine trial; 
• Risk reduction counseling; 
• Specimen collection, processing, and analysis; 
• Exploratory and ancillary studies and sub-studies, and 
• Destruction of specimens. 
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 135 Study Specific Procedures. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 97 of 151 
12.1 
Social impacts 
Participants in this study risk experiencing discrimination or other personal 
problems, resulting from the study participation itself or from the development of 
VISP. The HVTN CRS is obliged to provide advocacy for and assistance to 
participants regarding these negative social impacts associated with the vaccine 
trial. Mothers will be extensively counseled about these potential problems prior 
to and during study participation, and study staff will provide advocacy and 
assistance to any participants who may experience such problems. If HVTN CRS 
staff have questions regarding ways to assist a participant dealing with a social 
impact, a designated NIAID or HVTN Core representative can be contacted. 
In addition, the study site may participate in the VISP registry established by the 
HVTN. This registry allows trained staff to verify that an individual has received 
an HIV vaccine and therefore has the potential for VISP. The registry also 
supports poststudy HIV counseling and testing services for HIV vaccine 
recipients (see Section 9.6.2). 
Social harms are tabulated by the SDMC and are subjected to descriptive analysis. 
The goal is to reduce their incidence and enhance the ability of study staff to 
mitigate them when possible. 
Summary tables of social impact events will be generated weekly, and made 
available for review by the protocol chairs, protocol team leader, and the 
designated NIAID representative. 
12.2 
Emergency communication with study participants 
As in all clinical research, this study may generate a need to reach participants 
quickly to avoid imminent harm, or to report study findings that may otherwise 
concern their health or welfare. 
When such communication is needed, the CRS will request that its IRB/EC and 
any applicable RE expedite review of the message. If this review cannot be 
completed in a timeframe consistent with the urgency of the required 
communication, the site can contact the participant without IRB/EC approval if 
such communication is necessary to avoid imminent harm to the study participant. 
The CRS must notify the IRB/EC and any applicable RE of the matter as soon as 
possible. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 98 of 151 
13 
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 135 are described 
below. 
Protocol history and modifications 
Date: December 18, 2020 
Protocol version: 3.0 
Protocol modification: Full protocol amendment 2 
Item 1 
Revised in Section 3, Overview, Section 4, Background, Section 6, 
Statistical considerations, Section 8, Study product preparation and 
administration and Appendix A, Sample informed consent form: primary 
immunogenicity dose group, CH505TF gp120 dose for Groups 3 and 7 and 
group size for Groups 5 and 7 
Item 2 
Updated in Section 4.8.1, Clinical studies of CH505TF gp120 with GLA-SE 
adjuvant in adults, HVTN 115 and Section 6.1, Accrual and sample size 
calculations: safety data from HVTN 115 Part A 
Item 3 
Updated per Letter of Amendment 1 version 2.0, dated September 18, 2020 
Item 4 
Updated Section 14, Documents references and Appendix F, Procedures at 
HVTN CRS : web address and footnote cross-reference 
Item 5 
Updated Title page and Section 13, Version history: date, version number 
and contents of this amendment 
 
Date: September 18, 2020 
Protocol version: Version 2.0 
Protocol modification: Letter of Amendment 1 
Item 1 
Added in Section 7.1, Inclusion criteria: Mother, Section 9.2.1, Pre-
enrollment procedures, Mother, Appendix A, Sample informed consent 
form, Appendix C, Table of procedures (for sample informed consent form), 
Appendix E, Laboratory procedures: Mother: SARS-CoV-2 testing 
Item 2 
Updated in Section 3.1, Protocol Team: Membership 
Item 3 
Updated in Appendix A, Sample informed consent form and in Appendix B, 
Sample consent form for use of samples and information in other studies: 
contact information for South African Healthcare Products Regulatory 
Authority (SAHPRA) 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 99 of 151 
Date: January 24, 2020 
Protocol version: 2.0 
Protocol modification: Full protocol amendment 1 
Item 1 
Revised in Section 9.3, Enrollment and vaccination visits, infant and 
Appendix A, Sample informed consent form: time infant participant 
will be observed in clinic post administration of study product 
Item 2 
Revised in Section 8.3, Preparation of study products: syringe 
labelling 
Item 3 
Added in Section 8.5, Acquisition of study products: manufacturer of 
study products 
Item 4 
Revised in Appendix A, Sample informed consent form: text changes 
throughout to improve clarity 
Item 5 
Updated in Appendix A, Sample informed consent form, item 4: 
number of participants in HVTN 115 who received vaccine 
Item 6 
Updated in Appendix A, Sample Informed Consent Form and 
Appendix B, Sample informed consent for the use of samples and 
information in other studies: SAHPRA contact information 
Item 7 
Updated in Section 3.1, Overview: protocol team members and 
Statistical and data management center (SDMC) 
Item 8 
Corrected in Section 7.5.3, Discontinuing vaccination for a 
participant: method of body temperature measurement 
Item 9 
Added in Section 9.7.1, Assessment of systemic and local symptoms: 
erythema/redness as local symptom examples 
Item 10 Deleted in Section 9.6, HIV counseling and testing: reference to 
HVTN HIV testing algorithm 
Item 11 Added in Section 10.5.6, Antibody-dependent cellular cytotoxicity 
(ADCC) and Section 16, Literature cited: references to assays 
described 
Item 12 Updated in Appendices D, Laboratory procedures: Infant, and 
Appendix E, Laboratory procedures: Mother: cord blood volume, 
tube types, test location, and footnotes 
Item 13 Corrected in Appendix I, Infant visit windows and Appendix J, 
Mother visit windows: target day for visits 9 and 10 
Item 14 Added to Title page: “A non-IND study” 
Item 15 Added to Section 8.4, Administration: storage temperature for 
product in syringe 
Item 16 Updated in Section 14, Document references: URL for ICH 
Item 17  Updated Section 13, Version history: added amendment 1 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 100 of 151 
Date: July 30, 2019 
Protocol version: 1.0 
Protocol modification: 
 
Original Protocol 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 101 of 151 
14 
Document references (other than literature 
citations) 
Other documents referred to in this protocol, and containing information relevant 
to the conduct of this study, include: 
• Assessment of Understanding. Accessible through the HVTN protocol-
specific website. 
• DAIDS Policy on Enrolling Children (including Adolescents) in Clinical 
Research: Clinical Research Site Requirements Available at 
https://www.niaid.nih.gov/sites/default/files/enrollingchildrenrequirements.pd
f 
• Current CDC Guidelines: 
▪ Revised Recommendations for HIV Testing of Adults, Adolescents, and 
Pregnant Women in Health-Care Settings. Available at 
http://www.cdc.gov/mmwr/PDF/rr/rr5514.pdf  
▪ Revised Guidelines for HIV Counseling, Testing, and Referral. Available 
at http://www.cdc.gov/mmwr/preview/mmwrhtml/rr5019a1.htm 
• Division of AIDS (DAIDS) Clinical Research Policies and Standard 
Procedures Documents. Available at 
https://www.niaid.nih.gov/research/daids-clinical-research-policies-standard-
procedures 
• Division of AIDS Protocol Registration Manual. Available at 
https://www.niaid.nih.gov/sites/default/files/prmanual.pdf  
• Division of AIDS Table for Grading the Severity of Adult and Pediatric 
Adverse Events. Corrected Version 2.1, July 2017. Available at 
https://rsc.niaid.nih.gov/clinical-research-sites/daids-adverse-event-grading-
tables 
• The Manual for Expedited Reporting of Adverse Events to DAIDS. Version 
2.0, January 2010. Available at https://rsc.niaid.nih.gov/clinical-research-
sites/manual-expedited-reporting-adverse-events-daids 
• HVTN Certificate of Confidentiality. Accessible through the HVTN website. 
• HVTN 135 Special Instructions. Accessible through the HVTN protocol-
specific website. 
• HVTN 135 Study Specific Procedures. Accessible through the HVTN 
protocol-specific website. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 102 of 151 
• HVTN 135 Site Lab Instructions. Accessible through the HVTN protocol-
specific website. 
• HVTN Manual of Operations. Accessible through the HVTN website. 
• Dangerous Goods Regulations (updated annually), International Air Transport 
Association. Available for purchase at 
https://www.iata.org/publications/dgr/Pages/index.aspx 
• Lab assay algorithm (available upon request). 
• International Council on Harmonisation of Technical Requirements for 
Pharmaceuticals for Human Use (ICH) E6, Guideline for Good Clinical 
Practice: Section 4.8, Informed consent of trial subjects. Available at 
https://www.ich.org/page/efficacy-guidelines 
• Participants’ Bill of Rights and Responsibilities. Accessible through the 
HVTN website. 
• NIH Policy on Reporting Race and Ethnicity Data: Subjects in Clinical 
Research. Available at https://grants.nih.gov/grants/guide/notice-files/NOT-
OD-01-053.html 
• Pharmacy Guidelines and Instructions for DAIDS Clinical Trials Networks, 
July 2008. 
• Requirements for Source Documentation in DAIDS Funded and/or Sponsored 
Clinical Trials. Available at 
https://www.niaid.nih.gov/sites/default/files/daids-sourcedocpolicy.pdf  
• Title 21, Code of Federal Regulations, Part 50. Available at 
http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFR
Part=50 
• Title 45, Code of Federal Regulations, Part 46. Available at 
https://www.hhs.gov/ohrp/regulations-and-policy/regulations/45-cfr-
46/index.html 
• New EPI Vaccines Guidelines, National Department of Health, Republic of 
South Africa. Available at 
https://ndoh.dhmis.org/owncloud/index.php/s/R5cmdp0gY4Fa43Z?path=%2F
Chidrens%20Vaccines 
• Guidelines for Phase I clinical trials, The Association of the British 
Pharmaceutical Industry. Available at 
https://www.abpi.org.uk/publications/guidelines-for-phase-i-clinical-trials-
2018-edition/ 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 103 of 151 
• The Protection of Personal Information Act (POPIA, South Africa, 2013). 
Available at 
https://www.gov.za/sites/default/files/gcis_document/201409/3706726-
11act4of2013protectionofpersonalinforcorrect.pdf 
See Section 16 for literature cited in the background and statistics sections of this 
protocol. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 104 of 151 
15 
Acronyms and abbreviations 
 
Ab 
 
antibody 
Ad 
 
adenovirus 
ADCC 
 
Antibody-dependent cellular cytotoxicity 
ADCD 
 
Antibody dependent complement deposition 
ADCP 
 
Antibody-dependent cellular phagocytosis 
ADNP 
 
Antibody-dependent neutrophil phagocytosis 
AE 
 
adverse event 
AESI 
 
AE of special interest 
ALT 
 
alanine aminotransferase 
ANOVA  
analysis of variance 
ART 
 
antiretroviral therapy 
ARV 
 
antiretroviral 
AST 
 
aspartate aminotransferase 
AUC 
 
area under the curve 
AUC-MB  
area-under-the-magnitude-breadth 
AVEG 
 
AIDS Vaccine Evaluation Group 
β-HCG 
 
beta human chorionic gonadotropin 
BAMA 
 
Binding antibody multiplex assay 
BCR 
 
B cell receptor 
BMI 
 
body mass index 
bnAb 
 
broadly neutralizing antibody 
CAB 
 
Community Advisory Board 
cART 
 
combination anti-retroviral therapy 
CBC 
 
complete blood count 
CBMC 
 
Cord Blood Mononuclear Cells 
CD4bs 
 
CD4 binding site 
CDC 
 
US Centers for Disease Control and Prevention 
CFR 
 
Code of Federal Regulations 
CI 
 
confidence intervals 
CIOMS 
 
Council for International Organizations of Medical Sciences 
COMPASS 
Combinatorial Polyfunctionality analysis of Antigen-Specific 
T-cell Subsets 
CRF 
 
case report form 
CRPMC 
 
NIAID Clinical Research Products Management Center 
CRS 
 
clinical research site 
CTL 
 
cytotoxic T lymphocyte 
DAERS 
 
DAIDS Adverse Experience Reporting System 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 105 of 151 
DAIDS 
 
Division of AIDS (US NIH) 
DHHS 
 
US Department of Health and Human Services 
DSMB 
 
NIAID Data and Safety Monitoring Board 
EAE 
 
adverse events requiring expedited reporting to DAIDS 
EC 
 
Ethics Committee 
EIA 
 
enzyme immunoassay 
ELISA 
 
enzyme-linked immunosorbent assay 
ELISpot 
 
enzyme-linked immunospot 
Env 
 
HIV envelope protein 
EPI 
 
Expanded Program on Immunization 
FPR 
 
false positive rate 
Fred Hutch 
Fred Hutchinson Cancer Research Center 
 
GCP 
 
Good Clinical Practice 
GEE 
 
generalized estimating equation 
GLA-SE  
Glucopyranosyl Lipid A - stable emulsion 
GLP 
 
Good Laboratory Practices 
GPP 
 
Good Participatory Practices 
HAART 
 
highly active antiretroviral therapy 
HBsAg 
 
hepatitis B surface antigen 
HCV 
 
hepatitis C virus 
HEU 
 
HIV-1 exposed uninfected 
HIPAA 
 
Health Insurance Portability and Accountability Act 
HIV 
 
human immunodeficiency virus 
HLA 
 
human leukocyte antigen 
HSML-NICD 
HIV Sero-Molecular Laboratory, National Institute for 
Communicable Diseases 
HVTN 
 
HIV Vaccine Trials Network 
IA 
 
intermediate ancestor 
IB 
 
Investigator’s Brochure 
IBC 
 
Institutional Biosafety Committee 
ICF 
 
informed consent form 
ICH 
 
International Council on Harmonisation of Technical 
Requirements for Pharmaceuticals for Human Use 
ICS 
 
intracellular cytokine staining 
IFN-γ 
 
interferon gamma 
IM 
 
intramuscular 
IND 
 
Investigational New Drug 
IRB 
 
Institutional Review Board 
IUD 
 
intrauterine device 
LTFU 
 
loss to follow-up 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 106 of 151 
mAb 
 
monoclonal antibody 
MAR 
 
missing at random 
MCAR 
 
missing completely at random 
MIMOSA  
Mixture Models for Single-cell Assays 
MMR 
 
measles, mumps, and rubella 
MOP 
 
Manual of Operations 
MTCT 
 
mother to child transmission 
nAb 
 
neutralizing antibody 
NHP 
 
nonhuman primate 
NHP 
 
nonhuman primate 
NIAID 
 
National Institute of Allergy and Infectious Diseases (US NIH) 
NICD 
 
National Institute for Communicable Diseases (Johannesburg, 
South Africa) 
NIH 
 
US National Institutes of Health 
NZW 
 
New Zealand White 
OBA 
 
NIH Office of Biotechnology Activities 
OHRP 
 
US Office for Human Research Protections 
OPV 
 
oral polio vaccine 
PAB 
 
DAIDS Pharmaceutical Affairs Branch 
PBMC 
 
peripheral blood mononuclear cell 
PBS 
 
phosphate-buffered saline 
PCA 
 
Principal Component analysis 
PCR 
 
polymerase chain reaction 
PI 
 
Principal Investigator 
PSRT 
 
Protocol Safety Review Team 
PTE 
 
potential T -cell epitope 
PVMA 
 
Pediatric Vaccine Multiplex Assay 
RAB 
 
DAIDS Regulatory Affairs Branch 
RAC 
 
NIH Recombinant DNA Advisory Committee 
RE 
 
regulatory entity 
RSC 
 
DAIDS Regulatory Support Center 
SAE 
 
serious adverse event 
SAHPRA  
South African Healthcare Products Regulatory Authority 
SAIL-NICD 
South Africa Immunology Laboratory and National Institute 
for Communicable Diseases 
SAP 
 
Statistical Analysis Plan 
SCHARP  
Statistical Center for HIV/AIDS Research and Prevention 
SD 
 
standard deviation 
SDMC 
 
statistical and data management center 
SFC 
 
spot-forming cell 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 107 of 151 
SFU 
 
spot-forming unit 
SHIV 
 
simian-human immunodeficiency virus 
SIV 
 
simian immunodeficiency virus 
SMB 
 
Safety Monitoring Board 
SPT 
 
DAIDS Safety and Pharmacovigilance Team 
TB 
 
tuberculosis 
TF 
 
transmitted/founder 
UCA 
 
unmutated common ancestor 
UW-VSL  
University of Washington Virology Specialty Laboratory 
VISP 
 
Vaccine induced seropositivity 
VRC 
 
Vaccine Research Center (NIAID) 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 108 of 151 
16 
Literature cited 
1. UNAIDS. Ethical considerations in biomedical HIV prevention trials. 2007 
7/2007. 
2. The National Commission for the Protection of Human Subjects of 
Biomedical and Behavioral Research. The Belmont Report: Ethical Principles 
and Guidelines for the Protection of Human Subjects of Research. 1979 
4/18/1979. 
3. Council for International Organizations of Medical Sciences (CIOMS). 
International ethical guidelines for biomedical research involving human 
subjects. Bull Med Ethics. 2002(182):17-23. 
4. UNAIDS. Miles To Go. Global AIDS Update 2018. 
5. Mahy M, Stover J, Kiragu K, Hayashi C, Akwara P, Luo C, et al. What will it 
take to achieve virtual elimination of mother-to-child transmission of HIV? 
An assessment of current progress and future needs. Sex Transm Infect. 
2010;86 Suppl 2:ii48-55. 
6. Dellar RC, Dlamini S, Karim QA. Adolescent girls and young women: key 
populations for HIV epidemic control. J Int AIDS Soc. 2015;18(2 Suppl 
1):19408. 
7. Johnson LF, Stinson K, Newell ML, Bland RM, Moultrie H, Davies MA, et 
al. The contribution of maternal HIV seroconversion during late pregnancy 
and breastfeeding to mother-to-child transmission of HIV. J Acquir Immune 
Defic Syndr. 2012;59(4):417-25. 
8. Hoffman RM, Black V, Technau K, van der Merwe KJ, Currier J, Coovadia 
A, et al. Effects of highly active antiretroviral therapy duration and regimen 
on risk for mother-to-child transmission of HIV in Johannesburg, South 
Africa. J Acquir Immune Defic Syndr. 2010;54(1):35-41. 
9. Phillips T, Thebus E, Bekker LG, McIntyre J, Abrams EJ, Myer L. 
Disengagement of HIV-positive pregnant and postpartum women from 
antiretroviral therapy services: a cohort study. J Int AIDS Soc. 
2014;17:19242. 
10. Hofmann-Lehmann R, Vlasak J, Rasmussen RA, Jiang S, Li PL, Baba TW, et 
al. Postnatal pre- and postexposure passive immunization strategies: 
protection of neonatal macaques against oral simian-human 
immunodeficiency virus challenge. J Med Primatol. 2002;31(3):109-19. 
11. Mascola JR, Stiegler G, VanCott TC, Katinger H, Carpenter CB, Hanson CE, 
et al. Protection of macaques against vaginal transmission of a pathogenic 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 109 of 151 
HIV-1/SIV chimeric virus by passive infusion of neutralizing antibodies. Nat 
Med. 2000;6(2):207-10. 
12. Mascola JR, Haynes BF. HIV-1 neutralizing antibodies: understanding 
nature's pathways. Immunol Rev. 2013;254(1):225-44. 
13. Haynes BF, Kelsoe G, Harrison SC, Kepler TB. B-cell-lineage immunogen 
design in vaccine development with HIV-1 as a case study. Nat Biotechnol. 
2012;30(5):423-33. 
14. Liao HX, Lynch R, Zhou T, Gao F, Alam SM, Boyd SD, et al. Co-evolution 
of a broadly neutralizing HIV-1 antibody and founder virus. Nature. 
2013;496(7446):469-76. 
15. Saunders KO, Verkoczy LK, Jiang C, Zhang J, Parks R, Chen H, et al. 
Vaccine Induction of Heterologous Tier 2 HIV-1 Neutralizing Antibodies in 
Animal Models. Cell Rep. 2017;21(13):3681-90. 
16. Goo L, Chohan V, Nduati R, Overbaugh J. Early development of broadly 
neutralizing antibodies in HIV-1-infected infants. Nat Med. 2014;20(6):655-
8. 
17. Simonich CA, Williams KL, Verkerke HP, Williams JA, Nduati R, Lee KK, 
et al. HIV-1 Neutralizing Antibodies with Limited Hypermutation from an 
Infant. Cell. 2016;166(1):77-87. 
18. Muenchhoff M, Adland E, Karimanzira O, Crowther C, Pace M, Csala A, et 
al. Nonprogressing HIV-infected children share fundamental immunological 
features of nonpathogenic SIV infection. Sci Transl Med. 
2016;8(358):358ra125. 
19. Permar S, Levy O, Kollman TR, Singh A, De Paris K. Early Life HIV-1 
Immunization: Providing a Window for Protection Before Sexual Debut. 
AIDS Res Hum Retroviruses. 2018;34(10):823-7. 
20. Ditse Z, Muenchhoff M, Adland E, Jooste P, Goulder P, Moore PL, et al. 
HIV-1 Subtype C-Infected Children with Exceptional Neutralization Breadth 
Exhibit Polyclonal Responses Targeting Known Epitopes. J Virol. 
2018;92(17). 
21. Fouda GG, Cunningham CK, McFarland EJ, Borkowsky W, Muresan P, 
Pollara J, et al. Infant HIV-1 gp120 vaccination elicits robust and durable 
anti-V1V2 IgG responses and only rare Envelope-specific IgA responses. J 
Infect Dis. 2014. 
22. Chen ZJ, Wheeler CJ, Shi W, Wu AJ, Yarboro CH, Gallagher M, et al. 
Polyreactive antigen-binding B cells are the predominant cell type in the 
newborn B cell repertoire. Eur J Immunol. 1998;28(3):989-94. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 110 of 151 
23. Piatosa B, Birbach M, Siewiera K, Ussowicz M, Kalwak K, Drabko K, et al. 
Significant changes in the composition of the precursor B-cell compartment 
in children less than 2 years old. Cytometry B Clin Cytom. 2013;84(3):179-
86. 
24. Meyer-Bahlburg A, Andrews SF, Yu KO, Porcelli SA, Rawlings DJ. 
Characterization of a late transitional B cell population highly sensitive to 
BAFF-mediated homeostatic proliferation. J Exp Med. 2008;205(1):155-68. 
25. Haynes BF, Fleming J, St Clair EW, Katinger H, Stiegler G, Kunert R, et al. 
Cardiolipin polyspecific autoreactivity in two broadly neutralizing HIV-1 
antibodies. Science. 2005;308(5730):1906-8. 
26. Bonsignori M, Wiehe K, Grimm SK, Lynch R, Yang G, Kozink DM, et al. 
An autoreactive antibody from an SLE/HIV-1 individual broadly neutralizes 
HIV-1. J Clin Invest. 2014;124(4):1835-43. 
27. Schroeder KMS, Agazio A, Strauch PJ, Jones ST, Thompson SB, Harper MS, 
et al. Breaching peripheral tolerance promotes the production of HIV-1-
neutralizing antibodies. J Exp Med. 2017;214(8):2283-302. 
28. Mohr E, Siegrist CA. Vaccination in early life: standing up to the challenges. 
Curr Opin Immunol. 2016;41:1-8. 
29. Lee A, Shannon C, Amenyogbe N, Bennike T, Diray-Arce J, Idoko O, et al. 
Dynamic molecular changes during the first week of human life follow a 
robust developmental trajectory. Nature Communications. 2019;in press. 
30. Fera D, Schmidt AG, Haynes BF, Gao F, Liao HX, Kepler TB, et al. Affinity 
maturation in an HIV broadly neutralizing B-cell lineage through 
reorientation of variable domains. Proc Natl Acad Sci U S A. 
2014;111(28):10275-80. 
31. Bonsignori M, Zhou T, Sheng Z, Chen L, Gao F, Joyce MG, et al. Maturation 
Pathway from Germline to Broad HIV-1 Neutralizer of a CD4-Mimic 
Antibody. Cell. 2016;165(2):449-63. 
32. Coler RN, Baldwin SL, Shaverdian N, Bertholet S, Reed SJ, Raman VS, et 
al. A synthetic adjuvant to enhance and expand immune responses to 
influenza vaccines. PLoS One. 2010;5(10):e13677. 
33. Knudsen NP, Olsen A, Buonsanti C, Follmann F, Zhang Y, Coler RN, et al. 
Different human vaccine adjuvants promote distinct antigen-independent 
immunological signatures tailored to different pathogens. Sci Rep. 
2016;6:19570. 
34. Dagan R, Poolman J, Siegrist CA. Glycoconjugate vaccines and immune 
interference: A review. Vaccine. 2010;28(34):5513-23. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 111 of 151 
35. Phillips B, Van Rompay KKA, Rodriguez-Nieves J, Lorin C, Koutsoukos M, 
Tomai M, et al. Adjuvant-Dependent Enhancement of HIV Env-Specific 
Antibody Responses in Infant Rhesus Macaques. J Virol. 2018;92(20). 
36. Bemark M, Bergqvist P, Stensson A, Holmberg A, Mattsson J, Lycke NY. A 
unique role of the cholera toxin A1-DD adjuvant for long-term plasma and 
memory B cell development. J Immunol. 2011;186(3):1399-410. 
37. Li L, Honda-Okubo Y, Li C, Sajkov D, Petrovsky N. Delta Inulin Adjuvant 
Enhances Plasmablast Generation, Expression of Activation-Induced 
Cytidine Deaminase and B-Cell Affinity Maturation in Human Subjects 
Receiving Seasonal Influenza Vaccine. PLoS One. 2015;10(7):e0132003. 
38. Delgado MF, Coviello S, Monsalvo AC, Melendi GA, Hernandez JZ, Batalle 
JP, et al. Lack of antibody affinity maturation due to poor Toll-like receptor 
stimulation leads to enhanced respiratory syncytial virus disease. Nat Med. 
2009;15(1):34-41. 
39. van Haren SD, Ganapathi L, Bergelson I, Dowling DJ, Banks M, Samuels 
RC, et al. In vitro cytokine induction by TLR-activating vaccine adjuvants in 
human blood varies by age and adjuvant. Cytokine. 2016;83:99-109. 
40. Lewis DJ, Lythgoe MP. Application of "Systems Vaccinology" to Evaluate 
Inflammation and Reactogenicity of Adjuvanted Preventative Vaccines. J 
Immunol Res. 2015;2015:909406. 
41. Regules JA, Cicatelli SB, Bennett JW, Paolino KM, Twomey PS, Moon JE, 
et al. Fractional Third and Fourth Dose of RTS,S/AS01 Malaria Candidate 
Vaccine: A Phase 2a Controlled Human Malaria Parasite Infection and 
Immunogenicity Study. J Infect Dis. 2016;214(5):762-71. 
42. Fouda GG, Cunningham CK, McFarland EJ, Borkowsky W, Muresan P, 
Pollara J, et al. Infant HIV type 1 gp120 vaccination elicits robust and 
durable anti-V1V2 immunoglobulin G responses and only rare envelope-
specific immunoglobulin A responses. J Infect Dis. 2015;211(4):508-17. 
43. Phillips B, Fouda GG, Eudailey J, Pollara J, Curtis AD, 2nd, Kunz E, et al. 
Impact of Poxvirus Vector Priming, Protein Coadministration, and Vaccine 
Intervals on HIV gp120 Vaccine-Elicited Antibody Magnitude and Function 
in Infant Macaques. Clin Vaccine Immunol. 2017;24(10). 
44. Pegu P, Vaccari M, Gordon S, Keele BF, Doster M, Guan Y, et al. 
Antibodies with high avidity to the gp120 envelope protein in protection from 
simian immunodeficiency virus SIV(mac251) acquisition in an immunization 
regimen that mimics the RV-144 Thai trial. J Virol. 2013;87(3):1708-19. 
45. Lai L, Vodros D, Kozlowski PA, Montefiori DC, Wilson RL, Akerstrom VL, 
et al. GM-CSF DNA: An adjuvant for higher avidity IgG, rectal IgA, and 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 112 of 151 
increased protection against the acute phase of a SHIV-89.6P challenge by a 
DNA/MVA immunodeficiency virus vaccine. Virology. 2007;369(1):153-67. 
46. Xiao P, Patterson LJ, Kuate S, Brocca-Cofano E, Thomas MA, Venzon D, et 
al. Replicating adenovirus-simian immunodeficiency virus (SIV) 
recombinant priming and envelope protein boosting elicits localized, mucosal 
IgA immunity in rhesus macaques correlated with delayed acquisition 
following a repeated low-dose rectal SIV(mac251) challenge. J Virol. 
2012;86(8):4644-57. 
47. Adnan S, Reeves RK, Gillis J, Wong FE, Yu Y, Camp JV, et al. Persistent 
Low-Level Replication of SIVDeltanef Drives Maturation of Antibody and 
CD8 T Cell Responses to Induce Protective Immunity against Vaginal SIV 
Infection. PLoS Pathog. 2016;12(12):e1006104. 
48. Wang Y, Sundling C, Wilson R, O'Dell S, Chen Y, Dai K, et al. High-
Resolution Longitudinal Study of HIV-1 Env Vaccine-Elicited B Cell 
Responses to the Virus Primary Receptor Binding Site Reveals Affinity 
Maturation and Clonal Persistence. J Immunol. 2016;196(9):3729-43. 
49. Easterhoff D, Moody MA, Fera D, Cheng H, Ackerman M, Wiehe K, et al. 
Boosting of HIV envelope CD4 binding site antibodies with long variable 
heavy third complementarity determining region in the randomized double 
blind RV305 HIV-1 vaccine trial. PLoS Pathog. 2017;13(2):e1006182. 
50. Haynes BF, Kelsoe G, Harrison SC, Kepler TB. B-cell-lineage immunogen 
design in vaccine development with HIV-1 as a case study. Nat Biotechnol. 
2012;30(5):423-33. 
51. Haynes BF, Verkoczy L. AIDS/HIV. Host controls of HIV neutralizing 
antibodies. Science. 2014;344(6184):588-9. 
52. Williams WB, Zhang J, Jiang C, Nicely NI, Fera D, Luo K, et al. Initiation of 
HIV neutralizing B cell lineages with sequential envelope immunizations. 
Nat Commun. 2017;8(1):1732. 
53. Santini-Oliveira M, Coler RN, Parra J, Veloso V, Jayashankar L, Pinto PM, 
et al. Schistosomiasis vaccine candidate Sm14/GLA-SE: Phase 1 safety and 
immunogenicity clinical trial in healthy, male adults. Vaccine. 
2016;34(4):586-94. 
54. Treanor JJ, Essink B, Hull S, Reed S, Izikson R, Patriarca P, et al. Evaluation 
of safety and immunogenicity of recombinant influenza hemagglutinin 
(H5/Indonesia/05/2005) formulated with and without a stable oil-in-water 
emulsion containing glucopyranosyl-lipid A (SE+GLA) adjuvant. Vaccine. 
2013;31(48):5760-5. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 113 of 151 
55. Falloon J, Ji F, Curtis C, Bart S, Sheldon E, Krieger D, et al. A phase 1a, 
first-in-human, randomized study of a respiratory syncytial virus F protein 
vaccine with and without a toll-like receptor-4 agonist and stable emulsion 
adjuvant. Vaccine. 2016;34(25):2847-54. 
56. Falloon J, Talbot HK, Curtis C, Ervin J, Krieger D, Dubovsky F, et al. Dose 
Selection for an Adjuvanted Respiratory Syncytial Virus F Protein Vaccine 
for Older Adults Based on Humoral and Cellular Immune Responses. Clin 
Vaccine Immunol. 2017;24(9). 
57. Coler RN, Duthie MS, Hofmeyer KA, Guderian J, Jayashankar L, Vergara J, 
et al. From mouse to man: safety, immunogenicity and efficacy of a 
candidate leishmaniasis vaccine LEISH-F3+GLA-SE. Clinical & 
Translational Immunology [Internet]. 2015 6/10/2015 doi:10.1038/cti.2015.6; 
4:[e35 p.]. Available from: 
http://www.nature.com/cti/journal/v4/n4/full/cti20156a.html. 
58. Rerks-Ngarm S, Pitisuttithum P, Nitayaphan S, Kaewkungwal J, Chiu J, Paris 
R, et al. Vaccination with ALVAC and AIDSVAX to prevent HIV-1 
infection in Thailand. N Engl J Med. 2009;361(23):2209-20. 
59. Robb ML, Rerks-Ngarm S, Nitayaphan S, Pitisuttithum P, Kaewkungwal J, 
Kunasol P, et al. Risk behaviour and time as covariates for efficacy of the 
HIV vaccine regimen ALVAC-HIV (vCP1521) and AIDSVAX B/E: a post-
hoc analysis of the Thai phase 3 efficacy trial RV 144. Lancet Infect Dis. 
2012;12(7):531-7. 
60. Haynes BF, Gilbert PB, McElrath MJ, Zolla-Pazner S, Tomaras GD, Alam 
SM, et al. Immune-correlates analysis of an HIV-1 vaccine efficacy trial. N 
Engl J Med. 2012;366(14):1275-86. 
61. Yates NL, Liao HX, Fong Y, DeCamp A, Vandergrift NA, Williams WT, et 
al. Vaccine-induced Env V1-V2 IgG3 correlates with lower HIV-1 infection 
risk and declines soon after vaccination. Sci Transl Med. 
2014;6(228):228ra39. 
62. Gottardo R, Bailer RT, Korber BT, Gnanakaran S, Phillips J, Shen X, et al. 
Plasma IgG to linear epitopes in the V2 and V3 regions of HIV-1 gp120 
correlate with a reduced risk of infection in the RV144 vaccine efficacy trial. 
PLoS ONE. 2013;8(9):e75665. 
63. Zolla-Pazner S, Decamp AC, Cardozo T, Karasavvas N, Gottardo R, 
Williams C, et al. Analysis of V2 antibody responses induced in vaccinees in 
the ALVAC/AIDSVAX HIV-1 vaccine efficacy trial. PLoS ONE. 
2013;8(1):e53629. 
64. Tomaras GD, Ferrari G, Shen X, Alam SM, Liao HX, Pollara J, et al. 
Vaccine-induced plasma IgA specific for the C1 region of the HIV-1 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 114 of 151 
envelope blocks binding and effector function of IgG. Proc Natl Acad Sci U 
S A. 2013;110(22):9019-24. 
65. Cunningham CK, Wara DW, Kang M, Fenton T, Hawkins E, McNamara J, et 
al. Safety of 2 recombinant human immunodeficiency virus type 1 (HIV-1) 
envelope vaccines in neonates born to HIV-1-infected women. Clin Infect 
Dis. 2001;32(5):801-7. 
66. McFarland EJ, Johnson DC, Muresan P, Fenton T, Tomaras GD, McNamara 
J, et al. HIV-1 vaccine induced immune responses in newborns of HIV-1 
infected mothers. AIDS. 2006;20(11):1481-9. 
67. McGuire EP, Fong Y, Toote C, Cunningham CK, McFarland EJ, Borkowsky 
W, et al. HIV-Exposed Infants Vaccinated with an MF59/Recombinant 
gp120 Vaccine Have Higher-Magnitude Anti-V1V2 IgG Responses than 
Adults Immunized with the Same Vaccine. J Virol. 2018;92(1). 
68. Levy O, Goriely S, Kollmann TR. Immune response to vaccine adjuvants 
during the first year of life. Vaccine. 2013;31(21):2500-5. 
69. Kollmann TR, Levy O, Montgomery RR, Goriely S. Innate immune function 
by Toll-like receptors: distinct responses in newborns and the elderly. 
Immunity. 2012;37(5):771-83. 
70. Dowling DJ, Levy O. Pediatric Vaccine Adjuvants: Components of the 
Modern Vaccinologist's Toolbox. Pediatr Infect Dis J. 2015;34(12):1395-8. 
71. Borriello F, van Haren SD, Levy O. First International Precision Vaccines 
Conference: Multidisciplinary Approaches to Next-Generation Vaccines. 
mSphere. 2018;3(4). 
72. Lambert JS, McNamara J, Katz SL, Fenton T, Kang M, VanCott TC, et al. 
Safety and immunogenicity of HIV recombinant envelope vaccines in HIV-
infected infants and children. National Institutes of Health-sponsored 
Pediatric AIDS Clinical Trials Group (ACTG-218). J Acquir Immune Defic 
Syndr Hum Retrovirol. 1998;19(5):451-61. 
73. Johnson DC, McFarland EJ, Muresan P, Fenton T, McNamara J, Read JS, et 
al. Safety and immunogenicity of an HIV-1 recombinant canarypox vaccine 
in newborns and infants of HIV-1-infected women. J Infect Dis. 
2005;192(12):2129-33. 
74. Tameris MD, Hatherill M, Landry BS, Scriba TJ, Snowden MA, Lockhart S, 
et al. Safety and efficacy of MVA85A, a new tuberculosis vaccine, in infants 
previously vaccinated with BCG: a randomised, placebo-controlled phase 2b 
trial. Lancet. 2013;381(9871):1021-8. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 115 of 151 
75. Agresti A, Coull BA. Approximate is better than "exact" for interval 
estimation of binomial proportions. Am Stat. 1998;52(2):119-26. 
76. Lynch RM, Tran L, Louder MK, Schmidt SD, Cohen M, Dersimonian R, et 
al. The Development of CD4 Binding Site Antibodies During HIV-1 
Infection. J Virol. 2012;86(14):7588-95. 
77. Lydersen S, Fagerland MW, Laake P. Recommended tests for association in 
2 x 2 tables. Stat Med. 2009;28(7):1159-75. 
78. Hudgens MG. Estimating cumulative probabilities from incomplete 
longitudinal binary responses with application to HIV vaccine trials. Stat 
Med. 2003;22(3):463-79. 
79. Lachenbruch PA. Comparisons of two-part models with competitors. Stat 
Med. 2001;20(8):1215-34. 
80. Hughes JP. Mixed effects models with censored data with application to HIV 
RNA levels. Biometrics. 1999;55(2):625-9. 
81. Rotnitzky A, Robins J. Analysis of semi-parametric regression models with 
non-ignorable non-response. Stat Med. 1997;16(1-3):81-102. 
82. Rotnitzky A, Robins JM. Semiparametric regression estimation in the 
presence of dependent censoring. Biometrika. 1995;82(4):805-20. 
83. Gilbert PB, Sato A, Sun X, Mehrotra DV. Efficient and robust method for 
comparing the immunogenicity of candidate vaccines in randomized clinical 
trials. Vaccine. 2009;27(3):396-401. 
84. James G, Witten D, Hastie T, Tibshirani R. An introduction to statistical 
learning with applications in R. New York: R. Springer; 2013. 
85. Howie SR. Blood sample volumes in child health research: review of safe 
limits. Bull World Health Organ. 2011;89(1):46-53. 
86. Itell HL, McGuire EP, Muresan P, Cunningham CK, McFarland EJ, 
Borkowsky W, et al. Development and application of a multiplex assay for 
the simultaneous measurement of antibody responses elicited by common 
childhood vaccines. Vaccine. 2018;36(37):5600-8. 
87. Seaman M, Janes H, Hawkins N, Grandpre L, Devoy C, Giri A, et al. Tiered 
Categorization of a Diverse Panel of HIV-1 Env Pseudoviruses for 
Assessment of Neutralizing Antibodies. J Virol. 2010;84(3):1439-52. 
88. DeCamp A, Hraber P, Bailer RT, Seaman MS, Ochsenbauer C, Kappes J, et 
al. Global panel of HIV-1 Env reference strains for standardized assessments 
of vaccine-elicited neutralizing antibodies. J Virol. 2014;88(5):2489-507. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 116 of 151 
89. Pollara J, Hart L, Brewer F, Pickeral J, Packard BZ, Hoxie JA, et al. High-
throughput quantitative analysis of HIV-1 and SIV-specific ADCC-mediating 
antibody responses. Cytometry A. 2011;79(8):603-12. 
90. Fisher L, Zinter M, Stanfield-Oakley S, Carpp LN, Edwards RW, Denny T, 
et al. Vaccine-Induced Antibodies Mediate Higher Antibody-Dependent 
Cellular Cytotoxicity After Interleukin-15 Pretreatment of Natural Killer 
Effector Cells. Front Immunol. 2019;10:2741. 
91. Tay MZ, Liu P, Williams LD, McRaven MD, Sawant S, Gurley TC, et al. 
Antibody-Mediated Internalization of Infectious HIV-1 Virions Differs 
among Antibody Isotypes and Subclasses. PLoS Pathog. 
2016;12(8):e1005817. 
92. Ackerman ME, Moldt B, Wyatt RT, Dugast AS, McAndrew E, Tsoukas S, et 
al. A robust, high-throughput assay to determine the phagocytic activity of 
clinical antibody samples. J Immunol Methods. 2011;366(1-2):8-19. 
93. Worley MJ, Fei K, Lopez-Denman AJ, Kelleher AD, Kent SJ, Chung AW. 
Neutrophils mediate HIV-specific antibody-dependent phagocytosis and 
ADCC. J Immunol Methods. 2018;457:41-52. 
94. Perez LG, Martinez DR, deCamp AC, Pinter A, Berman PW, Francis D, et al. 
V1V2-specific complement activating serum IgG as a correlate of reduced 
HIV-1 infection risk in RV144. PLoS One. 2017;12(7):e0180720. 
95. Chung AW, Kumar MP, Arnold KB, Yu WH, Schoen MK, Dunphy LJ, et al. 
Dissecting Polyclonal Vaccine-Induced Humoral Immunity against HIV 
Using Systems Serology. Cell. 2015;163(4):988-98. 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 117 of 151 
Appendix A 
Sample informed consent form 
Title: A phase 1 clinical trial to evaluate the safety and immunogenicity of the 
HIV-1 CH505 transmitted/founder gp120 adjuvanted with GLA-SE in healthy, 
HIV-exposed uninfected infants 
HVTN protocol number: HVTN 135 
Site: [Insert site name] 
Thank you for your interest in our research study. Please read this consent form or 
ask someone to read it to you. If you decide to join the study, we will ask you to 
sign or make your mark on this form. We will offer you a copy to keep. We will 
ask you questions to see if we have explained everything clearly. You can also 
ask us questions about the study. 
Research is not the same as treatment or medical care. The purpose of a research 
study is to answer scientific questions. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 118 of 151 
Key Information  
These are some of the things you should know about this study: 
The HIV Vaccine Trials Network 
(HVTN) and PHRU are doing a study to 
test an HIV vaccine in babies. The study 
vaccine contains a protein called 
CH505TF gp120 and an adjuvant called 
GLA-SE. An adjuvant is a substance 
added to a vaccine to increase the 
body’s immune response. We want to 
see if the vaccine is safe to give to 
babies and does not make them too 
uncomfortable. We also want to see if 
babies’ immune systems respond to the 
study vaccine and in the same way that 
adults respond to the study vaccine.  
• About 38 uninfected babies and 
their HIV-infected mothers (aged 
18 and older) will be enrolled at 
one site, the PHRU clinic 
• About 14 visits over 2 ½ years 
• Babies will get 1 injection into 
the upper thigh at 5 separate 
visits over about 1 ½ years  
• Babies will get either the study 
vaccine or placebo (a substance 
that does not contain vaccine) 
• That is a 3-in-4 chance of your 
baby getting the study vaccine 
This study is divided into three parts: Part A, B and C. Parts A and B both include 
two groups and Part C contains 4 groups. 7 babies will take part in Part A of this 
study. After all 7 babies have joined the study and received 1 vaccination, we will 
decide whether or not to do Part B 
4 babies will join Part B. After the 4 babies in part B each receive 1 vaccine, we 
will determine if it is safe for these babies to continue receiving additional 
vaccines. After the 4 babies in Part B have all received 3 vaccinations, we will 
decide whether or not to do Part C. 
If we decide that the study vaccine is safe to give to babies in Part B, then we will 
do Part C. If we do Part C, 27 more babies will join. 
If you choose to join the study with your baby, here are some things that will 
happen: 
• We will request your medical records, including your HIV status 
• Take blood from you and your baby, including using blood from the 
umbilical cord that would otherwise be discarded 
• Physical exams of you and your baby 
• Test you for Hepatitis B, SARS-CoV-2 (if not in your medical records), 
your viral load, CD4 and T cell counts 
• Test your baby for HIV 
• Ask about how much breastmilk, formula or solid food your baby is eating 
There is no direct benefit to you or your baby from being in this study. Whether or 
not you and your baby join this study is up to you. You and your baby do not have 
to take part in this study. If you and your baby do not join this study, this will not 
affect the care that you and your baby receive at this clinic. 
Some general risks of vaccines include fever, chills, rash, aches and pains, nausea, 
headache, dizziness, and feeling tired. Vaccines can also cause pain, redness, 
swelling, or itching where your baby receives the injection. There may be other 
side effects that we don’t yet know about, even serious ones. 
The rest of this form provides a more complete description of this study. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 119 of 151 
About the study 
The HIV Vaccine Trials Network (HVTN) and PHRU are doing a study to test an 
HIV vaccine. HIV is the virus that causes AIDS. 
About 38 babies and their mothers (aged 18 and older) will take part in this study 
at one site, the PHRU clinic. The researcher in charge of this study at this clinic is 
Dr. Avy Violari. The United States (US) National Institutes of Health (NIH) is 
paying for the study. 
This study is divided into three parts: Part A, B and C. Parts A and B are divided 
into two groups. Seven babies will take part in Part A of this study. After all seven 
babies have joined the study and received one vaccination, we will decide 
whether or not to do Part B. We will be looking to see if the study vaccine is safe 
to give to babies in Part A. If we decide that the study vaccine is safe to give to 
babies in Part A, we will do Part B. Four babies will join Part B. After all four 
babies in Part B receive their first vaccination, we will decide whether to continue 
their vaccinations. Again, we will be looking to see if the study vaccine is safe to 
give to babies in Part B. After all four babies in Part B receive their third 
vaccination, we will decide whether or not to do Part C. If we decide that the 
study vaccine is safe to give to babies in Part B, then we will do Part C. Part C is 
divided into 4 groups. If we do Part C, 27 more babies will join. 
If you decide to join the study, you may be in any of these groups depending on 
when you joined the study. 
1. We are doing this study to answer several questions. 
• Is the study vaccine safe to give to babies? 
• Are babies able to take the study vaccine without becoming too 
uncomfortable? 
• How do babies’ immune systems respond to the study vaccine? (Your 
immune system protects you from disease.) Do babies respond in the same 
way that adults respond to the study vaccine?  
• How does getting the study vaccine effect babies’ response to other vaccines 
they may get early in life? 
2. The study vaccine cannot give babies HIV. 
The study vaccine is not made from actual HIV. It is impossible for the study 
vaccine to give babies HIV. Also, it cannot cause babies to give HIV to someone 
else. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 120 of 151 
3. We do not know if the study vaccine will decrease, increase, or not change 
babies’ risk of getting HIV if they are exposed to the virus. 
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. 
Studies of similar products involving thousands of adults were found to be safe 
and did not affect the risk of getting HIV. 
Several studies have tested whether HIV vaccines can reduce the risk of getting 
HIV from another person. In some studies, people who got the vaccine seemed to 
have the same risk of getting HIV as people who did not get the vaccine. In one 
study, people who got the vaccine seemed to have a lower risk of getting HIV 
than people who did not get the vaccine. In studies with a different vaccine, some 
people who got the vaccine had a higher risk of getting HIV than people who did 
not get the vaccine. This study differs from the studies in which people who got 
the vaccine had a higher or lower risk of getting HIV. The study staff can tell you 
about the differences. 
We do not know whether the vaccine in this study will affect babies’ risk of 
getting HIV. It’s very important for mothers to continue to take their ARVs to 
minimize the risk of HIV transmission through breast milk. We will tell you how 
you can reduce the risk of giving your baby HIV. 
4. This study vaccine is experimental. 
The study vaccine is called CH505TF gp120. From here on, we will call it CH505 
or the study vaccine. It is an experimental HIV vaccine. That means we do not 
know if the study vaccine will be safe to use in babies, or if it will work to prevent 
HIV infection. This study vaccine is used only in research studies. 
CH505 was developed by Duke Human Vaccine Institute and National Institute of 
Allergy & Infectious Disease (NIAID). The study vaccine has man-made pieces 
of protein that look like part of the protein found in HIV. A baby’s immune 
system might learn to recognize these proteins and prepare itself to fight HIV. 
This is called an immune response. 
The study vaccine has been given to about 36 adults in another study that is 
ongoing. In the ongoing study in adults, the study vaccine appears safe and people 
have been able to take it without becoming too uncomfortable. The study vaccine 
has not been given to babies before. In animal studies, the study vaccine appears 
safe. Even if something looks like it is safe or works in animals, it may not be true 
for people. 
The study vaccine is mixed with an adjuvant. An adjuvant is a substance that 
should help the immune system respond better. The adjuvant in this study is 
called GLA-SE. GLA-SE was developed by the Infectious Disease Research 
Institute (IDRI). 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 121 of 151 
General risks of vaccines: 
All vaccines can cause fever, chills, rash, aches and pains, nausea, headache, 
dizziness, fussiness, and feeling tired. Vaccines can also cause pain, redness, 
swelling, or itching where you got the injection. Most people can still do their 
planned activities after getting a vaccine. Rarely, people have side effects that 
limit their normal activities or make them go to the doctor. 
Rarely, a vaccine can cause an allergic reaction, including a rash, hives, or trouble 
breathing. Allergic reactions can be life-threatening. You should tell us if your 
baby has ever had a bad reaction to any injection or vaccine. 
Very rarely, a vaccine causes an autoimmune disease in a person, or makes an 
autoimmune disease worse. An autoimmune disease happens when your immune 
system attacks your own body, instead of attacking an infection. 
Risks of the study vaccine: 
This section lists the side effects we know about. There may be others that we 
don’t yet know about, even serious ones. We will tell you if we learn about any 
new side effects. 
The study vaccine has not been given to babies before, but has been given to 
adults in other studies. The most common complaints were pain or tenderness 
where they got the injection. In those studies, two people had reddening and 
hardening of their skin where they got the injection, but it went away in a few 
days. One of those people had a skin infection where they got the injection. It did 
not affect that person’s daily routine. That person took some medicine and it got 
better within 5 days. Two other people in those studies had bad headaches. Let the 
study staff know if your baby is fussier or is crying more than usual. 
The adjuvant, GLA-SE, has been tested in about 2,500 adults with vaccines for 
other diseases. The most common complaints were pain and tenderness at the 
injection site and feeling tired. GLA-SE is currently being tested in children and 
has not yet been tested in babies. We do not know how the babies will react to the 
study vaccine or adjuvant.  
Joining the study 
5. It is completely up to you whether or not to join the study with your baby. 
Take your time in deciding. If it helps, talk to people you trust, such as your 
doctor, friends or family. If you decide not to join this study, or if you and your 
baby leave it after you have joined, you and your baby’s other care at this clinic 
and the benefits or rights you would normally have will not be affected. 
If you and your baby join this study, your baby may not be allowed to join other 
HIV vaccine or HIV prevention studies now or in the future. You and your baby 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 122 of 151 
cannot be in this study while you are in another study where either of you get a 
study product. Being in more than one study may not be safe.  
Also during the study, your baby should not donate blood or tissue. 
If you choose not to join this study with your baby, you or your baby may be able 
to join another study. 
6. If you want to join the study with your baby, we will screen you and your 
baby to see if you are eligible. 
Screening involves a physical exam and a health history from both you and your 
baby. A physical exam may include, but is not limited to: 
• Checking your and your baby’s weight, temperature and blood pressure 
• Looking in your and your baby’s mouth and throat 
• Listening to your and your baby’s heart and lungs 
• Feeling your and your baby’s abdomen (stomach and liver) 
• Doing a genital exam on your baby (which includes touching that area on the 
outside) 
We will also do blood tests on both you and your baby. We will draw blood from 
your vein using a needle for these tests. After the baby is delivered, we will take 
blood samples from the part of the umbilical cord that is discarded. We will test 
your baby for HIV. The tests on your and your baby’s blood tell us about some 
aspects of you and your baby’s health, such as how healthy each of your kidneys, 
liver, and immune systems are. We may also test you for Hepatitis B and SARS-
CoV-2, test your viral load, CD4 and T cell counts. We will ask you about 
medications you and your baby are taking, including your antiretroviral 
medications.  
We will ask your doctor for your or your baby’s medical records, including your 
HIV status so that we can reduce the amount of tests that we need to do for the 
study. 
We will review the screening results with you. The screening results may show 
you and your baby are not eligible to join the study, even if you want to. If you 
and your baby are not eligible to join the study, any samples that have been taken 
will be destroyed.  
Site: adapt the following section so it is applicable to the care available at your site 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 123 of 151 
7. If we find that you or your baby 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 explain how we will help you get care 
elsewhere. For health problems that are unrelated to the study, we will not pay for 
care. 
Being in the study 
If you and your baby meet the study requirements and want to join, here is what 
will happen: 
8. You and your baby will come to the clinic for scheduled visits about [#] times 
over about two years. 
Site: 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 and your baby may have to come for more visits if either of you have a lab or 
health issue. 
We may contact you after the main study ends (for example, to tell you about the 
study results). 
9. We will give you [Site: Insert compensation] for each study visit you 
complete. 
This amount is to cover the costs of [Site: Insert text] 
You do not have to pay anything to be in this study. 
10. We will give your baby either the study vaccine or a placebo. 
Not all babies in this study will get the study vaccine. Some babies will get a 
placebo, a substance that does not contain vaccine. We will compare the results 
from babies who got the placebo with results from babies who got the study 
vaccine. In this study, the placebo is sterile salt water. 
Your baby has about a 3-in-4 chance of getting the study vaccine. Site: Modify the 
randomization metaphor in the next sentence as appropriate to your local culture. 
Whether your baby gets the study vaccine or the placebo is completely random, 
like flipping a coin. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 124 of 151 
We have no say in whether your baby gets the study vaccine or the placebo. We 
will not know which one your baby is getting, and neither will you. Only the 
pharmacist at this clinic will have this information while the study is going on. 
You will have to wait until everyone completes their final study visits to find out 
whether your baby got the study vaccine or the placebo. This could be several 
years. But, if your baby has a serious medical problem and you need to know 
what they got before the end of the study, we can tell you. 
11. We will give your baby the study products on a schedule. 
All babies will get one injection into the upper thigh at each of 5 separate visits. 
Your baby will be in one of three parts: Part A, B or C. Parts A and B are divided 
into two groups. Seven babies will take part in Part A of this study. Five of the 
seven babies will get the study vaccine and the lower dose of the adjuvant. The 
other two babies will get a placebo. After all seven babies have joined the study 
and received one vaccination, we will decide whether or not to do Part B. We will 
be looking to see if the study vaccine is safe to give to babies in Part A. If we 
decide that the study vaccine is safe to give to babies in Part A, we will do Part B.  
If we decide to do Part B, four more babies will join. Two of the four babies will 
get a dose of the study vaccine and a higher dose of the adjuvant. The other two 
babies will get a placebo. After all four babies in Part B receive their first 
vaccination, we will decide whether to continue their vaccinations. Again, we will 
be looking to see if the study vaccine is safe to give to babies in Part B. After all 
four babies in Part B have received three vaccinations, we will decide whether or 
not to open Part C. Again, we will be looking to see if the study vaccine is safe to 
give to babies in Part B. If we decide that the study vaccine is safe to give to 
babies in Part B, then we will do Part C.  
Part C is divided into 4 groups. If we decide to do Part C, 27 more babies will 
join. In Part C, 16 babies will get the study vaccine and the higher dose of the 
adjuvant. Also in Part C, five babies will get the lower dose of the study vaccine 
and the higher dose of the adjuvant. Six babies in Part C will get a placebo. 
 
Site: If a picture version of the injection schedule has been provided in a separate 
protocol appendix, you may insert it below in place of (or in addition to) the text 
version or give it as a separate document to volunteers if you believe it will be helpful 
to them. You are not required to do either. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 125 of 151 
 
 
 
Injection schedule in months  
Group 
# of 
babies 
Dose 
(protein/ 
adjuvant) 
0  
2  
4  
7 ½  
12 ½  
 
 
 
 
 
 
 
 
Part 
A 
 
 
 
 
 
 
 
1 
5 
 
Study vaccine 20 mcg /Low 
adjuvant 
2.5 mcg  
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
2 
2 
 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
 
 
 
 
 
 
 
 
Part 
B 
 
 
 
 
 
 
 
3 
2 
Study vaccine 20 mcg /High 
adjuvant  
5 mcg  
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
4 
2 
 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
 
 
 
 
 
 
 
 
Part 
C 
 
 
 
 
 
 
 
5 
16 
Study vaccine 20 mcg / 
High adjuvant 5 mcg  
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
6 
3 
 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
 
 
 
 
 
 
 
 
7 
5 
 
Low study vaccine 5 mcg 
/High adjuvant 
5 mcg  
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
Study 
Vaccine 
8 
3 
 
Placebo 
Placebo 
Placebo 
Placebo 
Placebo 
 
 
 
 
 
 
 
 
38 babies total (28 get vaccine and 10 get placebo) 
You and your baby will have to wait in the clinic for at least an hour after each 
injection to see if there are any problems. Then for that night and for 7 more days, 
you will need to keep track of how your baby is feeling and if they have any 
symptoms. Site: Customize the next sentence based on how you collect 
reactogenicity information. You will bring this information back to the clinic at 
your next visit. Within 3 days of each injection, we will also need to be in contact 
with you to ask how your baby is doing. Contact the clinic staff if you have any 
issues or concerns after your baby gets an injection. If they have a problem, we 
will continue to check on it until it goes away. 
If something happens to you and you are unable to take your baby to the study 
visits, a caregiver can bring your baby instead. 
12. In addition to giving your baby the study products, over the course of the 
trial, we will: 
• Do HIV testing of your baby, as well as counseling you on how you can avoid 
giving HIV to your baby 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 126 of 151 
• Do physical exams of your baby 
• Ask questions about you and your baby’s health, including medications each 
of you may be taking, and how much breastmilk, formula, or solid food your 
baby is eating 
• Ask questions about any personal problems or benefits you and your baby 
may have from being in the study 
• Take blood samples from both you and your baby by drawing blood from the 
vein using a needle to do various tests, including checking your viral load. If 
during the study your viral load goes above 400 copies per milliliter we will 
check to see if this is due to drug resistance. If this happens, we or your doctor 
may have to change your ART medication.  
When we take blood, the amount will depend on the lab tests we need to do and 
the baby’s weight. For your baby, it will be some amount between 2 mL and 27 
mL (1/2 of a teaspoon to just less than 3 tablespoons). For you, it will be some 
amount between 10 mL and 40 mL (2 teaspoons to 4 tablespoons). Your and your 
baby’s body will make new blood to replace the blood we take out. 
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. 
Site: Insert Appendix C, Table of procedures (for informed consent form) in this 
section. You may also distribute it as a separate sheet to your participants if that is 
helpful.  
We will be looking for side effects. We will review the results of these procedures 
and tests with you at your next visit, or sooner if necessary. If any of the results 
are important to you or your baby’s health, we will tell you. 
13. We would also like to collect stool samples and breast milk samples. 
We would like to collect a few small samples of your and your baby’s stool to 
look at the bacteria living in each of your stomachs. We would also like to collect 
two samples of your breast milk to look at the antibodies in it. We want to learn 
more about how a mother’s immune system affects her baby’s immune system. 
We would collect a total of eight stool samples from your baby at different visits, 
but we would collect only one stool sample from you. You and your baby can 
give stool samples without having any extra clinic visits. You can see the table 
above to find out which visits we would collect stool from you and your baby.  
We would collect your breast milk at screening and at the 4 ½ month visit. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 127 of 151 
If you agree, we will give you more information about how we will collect these 
samples. You do not have to give any of these samples if you choose. Or you can 
choose to give all of these samples and change your mind later. At the end of this 
consent form, we will ask if you allow us to collect these samples.  
14. We will counsel you about protecting your baby from HIV. 
We will talk with you about ways to keep your risk of giving your baby HIV low. 
15. The HVTN will test your and your baby’s samples for this study. 
We will send your and your baby’s samples (without your names) to labs 
approved by the HVTN for this study, which are located in the United States and 
South Africa. In rare cases, some of your and your baby’s samples may be sent to 
labs approved by the HVTN in other countries for research related to this study. 
Researchers may also do genetic testing related to this study on your and your 
baby’s samples. Your and your baby’s genes are passed to each of you from your 
birth parents. They affect how you and your baby looks and how your and your 
baby’s body works. The differences in people’s genes can help explain why some 
people get a disease while others do not. The genetic testing will only involve 
some of your and your baby’s genes, not all of your and your baby’s genes (your 
genomes). The researchers will study only the genes related to the immune system 
and HIV and those that affect how people get HIV. 
In some cases, researchers may take cells from your and your baby’s samples and 
grow more of them over time, so that they can continue to contribute to this study. 
These tests done on your and your baby’s samples are for research purposes, not 
to check your and your baby’s 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 and your baby’s samples are no longer needed for this study, the 
HVTN will continue to store them. 
16. We will do our best to protect your and your baby’s private information. 
Your and your baby’s study records and samples will be kept in a secure location. 
We will label all of your and your baby’s samples and most of the records with a 
code number, not your or your baby’s name or other personal information. 
However, it is possible to identify you or your baby, if necessary. We will not 
share your or your baby’s name with the lab that does the tests on the samples, or 
with anyone else who does not need to know your or your baby’s name. 
Site: Any change to the following boxed text requires approval from HVTN 
Regulatory Affairs. You can remove the box around the text.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 128 of 151 
We do need to share your baby’s name with the HVTN in case you or your baby 
needs proof in the future that your baby participated in an HIV vaccine study. The 
HVTN will keep your baby’s name in a secure file with these items:  
• The name of the study 
• Your baby’s age or date of birth 
• Your baby’s study ID number 
• What study product they received 
There are no HIV test results kept in this file. The HVTN will not share any 
information that could identify your baby without your agreement. The HVTN 
will remove your baby’s name from the file if you do not want it there. Once your 
baby is no longer a minor (at least 18 years old), they also can ask the HVTN to 
remove their name from the file if they do not want it there. 
Clinic staff will have access to your and your baby’s study records. Both of your 
records may also be reviewed by groups who watch over this study to see that we 
are protecting your and your baby’s rights, keeping you both safe, and following 
the study plan. These groups include: 
• The US National Institutes of Health (NIH) and its study monitors, 
• Any regulatory agency that reviews clinical trials, 
• The University of the Witwatersrand Human Research Ethics Committee, 
• South African Health Products Regulatory Authority (SAHPRA), 
• Duke Human Vaccine Institute (DHVI) and people who work for them, 
• The Infectious Disease Research Institute (IDRI) and people who work for 
them, 
• The HVTN and people who work for them, 
• The Safety Monitoring Board (SMB) and 
• The US Office for Human Research Protections (OHRP). 
All reviewers will take steps to keep your records private. 
We cannot guarantee absolute privacy. If you are found to 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:  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 129 of 151 
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] 
• COVID-19: At this clinic, we have to report COVID-19 test results to the 
National Institute of Communicable Diseases (NICD) and the national and 
provincial Department of Health (DoH) which includes your age and gender. 
The DoH may also require that we share additional demographic information 
about your COVID-19 test results to support contact tracing. 
• [Item 3] 
The results of this study may be published. No publication will use you or your 
baby’s name or identify either of you personally. 
We may share information from the study with other researchers. We will not 
share you or your baby’s name or information that can identify either of you. 
A description of this clinical trial will be available on 
http://www.ClinicalTrials.gov. This website will not include information that can 
identify you. At most, the website will include a summary of the results. You can 
search this website at any time. 
17. We will stop your baby’s injections if they get HIV.  
We will also take fewer samples, and we will help you get care and support for 
your baby. We may ask you to keep your baby in the study to complete other 
study procedures. We will encourage you to keep your baby in the study if you 
choose. We will discuss your baby’s study options with you. We will counsel you 
about your baby having HIV. Your baby's HIV care will be free of charge and 
will follow the standard of care in South Africa. This clinic is accredited by the 
Department of Health which means that if your baby gets HIV, your baby can 
receive ART and laboratory monitoring at this clinic.  
18. We may take you and your baby out of the study at any time.  
We may take you and your baby out of the study if: 
• you do not follow instructions, 
• we think that staying in the study might harm you or your baby, 
• you enroll your baby in a different research study where they get another 
study product, or 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 130 of 151 
• the study is stopped for any reason. 
Other Risks 
19. There are other risks to being in this study. 
This section describes the other risks and restrictions we know about. There may 
also be unknown risks, even serious ones. We will tell you if we learn anything 
new that may affect your willingness to stay in the study. 
Risks of routine medical procedures: 
In this study, we will do some routine medical procedures. These are taking blood 
and giving injections. These procedures can cause bruising, pain, fainting, 
soreness, redness, swelling, itching, a sore, bleeding, and (rarely) muscle damage 
or infection where your baby got the injection. Taking blood can cause a low 
blood cell count (anemia), making you or your baby feel tired. 
Personal problems/discrimination/testing HIV antibody positive: 
About 10 to 20% of people who join HVTN studies report personal problems or 
discrimination because of joining an HIV vaccine study. Family or friends may 
worry, get upset or angry, and treat you or your baby unfairly as a result. Rarely, a 
person has lost a job because the study took too much time away from work. 
The body makes antibodies to fight or prevent infection. As you know, HIV-
infected moms transfer antibodies to their babies which disappear over time. That 
is why we do special tests to find out whether your baby is HIV-infected or not. 
Most vaccines cause the body to make antibodies as a way of preventing 
infection. Your baby’s body may make antibodies to HIV because they received 
an HIV study vaccine. The study vaccine may also cause your baby to test 
positive on some types of HIV antibody tests, even if they do not have HIV. This 
is called vaccine-induced seropositivity (VISP). VISP means that after your baby 
gets the study vaccine, a routine HIV test done outside this clinic may say they 
have HIV, even if they don’t. For this reason, you should plan to get your baby 
HIV tested only at this clinic after your baby is 18 months old until it is 
confirmed they do not have VISP. Our tests can tell the difference between true 
HIV infection and a positive result that is caused by the study vaccine. 
If your baby has a positive test result caused by the study vaccine at any time, we 
can arrange free HIV testing for as long as your baby needs it. If this happens, we 
do not know how long your baby will test positive due to the study vaccine. If 
your baby receives a positive HIV test result and we determine it is because they 
have HIV, we will provide follow-up care for your baby. 
It is unlikely, but your baby could test negative at the end of the study and VISP 
some time later, even though they don’t have HIV. This could happen if different 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 131 of 151 
HIV tests come into use. We will give you a phone number to call for more 
information. 
Site: Modify the following paragraph if applicable. If someone believes your baby 
has HIV even if they do not, they could face discrimination and other problems. 
For example, in some countries, they could be denied medical or dental care, 
employment, insurance, a visa, or entry into the military. If your baby has a 
positive HIV antibody test caused by the study vaccine, they will not be able to 
donate blood or organs. Your family and friends may treat them differently. We 
will give you a brochure that tells you more about testing HIV positive because of 
an HIV vaccine, and how your baby can avoid some of these problems. 
Risks of embarrassment/anxiety/emotional stress: 
You may feel embarrassed when we ask about your HIV status and ARV usage. 
Also, waiting for your baby’s HIV test results or other health test results could 
make you feel anxious. You could feel worried if the test results show that your 
baby has HIV or if tests show that you have SARS-CoV-2. If you feel 
embarrassed or anxious, please tell us and we will try to help you. 
Risks of the nasal swab/throat swab procedure: 
If we test you for SARS-CoV-2, we will either use a nasal swab or throat swab to 
collect the specimen. The feeling of having a small, soft-tipped swab inserted into 
your nostril and twirled around may be a little uncomfortable, but it should not be 
painful. There is a small chance there could be some bleeding, but this is unlikely. 
Similarly, the feeling of having a small throat swab inserted into your throat may 
cause gagging and feel a little uncomfortable, but it should not be painful. 
Risks of disclosure of your personal information: 
We will take several steps to protect you and your baby’s personal information. 
Although the risk is very low, it is possible that your or your baby’s personal 
information could be given to someone who should not have it. If that happened, 
you or your baby could face discrimination, stress, and embarrassment. We can 
tell you more about how we will protect you and your baby’s personal 
information if you would like it. 
Risks of genetic testing: 
There may be some risks from tests of your or your baby’s genes. If others found 
out the results of these tests, they could treat you or your baby badly or 
unfairly. However, this is almost impossible because the results will not be given 
to the study staff or to you, and will not be in your or your baby’s study records. It 
would be illegal under South African law to share your or your baby’s personal 
information. 
Unknown risks: 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 132 of 151 
To prevent HIV transmission through breastfeeding, you should take your ART 
every day. In this study we are looking to see whether this vaccine approach, may 
in the future, protect against breast milk transmission. We do not know if the 
study vaccine will assist in the prevention of breast milk transmission. This 
vaccine regimen aims to encourage the baby’s immune system to produce potent 
antibodies. We do not know whether we will achieve this. We also do not know 
whether this study vaccine will prevent HIV infection later on in life. Should your 
baby get infected through breast milk, we do not know whether this study vaccine 
will influence the way your baby responds to HIV infection. Your baby should go 
onto ART if they become HIV infected. There are many ways for your baby to 
prevent HIV infection when they get older.  
Site: Any change to the following text requires approval from HVTN Regulatory 
Affairs at vtn.core.reg@hvtn.org.  
We do not know if getting these study vaccines will affect how your child will 
respond to any future approved HIV vaccine. It could be that a future HIV 
vaccine may not work as well for your child because he or she got the study 
vaccines. Currently, no HIV vaccine has been approved for use. We also do not 
know whether getting the study vaccine may affect the way your baby responds to 
other vaccines they may get early in life. This study may help answer that 
question.  
Benefits 
20. The study may not benefit you. 
We do not expect the study vaccine to benefit your baby 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 your baby avoid getting HIV. The lab tests and 
physical exams that you get while in this study may result in earlier detection and 
treatment of diseases you are not aware of. 
This study may help in the search for a vaccine to prevent HIV. However, if the 
study vaccine later becomes approved and sold, there are no plans to share any 
money with you or your baby. 
Your rights and responsibilities 
21. If you and your baby join the study, you both have rights and 
responsibilities. 
You and your baby have many rights that we will respect. You also have 
responsibilities. We list these in the Participant’s Bill of Rights and 
Responsibilities. We will give you a copy of it. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 133 of 151 
Leaving the study 
22. Tell us if you decide to leave the study. 
You and your baby are free to leave the study at any time and for any reason. 
Your and your baby’s care at this clinic and both of your legal rights will not be 
affected, but it is important for you to let us know. 
We will ask you and your baby to come back to the clinic one last time for a 
physical exam, and we may ask to take some blood samples. We will also ask 
about any personal problems or benefits you or your baby have experienced from 
being in the study. We believe these steps are important to protecting you and 
your baby’s health, but it is up to you whether to complete them. 
Injuries 
Site: Approval from HVTN Regulatory Affairs (at vtn.core.reg@hvtn.org) is needed 
for any change (other than those that the instructions specifically request or those 
previously approved by HVTN Regulatory Affairs) to the boxed text. You can remove 
the box around the text. 
23. If you or your baby get sick or injured during the study, contact us 
immediately. 
You and your baby’s health is important to us. (Site: adjust the following 2 
sentences if applicable to the care available at your site) We will tell you about 
the care that we can give here. For the care that we cannot provide, we will 
explain how we will help you and your baby get care elsewhere. 
If you or your baby become sick or injured in this study, the HVTN has a process 
to decide if it is related to the study product 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, our clinic has insurance to cover your medical treatment in the case 
of a study-related injury. We will follow the Association of the British 
Pharmaceutical Industry guidelines for payment of study-related injury. We can 
give you a copy of these guidelines. In rare cases, the insurance funds may not be 
enough. In this situation, some of the study product providers have agreed to pay 
medical costs for study-related injuries that are determined to be caused by their 
own study products. 
The HVTN has limited funds to pay medical costs that it determines are 
reasonable. (Site: insert locale- appropriate medical insurance language in the 
following sentence) If the injury is not study related, then you and/or your health 
insurance will be responsible for treatment costs. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 134 of 151 
Some injuries are not physical. For example, you or your baby 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 or your baby’s 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 you and your baby’s 
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 your baby has 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]. 
This study has been reviewed and approved by a committee called the 
[name of local IRB/EC]. If you have questions about your or your baby’s rights as 
a research participant, or problems or concerns about how you and your baby are 
being treated in this study, contact 
[name or title and telephone number of person on IRB/EC] , at the committee. 
The study has been structured in accordance with the Declaration of Helsinki (last 
updated October 2013) which deals with the recommendations guiding doctors in 
biomedical research involving human participants, the Ethics in Health Research: 
Principles, Structures and Processes Second Edition 2015, and Guidelines for 
Good Practice in the Conduct of Clinical Trials in Human Participants in South 
Africa. We can provide you with copies of these guidelines if you wish to review 
them. In addition, the recent Protection of Personal Information Act (POPIA) 
ensures that all South African institutions conducts themselves in a responsible 
manner when collecting, processing, storing and sharing another entity’s personal 
information by holding them accountable should they abuse or compromise your 
personal information in any way. 
If you want to leave this study, contact 
[name or title and telephone number of the investigator or other study staff]. 
You can reach a study staff member 24 hours a day at [telephone number]. 
If you have questions about this trial you should first discuss them with your 
doctor or the ethics committee (contact details as provided on this form). After 
you have consulted your doctor or the ethics committee and if they have not 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 135 of 151 
provided you with answers to your satisfaction, you should write to the South 
African Healthcare Products Regulatory Authority (SAHPRA) at: 
The Chief Executive Officer 
Dr Boitumelo Semete-Makokotlela 
South African Health Products Regulatory Authority 
Department of Health 
Private Bag X828 
PRETORIA 
0001 
Tel: (012) 842 7629/26 
e-mail: Boitumelo.Semete @sahpra.org.za 
Your permissions and signature  
25. In Section 13 of this form, we told you about optional additional samples that 
we would like to collect from you and your baby. Please write your initials or 
make your mark in the box next to the options you choose. Whatever you 
choose, the clinic keeps track of your decision. You can change your mind 
after signing this form. 
 
 
I agree to provide stool samples from me and my baby.  
 
 
 
I do not agree to provide stool samples from me and my baby. 
 
AND 
 
 
I agree to provide breast milk samples.  
 
 
 
I do not agree to provide breast milk samples. 
 
 
26. If you agree to join this study with your baby, 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. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 136 of 151 
• You feel that you understand what the study is about and what will happen to 
you and your baby 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 with your baby. 
You will not be giving up any of your or your baby’s rights by signing this 
consent form. 
 
 
 
 
 
 
 
Mother/Guardian’s name (print) 
 
Mother/Guardian’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 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 137 of 151 
Appendix B 
Sample consent form for use of samples and 
information in other studies 
Title: A phase 1 clinical trial to evaluate the safety and immunogenicity of the 
HIV-1 CH505 transmitted/founder gp120 adjuvanted with GLA-SE in healthy, 
HIV-exposed uninfected infants  
HVTN protocol number: HVTN 135 
Site: [Insert site name] 
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 and your baby’s 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. 
Key Information 
These are some of the things you should know about the use of your and your baby’s 
samples and information for other studies: 
• The extra samples will be labeled with a code number and some personal 
information. They will not be labeled with your or your baby’s name. The extra 
samples are stored in a secure place. At your request, the HVTN will destroy all 
your and your baby’s extra samples. You and your baby can still join the main 
study even if you do not agree to use of your and your baby’s extra samples in 
other studies. 
• Researchers may do genetic testing on your and your baby’s samples, which 
could include genome wide studies. It is unlikely, but these tests could show you 
or your baby may be at risk for certain diseases. In the very unlikely event that if 
others found out, this could lead to discrimination or other problems.  
• You will not be paid or otherwise benefit from allowing your and your baby’s 
extra samples to be used in other studies.   
The rest of this form gives more information about use of your and your baby’s extra 
samples for other studies. Please read it carefully.  
1. Do I have to agree? 
No. You are free to say yes or no, or to change your mind after you sign this form. 
At your request, we will destroy all extra samples that we have. Your decision 
will not affect your or your baby’s being in this study or have any negative 
consequences here. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 138 of 151 
2. Where are the samples stored? 
Extra samples are stored in a secure central place called a repository. Your and 
your baby’s samples will be stored in the HVTN repository in South Africa. 
3. How long will the samples be stored? 
There is no limit on how long your and your baby’s extra samples will be stored. 
[Site: Revise the previous sentence to insert limits if your regulatory authority 
imposes them.] 
4. Will I be paid for the use of my or my baby’s samples? 
No. Also, a researcher may make a new scientific discovery or product based on 
the use of your or your baby’s samples. If this happens, there is no plan to share 
any money with you or your baby. The researcher is not likely to ever know who 
you or your baby are. 
5. Will we benefit from allowing our 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 or your baby’s medical record. The studies 
are only being done for research purposes. 
6. Will the HVTN sell our samples and information? 
No, but the HVTN may share your or your baby’s samples with HVTN or other 
researchers. Once we share your or your baby’s samples and information, we may 
not be able to get them back. 
7. How do other researchers get our samples and information? 
When a researcher wants to use your or your baby’s 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 or your baby’s samples to 
the researcher’s location. 
8. What information is shared with HVTN or other researchers?  
The samples and information will be labeled with a code number. The HVTN 
keeps a key to the code but will not share this key with other researchers or with 
anyone else who does not need to know your or your baby’s name. Your and your 
baby’s names will not be part of the information. However, some information that 
we share may be personal, such as your and your baby’s race, ethnicity, gender, 
health information from the study, and HIV status. We may share information 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 139 of 151 
about the study product your baby received and how your baby’s body responded 
to the study product. 
9. What kind of studies might be done with my or my baby’s 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 or your baby’s samples. 
In some cases, researchers may take cells from your or your baby’s samples and 
grow more of them over time, so that they can continue to do research with them. 
If you agree, your or your baby’s samples could also be used for genome wide 
studies. In these studies, researchers will look at all of your or your baby’s 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 and your 
baby’s name and other personal information will not be included. Usually, no one 
would be able to look at your or your baby’s genome and link it to you or your 
baby as a person. However, if another database exists that also has information on 
your or your baby’s genome and your or your baby’s name, someone might be 
able to compare the databases and identify you or your baby. If others found out, 
it could lead to discrimination or other problems. The risk of this is very small. 
There may be other unknown risks. 
10. What are the risks of genetic testing? 
It is unlikely, but the genetic tests done on your or your baby’s samples could 
show you or your baby may be at risk for certain diseases. If others found out, it 
could lead to discrimination or other problems. However, it is almost impossible 
for you or others to know your test results from the genetic testing. The results are 
not part of your or your baby’s study records and are not given to you. 
11. Who will have access to my and my baby’s information in studies using our 
extra samples? 
People who may see your or your baby’s information are: 
• Researchers who use your or your baby’s extra samples and information for 
other research 
• Government agencies that fund or monitor the research using your or your 
baby’s extra samples and information 
• The researcher’s Institutional Review Board or Ethics Committee 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 140 of 151 
• 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 and your baby’s information. 
The results of any new studies that use your or your baby’s extra samples and 
information may be published. No publication will use your or your baby’s name 
or identify you or your baby personally. 
Questions  
12. If you have questions or problems about allowing your or your baby’s 
samples and information to be used in other studies, use the following 
important contacts. 
If you have questions about the use of your or your baby’s samples or information 
or if you want to change your mind about their use, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you think you or your baby may have been harmed because of studies using 
your or your baby’s samples or information, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you have questions about your or your baby’s rights as a research participant, 
contact [name or title and telephone number of person on IRB/EC . 
The study has been structured in accordance with the Declaration of Helsinki (last 
updated October 2013) which deals with the recommendations guiding doctors in 
biomedical research involving human participants, the Ethics in Health Research: 
Principles, Structures and Processes Second Edition 2015, and Guidelines for 
Good Practice in the Conduct of Clinical Trials in Human Participants in South 
Africa. We can provide you with copies of these guidelines if you wish to review 
them. 
You can reach a study staff member 24 hours a day at [telephone number]. 
If you have questions about this trial you should first discuss them with your 
doctor or the ethics committee (contact details as provided on this form). After 
you have consulted your doctor or the ethics committee and if they have not 
provided you with answers to your satisfaction, you should write to the South 
African Healthcare Products Regulatory Authority (SAHPRA) at: 
The Chief Executive Officer 
Dr Boitumelo Semete-Makokotlela 
South African Health Products Regulatory Authority 
Department of Health 
Private Bag X828 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 141 of 151 
PRETORIA 
0001 
Tel: (012) 842 7629/26 
e-mail: Boitumelo.Semete @sahpra.org.za 
 
13. Please choose only one of the options below and write your initials or make 
your mark in the box next to it. Whatever you choose, the HVTN keeps track 
of your choice about how your or your baby’s samples and information can 
be used. You can change your mind after signing this form. 
 
 
I allow my and my baby’s 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 and my baby’s cells growing over time. 
OR 
 
 
I agree to the option above and also to allow my and my baby’s extra samples and 
information to be used in genome wide studies. 
 
OR 
 
 
I do not allow my and my baby’s extra samples to be used in any other studies. This 
includes not allowing genetic testing, growing more of my and my baby’s cells, or 
genome wide studies.  
 
 
 
 
 
 
 
 
 
 
Mother/ Guardian’s name (print) 
 
Mother/ Guardian’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 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 142 of 151 
Appendix C 
Table of procedures (for sample informed consent form) 
 
 
 
 
Time after first injection visit 
 
Procedures for 
Infant 
Screening 
visit(s) 
First 
injection 
visit 
1 day 
2 
weeks 
2 
months 
2 ½ 
months 
4 
months 
4 ½ 
months 
8 
months 
8½ 
months 
12 ½ 
months 
13 
months 
17 
months 
2 years + 2 
weeks 
Injection 
 
√ 
 
 
√ 
 
√ 
 
√ 
 
√ 
 
 
 
Medical history 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Complete physical 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
√ 
Brief physical 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
Blood drawn 
√ 
√ 
√ 
√ 
 
√ 
 
√ 
 
√ 
√ 
√ 
√ 
√ 
Cord blood 
collection 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Stool collection 
 
√ 
 
√ 
√ 
 
√ 
 
√ 
 
√ 
√ 
 
√ 
 
 
 
 
 
 
Time after first visit 
Procedures for 
Mother 
Screening 
visit(s) 
First visit 
1 
day 
2 
weeks 
2 
months 
2 ½ 
months 
4 
months 
4 ½ 
months 
8 
months 
8½ 
months 
12 ½ 
months 
13 
months 
17 
months 
2 years + 2 
weeks 
Medical history 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Complete Physical 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Questionnaire 
 
 
 
 
√ 
 
 
 
√ 
 
 
 
 
√ 
Blood drawn 
√ 
√ 
 
 
 
√ 
 
√ 
 
√ 
 
√ 
 
 
Nasal or throat 
swab* 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Breastmilk 
Collection 
 
√ 
 
 
 
 
 
√ 
 
 
 
 
 
 
Stool collection 
 
√ 
 
 
 
 
 
 
 
 
 
 
 
 
*Swab for SARS-CoV-2 testing will only be collected if testing results are not already available in your medical records.  
Not shown in this table is a time after all study participants have completed their last scheduled visit when you can find out what products your baby received.  

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 143 of 151 
Appendix D 
Laboratory procedures: Infant 
 
Visit:
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Day:
D0
D1
D14
D56
D70
D112
D126
D224
D238
D379
D393
D516
D743
Week:
W0
W2
W8
W10
W16
W18
W32
W34
W54
W56
W74
W106
Month:
M0
M0.5
M2
M2.5
M4
M4.5
M8
M8.5
M12.5
M13
M17
M24.5
VAC1
VAC2
VAC3
VAC4
VAC5
Procedure
Ship to1
Tube4
Tube size 
(vol. 
capacity)4
CH505TFpg120 
+GLA-SE or 
Placebo
CH505TFpg120 
+GLA-SE or 
Placebo
CH505TFpg120 
+GLA-SE or 
Placebo
CH505TFpg120 
+GLA-SE or 
Placebo
CH505TFpg120 
+GLA-SE or 
Placebo
Total
BLOOD COLLECTION
Diagnostics
Screening HIV Test
Local lab
Local lab
EDTA
1mL
1
—
—
—
—
—
—
—
—
—
—
—
—
—
1.0
HIV diagnostics8,12
HSML-NICD
HSML-NICD
EDTA
5mL
—
—
—
—
—
—
—
—
—
—
—
—
5
10
15.0
Safety labs9
CBC/Diff
Local lab
Local lab
EDTA
0.5mL 
0.5
—
—
0.5
—
0.5
—
0.5
—
0.5
—
0.5
—
0.5
3.5
Chemistry panel5
Local lab
Local lab
SST
0.8mL
0.8
—
—
0.8
—
0.8
—
0.8
—
0.8
—
0.8
—
0.8
5.6
Immunogenicity assays
Humoral assays
Pediatric Vaccine Multiplex Assay
BARC
HVTN Labs
SST
2mL
—
—
—
—
—
—
—
—
—
2
—
2
—
2
6.0
Binding Ab
BARC
HVTN Labs
SST
2mL
—
—
—
—
—
—
—
—
—
2
2
2
—
2
8.0
HIV-1 neutralizing Ab
BARC
HVTN Labs
SST
2mL
—
—
—
—
—
—
—
—
—
2
—
2
—
2
6.0
Ab Effector Function Assays 10
BARC
HVTN Labs
SST
2mL
—
—
—
—
—
—
—
—
—
—
—
2
—
2
4.0
Ab Avidity
BARC
HVTN Labs
SST
2mL
—
—
—
—
—
—
—
—
—
y
y
y
—
y
0.0
Cellular assays
Ag-Specific B-Cell Phenotyping
BARC
HVTN Labs
NaHep
2mL
—
—
—
4
—
6
—
6
—
4
—
6
—
6
32.0
B-cell lineage
BARC
HVTN Labs
NaHep
2mL
—
—
—
—
—
—
—
z
—
—
—
z
—
—
0.0
Systems Biology Assays - Venous Blood
BARC
Non HVTN Labs
TBD
TBD
211
—
2
1
—
1
—
1
—
1
—
1
—
1
8.0
In Vitro Vaccine Modeling - Cord Blood
BARC
Non HVTN Labs
TBD
TBD
206
—
—
—
—
—
—
—
—
—
—
—
—
—
0.0
Specimen storage
Serum - Cord Blood
BARC
SST
8.5mL
8.56
—
—
—
—
—
—
—
—
—
—
—
—
—
Plasma - Cord Blood
BARC
NaHep
10mL
v
 CBMC
BARC
NaHep
10mL
606
—
—
—
—
—
—
—
—
—
—
—
—
—
0.0
Serum - Venous Blood
BARC
SST
2mL
—
2
—
—
—
—
—
—
—
—
4
—
—
—
6.0
Plasma - Venous Blood
BARC
NaHep
2mL
—
—
—
w
—
w
—
w
—
w
—
w
—
w
Visit total
3.3
2.0
2
6.3
0
8.3
0
8.3
0
12.3
6
16.3
5
26.3
96.1
30-Day total7
4.3
5.3
7.3
13.6
0.0
8.3
0
8.3
0
12.3
6
22.3
5
26.3
STOOL COLLECTION (OPTIONAL)
      Stool 
BARC
HVTN Labs
—
X
—
X
X
—
X
—
X
—
X
X
—
X
Assay location2
Screening 
visit3
 
 
1BARC = Bio Analytical Research Corporation South Africa (Pty) Ltd (Johannesburg, South Africa); HSML-NICD = HIV Sero-Molecular Laboratory–National Institute for Communicable Diseases 
(Johannesburg, South Africa) 
2HVTN Laboratories include: Fred Hutchinson Cancer Research Center (Seattle, Washington, USA); Duke University Medical Center (Durham, North Carolina, USA); Duke Human Vaccine Institute 
(Durham, North Carolina, USA); South African Immunology Laboratory-National Institute for Communicable Diseases (SAIL-NICD, Johannesburg, South Africa) 
Non HVTN Laboratories include: Precision Vaccines Program, Boston Children’s Hospital/Harvard Medical Center (Boston, Massachusetts, USA); University of British Columbia (Vancouver, 
Canada) 
3Screening may occur over the course of several contacts/visits up to and including Day 0, prior to vaccination.  
4Local labs may assign appropriate alternative tube types for locally performed tests. Use of alternate blood collection tubes for immunogenicity or storage specimens must be approved by the HVTN 
laboratory program.  
5Chemistry panels are defined in section 9.2.2 (pre-enrollment) and section 9.4 (postenrollment).  
6Speicmens for In Vitro Vaccine Modeling - Cord Blood, and Serum - Cord Blood and CBMC specimen storage will be collected from umbilical cord blood at delivery. 
7The 30-day total blood volume does not include up to 1mL collection for HIV diagnostics following local standard of care guidelines and any additional HIV testing based on mother's viral load. 
However, the 30-day blood draw limit of 5% of total blood volume is not exceeded at any visit by the collection of blood for HIV diagnostic testing. 
8At an early termination visit for a withdrawn or terminated infant who is not HIV-infected (see Section 9.9), blood should be drawn for HIV diagnostic testing. Refer to the HVTN 135 SSP for more 
information. If the infant has a confirmed diagnosis of HIV infection, do not collect blood for HIV diagnostic testing (see Section 9.10). 
9For participants with confirmed diagnosis of HIV infection, only specimens required for protocol-specified safety laboratory tests will be collected. 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 144 of 151 
10Ab Effector Function Assays may include FcγR binding assay, ADCC, ADCP, ADNP and ADCD. 
11Specimen for System Biology Assays - Venous Blood will be collected within the first 24 hours after birth, prior to vaccination.  
12Additional HIV diagnostic testing may be performed based on mother's viral load. 
v=NaHep plasma - Cord Blood aliquots will be harvested for storage during CBMC processing; no separate blood draw is needed. 
w=NaHep plasma - Venous Blood aliquots will be harvested for storage during PBMC processing; no separate blood draw is needed. 
y=SST collected for Binding Ab assay will also cover specimen needs for Ab avidity assay; no separate blood draw is needed. 
z=NaHep collected for Ag-Specific B-Cell Phenotyping will also cover specimen needs for B-cell lineage; no separate blood draw is needed. 
 

HVTN 135 Version 3.0 / December 18, 2020 
HVTN135_v3.0_Final.docx / Page 145 of 151 
Appendix E 
Laboratory procedures: Mother 
 
 
 
1BARC = Bio Analytical Research Corporation South Africa (Pty) Ltd (Johannesburg, South Africa); HSML-NICD = HIV Sero-Molecular Laboratory–National Institute for Communicable Diseases 
(Johannesburg, South Africa) 
2HVTN Laboratories include: Fred Hutchinson Cancer Research Center (Seattle, Washington, USA); Duke University Medical Center (Durham, North Carolina, USA). 
Non HVTN Laboratories include: Precision Vaccines Program, Boston Children’s Hospital/Harvard Medical Center (Boston, Massachusetts, USA); University of British Columbia (Vancouver, 
Canada) 
3Screening may occur over the course of several contacts/visits up to and including D0, prior to infant vaccination.  
4Local labs may assign appropriate alternative tube types for locally performed tests. 
5Collect specimens for specified testing only if results are not available through medical records. See protocol section 9.2 for more information.  
6Specimen for System Biology Assays will be collected within the first 24 hours after birth.  
7 Local labs may assign appropriate alternative specimen type for SARS-CoV-2 testing. 
8Optional breastmilk collection will be obtained within 7 days after delivery but no sooner than 48 hours after delivery.  
y=EDTA blood for HIV Genotypic Antiretroviral Resistance Testing - Reflex assay  will only be collected if HIV PCR viral load is ≥400 copies/mL at specified visits. Refer to the HIV Testing SSP 
for additional information.   
Visit:
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Day:
D0
D1
D14
D56
D70
D112
D126
D224
D238
D379
D393
D516
D743
Week:
W0
W2
W8
W10
W16
W18
W32
W34
W54
W56
W74
W106
Month:
M0
M0.5
M2
M2.5
M4
M4.5
M8
M8.5
M12.5
M13
M17
M24.5
Procedure
Ship to1
Assay location2
Tube4
Tube size 
(vol. 
capacity)4
Total
BLOOD COLLECTION
Screening/Diagnostic
Screening HIV EIA
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
—
—
CD4+T Cell Count
Local lab
Local lab
EDTA
5mL
55
—
—
—
—
—
—
—
—
—
—
—
—
—
0
HBsAg
Local lab
Local lab
SST
5mL
55
—
—
—
—
—
—
—
—
—
—
—
—
—
0
HIV PCR Viral Load
Local lab
Local lab
EDTA
10mL
10
—
—
—
—
10
—
10
—
10
—
10
—
—
40
 HIV Genotypic Antiretroviral Resistance 
Testing 
Local lab
Local lab
EDTA
10mL
—
—
—
—
—
y
—
y
—
y
—
y
—
—
0
ARV detection by dried blood spots
BARC
HVTN Labs
EDTA
2mL
—
—
—
—
—
2
—
2
—
2
—
2
—
—
8
Immunogenicity assays
Humoral assays
          Binding Ab
BARC
HVTN Labs
SST
8.5mL
—
8.5
—
—
—
—
—
—
—
8.5
—
8.5
—
—
26
Systems Biology Assays
BARC
Non HVTN Labs
TBD
TBD
26
—
—
—
—
—
—
—
—
—
—
—
—
0
Visit total
27
8.5
0
0
0
12
0
12
0
20.5
0
20.5
0
0
101
56-Day total
27
35.5
35.5
35.5
35.5
12
12
24
24
20.5
0
20.5
0
0
NASOPHARYNGEAL/OROPHARYNGEAL SWAB COLLECTION7
SARS-CoV-2 RNA PCR
Local lab
Local Lab
X5
BREASTMILK COLLECTION (OPTIONAL)
 Breastmilk
BARC
HVTN Labs
—
X8
—
—
—
—
—
X
—
—
—
—
—
—
STOOL COLLECTION (OPTIONAL)
      Stool 
BARC
HVTN Labs
—
X
—
—
—
—
—
—
—
—
—
—
—
—
Screening 
visit3

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 146 of 151 
Appendix F 
Procedures at HVTN CRS: Infant 
 
Visit: 
011 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
14 
Post 
Day: 
 
D0 
D1 
D14 
D56 
D70 
D112 
D126 
D224 
D238 
D379 
D393 
D516 
D743 
 
Week 
 
W0 
 
W2 
W8 
W10 
W16 
W18 
W32 
W34 
W54 
W56 
W74 
W106 
 
Month: 
 
M0 
 
M0.5 
M2 
M2.5 
M4 
M4.5 
M8 
M8.5 
M12.5 
M13 
M17 
M24.5 
 
Procedure 
Scr. 
VAC1 
 
 
VAC2 
 
VAC3 
 
VAC4 
 
VAC5 
 
 
 
 
Study procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history  
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Feeding history 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Complete physical exam 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
— 
Abbreviated physical exam 
— 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
— 
Obtain demographics (for 
infant) 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Randomize 
— 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Concomitant medications 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Intercurrent illness/adverse 
experience 
— 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
HIV assessment  
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
X 
— 
Confirm HIV assessment 
results provided to mother 
— 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
X 
Vaccination procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Vaccination 
— 
X 
— 
— 
X 
— 
X 
— 
X 
— 
X 
— 
— 
— 
— 
Reactogenicity assessments2 
— 
X 
— 
— 
X 
— 
X 
— 
X 
— 
X 
— 
— 
— 
— 
Specimen collection3  
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Poststudy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Unblind participant 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
 
1 Screening may occur over the course of several contacts/visits up to and including day 0 prior to vaccination. 
2 Reactogenicity assessments performed daily for at least 3 days postvaccination (see Section 9.7). 
3 For specimen collection requirements, see Appendix D, Laboratory procedures- Infant. 
 

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 147 of 151 
Appendix G 
Procedures at HVTN CRS: Mother  
 
Visit: 
011 
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
14 
Post 
Day: 
 
D0 
D1 
D14 
D56 
D70 
D112 
D126 
D224 
D238 
D379 
D393 
D516 
D743 
 
Week: 
 
W0 
 
W2 
W8 
W10 
W16 
W18 
W32 
W34 
W54 
W56 
W74 
W106 
 
Month: 
 
M0 
 
M0.5 
M2 
M2.5 
M4 
M4.5 
M8 
M8.5 
M12.5 
M13 
M17 
M24.5 
 
Procedure 
Scr. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study procedures 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Assessment of understanding 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Complete Physical Exam 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Signed protocol consent 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Medical history  
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Risk reduction counseling2 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Confirm eligibility 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Obtain demographics (for mother) 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Social impact assessment 
— 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Outside testing of baby 
questionnaire 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
— 
Belief questionnaire 
— 
— 
— 
— 
X 
— 
— 
— 
X 
— 
— 
— 
— 
X 
 
Concomitant medications 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
Specimen Collection 3 
X 
X 
— 
— 
— 
X 
— 
X 
— 
X 
— 
X 
— 
— 
— 
Poststudy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Provide unblinding information 
 to mother 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
 
1 Screening may occur over the course of several contacts/visits up to and including day 0. 
2 Risk reduction counseling, see Section 9.6 and HVTN 135 SSP. 
3 For specimen collection requirements, see Appendix E, Laboratory procedures- Mother. 
 

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 148 of 151 
Appendix H 
Adverse events of special interest 
AEs 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 135 Study Specific Procedures. 
Gastrointestinal disorders 
Liver disorders 
Metabolic diseases 
• 
Celiac disease 
• 
Crohn’s disease 
• 
Ulcerative colitis 
• 
Ulcerative proctitis 
• 
Autoimmune cholangitis 
• 
Autoimmune hepatitis 
• 
Primary biliary cirrhosis 
• 
Primary sclerosing cholangitis 
• 
Addison’s disease 
• 
Autoimmune thyroiditis 
(including Hashimoto 
thyroiditis) 
• 
Diabetes mellitus type I 
• 
Grave’s or Basedow’s 
disease 
Neuroinflammatory disorders 
Musculoskeletal disorders 
Skin disorders 
• 
Acute disseminated 
encephalomyelitis, including site 
specific variants (eg, non-infectious 
encephalitis, encephalomyelitis, 
myelitis, myelo radiculomyelitis) 
• 
Cranial nerve disorders, included 
paralyses/paresis (eg, Bell’s palsy) 
• 
Guillain-Barré syndrome, including 
Miller Fisher syndrome and other 
variants 
• 
Immune-mediated peripheral 
neuropathies and plexopathies, 
including chronic inflammatory 
demyelinating polyneuropathy, 
multifocal motor neuropathy and 
polyneuropathies associated with 
monoclonal gammopathy 
• 
Multiple sclerosis 
• 
Narcolepsy 
• 
Optic neuritis 
• 
Transverse Myelitis 
• 
Antisynthetase syndrome 
• 
Dermatomyositis 
• 
Juvenile chronic arthritis 
(including Still’s disease) 
• 
Mixed connective tissue disorder 
• 
Polymyalgia rheumatic 
• 
Polymyositis 
• 
Psoriatic arthropathy 
• 
Relapsing polychondritis 
• 
Rheumatoid arthritis 
• 
Scleroderma, including diffuse 
systemic form and CREST 
syndrome 
• 
Spondyloarthritis, including 
ankylosing spondylitis, reactive 
arthritis (Reiter’s Syndrome) and 
undifferentiated 
spondyloarthritis 
• 
Systemic lupus erythematosus 
• 
Systemic sclerosis 
• 
Alopecia areata 
• 
Autoimmune bullous skin 
diseases, including 
pemphigus, pemphigoid 
and dermatitis 
herpetiformis 
• 
Cutaneous lupus 
erythematosus 
• 
Erythema nodosum 
• 
Morphoea 
• 
Lichen planus 
• 
Psoriasis 
• 
Sweet’s syndrome 
• 
Vitiligo 
Vasculitides 
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 
• 
Antiphospholipid syndrome 
• 
Autoimmune hemolytic anemia 
• 
Autoimmune glomerulonephritis (including IgA nephropathy, 
glomerulonephritis rapidly progressive, membranous 
glomerulonephritis, membranoproliferative glomerulonephritis, and 
mesangioproliferative glomerulonephritis) 
• 
Autoimmune myocarditis/cardiomyopathy 
• 
Autoimmune thrombocytopenia 
• 
Goodpasture syndrome 
• 
Idiopathic pulmonary fibrosis 
• 
Pernicious anemia 
• 
Raynaud’s phenomenon 
• 
Sarcoidosis 
• 
Sjögren’s syndrome 
• 
Stevens-Johnson syndrome 
• 
Uveitis 
 

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 149 of 151 
Appendix I 
Infant visit windows 
 
Visit 
Number 
Visit Type 
Lower 
Allowable 
Window  
Lower Target 
Day 
Target 
Day1 
Upper Target 
Day 
Upper Allowable 
Window  
01.0 
Screening 
-5 
- 
- 
- 
+0 
02.0 
Enrollment/Vaccination 1 
- 
- 
0 
- 
- 
03.0 
Follow-up 
- 
- 
1 
+2 
- 
04.0 
2 Weeks Post-Vaccination 
- 
-4 
14 
+4 
+7 
05.0 
Vaccination 2 
- 
-9 
56 
+9 
+14 
06.0 
2 Weeks Post-Vaccination 
- 
-4 
70 
+4 
+7 
07.0 
Vaccination 3 
-21 
-14 
112 
+14 
+21 
08.0 
2 Weeks Post-Vaccination 
Primary Immunogenicity 
- 
-4 
126 
+4 
+7 
09.0 
Vaccination 4 
-28 
-14 
224 
+14 
+28 
10.0 
2 Weeks Post-Vaccination 
- 
-4 
238 
+4 
+7 
11.0 
Vaccination 5 
-28 
-14 
379 
+14 
+28 
12.0 
2 Weeks Post-Vaccination 
Primary Immunogenicity 
- 
-4 
393 
+4 
+7 
13.0 
Follow-up 
- 
-14 
516 
+14 
+21 
14.0 
Final Visit 
-28 
-14 
743 
+14 
+28 
1Target dates are relative to Day 0 (Enrollment), with the exception of postvaccination visits 
4.0, 6.0, 8.0, 10.0, and 12.0 which are relative to the prior vaccination visit. 
 

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 150 of 151 
Appendix J 
Mother visit windows 
 
Visit 
Number 
Visit Type 
Lower 
Allowable 
Window  
Lower Target 
Day 
Target 
Day1 
Upper Target 
Day 
Upper Allowable 
Window  
01.0 
Screening 
-273 
- 
- 
- 
+0 
02.0 
Enrollment/Infant 
Vaccination 
- 
- 
0 
- 
- 
03.0 
Follow-up 
- 
- 
1 
+2 
- 
04.0 
Follow-up 
- 
-4 
14 
+4 
+7 
05.0 
Follow-up 
- 
-9 
56 
+9 
+14 
06.0 
Specimen Collection 
- 
-4 
70 
+4 
+7 
07.0 
Follow-up 
-21 
-14 
112 
+14 
+21 
08.0 
Specimen Collection  
- 
-4 
126 
+4 
+7 
09.0 
Follow-up 
-28 
-14 
224 
+14 
+28 
10.0 
Specimen Collection 
- 
-4 
238 
+4 
+7 
11.0 
Follow-up 
-28 
-14 
379 
+14 
+28 
12.0 
Specimen Collection 
- 
-4 
393 
+4 
+7 
13.0 
Follow-up 
- 
-14 
516 
+14 
+21 
14.0 
Final Visit 
-28 
-14 
743 
+14 
+28 
1Target dates are relative to Day 0 (Enrollment), with the exception of visits 4.0, 6.0, 8.0, 10.0, 
and 12.0 which are relative to the prior visit. 
 
 
 

 
HVTN 135 Version 3.0 / December 18, 2020 
 
HVTN135_v3.0_Final.docx / Page 151 of 151 
Appendix K 
Protocol Signature Page 
 
A phase 1 clinical trial to evaluate the safety and immunogenicity of the HIV-1 
CH505 transmitted/founder gp120 adjuvanted with GLA-SE in healthy, HIV-
exposed uninfected infants 
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 Conference on Harmonization Guideline for Good Clinical 
Practice (E6); Institutional Review Board/Ethics Committee determinations; all 
applicable in-country, state, and local laws and regulations; and other applicable 
requirements (eg, 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 135 
DAIDS Protocol Version: Version 3.0 
Protocol Date: December 18, 2020