A Phase 1 Clinical Trial to Evaluate the Safety and Immunogenicity of an HIV-1 gp41 MPER-656 Liposome Vaccine in Healthy, HIV-uninfected Adult Participants
Phase: PHASE1
Conditions: HIV Infections
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

PROTOCOL 
HVTN 133 
A phase 1 clinical trial to evaluate the safety and 
immunogenicity of an HIV-1 gp41 MPER-656 liposome 
vaccine in healthy, HIV-uninfected adult participants 
DAIDS DOCUMENT ID 12047 
IND [IND # OR DELETE] HELD BY DAIDS 
CLINICAL TRIAL SPONSORED BY 
Division of AIDS (DAIDS) 
National Institute of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
Department of Health and Human Services (DHHS) 
Bethesda, Maryland, USA 
STUDY PRODUCT(S) PROVIDED BY 
International AIDS Vaccine Initiative (IAVI) 
New York, New York, USA 
March 13, 2019 
Final 
HVTN 133 
Version 1.0 

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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 ...........................................................................8 
2.6 
Protect privacy/confidentiality .......................................................................8 
3 Overview ......................................................................................................................10 
3.1 
Protocol Team ..............................................................................................13 
4 Background ..................................................................................................................14 
4.1 
Rationale for trial concept ............................................................................14 
4.2 
GTH1-656 MPER peptide liposome ............................................................15 
4.3 
Alhydrogel® adjuvant ...................................................................................16 
4.4 
Trial design rationale ....................................................................................16 
4.5 
Plans for future product development and testing ........................................17 
4.6 
Preclinical safety studies ..............................................................................18 
4.7 
Preclinical immunogenicity studies .............................................................20 
4.8 
Clinical studies .............................................................................................24 
4.9 
Potential risks of study products and administration ...................................26 
5 Objectives and endpoints .............................................................................................27 
5.1 
Primary objectives and endpoints ................................................................27 
5.2 
Secondary objectives and endpoints ............................................................27 
5.3 
Exploratory objectives ..................................................................................28 
6 Statistical considerations ..............................................................................................29 
6.1 
Accrual and sample size calculations ...........................................................29 
6.2 
Randomization .............................................................................................32 
6.3 
Blinding ........................................................................................................32 
6.4 
Statistical analyses ........................................................................................33 
7 Selection and withdrawal of participants .....................................................................39 
7.1 
Inclusion criteria ...........................................................................................39 
7.2 
Exclusion criteria ..........................................................................................42 
7.3 
Participant departure from vaccination schedule or withdrawal ..................45 
8 Study product preparation and administration .............................................................48 
8.1 
Vaccine regimen ...........................................................................................48 
8.2 
Study product formulation ...........................................................................48 
8.3 
Preparation of study products .......................................................................49 
8.4 
Administration ..............................................................................................51 
8.5 
Acquisition of study products ......................................................................52 
8.6 
Pharmacy records .........................................................................................52 
8.7 
Final disposition of study products ..............................................................52 
9 Clinical procedures ......................................................................................................53 

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9.1 
Informed consent ..........................................................................................53 
9.2 
Pre-enrollment procedures ...........................................................................55 
9.3 
Enrollment and vaccination visits ................................................................56 
9.4 
Follow-up visits ............................................................................................58 
9.5 
AESI health contact ......................................................................................59 
9.6 
HIV counseling and testing ..........................................................................60 
9.7 
Contraception status .....................................................................................62 
9.8 
Urine testing .................................................................................................62 
9.9 
Assessments of reactogenicity .....................................................................63 
9.10 Visit windows and missed visits ..................................................................64 
9.11 Early termination visit ..................................................................................65 
9.12 Pregnancy .....................................................................................................65 
9.13 HIV infection during the study .....................................................................65 
10 Laboratory ....................................................................................................................66 
10.1 HVTN CRS laboratory procedures ..............................................................66 
10.2 Total blood volume ......................................................................................66 
10.3 Primary immunogenicity timepoints ............................................................66 
10.4 Endpoint assays: cellular ..............................................................................66 
10.5 Endpoint assays: humoral .............................................................................67 
10.6 Lab assay algorithm .....................................................................................68 
10.7 Exploratory studies .......................................................................................68 
10.8 Specimen storage and other use of specimens .............................................69 
10.9 Biohazard containment .................................................................................69 
11 Safety monitoring and safety review ...........................................................................70 
11.1 Safety monitoring and oversight ..................................................................70 
11.2 Safety reporting ............................................................................................71 
11.3 Safety reviews ..............................................................................................74 
11.4 Safety pause and prompt PSRT AE review..................................................74 
11.5 Review of cumulative safety data ................................................................75 
11.6 Study termination .........................................................................................76 
12 Protocol conduct ..........................................................................................................77 
12.1 Social impacts ..............................................................................................78 
12.2 Emergency communication with study participants ....................................78 
13 Version history .............................................................................................................79 
14 Document references (other than literature citations) ..................................................80 
15 Acronyms and abbreviations........................................................................................82 
16 Literature cited .............................................................................................................85 
Appendix A Sample informed consent form ....................................................................89 
Appendix B Approved birth control methods for persons assigned female sex at birth 
(for sample informed consent form) ...........................................................107 
Appendix C Sample consent form for use of samples and information in other studies 108 
Appendix D Table of procedures (for sample informed consent form) ..........................112 

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Appendix E Laboratory procedures ................................................................................113 
Appendix F Procedures at HVTN CRS ..........................................................................115 
Appendix G HVTN low risk guidelines for the US ........................................................117 
Appendix H Adverse events of special interest ..............................................................119 
Appendix I HVTN 133 Visit Windows .........................................................................120 
Appendix J Protocol Signature Page .............................................................................121 
 

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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. 
• HVTN clinical trial staff counsel study participants routinely on how to 
reduce HIV risk. Participants who become HIV-infected during the trial are 
provided counseling on notifying their partners and about HIV infection 
according to local guidelines. Staff members will also counsel them about 
reducing their risk of transmitting HIV to others. 
• Participants who become HIV-infected during the trial are referred to medical 
practitioners to manage their HIV infection and to identify potential clinical 
trials they may want to join. If a program for antiretroviral therapy (ART) 
provision is not available at a site and ART is needed, a privately established 
fund will be used to pay for access to treatment to the fullest extent possible. 
• The HVTN provides training so that all participating sites similarly ensure fair 
participant selection, protect the privacy of research participants, and obtain 
meaningful informed consent. During the study, participants will have their 
wellbeing monitored, and to the fullest extent possible, their privacy 
protected. Participants 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. 

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• 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. 

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2 
IRB/EC review considerations 
US Food and Drug Administration (FDA) and other 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. 
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 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 and HIV risk reduction counseling; (e) providing HIV risk 
reduction counseling and checking on contraception use (for persons assigned 
female at birth); and (f) 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 
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. 

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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 subject or the subject’s 
legally authorized representative 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 protocol specifies that informed consent must be obtained before any study 
procedures are initiated and assessed throughout the trial (see Section 9.1). Each 
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. 
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 9). Safety is 
monitored daily by HVTN Core and routinely by the HVTN 133 Protocol Safety 
Review Team (PSRT). In addition, the HVTN Safety Monitoring Board (SMB) 
periodically reviews study data. 
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 

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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 the 
United States, research participants in HVTN protocols are protected by a 
Certificate of Confidentiality from the US NIH, which can prevent disclosure of 
study participation even when that information is requested by subpoena. 
Participants are told of the use and limits of the certificate in the study consent 
form. 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. 
Each 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. 

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3 
Overview 
Title  
A phase 1 clinical trial to evaluate the safety and immunogenicity of an HIV-1 
gp41 MPER-656 liposome vaccine in healthy, HIV-uninfected adult participants 
Primary objective(s) 
Primary objective 1 
• To evaluate the safety and tolerability of different doses of a prime-boost 
regimen of MPER-656 liposomes in HIV-uninfected healthy adults 
Primary endpoint 1 
• Local and systemic reactogenicity signs and symptoms, laboratory measures 
of safety, and adverse events (AEs) and serious adverse event (SAEs) 
Primary objective 2 
• To evaluate binding antibody responses elicited by different doses of the 
MPER-656 liposome vaccine 
Primary endpoint 2 
• MPER-peptide-specific immunoglobulin G (IgG) binding antibody (Ab) 
responses as assessed by binding Ab multiplex assay 2 weeks after the 3rd and 
4th vaccination with MPER-656 liposome vaccine 
Study products and routes of administration 
• MPER-656 liposomes at 0.5 mg/mL with 500 mcg alum (from 5 mg/mL 
suspension) for two 0.5 mL doses in two sites (total dose of 500 mcg in 1 mL) 
administered by IM injection in the deltoid muscles (one injection into each 
deltoid) 
• MPER-656 liposomes at 2 mg/mL with 500 mcg alum (from 5 mg/mL 
suspension) for two 0.5 mL doses in two sites (total dose of 2000 mcg in 1 
mL) administered by IM injection in the deltoid muscles (one injection into 
each deltoid) 
• Sodium Chloride for Injection USP, 0.9% administered for two 0.5 mL doses 
in two sites by IM injection in the deltoid muscles (one injection into each 
deltoid) 

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Table 3-1 Schema 
Study arm 
N 
Month 0 
 (Day 0) 
Month 2 
(Day 56) 
Month 6 
(Day 168) 
Month 12 
(Day 364) 
 
 
 
 
 
 
Group 1 
5 
500 mcg 
peptide 
500 mcg 
peptide 
500 mcg 
peptide 
500 mcg 
peptide 
 
 
 
 
 
 
 
1 
Placebo 
placebo 
placebo 
placebo 
 
 
 
 
 
 
 
 
 
 
 
 
Group 2 
15 
2000 mcg 
peptide 
2000 mcg 
peptide 
2000 mcg 
peptide 
2000 mcg 
peptide 
 
 
 
 
 
 
 
3 
Placebo 
placebo 
placebo 
placebo 
 
 
 
 
 
 
Total  
24 
(20/4) 
 
 
 
 
Note: 
Safety data for Group 1 through 2 weeks after the first vaccination will inform 
whether to proceed with escalation to the higher dose (Group 2). 
Participants 
24 healthy, HIV-1–uninfected volunteers aged 18 to 50 years; 20 vaccinees, 4 
placebo recipients 
Design 
Multicenter, randomized, dose escalation, placebo-controlled, double-blind trial 
Duration per participant 
18 months of scheduled clinic visits per participant (main study), followed by an 
adverse event of special interest (AESI) health contact at month 24 
Estimated total study duration 
30 months (includes enrollment, planned safety holds, follow-up, and AESI health 
contact) 
Investigational New Drug (IND) sponsor 
DAIDS, NIAID, NIH, DHHS (Bethesda, Maryland, USA) 
Study product provider 
• MPER-656 liposomes: International AIDS Vaccine Initiative, (IAVI, New 
York, New York, USA) 

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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 
University of Washington Virology Specialty Laboratory (UW-VSL) (Seattle, 
Washington, USA) 
Endpoint assay laboratories 
• Duke University Medical Center (Durham, North Carolina, USA) 
• Fred Hutch/University of Washington (Seattle, Washington, USA) 
Study sites 
HVTN Clinical Research Sites (HVTN CRSs) in the United States to be specified 
in the Site Announcement Memo 
Safety monitoring 
HVTN 133 PSRT; HVTN Safety Monitoring Board (SMB) 

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3.1 
Protocol Team 
Protocol leadership 
Chair 
Lindsey Baden 
Brigham & Women’s 
Hospital 
617-732-6801 
lbaden@bwh.harvard.edu 
Statistician 
Paul Edlefsen 
SCHARP, Fred Hutch 
206-667-4086 
pedlefsen@gmail.com 
Co-chair 
Nathan Erdmann 
University of Alabama 
205-975-1690 
nerdmann@uab.edu 
Medical officer 
Edith Swann 
DAIDS, NIAID 
240-627-3035 
swanne@niaid.nih.gov 
Protocol Team 
leader 
Cecilia Morgan 
HVTN Core, Fred Hutch 
206-667-5875 
cmorgan@fredhutch.org 
Laboratory lead 
David Montefiori 
HVTN Laboratory Program 
919-684-5278 
David.Montefiori@duke.edu 
Other contributors to the original protocol 
Core medical monitor 
Nicole Grunenberg 
HVTN Core, Fred 
Hutch 
Clinical safety specialist 
Megan Jones 
HVTN Core, Fred Hutch 
Study product developer 
representatives 
Tony Moody 
Duke University 
Clinical trials manager 
Carrie Sopher 
HVTN Core, Fred Hutch 
 
Zachary Sagawa 
IDRI 
Clinical data manager 
Kris Donaty 
SCHARP, Fred Hutch 
Laboratory center 
representative 
Lisa Sanders 
HVTN Laboratory 
Program, Fred Hutch 
SDMC Associate director 
of lab science 
April Randhawa 
SCHARP, Fred Hutch 
Regulatory affairs 
associate 
Meg Brandon 
HVTN Core, Fred 
Hutch 
DAIDS Project Officers 
Shyam Rele 
DAIDS, NIAID 
Clinic coordinators 
Pam Cunningham 
Birmingham CRS 
 
Chris Hamlin 
DAIDS, NIAID 
Community Advisory 
Board (CAB) members 
Courtney Schopke 
Boston CAB 
Protocol development 
manager 
Ramey Fair 
HVTN Core, Fred Hutch 
 
Edward Jackson 
Birmingham CAB 
Community engagement 
unit representative 
Stephaun Wallace  
HVTN Core, Fred Hutch 
DAIDS protocol 
pharmacist 
Kelly Parsons 
DAIDS, NIAID 
240-669-5721 
Community 
educator/recruiter 
Brianna Patterson 
Birmingham CRS 
 
Oladapo Alli 
DAIDS, NIAID 
240-627-3593 
Technical editor 
Richa Chaturvedi 
HVTN Core, Fred Hutch 
Statistical research 
associate 
Mengshu Shao 
SCHARP, Fred 
Hutch 
 
 
 
 
 
 

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4 
Background 
4.1 
Rationale for trial concept 
Elicitation of broadly neutralizing antibodies (bnAbs) against the HIV-1 envelope 
(Env) is likely to be central to a successful protective HIV-1 vaccine (4). There 
are multiple conserved sites on Env that are targets for bnAbs but no current 
vaccination regimens elicit these kinds of antibodies. A subset of chronically 
infected subjects make bnAbs, and this has allowed identification of several 
characteristics shared by all bnAbs (5).  
One highly conserved and well-studied vaccine target in Env is gp41 near the 
viral membrane proximal external region (MPER) where several bnAbs bind (6). 
One of these bnAbs, 2F5, has been shown to protect against vaginal 
simian/human immunodeficiency virus (SHIV) transmission (7) when given 
passively suggesting that, if induced in sufficient titers, such bnAbs may be 
effective for prevention of HIV-1 infection. The MPER of gp41 is a highly 
conserved region, rich in aromatic residues, and its role in HIV-1 fusion is evident 
from studies showing that mutation of tryptophan residues in the MPER inhibits 
cell fusion and viral infectivity (8). However, MPER-specific bnAbs are rarely 
made in natural HIV-1 infection (9) and as yet are not made following HIV-1 
envelope protein vaccination (10, 11).  
Immunological tolerance mechanisms (12, 13), which cause profound clonal 
deletion of B cells expressing autoreactive bnAbs like 2F5 (14), have been 
postulated to limit induction of MPER gp41 bnAbs. Verkoczy and Haynes have 
recently demonstrated in mice that although most precursors of MPER bnAbs are 
deleted at the first tolerance checkpoint in bone marrow, a small population (5%) 
of bnAb precursors emigrate from the bone marrow and exist in peripheral 
immune organs as anergic B cells that can be activated by an appropriate 
immunogen to produce 2F5 bnAbs in plasma (15). These residual anergic clones 
are competent to undergo class-switching, activation and expansion in response to 
immunization resulting in potent serum IgG neutralizing responses. Thus, in 
principle, overcoming tolerance of self-reactive bnAb-expressing B cells by 
immunization is a feasible part of an antibody-based HIV-1 vaccine strategy. 
Despite these data showing tolerance control of this class of antibodies, there are 
data that suggest it might be possible to elicit such antibodies by vaccination. 
While most individuals develop bnAbs only after several years of infection, 
Sather et al. described two HIV-1–infected persons who developed MPER-
directed bnAb activity one year after infection (16) suggesting that it may be 
possible to develop MPER-directed bnAb activity relatively quickly compared 
with other bnAb epitopes (17). Rhesus macaques immunized using MPER 
peptides displayed on liposomes (described in Section 4.2) developed antibodies 
that recognized the 2F5-epitope (18, 19), and monoclonal antibodies isolated from 
these macaques demonstrated a maturation pattern similar to that of 2F5-including 
broad neutralization in an assay using TZM-bl cells that express CD64 (FcγR1) 

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(19). Thus, MPER peptide liposomes are an immunogen that have been shown to 
initiate the development of 2F5-like bnAb precursors in rhesus macaques, 
providing a rationale for the testing of this immunogen in human participants. 
This study will determine if 2F5-like bnAb precursors can be elicited in humans 
utilizing an MPER peptide thus opening the path to exploiting B cell lineage 
development as a mechanism to elicit bnAbs. 
4.2 
GTH1-656 MPER peptide liposome 
Rationale for gp41 MPER peptide liposome immunogen design 
Data suggest that MPER residues are oriented or presented differently on a lipid 
bi-layer compared to free peptides (20-22). MPER peptides alone as immunogens 
generally are ineffective for the induction of bnAbs (23). MPER peptides in 
micelles or in liposomes with membrane anchor tags are likely to be less flexible 
and more likely to adopt a relatively ordered conformation (20, 21). It has been 
shown that stable scaffold structures of MPER peptides can induce antibody 
responses that target the 2F5-bound peptide conformation (23). Studies using 
synthetic peptide-liposome conjugates have shown that membrane anchoring is 
important for the presentation of 2F5 epitope structure (24); the N and C termini 
of the gp41 MPER are differentially exposed on the membrane; and the stable 
docking of monoclonal antibodies (mAbs) 2F5 to peptide-liposomes is influenced 
by the extent of membrane immersion of the binding epitopes (21). 
 
Figure 4-1 Design of the MPER peptide liposome 
Figure 4-1 shows the general design of the MPER peptide-liposome. The MPER-
656 liposomes immunogen includes the relevant gp41 MPER component and a 

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lipid component to induce membrane binding. The MPER-656 peptide sequence 
(656NEQELLELDKWASLWNWFNITNWLWYIK683) is derived from the 
membrane proximal region of HIV-1 gp41 and includes the core epitopes of the 
broadly neutralizing mAbs 2F5 (662ELDKWAS), 4E10 (671NWFNIT), and 10E8 
(671NWF676T683K). The MPER-656 peptide was synthesized with a C-terminal 
hydrophobic membrane anchor tag (YKRWIILGLNKIVRMYS) which facilitates 
anchoring of the peptide to synthetic liposomes and presentation of the optimal 
peptide conformation for MPER bnAb binding, while occluding the binding of 
non-neutralizing MPER antibodies (21, 24). The gp41 MPER-656 peptide 
liposome immunogen has been shown to be able to induce triggering of anergic 
2F5 bnAb cells in 2F5 knock-in mice, resulting in high levels of plasma bnAb. 
MPER-656 peptide adjuvanted liposomes were constructed as described (21, 24, 
25). 
4.3 
Alhydrogel® adjuvant 
Alhydrogel® adjuvant, aluminum hydroxide (alum), is a commonly used vaccine 
adjuvant and will be administered with this peptide vaccine. 
4.4 
Trial design rationale 
This study will evaluate the safety and immunogenicity of a novel gp41 MPER 
peptide-liposome vaccine in healthy HIV-1–uninfected adults. The study will 
evaluate 2 different dose levels of the vaccine (500 and 2,000 mcg) in a dose 
escalation trial. Safety and tolerability will be assessed after the first vaccination 
and safety parameters met before advancing to the higher dose level. Vaccination 
will be given by needle and syringe intramuscularly. Each dose will be split in 
two and given at 2 separate injection sites. 
Preclinical testing of this and other HIV-1 vaccine candidates suggests that 
sequential immunization with immunogens, perhaps over a prolonged period of 
time, may be required to induce bnAb production. This vaccine will be given at 4 
timepoints over a period of 12 months to determine the degree to which MPER-
directed bnAb precursors can be elicited and matured by this vaccine candidate. 
Safety, tolerability, and immunogenicity data will be collected after each 
vaccination. 
Preclinical trials have given up to 2 mg of peptide-liposome vaccine without 
significant toxicity. However, a human mAb targeting the epitope included in this 
vaccine has been shown to react with cardiolipin histones and centromere B 
autoantigens (26). Participants in this study will be monitored for vaccine-induced 
toxicities. This vaccine is designed to elicit 2F5-like bnAb precursors, and human 
trials of passive immunization with up to 5 grams of 2F5 antibody did not result 
in clinical prolongation of clotting times (activated partial thromboplastin time 
[aPTT], prothrombin time [PT] and dilute Russell’s viper venom time [DRVVT]) 
and no binding to cardiolipin was seen in clinical samples. In human studies 

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administering combinations of mAbs 2F5, 4E10, and 2G12, slight rises in binding 
to phosphatidylserine and cardiolipin were seen in plasma along with a mild, 
transient prolongation of aPTT in 4/12 participants (27). Because of this possible 
autoreactivity, volunteers will be screened for pre-existing autoimmune disorders 
and safety labs will include evaluation of clotting times as well as induction of 
anti-cardiolipin antibody and anti-nuclear antibody. The full MPER neutralizing 
antibody (nAb) epitope, ELDKWA, is present in the host tryptophan catabolism 
pathway enzyme kynureninase (KYNU). It is not expected that antibodies against 
this enzyme will have clinically apparent adverse effects, and studies in mice and 
non-human primates (NHPs) have shown that immunization with MPER peptide 
liposome vaccine did not alter tryptophan metabolism (18). 
4.4.1 
Dose selection 
Based on preclinical studies (see Section 4.7 below) two doses have been selected 
to be given in a 4-dose regimen over 12 months. This will allow assessment of a 
dose response, including up to the maximum studied in the NHP model, to 
determine if bnAbs can be elicited via this approach. 
4.4.2 
Schedule 
At present it is not known what the optimal vaccine interval is for eliciting HIV-1 
bnAbs. Natural history studies show that most persons who develop breadth do so 
only after years of infection (28, 29), and to date no vaccine candidate has reliably 
elicited bnAbs, making the rational choice of a vaccine interval challenging. Data 
from individuals who have been found to make MPER-directed bnAbs vary, with 
some individuals making bnAbs after only 1 year of infection (16) while others 
appear to have required a longer period of time (30). Trials with Env immunogens 
that did not elicit bnAbs but that were associated with protection (ie, RV144 and 
the follow-on RV305 study) suggest that immunogenicity and lineage maturation 
may be enhanced/improved when boosting occurs after a prolonged rest (31). The 
length of time between immunizations to increase bnAb development is not 
known and must be inferred based on other analyses such as the expansion of B 
cell lineages or B cell precursors of bnAb lineages. For these reasons, a longer 
interval of 4 months (vaccine #3) and 6 months (vaccine #4) is proposed. 
4.4.3 
Control 
Placebo used will be Sodium Chloride for Injection USP, 0.9% 
4.5 
Plans for future product development and testing 
These data will inform our ability to elicit bnAbs via vaccination, and in 
particular, this trial will determine whether this immunogen can elicit and expand 
MPER-directed bnAb precursors in humans. Success in this trial would include 
the elicitation of neutralizing antibodies directly or the initiation of bnAb lineages 
that could be further matured using additional vaccine strategies. If successful, an 

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important avenue of HIV-1 vaccine development will be opened up and follow-on 
studies to increase the efficiency of antibody elicitation and breadth of the 
immune response will be pursued. 
4.6 
Preclinical safety studies 
4.6.1 
Evaluation of kynureninase cross-reactivity 
4.6.1.1 
Mouse #059 
The HIV MPER gp41 bnAb epitope ELDKWA is present in enzyme 
kynureninase (KYNU), which is in the tryptophan catabolism pathway and the 
2F5 bnAb cross-reacts in vitro with monomeric KYNU (18). Because MPER-
peptide liposomes are designed to elicit 2F5-like antibodies which can cross-react 
with KYNU, it is possible that immunization with this vaccine will have an 
autoimmune effect targeting KYNU enzyme activity. This risk of autoimmunity 
was assessed in 2F5 knock-in (KI) mice which were engineered to express the 
broadly and potently neutralizing mature HIV-1 antibody 2F5 that binds to gp41 
MPER (15). MPER-656 liposomes were administered in combination with 
Glucopyranosyl Lipid Adjuvant (GLA: a synthetic toll-like receptor 4 agonist, 
that has been used as an adjuvant in investigational vaccine studies) or alum (19). 
In addition to MPER binding antibodies, KYNU-reactive antibodies were 
detected by enzyme-linked immunosorbent assay (ELISA). In an in vitro 
enzymatic assay of KYNU activity, serum from all MPER-656 liposome-
immunized mouse groups had no impact on KYNU activity. Additionally, 
immunization had no effect on levels of tryptophan, kynurenine and kynurenic 
acid in serum and the brain measured by liquid chromatography-mass 
spectrometry. Histopathology of brain and spleen tissue revealed no pathological 
changes as a result of immunization (19). These results indicated KYNU-reactive 
antibodies elicited by MPER-656 liposomes had no impact on the KYNU 
metabolic pathway metabolites in serum or brain nor had any impact on brain or 
spleen histopathology. KYNU reactivity was further evaluated in rhesus 
macaques as described in Section 4.6.1.2. 
4.6.1.2 
NHP #204/#205 
The effect of MPER-656 liposome immunization on KYNU activity was also 
evaluated in NHP studies 204 and 205. Immunogenicity outcomes are 
summarized in Section 4.7.1.4. While KYNU-reactive, MPER gp41-reactive 
antibodies were elicited by MPER-656 liposomes, those antibodies had no 
detectable impact on in vitro KYNU activity, and histopathology of spleen tissue 
was normal in all groups. 4E10-like antibodies, which would pose a greater risk 
of autoreactive pathogenicity, were not detected in 4E10 blocking assays, 
indicating that these antibodies are not elicited by MPER-656 liposomes. Rhesus 
macaques in Studies 204 and 205 were also monitored for changes in 
comprehensive clinical chemistry and hematology assays. No significant 
abnormalities deemed related to study treatment were observed, although mild 

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anemia was noted throughout the study in all groups, as well as transient low 
phosphorous measurements (19). The anemia was likely a result of the blood 
draw volumes and the physiological stress of study procedures, despite adherence 
to safe blood volume limits. In combination with the 2F5 mouse data in Section 
4.6.1.1, these data suggest that MPER-binding antibodies that are KYNU cross-
reactive will not impair host tryptophan metabolism in human subjects. 
4.6.2 
NHP #208 
A dose study in rhesus macaques of MPER-656 liposomes mixed with alum 
(Alhydrogel®) tested the three vaccine doses planned for the clinical study. 
Immunogenicity results are summarized in Section 4.7.1.5. There were no adverse 
events in this study thought to be related to the vaccine, although 2 of 20 animals 
were anemic at all timepoints after the initial immunization, both of which were in 
the empty liposome and alum control group. As with NHP #204/205, the anemia 
was likely the result of blood draws and the stress of study procedures. One 
animal was necropsied at week 11 due to more profound anemia (total 
hemoglobin of 4.9 mg/dL) and found to have erythroid hypoplasia on bone 
marrow but with no obvious association to the study treatment. As with NHP 
#204/#205, autoreactive 4E10-blocking antibodies were not detected. These 
results support the safety of the use of MPER-656 liposomes with alum at the 
planned clinical dose range. 
4.6.3 
GLP Rabbit Toxicology Study CRL505285 
A GLP toxicology study in New Zealand White Rabbits was designed to 
determine the potential toxicity and local tolerance of MPER-656 liposomes when 
administered by IM injection with alum once every two weeks for 10 weeks (total 
6 injections) and to evaluate the potential reversibility of any findings. Three 
groups of twenty animals were dosed as indicated in Table 4-1. 
Table 4-1 Repeat dose toxicity schedule 
Gp 
Test article 
Dosing Days 
Animals/Group 
Primary 
Necropsy 
SD 74 
Recovery 
Necropsy 
SD 92 
1 
Vehicle control 
1, 15, 29, 43, 57, 71 
5M/5F 
5M/5F 
2 
2 mg peptide liposome 
+ 500 mcg alum* 
1, 15, 29, 43, 57, 71 
5M/5F 
5M/5F 
3 
0.2 mg peptide 
liposome + 500 mcg 
alum* 
1, 15, 29, 43, 57, 71 
5M/5F 
5M/5F 
The toxicity study demonstrated an acceptable safety profile for use in human 
subjects at the highest dose planned for clinical use. 

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4.7 
Preclinical immunogenicity studies 
Table 4-2 Summary of preclinical immunogenicity studies 
Study 
number 
Animal 
N/group 
Dose (mg) 
groups 
Route 
Schedule (wks) 
VMU 
#029 
2F5 Mature KI 
mice 
3 
0.025 
IP 
0, 2, 4, 6, 8, 10 
VMU 
#048 
2F5 Mature UA 
mice 
4 
0.025 
IP 
0, 2, 4, 6, 8, 10 
NHP #22 
Rhesus 
macaques 
4 
0.35 
mucosal; 
1.0 IM 
mucosal; 
IM 
JRFL gp140 prime and boost with 
MPER-liposomes: mucosal at wks 
12, 16, 20, 24 and IM. at wks 41, 
45, 50, 88, 92, 129, 135, 137, 142, 
148 
NHP #204 
Rhesus 
macaques 
8 
1.0 
IM 
0, 6, 12, 18, 24, 30 
NHP #205 
Rhesus 
macaques 
7 
1.0 
IM 
0, 6, 12, 18, 24, 30 
NHP #208 
 
4 
2.0 
IM 
0, 5, 8 
R25 
Rabbits 
3 
2.0 
IM 
0, 3, 6, 9, 12 
IM = intramuscular, IP = intraperitoneal.  
4.7.1 
Immunogenicity of HIV-1 MPER peptide-liposome 
4.7.1.1 
Study VMU #029 
Initial proof-of-concept data were generated using a prototype of the MPER-656 
liposomes formulation made at the Duke Human Vaccine Institute. 2F5 KI mice 
were engineered to express the broadly and potently neutralizing mature HIV-1 
antibody 2F5 that binds to gp41 MPER and is also autoreactive (15). In these 
mice, the majority of the 2F5 B cells are deleted in the bone marrow, leaving a 
small population of anergic B cells in the periphery (19). The 2F5 KI model has 
provided a system to determine if an MPER-656 liposome designed to optimally 
bind to the 2F5 antibody can selectively target and/or reverse functional silencing 
of residual autoreactive B-cell subsets with B-cell receptor (BCR) dual reactivity 
(ie, lipid/MPER) required for 2F5's neutralization ability (32). 
In the first proof-of-concept study, 2F5 mature heavy chain variable region (VH) 
+ light chain variable region (VL) KI mice received an HIV-1 JRFL gp140 Env 
prime followed by 6 biweekly immunizations with MPER-656 liposomes 
manufactured with monophosphoryl lipid A (MPLA) and mixed with the 
adjuvants Emulsigen and CpG before injection. Class-switching of 2F5 bnAb B 
cells was observed, and high levels of plasma 2F5 antibody were produced in 
mature 2F5 KI mice, thus demonstrating that when B cells with bnAb BCRs are 
present in sufficient numbers, the MPER-656 liposome vaccine is antigenic and 
immunogenic for their activation (15). Despite the immunological tolerance that 
results in deletion of the majority of 2F5-recognizing B cells in bone marrow 

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from 2F5 KI mice, the small population of 2F5 BCR-bearing B cells that emerge 
as anergic B cells can be activated to produce 2F5 bnAbs in plasma, and are 
competent to undergo class-switching, activation and expansion in response to 
immunization resulting in potent serum IgG neutralizing responses (15). 
Thus, in principle, overcoming tolerance of self-reactive bnAb-expressing B cells 
by immunization is a feasible part of an antibody-based HIV-1 vaccine strategy. 
Hence, a major goal of the human clinical trial of MPER-656 liposomes is to 
determine if the germline-targeting immunogen can expand MPER-targeted bnAb 
precursors in humans. 
4.7.1.2 
VMU #048 
In a second study, a related strain of 2F5 KI mice were used, that expressed the 
VH and VL of the germline unmutated ancestor (UA) of the 2F5 antibody. These 
2F5-UA mice manifested even greater B cell deletion than the 2F5-mature VH + 
VL KI mice (32). After six immunizations of these 2F5-UA mice with MPER-656 
liposomes, the remaining anergic B cells were capable of being activated by 
germline-binding immunogens to make gp41-reactive immunoglobulin M (IgM), 
but experienced limited class-switching to IgG and limited somatic mutations 
required for neutralizing activity (32). This study demonstrated that immunization 
with MPER-656 liposomes can at least partially rescue anergic bnAb precursors 
by inducing activation and expansion of the population of 2F5 UA BCR-bearing 
naïve B cells. 
4.7.1.3 
NHP Study #22 
An early study in rhesus macaques evaluated the ability of MPER-656 liposomes 
to initiate gp41-specific B cell lineages. This study utilized an immunization 
regimen of a prime with vaccinia virus-expressed HIV-1 JRFL gp140 Env and a 
combination of HIV-1 JRFL gp140 Env protein and MPER-656 liposome boosts, 
using MPLA, R848 and ODN10103 (CpG) as the MPER-656 liposome adjuvant, 
followed by multiple MPER-656 liposome boosts as described in Zhang et al, 
2016 (19). Immunized macaques made B cell clonal lineages targeted to the 2F5 
bnAb epitope, but 2F5-like antibodies were either deleted or did not attain 
sufficient affinity for gp41-lipid complexes to achieve the neutralization potency 
of 2F5 (19). This limitation of neutralization potency was shown to be due to 
MPER-reactive B cell control by tolerance mechanisms preventing mutations to 
generate the needed hydrophobic HCDR3 regions needed for bnAb breadth (19). 
4.7.1.4 
NHP Study #204 and #205 
Two NHP studies were designed to evaluate the effects of the adjuvant regimens 
(GLA or alum) on the immunogenicity of the MPER-656 liposome product. NHP 
204 tested the immunogenicity of the MPER-656 liposome product formulated 
with GLA or GLA plus alum (n=8 per group), and NHP 205 tested the MPER-
656 liposome vaccine with alum alone (n=7), as described by Bradley et al. (18). 
Rhesus macaques were immunized with 1 mg of MPER-656 liposomes six times, 

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six weeks apart, formulated with either 25 mcg of GLA, or 500 mcg alum, or a 
combination of both adjuvants. 
After 2 immunizations in rhesus macaques, MPER-656-specific antibodies were 
present in all animals in the alum alone and GLA plus alum arms but not in the 
GLA-alone adjuvant arm. Sequential vaccinations did not further boost the 
immune response above that observed following the 2nd injection, perhaps due to 
hyperimmunization at 6-week intervals (18), a period shorter than that proposed 
for the current study.  
These NHP studies demonstrated that the MPER peptide contained within 
liposomes was antigenic and can be used to generate antibodies against the 2F5 
epitope in immunized NHPs after >1 immunization. The data also support the use 
of alum as an adjuvant in this phase 1 clinical study. A neutralizing antibody 
response was not detected in these NHPs, likely due to the immune tolerance 
control mechanisms indicated above for NHP 22 study (Section 4.7.1.3), but data 
from that study did indicate that the vaccine regimen could begin the maturation 
process for 2F5-like antibodies, and this phase 1 study will determine whether the 
same initiation of bnAb lineages can occur in humans. 
4.7.1.5 
NHP Study #208 
A dose study was performed in Rhesus macaques (NHP 208) to test three vaccine 
doses of MPER-656 liposomes mixed with alum (Alhydrogel®). This study 
covered the dose range used for the preclinical GLP toxicity study and the 
proposed HVTN clinical study (0.2, 1.0, and 2.0 mg MPER-656 liposomes, all 
with 0.5 mg alum adjuvant). There were also 2 control groups—2.0 mg MPER 
liposomes without alum, and empty liposomes plus 0.5 mg alum (N=4/ group). 
All animals were immunized at weeks 0, 4, and 8 (three immunizations) and bled 
on the immunization days, two weeks after each immunization, and a final bleed 
was collected at 12 weeks (four weeks after the last immunization) for antibody 
binding titers. Plasma antibody binding results at the week 12 timepoint showed 
that adjuvant was required to achieve the highest MPER-binding titers in this 
study, but there was no significant difference in MPER antibody titers between 
the tested doses (Figure 4-2). 

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Figure 4-2 Alum adjuvant significantly enhances MPER-specific antibodies in rhesus 
macaques. At week 12, four weeks after the final immunization, MPER-binding plasma antibodies 
were detected by ELISA. Data are shown as the log-transformed area under the curve (AUC) 
values with each data point representing a single animal. Alum significantly enhanced antibodies 
levels at all MPER-peptide liposome doses, and there were no significant differences between the 
three doses (P < 0.05; Wilcoxon Mann-Whitney compared to no adjuvant). 
There is only a 4-fold difference between the lowest and highest doses planned 
for the clinical trial. There was no significant difference in MPER-specific 
binding antibodies tested at week 12 between any of the treatment groups in this 
NHP study.  
4.7.1.6 
Rabbit study R25 
An early immunogenicity study was performed in rabbits to confirm relevance of 
this animal model for use in the toxicity study. Only the high dose was used in 
this study, 2.0 mg of MPER-656 liposomes containing 1.0 mg alum. The data 
demonstrated that the MPER-656 liposome vaccine was immunogenic, supporting 
the rabbit as an appropriate model for the GLP toxicity study, and no safety 
concerns were observed. 
Three rabbits received an IM immunization every 2 weeks (for a total of 5 
immunizations) with 2 mg of peptide in MPER-656 liposome vaccine formulated 
with alum (1.0 mg of total aluminum). Bleeds were taken 10 days after each 
immunization. Results (measured by direct-binding ELISA) for longitudinal 
binding of serum antibodies from R25 rabbits to the MPER-656 (MPER 656-
GTH1) and the 2F5 bnAb epitope (sp62 peptide 
QQEKNEQELLELDKWASLWN) are shown in Figure 4-3. 

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Figure 4-3 Longitudinal binding by direct-binding ELISA of serum antibodies from R25 
rabbits to the MPER-peptide (MPER656-GTH1) and the 2F5 bnAb epitope (sp62). Animals 
were immunized every 3 weeks, as indicated by arrows. AUC: area under the curve. Horizontal 
lines indicate the geometric mean. 
Immunization with 2 mg of MPER-656 liposome vaccine formulated with alum 
(1.0 mg total aluminum) induced antibodies in rabbits that bound to the GTH1-
656 MPER peptide and the 2F5 bnAb epitope peptide sp62. Sequential 
vaccinations did not further boost the immune response above that observed 
following the 1st injection, possibly due to hyperimmunization at 3-week 
intervals. 
4.8 
Clinical studies 
4.8.1 
Rationale for human trials 
Preclinical trials have shown that immunization with HIV-1 MPER peptide-
liposome vaccines can initiate bnAb lineages against HIV-1 gp41 (15) in mice 
and non-human primate models (25). The purpose of this trial will be to determine 
if MPER-directed bnAb precursors can be expanded in humans. The primary 
objective of this phase 1 study will be to determine the safety of the MPER 
peptide liposome vaccine in healthy adults. In previous studies with small animals 
(guinea pigs) an MPER peptide immunogen dose of 100 mcg per animal induced 
antibodies that bound to peptides that included the bnAb 2F5 epitope (25). A 
higher immunogen dose of 1 mg administered to NHP induced antibodies that 
mapped to the core (664DKW) 2F5 epitope and also bound avidly to the 
conformationally relevant gp41-inter and 2F5 scaffold proteins (25). Thus, in 
human vaccinees, we will evaluate doses of 0.5 mg and 2 mg of peptide in a dose 
escalation fashion. Preclinical trials utilizing doses as high as 2 mg peptide in 
liposomes have shown no adverse effects. This study will evaluate two doses over 
a longer period of time (4 immunizations over a 12-month period). Safety and 

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tolerability will continue to be the primary endpoints but immunogenicity, 
including binding antibody that maps to the 2F5-epitope, expansion of antibodies 
with characteristics of MPER-directed bnAb B-cell lineages, and evaluation of 
sera for the production of bnAbs will be important endpoints. 
The MPER peptide included in the vaccine includes epitopes for both 2F5 and 
4E10 bnAbs. Despite the inclusion of the 4E10 epitope, antibodies to this area of 
the peptide have not been elicited in any preclinical trials (15). This is felt to be 
due to the fact that the 4E10 epitope is located adjacent to the anchoring tag and 
thus is likely not well presented. For this reason, the immunological endpoints in 
this study will be directed to the 2F5-epitope. As noted in Section 4.4, some 
MPER bnAbs including 2F5 and 4E10 monoclonal antibodies (mAbs) have 
autoreactivity. The preclinical data presented above demonstrate that this 
immunogen does not elicit pathological antibodies supporting the safe use of the 
MPER-656 liposome vaccine in humans. We do not anticipate problems with 
those kinds of antibodies in this trial, but our monitoring plan is designed to 
capture those problems should they arise. 
Development of bnAbs appears to be a lengthy process in HIV-1–infected 
persons, and vaccines to elicit bnAbs will likely require sequential immunogens 
over a long period of time. This trial will evaluate whether MPER peptide 
liposomes can expand 2F5-epitope directed bnAb precursor B cell pools in 
humans, a critical first step in the development of a vaccine to elicit MPER-
directed bnAbs. 

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4.9 
Potential risks of study products and administration 
Table 4-3 Summary of potential risks of study products and administration 
Common 
• 
Mild to moderate injection site pain, tenderness, erythema, or 
swelling/induration/edema 
• 
Malaise/fatigue, myalgia, or headache in the first few days following 
injection 
• 
A vaccine-induced positive HIV antibody test result 
Less common 
• 
Severe injection site pain or tenderness 
• 
Fever, chills, flu-like syndrome, arthralgia, rash, nausea, or dizziness in the 
first few days following injection  
• 
Vasovagal reaction/lightheadedness/dizziness related to the injection 
procedure 
• 
Transient changes in clinical laboratory values 
• 
Injection site hematoma, bruising/ecchymosis, other transient lesions, 
itching, or bleeding related to the injection procedure 
Uncommon or rare 
• 
Severe localized injection site reaction, such as sterile abscess or secondary 
bacterial infection  
• 
Allergic reaction, including rash, urticaria, angioedema, bronchospasm, or 
anaphylaxis 
• 
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 
• 
Effects on the fetus and on pregnancy 

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5 
Objectives and endpoints 
5.1 
Primary objectives and endpoints 
Primary objective 1:  
To evaluate the safety and tolerability of different doses of a prime-boost regimen 
of MPER-656 liposomes in HIV-uninfected healthy adults 
Primary endpoint 1:  
• Local and systemic reactogenicity signs and symptoms, laboratory measures 
of safety, and AEs and SAEs 
Primary objective 2:  
To evaluate binding antibody responses elicited by different doses of the MPER-
656 liposome vaccine 
Primary endpoint 2:  
• MPER-peptide-specific IgG binding Ab responses as assessed by binding Ab 
multiplex assay 2 weeks after the 3rd and 4th vaccination with MPER-656 
liposome vaccine  
5.2 
Secondary objectives and endpoints 
Secondary objective 1: 
To assess other vaccine elicited antibodies including the mapping of MPER-
specific binding antibody responses and lipid-specific antibody responses 
Secondary endpoint 1: 
• MPER-peptide specific antibody responses as assessed by binding Ab 
multiplex assay and/or other binding assays using an array of antigens that 
identify epitope- and lipid-specific responses to the vaccine 2 weeks after the 
3rd and 4th vaccinations with MPER-656 liposomes 
Secondary objective 2: 
To evaluate the ability of different doses of the vaccine regimen to elicit HIV-
specific nAbs 
Secondary endpoint 2: 

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• nAb responses against HIV-1 viral isolates 2 weeks after the 3rd and 4th 
vaccinations with MPER-656 liposomes 
5.3 
Exploratory objectives 
Exploratory objective 1: 
To determine the frequency and phenotype of MPER-specific B cells 
Exploratory objective 2: 
To isolate single B cells with desired specificities and determine lineage 
characteristics. Monoclonal antibodies may be evaluated for binding and 
neutralization including tier 2 virus bnAb activity and 2F5 and 10E8 binding site 
binding. Serum antibodies may be evaluated as well. 
Exploratory objective 3: 
To evaluate HIV-specific T-cell responses induced by the MPER-656 liposome 
vaccine 
Exploratory objective 4: 
To assess vaccine-induced blood follicular helper T-cell (Tfh) responses 
Exploratory objective 5: 
To further evaluate immunogenicity of each vaccine regimen, additional 
immunogenicity assays may be performed, including on samples from other 
timepoints, based on the HVTN Laboratory Assay Algorithm. 
Exploratory objective 6: 
To conduct analyses related to furthering the understanding of HIV, immunology, 
vaccines, and clinical trial conduct. 

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6 
Statistical considerations 
6.1 
Accrual and sample size calculations 
Recruitment will target enrolling 24 healthy, HIV-uninfected adult participants 
aged 18 to 50 years old at low risk of HIV infection in regions where clade B is 
the predominant clade. 
Enrollment will be concurrent with receiving the first study vaccination, thus all 
participants will provide some safety data. For immunogenicity analyses, 
however, it is possible that data may be missing for various reasons such as 
participants terminating from the study early, problems in shipping specimens, or 
low cell viability of processed peripheral blood mononuclear cells (PBMCs). 
Immunogenicity data from 11 phase 1 and 1 phase 2a HVTN trials, which began 
enrolling after June 2005 (data as of June 2011), indicate that 10% is a reasonable 
estimate for the rate of missing data. For this reason, the sample size calculations 
in Section 6.1.2 account for 10% 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 Group 1 vaccine arm (n = 5), there is at least a 90% 
chance of observing at least one event, if the true rate is 37% or more, and there is 
at least a 90% chance of observing no events, if the true rate is 1% or less. In the 
Group 2 vaccine arm (n = 15), there is at least a 90% chance of observing at least 
1 event if the true rate of such an event is 14.3% or more, and there is at least a 
90% chance of observing no events if the true rate is 0.6% or less. In all vaccine 
arms of the study combined (n = 20), there is at least a 90% chance of observing 
at least one event, if the true rate is 10.9% or more, and there is at least a 90% 
chance of observing no events, if the true rate is 0.5% or less. 
As a reference, in HVTN vaccine trials from April 2008 through March 2018 
conducted in the US, about 1.7% of participants who received placebos 
experienced an SAE. 
Binomial probabilities of observing 0, 1 or more, and 2 or more events among 
arms of size 15 and 5 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. 

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Table 6-1 Probability of observing 0 events, 1 or more events, and 2 or more events, among 
arms of size 15 and 5, for different true event rates 
True event rate (%) 
Pr(0/15) 
Pr(1+/15) 
Pr(2+/15) 
Pr(0/5) 
Pr(1+/5) 
Pr(2+/5) 
1 
86 
14 
1 
95.1 
4.9 
0.1 
5 
46.3 
53.7 
17.1 
77.4 
22.6 
2.3 
10 
20.6 
79.4 
45.1 
59 
41 
8.1 
20 
3.5 
96.5 
83.3 
32.8 
67.2 
26.3 
30 
0.5 
99.5 
96.5 
16.8 
83.2 
47.2 
40 
0 
100 
99.5 
7.8 
92.2 
66.3 
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 a particular observed rate. Calculations are 
done using the score test method (33). If none of the 20 participants receiving a 
vaccine regimen experience a safety event, the 95% 2-sided upper confidence 
bound for the true rate of such events in the total vaccinated population is 16%. 
For the Group 1 vaccine arm (n = 5), the 2-sided upper confidence bound for this 
rate is 43%. For the Group 2 vaccine arm (n = 15), the 2-sided upper confidence 
bound for this rate is 20%. 
Table 6-2 Two-sided 95% confidence intervals for the probability of observing a safety event 
based on observing a particular rate of safety endpoints for arms of size 5, 15 and 20 
Observed event rate 
95% Confidence interval (%) 
0/5 
[0, 43.4] 
1/5 
[3.6, 62.4] 
2/5 
[11.8, 76.9] 
 
 
0/15 
[0, 20.4] 
1/15 
[1.2, 29.8] 
2/15 
[3.7, 37.9] 
 
 
0/20 
[0, 16.1] 
1/20 
[0.9, 23.6] 
2/20 
[2.8, 30.1] 
6.1.2 
Sample size calculations for immunogenicity 
The main goals of this trial regarding immunogenicity outcomes involve a 
preliminary estimation of rates of “response” (defined as binding response 
positivity to the 2F5 epitope) based on data from immune assays among 
vaccinees. To address binding antibody endpoints, the analysis will descriptively 
summarize binding response positivity call rates, and for the comparison of Group 
1 vs 2, test superiority of the rate of responses targeting the 2F5 binding site by 
Barnard’s exact test. No adjustment for multiple comparisons will be 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 

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and the sample size. Two-sided 95% confidence intervals for the response rate 
based on observing a particular rate of responses in the vaccinees is shown in 
Table 6-3. Calculations are done using the score test method (33). The n = 4, n = 
13, and n = 18 (for the two arms combined) assume a 10% 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 = 4, n = 13, and n = 18) 
No. of responses 
Observed response rate (%) 
95% Confidence interval 
1/4 
25 
[4.6, 69.9] 
2/4 
50 
[15, 85] 
3/4 
75 
[30.1, 95.4] 
 
 
 
1/13 
7.7 
[1.4, 33.3] 
3/13 
23.1 
[8.2, 50.3] 
5/13 
38.5 
[17.7, 64.5] 
7/13 
53.8 
[29.1, 76.8] 
9/13 
69.2 
[42.4, 87.3] 
 
 
 
3/18 
17 
[5.8, 39.2] 
6/18 
33 
[16.3, 56.3] 
9/18 
50 
[29, 71] 
12/18 
67 
[43.7, 83.7] 
15/18 
83 
[60.8, 94.2] 
An alternative approach is to consider the probability of observing 0, 1 or more, 
and 50% or more responses for groups of sizes 4, 13 and 18; presented in Table 
6-4 for a range of possible true response rates. If the true rate is 20%, then there is 
a 59% chance of observing any responses in the Group 1 vaccine arm, a 94.5% 
chance of observing any responses in the Group 2 vaccine arm and a 98% chance 
of observing at least one response across the two arms. In all cases, the probability 
of observing 50% or more positive responses is in excess of 80% if the true 
positivity rate is at least 60%, but when the true rate is below 40%, there is at least 
a 50% chance of observing fewer than half respond. If the true rate is 50%, then 
there is a 59% chance of observing at least half (2) respond in the n = 4 Group 1 
vaccine arm, a 71% chance of observing at least half (6, rounding down) respond 
in the n = 13 Group 2 vaccine arm and a 59% chance of observing at least half (9) 
respond across the two arms combined (n = 18). 

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Table 6-4 Probability of observing 0 events, 1 or more events, and 50% or more events, 
among groups of size 4, 13 and 18, for different true event rates 
True rate (%) 
Pr(0/4) 
Pr(1+/4) 
Pr(2+/4) 
Pr(0/13) 
Pr(1+/13) 
Pr(6+/13) 
Pr(0/18) 
Pr(1+/18) 
Pr(9+/18) 
10 
65.6 
34.4 
5.2 
25.4 
74.6 
0.1 
15 
85 
0 
20 
41 
59 
18.1 
5.5 
94.5 
3 
1.8 
98.2 
0.4 
30 
24 
76 
34.8 
1 
99 
16.5 
0.2 
99.8 
6 
40 
13 
87 
52.5 
0.1 
99.9 
42.6 
0 
100 
26.3 
50 
6.2 
93.8 
68.8 
0 
100 
70.9 
0 
100 
59.3 
60 
2.6 
97.4 
82.1 
0 
100 
90.2 
0 
100 
86.5 
70 
0.8 
99.2 
91.6 
0 
100 
98.2 
0 
100 
97.9 
80 
0.2 
99.8 
97.3 
0 
100 
99.9 
0 
100 
99.9 
90 
0 
100 
99.6 
0 
100 
100 
0 
100 
100 
As shown in Table 6-5, there is limited power for a formal comparison of 
immunogenicity response rates between vaccine arms of size n = 13 vs n = 4. For 
either 80% or 90% power, the sizes of differences that the trial is powered to 
detect are fairly large. These calculations use a Fisher’s exact 2-sided test with a 
Type I error rate of 0.05. 
Table 6-5 Power for comparison of response rates between 2 arms (n1 = 13, n2 = 4) 
 
Minimum true response rate in Arm 2 in order to detect a 
difference 
True response rate Arm 1 
(%) 
80% power 
90% power 
0 
89 
97 
1 
90 
97 
2 
91 
98 
3 
92 
99 
4 
92 
99 
5 
93 
100 
6.2 
Randomization  
A participant’s randomization assignment will be computer generated and 
provided to the HVTN CRS pharmacist through a web-based randomization 
system. At each institution, the pharmacist with primary responsibility for 
dispensing study products is charged with maintaining security of the treatment 
assignments (except in emergency situations as specified in the HVTN 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 control). Study product 
assignments are accessible to those HVTN CRS pharmacists, DAIDS protocol 
pharmacists and contract monitors, and SDMC staff who are required to know 

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this information in order 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 
in order 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 Principal Investigator (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 
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. 

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6.4.3 
Safety/tolerability analysis 
Since enrollment is concurrent with receiving the first vaccination, all participants 
will have received at least 1 vaccination and therefore will provide some safety 
data. 
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 
systemic symptoms will be calculated. Wilcoxon rank sum tests will be used to 
test for differences in severity between arms. 
6.4.3.2 
AEs and SAEs 
AEs will be summarized using Medical Dictionary for Regulatory Activities 
(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 course of 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 11.2.2) will be tabulated by treatment arm for each 
postvaccination timepoint. Reportable clinical laboratory abnormalities without an 

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associated clinical diagnosis will also be included in the tabulation of AEs 
described above. 
6.4.3.4 
Dose Escalation 
All available safety data through 2 weeks after the first vaccination for Group 1 
participants will be evaluated by the PSRT to inform whether to proceed with 
escalation to the higher dose (Group 2). 
6.4.3.5 
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. 
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 (33). 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 of any 2 vaccine arms, with a significant difference declared if the 2-sided p-
value is ≤ 0.05. In general Barnard’s is preferred since under most circumstances 
it is more powerful than Fisher’s (34).  
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 (35). 

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For quantitative assay data (eg, binding antibody multiplex assay [BAMA]), 
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.  
Based upon previous HVTN trials, missing 10% of immunogenicity results for a 
specific assay is common due to study participants terminating from the study 
early, problems in shipping specimens, or low cell viability of processed 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 data [ie, the observed data are just a 
random sample of all the potential data]). 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 are 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 with methods such as targeted minimum loss-based estimation 
(TMLE), using parametric generalized linear models fit by maximum likelihood 
based on appropriate modeling assumptions for the endpoint and adjusted using 
covariate adjustment, weighting methods or combined with imputation of missing 
data. 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 only depends upon the observed responses and upon observed 
covariates, but not upon any unobserved factors. Thus, this assumption is less 
stringent than the MCAR assumption. 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 (36) will 
be used, because they accommodate the censoring. In addition, secondary 
analyses of repeated immunogenicity measurements may be done using weighted 
generalized estimating equation (GEE) (37) methods, which are valid under 
MAR. All of the models described above in this paragraph will include as 
covariates all available baseline predictors of the missing outcomes.  

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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 (eg, intracellular cytokine staining [ICS]), 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 (38). 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, Env-specific CD4+ T cell polyfunctionality 
score, Env-specific CD8+ T cell total magnitude) 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. Two choices are to compare the M-B curves 
among vaccine arms, as follows: a nonparametric Wilcoxon rank sum test on the 

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subject-specific area-under-the-magnitude-breadth (AUC-MB) or a Kolmogorov-
Smirnov type test on the 2 group-average M-B curves. Simulations can be used to 
obtain 2-sided p-values for the latter test. Second, a weighted-average score-like 
variable may be constructed to account for the correlations between analytes as an 
integrate magnitude of responses to multiple analytes. Similar group comparison 
methods described in the first approach may be adopted. Details of either 
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, HVTN 133 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 HVTN 133 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 have 
completed the corresponding primary immunogenicity visit 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 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 de-identified publication 
identification numbers (PubIDs) and not participant identification numbers 
(PTIDs) for individual responses. Distribution of reports will be limited to those 
with a need to know for the purpose of informing future trial-related decisions. 
The HVTN leadership must approve any other requests for HVTN 
immunogenicity analyses prior to the end of the scheduled follow-up visits. 

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7 
Selection and withdrawal of participants 
Participants will be healthy, HIV-uninfected (seronegative) adults who 
comprehend the purpose of the study and have provided written informed consent. 
Volunteers will be recruited and screened; those determined to be eligible, based 
on the inclusion and exclusion criteria, will be enrolled in the study. 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 
volunteer’s overall fitness for trial participation. Some volunteers may not be 
appropriate for enrollment even if they meet all inclusion/exclusion criteria. 
Medical, psychiatric, occupational, or other conditions may make evaluation of 
safety and/or immunogenicity difficult, and some volunteers may be poor 
candidates for retention. 
Determination of eligibility, taking into account all inclusion and exclusion 
criteria, must be made within 56 days prior to enrollment unless otherwise noted 
in Sections 7.1 and 7.2. 
7.1 
Inclusion criteria 
General and Demographic Criteria 
1. Age of 18 to 50 years 
2. Access to a participating HVTN CRS and willingness to be followed for the 
planned duration of the study 
3. Ability and willingness to provide informed consent 
4. Assessment of understanding: volunteer demonstrates understanding of this 
study; completes a questionnaire prior to first vaccination with verbal 
demonstration of understanding of all questionnaire items answered incorrectly 
5. Agrees not to enroll in another study of an investigational research agent while 
in this study 
6. Good general health as shown by medical history, physical exam, and screening 
laboratory tests 
HIV-Related Criteria: 
7. Willingness to receive HIV test results 

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8. Willingness to discuss HIV infection risks and amenable to HIV risk reduction 
counseling  
9. Assessed by the clinic staff as being at “low risk” for HIV infection and 
committed to maintaining behavior consistent with low risk of HIV exposure 
through the last required protocol clinic visit (see Appendix F) 
Laboratory Inclusion Values 
Hemogram/Complete blood count (CBC) 
10. Hemoglobin ≥ 11.0 g/dL for volunteers who were assigned female sex at birth, ≥ 
13.0 g/dL for volunteers who were assigned male sex at birth. For transgender 
participants who have been on hormone therapy for more than 6 consecutive 
months, determine hemoglobin eligibility based on the gender with which they 
identify (ie, a transgender female who has been on hormone therapy for more than 
6 consecutive months should be assessed for eligibility using the hemoglobin 
parameters for persons assigned female sex at birth) 
11. White blood cell count = 2,500 to 12,000 cells/mm3 with normal differential, or 
differential approved by Investigator of Record (IoR) as not clinically significant 
12. Total lymphocyte count ≥ 650 cells/mm3 with normal differential, or differential 
approved by IoR as not clinically significant 
13. Remaining differential either within institutional normal range or with site 
physician approval 
14. Platelets = 125,000 to 550,000 cells/mm3 
Chemistry 
15. Chemistry panel: alanine aminotransferase (ALT) < 1.25 times the 
institutional upper limit of normal; creatinine ≤ 1.1 times the institutional upper 
limit of normal 
Clotting and autoantibodies 
16. Anticardiolipin IgG antibodies below the upper limit of normal 
17. Negative antinuclear antibodies 
Virology 
18. Negative HIV-1 and -2 blood test: US volunteers must have a negative FDA-
approved enzyme immunoassay (EIA) 
19. Negative Hepatitis B surface antigen (HBsAg)  

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20. Negative anti-Hepatitis C virus antibodies (anti-HCV), or negative HCV 
polymerase chain reaction (PCR) if the anti-HCV is positive 
Urine 
21. Normal urine: 
• Negative or trace urine protein, and 
• Negative, trace, or 1+ blood urine hemoglobin (if +1 hemoglobin is present on 
dipstick, a microscopic urinalysis with red blood cells levels within 
institutional normal range) 
Reproductive Status 
22. Volunteers who were assigned female sex at birth: negative serum or urine beta 
human chorionic gonadotropin (β-HCG) pregnancy test at screening (ie, prior to 
randomization) and prior to study product administration on the day of study 
product administration.. Persons who are NOT of reproductive potential due to 
having undergone hysterectomy or bilateral oophorectomy (verified by medical 
records), are not required to undergo pregnancy testing. 
23. Reproductive status: A volunteer who was assigned female sex at birth: 
• Must agree to use effective contraception for sexual activity that could lead to 
pregnancy from at least 21 days prior to enrollment through the last required 
protocol clinic visit 
• Effective contraception is defined as using the following methods: 
▪ Condoms (male or female) with or without a spermicide, 
▪ Diaphragm or cervical cap with spermicide, 
▪ Intrauterine device (IUD), 
▪ Hormonal contraception,  
▪ Tubal ligation, or 
▪ Any other contraceptive method approved by the HVTN 133 PSRT, 
▪ Successful vasectomy in any partner assigned male sex at birth 
(considered successful if a volunteer reports that a male partner has [1] 
documentation of azoospermia by microscopy, or [2] a vasectomy more 
than 2 years ago with no resultant pregnancy despite sexual activity 
postvasectomy); 
• Or not be of reproductive potential, such as having reached menopause (no 
menses for 1 year) or having undergone hysterectomy, or bilateral 
oophorectomy, 

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• Or be sexually abstinent. 
24. Volunteers who were assigned female sex at birth must also agree not to seek 
pregnancy through alternative methods, such as artificial insemination or in 
vitro fertilization until after the last required protocol clinic visit 
7.2 
Exclusion criteria 
General 
1. Blood products received within 120 days before first vaccination  
2. Investigational research agents received within 30 days before first vaccination 
3. Body mass index (BMI) ≥ 40; or BMI ≥ 35 with 2 or more of the following: age 
> 45, systolic blood pressure > 140 mm Hg, diastolic blood pressure > 90 mm Hg, 
current smoker, known hyperlipidemia  
4. Intent to participate in another study of an investigational research agent or 
any other study that requires non-HVTN HIV antibody testing during the planned 
duration of the HVTN 133 study  
5. Pregnant or breastfeeding 
6. Active duty and reserve US military personnel 
Vaccines and other Injections 
7. HIV vaccine(s) received in a prior HIV vaccine trial. For volunteers who have 
received control/placebo in an HIV vaccine trial, the HVTN 133 PSRT will 
determine eligibility on a case-by-case basis. 
8. Previous receipt of monoclonal antibodies (mAbs), whether licensed or 
investigational; the HVTN 133 PSRT will determine eligibility on a case-by-case 
basis. 
9. Non-HIV experimental vaccine(s) received within the last 1 year in a prior 
vaccine trial. Exceptions may be made by the HVTN 133 PSRT for vaccines that 
have subsequently undergone licensure by the FDA. For volunteers who have 
received control/placebo in an experimental vaccine trial, the HVTN 133 PSRT 
will determine eligibility on a case-by-case basis. For volunteers who have 
received an experimental vaccine(s) greater than 1 year ago, eligibility for 
enrollment will be determined by the HVTN 133 PSRT on a case-by-case basis. 
10. Live attenuated vaccines received within 30 days before first vaccination or 
scheduled within 14 days after injection (eg, measles, mumps, and rubella 

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[MMR]; oral polio vaccine [OPV]; varicella; yellow fever; live attenuated 
influenza vaccine) 
11. Any vaccines that are not live attenuated vaccines and were received within 14 
days prior to first vaccination (eg, tetanus, pneumococcal, Hepatitis A or B) 
12. Allergy treatment with antigen injections within 30 days before first 
vaccination or that are scheduled within 14 days after first vaccination  
Immune System  
13. Immunosuppressive medications received within 168 days before first 
vaccination (Not exclusionary: [1] corticosteroid nasal spray; [2] inhaled 
corticosteroids; [3] topical corticosteroids for mild, uncomplicated dermatologic 
condition; or [4] a single course of oral/parenteral prednisone or equivalent at 
doses < 60 mg/day and length of therapy < 11 days with completion at least 30 
days prior to enrollment) 
14. Serious adverse reactions to vaccines or to vaccine components including 
history of anaphylaxis and related symptoms such as hives, respiratory difficulty, 
angioedema, and/or abdominal pain. (Not excluded from participation: a 
volunteer who had a nonanaphylactic adverse reaction to pertussis vaccine as a 
child.) 
15. Immunoglobulin received within 60 days before first vaccination (for mAb see 
criterion 8 above) 
16. Autoimmune disease, current or history, (Not exclusionary: mild, well-
controlled psoriasis)  
17. AESIs: Volunteers who currently have, or have a history of any condition that 
could be considered an AESI for the products administered in this protocol 
(representative examples are listed in Appendix H) 
18. Immunodeficiency 
Clinically significant medical conditions  
19. Clinically significant medical condition, physical examination findings, 
clinically significant abnormal laboratory results, or past medical history with 
clinically significant implications for current health. A clinically significant 
condition or process includes but is not limited to: 
• A process that would affect the immune response, 
• A process that would require medication that affects the immune response, 
• Any contraindication to repeated injections or blood draws, 

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• A condition that requires active medical intervention or monitoring to avert 
grave danger to the volunteer’s health or well-being during the study period, 
• A condition or process for which signs or symptoms could be confused with 
reactions to vaccine, or 
• Any condition specifically listed among the exclusion criteria below. 
20. Any medical, psychiatric, occupational, or other condition that, in the 
judgment of the investigator, would interfere with, or serve as a contraindication 
to, protocol adherence, assessment of safety or reactogenicity, or a volunteer’s 
ability to give informed consent 
21. Any contraindication that would preclude injections into both left and right 
deltoids 
22. Psychiatric condition that precludes compliance with the protocol. 
Specifically excluded are persons with psychoses within the past 3 years, ongoing 
risk for suicide, or history of suicide attempt or gesture within the past 3 years. 
23. Current anti-tuberculosis (TB) prophylaxis or therapy 
24. Asthma exclusion criteria: 
Asthma other than mild, well-controlled asthma. (Symptoms of asthma severity 
as defined in the most recent National Asthma Education and Prevention Program 
(NAEPP) Expert Panel report).  
 
Exclude a volunteer who: 
• Uses a short-acting rescue inhaler (typically a beta 2 agonist) daily, or 
• Uses moderate/high dose inhaled corticosteroids, or 
• In the past year has had either of the following: 
▪ Greater than 1 exacerbation of symptoms treated with oral/parenteral 
corticosteroids; 
▪  Needed emergency care, urgent care, hospitalization, or intubation for 
asthma. 
25. Diabetes mellitus type 1 or type 2. (Not exclusionary: type 2 cases controlled 
with diet alone or a history of isolated gestational diabetes.) 
26. Thyroidectomy, or thyroid disease requiring medication during the last 12 
months (Not exclusionary: well-controlled non-autoimmune thyroid disease) 
27. Hypertension: 

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• If a person has been found to have elevated blood pressure or hypertension 
during screening or previously, exclude for blood pressure that is not well 
controlled. Well-controlled blood pressure is defined in this protocol as 
consistently < 140 mm Hg systolic and ≤ 90 mm Hg diastolic, with or without 
medication, with only isolated, brief instances of higher readings, which must 
be ≤ 150 mm Hg systolic and ≤ 100 mm Hg diastolic. For these volunteers, 
blood pressure must be ≤ 140 mm Hg systolic and ≤ 90 mm Hg diastolic at 
enrollment. 
• If a person has NOT been found to have elevated blood pressure or 
hypertension during screening or previously, exclude for systolic blood 
pressure ≥ 150 mm Hg at enrollment or diastolic blood pressure ≥ 100 mm Hg 
at enrollment. 
28. Bleeding disorder (eg, factor deficiency, coagulopathy, or platelet disorder 
requiring special precautions) 
29. Malignancy (Not excluded from participation: Volunteer who has had 
malignancy excised surgically and who, in the investigator’s estimation, has a 
reasonable assurance of sustained cure, or who is unlikely to experience 
recurrence of malignancy during the period of the study) 
30. Seizure disorder: History of seizure(s) within past three years. Also exclude if 
volunteer has used medications in order to prevent or treat seizure(s) at any time 
within the past 3 years. 
31. Asplenia: any condition resulting in the absence of a functional spleen 
32. History of generalized urticaria, angioedema, or anaphylaxis. (Not 
exclusionary: angioedema or anaphylaxis to a known trigger with at least 5 years 
since last reaction to demonstrate satisfactory avoidance of trigger.) 
7.3 
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. 
7.3.1 
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 
• Within 30 days prior to any study injection 

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▪ Receipt of live attenuated vaccines  
▪ Receipt of allergy treatment with antigen injections 
• Within 14 days prior to any study injection 
▪ Receipt of any vaccines that are not live attenuated vaccines (eg, 
pneumococcal)  
• Prevaccination abnormal vital signs or clinical symptoms that may mask 
assessment of vaccine reaction. 
Vaccinations should not be administered outside the visit window period. 
In order to avoid vaccination delays and missed vaccinations, participants who 
plan to receive licensed vaccines or allergy treatments should be counseled to 
schedule receipt of these substances, when possible, outside the intervals 
indicated above. The effects of these substances on safety and immunogenicity 
assessments and their interactions with study vaccines are unknown. Therefore, if 
circumstances allow, these substances should also be avoided in the 2-week 
interval between a study vaccination and completion of the 2 weeks 
postvaccination follow-up visit.  
7.3.2 
Participant departure from vaccination schedule 
Every effort should be made to follow the vaccination schedule per the protocol. 
If a participant misses a vaccination and the visit window period for the 
vaccination has passed, that vaccination cannot be given. The participant should 
be asked to continue study visits. The participant should resume the vaccination 
schedule with the next vaccination unless there are circumstances that require 
further delay or permanent discontinuation of vaccination (see Sections 7.3.1 and 
7.3.3). 
7.3.3 
Discontinuing vaccination for a participant 
Under certain circumstances, an individual participant’s vaccinations will be 
permanently discontinued. Specific events that will result in stopping a 
participant’s vaccination schedule include: 
• 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 HVTN 133 PSRT) 
• Clinically significant condition (ie, 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: 
▪ Pregnancy (regardless of outcome) 
▪ HIV infection 

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▪ 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 with the exception of fever, 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 
▪ Clinically significant type 1 hypersensitivity reaction associated with 
study vaccination. Consultation with the HVTN 133 PSRT is required 
prior to subsequent vaccinations following any type 1 hypersensitivity 
reaction associated with study vaccination 
• Investigator determination in consultation with Protocol Team leadership (eg, 
for repeated nonadherence to study staff instructions) 
Participants 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 133 SSP). 
Participants diagnosed with HIV infection during the study should be encouraged 
to participate in follow-up visits as indicated in Section 9.13. 
7.3.4 
Participant termination from the study 
Under certain circumstances, an individual participant may be terminated from 
participation in this study. Specific events that will result in early termination 
include: 
• Participant refuses further participation, 
• Participant relocates and remote follow-up or transfer to another HVTN CRS 
is not possible, 
• HVTN CRS determines that the participant is lost to follow-up, or 
• Investigator decides, in consultation with Protocol Team leadership, to 
terminate participation (eg, if participant exhibits inappropriate behavior 
toward clinic staff). 
• Any condition where termination from the study is required by applicable 
regulations. 

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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. 
Group 1 
Treatment 1 (T1): MPER-656 liposomes, 500 mcg, admixed with Aluminum 
Hydroxide Suspension, to be administered as two 0.5 mL doses intramuscularly 
(IM) at months 0, 2, 6, and 12. 
Or 
Control 1 (C1): Placebo for MPER-656 liposomes (Sodium Chloride for Injection 
USP, 0.9%) to be administered as two 0.5 mL doses IM at months 0, 2, 6, and 12. 
Group 2 
Treatment 2 (T2): MPER-656 liposomes, 2000 mcg, admixed with Aluminum 
Hydroxide Suspension, to be administered as two 0.5 mL doses IM at months 0, 
2, 6, and 12. 
Or 
Control 2 (C2): Placebo for MPER-656 liposomes (Sodium Chloride for Injection 
USP, 0.9%) to be administered as two 0.5 mL doses IM at months 0, 2, 6, and 12. 
8.2 
Study product formulation 
8.2.1 
MPER-656 Liposomes 
MPER-656 liposomes will be provided as a frozen liquid in 2 mL borosilicate 
glass vials. Each vial contains 1.1 mL of MPER liposomes at a nominal 
concentration of 2.2 mg/mL. MPER-656 liposomes are a homogenous translucent 
off-white liquid with consistent visual density and without visible phase 
separation. MPER liposomes are formulated as a sterile, aqueous buffered 
solution that is filled in single-use glass vials for IM injection.  

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The product is stored at -25°C to -10°C. The study product is described in further 
detail in the IB. 
8.2.2 
Aluminum Hydroxide Suspension (Alhydrogel®) 
The Aluminum Hydroxide Suspension will be provided in 3 mL glass vials. Each 
vial contains 0.7 mL fill volume at a concentration of 5 mg/mL aluminum. The 
Aluminum Hydroxide Suspension appears as an opaque, white gelatinous 
precipitate in aqueous suspension.  
The product is stored refrigerated at 2° to 8° C. The study product is described in 
further detail in the MPER-656 liposomes delivered with Alhydrogel® IB. 
8.2.3 
Placebo for MPER-656 Liposomes (Sodium Chloride for Injection 
USP, 0.9%) 
Sodium Chloride for Injection USP, 0.9% will be used as the placebo for MPER-
656 liposomes. The product must be stored as directed by the manufacturer. 
8.3 
Preparation of study products 
Pharmacists should keep in mind that the preparation instructions below are 
considered medium risk per USP 38 General Chapter Physical Tests / <797> 
Pharmaceutical Compounding - Sterile, and should follow the requirements of 
their country, their institution, and their pharmacy regulatory authority regarding 
these procedures. 
Any unused portion of study product will not be used for another participant. 
Empty vials, unused portion of entered vials, or unused prepared study product 
should be discarded in a biohazard container and disposed of in accordance with 
institutional or pharmacy policy. 
8.3.1 
MPER-656 Liposomes, 500 mcg, admixed with Aluminum Hydroxide 
Suspension (Group 1) 
1. Remove one vial of MPER-656 Liposomes from freezer and equilibrate to 
room temperature for approximately 30-60 minutes.  
2. Remove one vial of Aluminum Hydroxide Suspension from refrigerator and 
equilibrate to room temperature for at least 5 minutes.  
3. Using aseptic technique, withdraw 0.5 mL MPER-656 Liposomes and 
dispense into a sterile empty glass vial. 
4. Using aseptic technique, withdraw 1.4 mL Sodium Chloride for Injection 
USP, 0.9% and dispense into the vial from Step 3. 

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5. Resuspend Aluminum Hydroxide Suspension by inverting the vial for 30-60 
seconds. 
6. Using aseptic technique, withdraw 0.2 mL of Aluminum Hydroxide 
Suspension and inject it into the vial from Step 4. Gently swirl the vial to mix. 
Allow formulation to incubate at room temperature for 60 minutes with 
intermittent gentle swirling or inverting of vial every 15-20 minutes. 
7. Using aseptic technique, withdraw 0.5 mL of the mixed preparation into a 1 
mL syringe with a 23-25 gauge needle attached. Repeat this step to prepare a 
second syringe. Prior to administration, gently invert syringes to resuspend 
admixed product. 
8. The admixed product is stable at 2-8° C for four hours, after the 60 minute 
incubation is complete. Each admixed MPER-656 liposomes and alum dose 
should be kept refrigerated or on ice and injected within four hours after 
preparation. The admixed product should be kept on ice during transportation 
from the pharmacy to the clinic and up until the point of administration. 
8.3.2 
MPER-656 Liposomes, 2000 mcg, admixed with Aluminum Hydroxide 
Suspension (Group 2) 
1. Remove one vial of MPER-656 Liposomes from freezer and equilibrate to 
room temperature for approximately 30-60 minutes.  
2. Remove one vial of Aluminum Hydroxide Suspension from refrigerator and 
equilibrate to room temperature for at least 5 minutes.  
3. Resuspend Aluminum Hydroxide Suspension by inverting the vial for 30-60 
seconds. 
4. Using aseptic technique, withdraw 0.1 mL of Aluminum Hydroxide 
Suspension and inject it into the MPER-656 Liposomes vial. Gently swirl the 
vial to mix. Allow formulation to incubate at room temperature for 60 minutes 
with intermittent gentle swirling or inverting of vial every 15-20 minutes.  
5. Using aseptic technique, withdraw 0.5 mL of the mixed preparation into a 1 
mL syringe with a 23-25 gauge needle attached. Repeat this step to prepare a 
second syringe. Prior to administration, gently invert syringes to resuspend 
admixed product.  
6. The admixed product is stable at 2-8° C for four hours, after the 60 minute 
incubation is complete. Each admixed MPER-656 liposomes and alum dose 
should be kept refrigerated or on ice and injected within four hours after 
preparation. The admixed product should be kept on ice during transportation 
from the pharmacy to the clinic and up until the point of administration. 

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8.3.3 
Placebo for MPER-656 Liposomes (Sodium Chloride for Injection 
USP, 0.9%) (Control 1 and Control 2) 
1. Remove one vial of Sodium Chloride for Injection USP, 0.9% from storage. 
2. Using aseptic technique, withdraw 0.5 mL of Sodium Chloride for Injection 
USP, 0.9% into a 1 mL syringe with a 23-25 gauge needle attached. Repeat 
this step to make a second syringe. 
3. The placebo should be kept on ice during transportation from the pharmacy to the 
clinic and up until the point of administration 
8.3.4 
Labeling of Study Product 
Label the study product as follows and apply an overlay/tape to each syringe to 
ensure blinding is maintained:  
• Participant identifier(s) 
• HVTN 133 Study Product or Placebo 
• Final volume (mL) 
• Route (IM) 
• Beyond use date and time 
• Any additional information required by jurisdiction 
8.4 
Administration 
All injections are to be given IM in the deltoid, using standard IM injection 
technique. As there are two injections per visit, one injection will be given in the 
right deltoid and the other injection will be given in the left deltoid. At sites where 
registered pharmacists are legally authorized to administer injections, the HVTN 
CRS may choose to have the pharmacist administer vaccinations.  
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.  
If an injection is administered in the contralateral deltoid due to a medical 
contraindication, the appropriate study staff should document this clearly. Under 

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this circumstance, this is NOT a protocol violation. Two injections administered 
into the same deltoid should be documented in the participant’s study record. 
Any administrator of study product will be blinded to the individual participant's 
treatment assignment. 
8.5 
Acquisition of study products 
MPER-656 liposome will be provided by the International AIDS Vaccine 
Initiative (IAVI, New York, New York, USA). 
Aluminum Hydroxide Suspension will be provided by the NIH, NIAID, Vaccine 
Research Center (VRC, Bethesda, MD).  
Sodium Chloride for Injection USP, 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. 
8.6 
Pharmacy records 
The HVTN CRS pharmacist is required to maintain complete records of all study 
products. 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 US clinical research sites, all unused study products must be returned to the 
CRPMC after the study is completed or terminated unless otherwise instructed by 
the study sponsor. For non-US 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. 

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9 
Clinical procedures 
The schedule of clinical procedures is shown in Appendix D and Appendix F. 
9.1 
Informed consent 
Informed consent is the process of working with participants so that they fully 
understand what will and may happen to them while participating in a research 
study. The HVTN informed consent form (ICF) documents that a participant (1) 
has been informed about the potential risks, benefits, and alternatives to 
participation, and (2) is willing to participate in an HVTN study. Informed 
consent encompasses all written or verbal study information HVTN CRS staff 
provide to the participant, before and during the trial. HVTN CRS staff will 
obtain informed consent of participants 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 participants’ 
decisions to stay in the trial, this information will be shared with trial participants. 
If necessary, participants will be asked to sign revised ICFs. 
An HVTN CRS may employ recruitment efforts prior to the participant 
consenting. For example, some HVTN CRSs use a telephone script to prescreen 
people before they come to the clinic for a full screening visit. Participants must 
sign a screening or protocol-specific consent before any procedures are performed 
to determine eligibility. HVTN CRSs must submit recruitment and prescreening 
materials to their IRB/EC and any applicable RE for human subjects protection 
review and approval. 
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 prior to implementation at a site.” 
CRSs are responsible for knowing the requirements of their applicable REs. 
9.1.1 
Screening consent form 
Without a general screening consent, screening for a specific study cannot take 
place until the site receives protocol registration from the DAIDS RSC Protocol 
Registration Office. 
Some HVTN CRSs have approval from their IRB/EC and any applicable RE to 
use a general screening consent form that allows screening for an unspecified HIV 
vaccine trial. In this way, HVTN CRS staff can continually screen potential 
participants and, when needed, proceed quickly to obtain protocol-specific 

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enrollment consent. Sites conducting general screening or prescreening approved 
by their IRB/EC and any applicable RE may use the results from this screening to 
determine eligibility for this protocol, provided the tests are conducted within the 
time periods specified in the eligibility criteria. 
9.1.2 
Protocol-specific consent forms 
The protocol-specific consent forms describe the study products to be used and all 
aspects of protocol participation, including screening and enrollment procedures. 
A sample protocol-specific 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 C. 
Each 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 C. 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. 
Study sites are strongly encouraged to have their local CABs review their sites-
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 ICFs include instructions for developing specific content. 
Regarding protocol registration, sites should follow procedures outlined in the 
current version of the DAIDS Protocol Registration Manual. 
9.1.3 
Assessment of Understanding 
Study staff are responsible for ensuring that participants fully understand the 
study before enrolling them. This process involves reviewing the ICF with the 
participant, allowing time for the participant to reflect on the procedures and 
issues presented, and answering all questions completely. 
An Assessment of Understanding is used to document the participant’s 
understanding of key concepts in this HIV vaccine trial. The participant must 
complete the Assessment of Understanding before enrollment. Staff may provide 
assistance in reading and understanding the questions and responses, if necessary. 
Participants must verbalize understanding of all questions answered incorrectly. 

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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 participant has signed either a 
screening or protocol-specific consent document 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 must 
be assessed within 56 days before enrollment, unless otherwise specified in the 
eligibility criteria (or below in this section). 
After the appropriate informed consent has been obtained and before enrollment, 
the following procedures are performed: 
• Medical history, documented in the case history record 
• Assessment of whether the volunteer is at low risk for HIV infection (see 
Appendix G). 
• 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 volunteer is taking, including 
prescription and nonprescription drugs, vitamins, topical products, 
alternative/complementary medicines (eg, herbal and health food 
supplements), recreational drugs, vaccinations, and allergy shots 
• Laboratory tests, including: 
▪ Screening HIV test 
▪ HBsAg 
▪ Anti-HCV abs 
▪ Syphilis test 
▪ CBC with differential 
▪ Chemistry panel (ALT, AST, alkaline phosphatase, creatinine) 

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▪ Urine dipstick  
▪ Urine or serum pregnancy test (volunteers who were born female); Persons 
who are NOT of reproductive potential due to having undergone total 
hysterectomy or bilateral oophorectomy (verified by medical records), are 
not required to undergo pregnancy testing 
▪ Anticardiolipin IgG antibodies  
▪ Antinuclear antibodies 
• Administration of behavioral risk assessment questionnaire 
• 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) 
• Counseling on HIV testing and risk reduction, performed in compliance with 
the US Centers for Disease Control and Prevention (CDC)’s current 
guidelines or other local guidelines for HIV counseling, testing, and referral as 
described in Section 9.6 
• Contraception status assessment. A pregnant or breastfeeding person may not 
be enrolled in this trial. Specific criteria and assessment of contraception and 
pregnancy status are described in study inclusion criteria. Discussion of 
contraception status includes advising a participant who was assigned female 
sex at birth and who reports no current sexual activity that could lead to that 
participant becoming pregnant to have a plan to begin adequate birth control. 
This plan would be put to use if, during the study, the participant becomes 
sexually active in a way that could lead to that participant becoming pregnant. 
9.2.1 
Use of screening results from another HVTN study 
If a participant screens for an HVTN study at the same HVTN CRS but then does 
not join that study, screening results from that effort may be applied to the 
screening for this protocol, as long as the screening was done under participant 
consent, the participant has signed a consent form to begin screening for this 
study, and the tests were conducted within the time periods specified in the 
eligibility criteria (see Sections 7.1 and 7.2). 
9.3 
Enrollment and vaccination visits 
Once a volunteer has consented to trial participation and is found to meet all 
eligibility criteria (see Sections 7.1 and 7.2), the HVTN CRS requests the 
randomization assignment via a Web-based randomization system. Enrollment is 
simultaneous with first vaccination. In general, the time interval between 

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randomization and enrollment should not exceed 4 working days. However, 
circumstances may require a participant’s enrollment visit to be changed. This 
may exceed the 4-day randomization time limit. 
At all vaccination visits, the following procedures are performed before 
vaccination: 
• Abbreviated physical examination, including weight, vital signs, and a 
symptom-directed evaluation by history and/or appropriate physical exam 
based on participant self-reported symptoms or complaints; 
• 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 
• Urine or serum pregnancy test (for participants who were assigned female sex 
at birth). Persons who are NOT of reproductive potential due to having 
undergone hysterectomy or bilateral oophorectomy (verified by medical 
records), are not required to undergo pregnancy testing. For pregnant 
participants, see Section 9.12. 
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. 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 participant is given the Participant Diary and is instructed on how to 
complete it. The site will make arrangements to be in contact with the participant 
during the reactogenicity period (as described in Section 9.9). 
The following procedures will be performed at all vaccination visits. These 
procedures may be performed prior to or following vaccination: 
• Risk reduction counseling (as described in Section 9.6); 
• Contraception status assessment (as described in Section 9.2 and 9.7). During 
follow-up in persons who are confirmed pregnant, contraception status 
assessment is not required; and 
• Assessment of new or unresolved social impacts (site staff will ask participant 
about the status of any unresolved social impacts and if s/he has experienced 
any new social impacts as a result of the trial participation). 

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Additional procedures will be performed at scheduled visits as specified in 
Appendix E and Appendix F: 
• Administration of behavioral risk assessment questionnaire  
• Administration of the social impact assessment questionnaire (types of 
impacts assessed involve personal relationships, health insurance, life 
insurance, educational or employment opportunities, housing, immigration, or 
travel); 
• Administration of a questionnaire that asks the participant about any HIV 
testing he or she may have received outside of the study. Participants will also 
be asked whether they believe they received the active vaccine or the control; 
• 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; 
• Confirm that participants received HIV test results from previous visit. If not, 
provide test results and post-test counseling as appropriate; and 
• Specimen collection (should be completed prior to vaccination) 
9.4 
Follow-up visits 
The following procedures are performed at all scheduled follow-up visits: 
• Risk reduction counseling (as described in Section 9.6); 
• Contraception status assessment (as described in Section 9.2 and 9.7). During 
follow-up in persons who are confirmed pregnant, contraception status 
assessment is not required; and 
• Assessment of new or unresolved social impacts (site staff will ask participant 
about the status of any unresolved social impacts and if s/he has experienced 
any new social impacts as a result of the trial participation); 
• Assessment of new or continuing concomitant medications (as described in 
Section 9.2); and 
• Assessment of new or unresolved AEs/intercurrent illnesses. 
Additional procedures will be performed at scheduled follow-up visits as 
specified in Appendix F: 
• Administration of the social impact assessment questionnaire (types of 
impacts assessed involve personal relationships, health insurance, life 

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insurance, educational or employment opportunities, housing, immigration, or 
travel); 
• Administration of a questionnaire that asks the participant about any HIV 
testing he or she may have received outside of the study. Participants will also 
be asked whether they believe they received the active vaccine or the control; 
• Behavioral risk assessment 
• 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; 
• Confirm that participants received HIV test results from previous visit. If not, 
provide test results and post-test counseling as appropriate; 
• Abbreviated physical examination including weight, vital signs, and a 
symptom-directed evaluation by history and/or appropriate physical exam 
based on participant self-reported symptoms or complaints; 
• Complete physical examination, including weight, vital signs, and clinical 
assessments of head, ears, eyes, nose, and throat; neck; lymph nodes; heart; 
chest; abdomen; extremities; neurological function; and skin; 
• Specimen collection; 
• Clinical laboratory tests including: 
▪ CBC with differential, 
▪ Chemistry panel (see Section 9.2), and 
▪ Urine dipstick (urinalysis if appropriate; see Section 9.8); and 
• Urine or serum pregnancy test (for participants who were assigned female sex 
at birth). Persons who are NOT of reproductive potential due to having 
undergone hysterectomy or bilateral oophorectomy (verified by medical 
records), are not required to undergo pregnancy testing. During follow-up in 
persons who are confirmed pregnant, pregnancy testing is not required, unless 
clinically indicated. 
• AESI health contact (see HVTN 133 SSP). 
9.5 
AESI health contact 
CRS staff will contact study participants 12 months after last vaccination to 
collect the information listed below. Clinic visits will only be required if HIV 

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confirmatory testing is necessary (see Section 9.6.1); however, a clinic visit may 
be arranged for other reasons (eg, AESI assessment and referral). 
• Confirmation of vital status; if deceased, attempt to learn cause and date of 
death; 
• If participant is alive, record the following events: 
▪ New AEs related to study product(s) 
▪ AEs of special interest (AESI, see Section 11.2.2). A sample list of AESI 
is provided in Appendix H. AESI are reported regardless of relationship to 
study product(s); 
▪ New diagnosis of HIV infection; and 
▪ Pregnancies and outcomes, including congenital anomalies/birth defects. 
9.5.1 
Interim contacts 
CRSs may report safety information obtained at a contact other than the annual 
contact. These contacts are reported as interim visits. 
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. HIV testing will be 
performed in accordance with the current HVTN HIV testing algorithm following 
enrollment. 
Participants will be counseled routinely during the trial on the avoidance of HIV 
infection and on the potential negative social impacts of testing antibody positive 
due to the vaccine. They will also be counseled on the risks of HIV antibody 
testing outside of the HVTN CRSs and will be discouraged from doing so during 
study participation and/or during any period of vaccine-induced positive serology. 
Study staff will take particular care to inform study participants of the likelihood 
of routine HIV testing being offered or performed outside the study CRS at 
emergency rooms, clinics, and medical offices. Such testing has become more 
likely due to the CDC’s revised guidelines for HIV counseling and testing, as well 
as policy changes in many countries to make HIV testing more frequent and 
routine. CRS staff should inform participants of their right to opt out of HIV 
testing outside the study site. CRS staff should inform study participants if local 
and/or state policies and regulations permit medical providers to perform HIV 
testing without first informing patients. If this is the case, then CRS staff should 
advise study participants that they may decline testing preemptively. CRS staff 
should also inform participants if positive results must be reported to local public 
health authorities. CRS staff should also inform participants of the need to 
maintain study blinding by getting HIV testing only at the study CRS. CRS staff 

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should provide participants with CRS contact information and should encourage 
participants to ask medical providers to contact the CRS. The CRS can verify that 
the participant is in an HIV vaccine clinical trial and should only be tested at the 
study CRS. 
Potential participants identified as being HIV-infected during screening are not 
enrolled. Potential and enrolled participants identified as being HIV-infected will 
be referred for medical treatment, counseling, and management of the HIV 
infection. Participants who are found to be HIV-infected after enrollment will not 
receive any additional study product but will continue to be followed in the study 
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 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: 
• Participants 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. 
• HIV testing will be performed at multiple timepoints throughout the study 
(see Appendix E and Appendix F). The Laboratory Program (or approved 
diagnostic laboratory) will follow the HVTN HIV testing algorithm (see 
HVTN 133 SSP), which is able to distinguish vaccine-induced antibody 
responses from actual HIV infections. 
• All participants 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 participants 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. 

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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) (see Section 9.6.1). In order to provide 
poststudy HIV testing to distinguish between VISP and HIV infection, and to 
mitigate potential 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 they request that their names be removed. 
9.7 
Contraception status 
Contraception status is assessed and documented at every scheduled clinic visit 
for a participant who was assigned female sex at birth and who is sexually active 
in a way that could cause that participant to become pregnant. Prior to enrollment 
and throughout the study, staff will ask participants to verbally confirm their use 
of adequate contraceptive methods. A participant who was assigned female sex at 
birth and is sexually active in a way that could cause that participant to become 
pregnant should be reminded at all scheduled clinic visits of the importance of 
using contraception and should be referred to specific counseling, information, 
and advice as needed. (specific contraception requirements are listed in Section 
7.1). This reminder should be documented in the participant’s study record. 
Self-reported infertility—including having reached menopause (no menses for 1 
year) or having undergone hysterectomy, bilateral oophorectomy, or tubal 
ligation—must be documented in the participant’s study record. 
9.8 
Urine testing 
Dipstick testing may be performed in the clinic or the lab, as long as the required 
elements (glucose, protein, and hemoglobin) are tested. The examination is 
performed on urine obtained by clean catch. 
If the screening dipstick is transiently abnormal due to non-urinary bleeding (eg, 
menstruation) or infection, document this issue in the participant’s source 
documentation. For infection, provide appropriate treatment and/or referral and 
document this in the participant's chart. Following resolution, repeat the dipstick 
and, if within the eligibility limits specified in the protocol, the participant may be 
enrolled. 

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Follow-up visit dipstick testing should be deferred if a participant is experiencing 
non-urinary bleeding (eg, menstruation), but should be performed as soon as 
possible. If a follow-up visit dipstick is abnormal due to a participant’s non-
urinary bleeding (eg, menstruation), document in the comment section of the case 
report form (CRF), and repeat the dipstick once the participant is no longer 
experiencing non-urinary bleeding. A micro-urinalysis is not required. If a follow-
up visit dipstick or micro-urinalysis is abnormal due to infection, provide 
appropriate treatment and/or referral and document this in the participant's source 
documentation. See the Urine Testing MOP for further details. 
9.9 
Assessments of reactogenicity 
For all participants, baseline assessments are performed before and reactogenicity 
assessments are performed 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. Participants are instructed to record 
symptoms using a Participant Diary. Contacts between the participant and the site 
staff should take place at least once between 1-3 days postvaccination. In general, 
a participant who self-reports any postvaccination reaction greater than mild is 
seen by a clinician within 48 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. 

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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 
participant 
1-7b 
Between 12:00am and 11:59pm on the 
respective day 
HVTN CRS clinician or 
participant 
a Day of vaccination 
b New or unresolved reactogenicity symptoms present on day 7 are followed until resolution 
9.9.1 
Assessment of systemic and local symptoms 
Systemic symptoms include increased body temperature, malaise and/or fatigue, 
myalgia, headache, chills, arthralgia, and nausea. Local symptoms include pain 
and/or tenderness at the injection site. The daily maximum severity reached for 
each symptom during the assessment period is reported.  
Body temperature is measured by oral or infrared thermometry. All temperatures 
must be measured by nonaxillary thermometry. This includes temperatures taken 
in the clinic, as well as temperatures taken by participants during the 
reactogenicity period. 
Temperature is reported in degrees Celsius. If temperature is measured in 
Fahrenheit, the conversion to Celsius should be documented in the participant’s 
chart note. A measurement is taken once daily during the assessment period and 
should be repeated if participant is feeling feverish. 
9.9.2 
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 5 cm are followed daily; otherwise, the frequency of 
follow-up is based on clinician judgment. See HVTN 133 SSP for details. 
9.10 
Visit windows and missed visits 
Visit windows are included in Appendix I. Visits conducted outside of the 
allowable visit windows are considered protocol deviations. Further information 
about visit windows are described in the HVTN 133 SSP. If a missed visit is one 
that required safety assessments or local safety labs, HVTN CRS staff should 
attempt to bring the participant in for an interim visit as soon as possible. 
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 

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clinically indicated. HIV testing may be performed as deemed appropriate by the 
study staff. Blood samples for immunogenicity assays are not typically collected 
at interim visits. 
If a missed visit required vaccination, please refer to Section 7.3.2 and Section 
7.3.3 for resolution. 
9.11 
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 urine dipstick, CBC with differential, chemistry panel, 
antinuclear antibodies, anticardiolipin IgG antibodies), pregnancy testing (note: 
for persons who are confirmed pregnant, pregnancy testing is not required, unless 
clinically indicated), social impact assessment, and HIV test. For participants who 
have a confirmed diagnosis of HIV infection, see Section 9.13. 
9.12 
Pregnancy 
If a participant becomes pregnant during the course of the study, no more 
injections of study product will be given, but remaining visits and study 
procedures should be completed unless medically contraindicated or applicable 
regulations require termination from the study. During follow-up in persons who 
are confirmed pregnant, pregnancy testing is not required, unless clinically 
indicated. If the participant terminates from the study prior to the pregnancy 
outcome, the site should make every effort to keep in touch with the participant in 
order to ascertain the pregnancy outcome. Pregnancies and pregnancy outcomes 
will be reported. If the participant is no longer pregnant, refer to Section 7.3.3. 
9.13 
HIV infection during the study 
If a participant becomes HIV-infected during the course of the study, no 
additional study product will be administered. Participants will be encouraged to 
continue scheduled study visits for up to 18 weeks following their last study 
product administration. Follow-up duration for participants diagnosed with HIV 
infection may be adjusted in consultation with the CRS investigator and the 
HVTN 133 PSRT (eg, to avoid interference with participant initiation of HIV 
treatment). At postinfection follow-up visits, only specimens required for 
protocol-specified safety laboratory tests, urinalysis and pregnancy tests will be 
collected (note: for persons who are confirmed pregnant, pregnancy testing is not 
required, unless clinically indicated); in addition, some clinic procedures may be 
modified or discontinued (see Appendix E and Appendix F). 

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10 
Laboratory 
10.1 
HVTN CRS laboratory procedures 
The HVTN 133 Site Lab Instructions and 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 E. For tests performed 
locally, the local lab may assign appropriate tube types. 
Of note, all assays described below are performed as research assays 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 E. Not shown is any 
additional blood volume that would be required if a safety lab needs to be 
repeated, or if a serum pregnancy test needs to be performed. The additional 
blood volume would likely be minimal. The total blood volume drawn for each 
participant will not exceed 500 mL in any 56-day (8-week) period. 
10.3 
Primary immunogenicity timepoints 
The primary immunogenicity timepoints in this study occur 2 weeks after the 
third and fourth vaccinations. Endpoint assays may be performed on specimens 
collected from participants at the primary immunogenicity timepoints and 
samples collected at baseline and other timepoints; the schedule is shown in 
Appendix E. 
10.4 
Endpoint assays: cellular 
10.4.1 
Flow cytometry: intracellular cytokine staining (ICS) 
Flow cytometry will be used to examine vaccine-specific CD4+ and CD8+ T-cell 
responses following stimulation of PBMCs with synthetic HIV peptides that span 
the proteins encoded by the vaccine. ICS parameters will include cytokines such 
as interferon (IFN)-γ, interleukin (IL)-2, and tumor necrosis factor (TNF)-α, and 
may include other cytokines (such as cytokines relevant to Th2 and Th17 
responses) to identify T cells of specific functionality. Data will be reported as 
percentages of CD4+ or CD8+ T cells responding to a specific peptide pool. 

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Additional cell surface markers, cytokines, or functional markers may also be 
analyzed. 
10.4.2 
Flow cytometry: antigen-specific B cell phenotyping assay 
Antigen-specific B cells induced by vaccination will be identified and 
characterized using fluorescently-labeled recombinant protein probes in 
combination with a flow cytometry phenotyping panel. In particular, MPER-
specific B cells will be enumerated and may be further characterized for 
expression of memory, activation, inhibitory or other markers by protein and/or 
gene expression. B cells may also be sorted for further analysis by BCR 
sequencing or gene expression analysis. 
10.4.3 
B-cell lineage analysis 
MPER-specific B cells may be single cell or bulk sorted. Individual MPER-
reactive memory B cells or plasmablasts may be sorted into individual wells of 
culture plates, expanded in short-term cultures and screened for binding or in vitro 
neutralization before PCR and VH and VL sequencing. VH and VL genes may be 
amplified and cloned into an antibody expression backbone and tested for Env 
binding and HIV neutralization. Finally, 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-cells or bulk sorted MPER-specific B cells. 
10.4.4 
Peripheral T follicular helper cells (pTfh) 
Flow cytometry may be used to identify and phenotype T cells in the 
peripheral blood. pTfh may be characterized based on expression of lineage 
markers including CXCR5, PD-1, and ICOS on CD4+ T cells. In addition, 
serum may be used to determine the level of CXCL13 in circulation to 
monitor the germinal center reaction upon vaccination. The flow cytometry 
panels might also include additional markers. 
10.5 
Endpoint assays: humoral 
10.5.1 
Binding antibody multiplex assay (BAMA) 
HIV-1–specific IgG antibodies to MPER epitopes will be assessed on serum 
samples from study participants taken at the primary immunogenicity timepoints 
and baseline by Binding Antibody Multiplex Assay (BAMA). In addition, HIV-1–
specific total binding IgA antibodies and binding to IgG subclasses (IgG1, IgG2, 
IgG3, and IgG4) may also be assessed. Specimens from other timepoints may also 
be assayed based on the results of the initial assays. 

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10.5.2 
Lipid binding assays 
Binding antibodies to lipid antigens will be assessed by binding assays using 
serum samples from study participants taken at baseline and at the primary 
immunogenicity timepoints. Specimens from other timepoints may also be 
assayed based on the results of the initial assays. 
10.5.3 
Antibody avidity 
MPER epitopes that map specificity and kynureninase may be included as part of 
the binding and avidity index measurements (BAMA-AI). Biolayer 
Interferometry (BLI) and/or Surface Plasmon Resonance (SPR) technologies may 
also be utilized to define antibody avidity to the MPER and MPER-liposomes. 
10.5.4 
Neutralizing antibody assay 
HIV-1–specific nAb assays will be performed on serum samples from study 
participants taken at the primary immunogenicity timepoints. Specimens from the 
baseline and 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 and TZM-bl/FcRI assays will test 
neutralization of three clade B strains (HXB2, W61D.TCLA.71 and SC422661.8) 
that are highly neutralization-sensitive to antibodies induced in immunized 2F5 
knock-in mice. The global panel and/or clade-specific panels may be used to 
assess Tier 2 neutralization breadth in greater detail (39, 40). 
10.6 
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. 
10.7 
Exploratory studies 
Samples may be used for other testing and research related to furthering the 
understanding of HIV immunology or vaccines. In addition, cryopreserved 
samples may be used to perform additional assays to support standardization and 
validation of existing or newly developed methods.  

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10.8 
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.  
Other use of specimens is defined as studies not covered by the protocol or the 
ICF for the main study (see Appendix C).  
This research may relate to HIV, vaccines, the immune system, and other 
diseases. This could include genetic testing and, potentially, genome-wide studies. 
This research is done only to the extent authorized in each study site’s ICF, 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 CRS’s IRBs/ECs/REs if required.  
As part of consenting for the study, participants document their initial decision to 
allow or not allow their specimens to be used in other research, and they may 
change their decision at any time. The participant’s initial decision about other 
use of their specimens, and any later change to that decision, is recorded by their 
CRS in a web-based tool that documents their current decisions for other use of 
their specimens. The HVTN will only allow other research to be done on 
specimens from participants who allow such use.  
CRSs must notify HVTN Regulatory Affairs if institutional or local governmental 
requirements pose a conflict with or impose restrictions on specimen storage or 
other use of specimens. 
10.9 
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. 

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11 
Safety monitoring and safety review 
11.1 
Safety monitoring and oversight 
11.1.1 
HVTN 133 PSRT 
The HVTN 133 PSRT is composed of the following members: 
• DAIDS medical officer representative 
• Protocol chair and cochair 
• Protocol Team leader 
• Core medical monitor 
• Clinical safety specialist 
The clinician members of HVTN 133 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 
133 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 during the main study, as defined in Section 3. 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 133 PSRT. 
Study sites 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: 

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• Maintaining a central database management system for HVTN clinical data; 
• Providing reports of clinical data to appropriate groups such as the HVTN 133 
PSRT and HVTN SMB (see Section 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 133 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 133 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, 
urinalysis, 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). All AEs are graded according to the Division of AIDS 

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(DAIDS) Table for Grading the Severity of 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: 
• Unintentional Weight Loss is required to be reported as an AE only if it is 
considered to be potentially deleterious to the participant’s health (see HVTN 
133 SSP); 
• Injection Site Erythema or Redness and Injection Site Induration or Swelling 
will not consider surface area and interference with usual social and functional 
activities such that: 
▪ Grade 1 is: 2.5 to < 5 cm in diameter; 
▪ Grade 2 is: ≥ 5 to < 10 cm in diameter; 
▪ Grade 3 is: ≥ 10 cm in diameter OR Ulceration OR Secondary infection 
OR Phlebitis OR Sterile abscess OR Drainage; 
▪ Grade 4 is: Potentially life-threatening consequences (eg, abscess, 
exfoliative dermatitis, necrosis involving dermis or deeper tissue); 
• Creatinine is required to be reported as an AE only if it is gradable per the 
increase from local lab ULN parameter. Do not grade elevated creatinine 
based on the change from the baseline parameter. 
During the main study period (see Section 3), 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 to DAIDS (see Section 
11.2.3) and (2) if the AE meets the criteria for a safety pause/prompt AE review 
(see Section 11.3), and (3) if the AE is listed as an AESI. A list of AESI to be 
reported in this protocol is provided in Appendix H. 
After the main study period, report the subset of AEs bulleted in Section 9.5 until 
the health contact (see Section 3) is complete. 
Sites are expected to notify HVTN clinical safety staff of any serious safety 
concern requiring their attention (Table 11-1). Telephone numbers and email 
addresses are found on the protocol home page on the HVTN Members’ site 
(https://members.hvtn.org/protocols/hvtn133). 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 11-1). If email service is not available, the CRS 
should notify clinical safety staff of the event by telephone, and then submit 
CRFs. 

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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: 
• MPER-656 liposomes with 500 mcg alum 
• Sodium Chloride for Injection USP, 0.9% 
While the participant is in the main study reporting period (see Section 3), the 
SAE Reporting Category will be used. 
After completion of the main study period through the completion of the AESI 
health contact (see Section 3) the Suspected, Unexpected Serious Adverse 
Reactions (SUSAR) Reporting Category will be used. 
After the participant has completed the AESI health contact and is off study, sites 
must report SUSARs if the study site staff becomes aware of the events on a 
passive basis (eg, from publicly available information). 
The NIAID/DAIDS will report all unexpected SAEs related to the study products 
observed in this clinical trial to the FDA in accordance with 21 CFR 312.32 (IND 
Safety Reports). 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 FDA based on the blinded 
attribution assessment. 

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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 considerations for dose escalation 
In addition to monitoring participant safety throughout the study period, the 
HVTN 133 PSRT will review cumulative safety data available on all participants 
in Group 1 up to and including the 2-week visit after the first vaccination to 
determine whether dose escalation may occur. The HVTN 133 PSRT may consult 
with the HVTN SMB on an ad hoc basis for these evaluations. 
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 133 PSRT AE 
review are summarized in Table 11-1. Vaccinations may be suspended for safety 
concerns other than those described in the table, or before pause rules are met, if, 
in the judgment of the HVTN 133 PSRT, participant safety may be threatened. 
Criteria for an individual participant’s departure from the schedule of vaccinations 
are listed in Section 7.3. 
Table 11-1 AE notification and safety pause/AE review rules 
Event and 
relationship to study 
products 
Severity 
HVTN CRS actiona 
HVTN Core action 
SAE, related 
Grade 5 or 
Grade 4 
Phone immediately, email 
and submit forms 
immediately 
Immediate pause 
SAE, not related 
Grade 5 
Phone immediately, email 
and submit forms 
immediately 
Immediate PSRT notification 
SAE, related 
Grade 3, 2, 
or 1 
Email and submit forms 
immediately 
Immediate PSRT notification and 
prompt PSRT AE review to 
consider pause 
AEb, related 
Grade 4 or 
3 
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/hvtn133). 
b Does not include the following Grade 3 subjective reactogenicity symptoms: injection site pain, tenderness, 
fatigue/malaise, myalgia, arthralgia, chills, headache, nausea (unless IV rehydration required). 
For all safety pauses, HVTN Core notifies the HVTN 133 PSRT, HVTN 
Regulatory Affairs, DAIDS Pharmaceutical Affairs Branch (PAB), DAIDS 

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Regulatory Affairs Branch (RAB), DAIDS Safety and Pharmacovigilance Team 
(SPT), and participating HVTN CRSs. When an immediate safety pause is 
triggered, HVTN Core notifies the SMB. 
Once a trial is paused, the HVTN 133 PSRT reviews safety data and decides 
whether the pause can be lifted or permanent discontinuation of vaccination is 
appropriate, consulting the SMB if necessary. HVTN Core notifies the 
participating HVTN CRSs, HVTN Regulatory Affairs, DAIDS PAB, DAIDS 
RAB, and DAIDS SPT of the decision regarding resumption or discontinuation of 
study vaccinations. Based on the HVTN 133 PSRT assessment, DAIDS RAB 
notifies the FDA as needed. 
If an immediate HVTN 133 PSRT notification or prompt HVTN 133 PSRT AE 
review is triggered, HVTN Core notifies the HVTN 133 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 133 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. 
In addition, all other AEs are reviewed routinely by the HVTN 133 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 133 PSRT AE review criteria. 
11.5.2 
Weekly review 
During the injection phase of the trial, the HVTN 133 PSRT reviews clinical 
safety reports on a weekly basis and conducts calls to review the data as 

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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 133 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 133 PSRT, 
a pertinent national regulatory authority, the FDA, NIH, Office for Human 
Research Protections (OHRP), or study product developer(s). In addition, the 
conduct of this study at an individual HVTN CRS may be terminated by the 
determination of the IRB/EC and any applicable RE. 

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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 133 SSP. 

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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. 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. 
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. 

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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 133 are described 
below. 
Protocol history and modifications 
Date: March 13, 2019 
Protocol version: 1.0 
Protocol modification: 
 
Original protocol 

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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. 
• 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 133 Special Instructions. Accessible through the HVTN protocol-
specific website. 
• HVTN 133 Study Specific Procedures. Accessible through the HVTN 
protocol-specific website. 
• HVTN 133 Site Lab Instructions. Accessible through the HVTN protocol-
specific website. 
• HVTN Manual of Operations. Accessible through the HVTN website. 

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• Dangerous Goods Regulations (updated annually), International Air Transport 
Association. Available for purchase at 
http://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 
http://www.ich.org/products/guidelines/efficacy/article/efficacy-
guidelines.html 
• Participants’ Bill of Rights and Responsibilities. Accessible through the 
HVTN website. 
• NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic 
Acid Molecules. Available at http://osp.od.nih.gov/office-biotechnology-
activities/biosafety/nih-guidelines 
• 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  
See Section 16 for literature cited in the background and statistics sections of this 
protocol. 

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15 
Acronyms and abbreviations 
Ab 
 
antibody 
AE 
 
adverse event 
AESI 
 
adverse event of special interest 
ALP 
 
alkaline phosphatase 
ALT 
 
alanine aminotransferase 
aPTT 
 
activated partial thromboplastin time 
ART 
 
antiretroviral therapy 
AST 
 
aspartate aminotransferase 
AUC 
 
area under the curve 
AUC-MB  
area-under-the-magnitude-breadth 
β-HCG 
 
beta human chorionic gonadotropin 
BAMA 
 
binding antibody multiplex assay 
BCR 
 
B-cell receptor 
BMI 
 
body mass index 
bnAb 
 
broadly neutralizing antibodies 
CAB 
 
Community Advisory Board 
CBC 
 
complete blood count 
CDC 
 
US Centers for Disease Control and Prevention 
CFR 
 
Code of Federal Regulations 
CI 
 
confidence intervals 
CIOMS 
 
Council for International Organizations of Medical Sciences 
CRF 
 
case report form 
CRPMC 
 
NIAID Clinical Research Products Management Center 
CRS 
 
clinical research site 
DAERS 
 
DAIDS Adverse Experience Reporting System 
DAIDS 
 
Division of AIDS (US NIH) 
DHHS 
 
US Department of Health and Human Services 
DRVVT 
 
dilute Russell’s viper venom time 
EAE 
 
expedited adverse event 
EC 
 
Ethics Committee 
EIA 
 
enzyme immunoassay 
ELISA 
 
enzyme-linked immunosorbent assay 
Env 
 
HIV-1 envelope 
FDA 
 
US Food and Drug Administration 
Fred Hutch 
Fred Hutchinson Cancer Research Center 
GCP 
 
Good Clinical Practice 
GEE 
 
generalized estimating equation 
GLA 
 
Glucopyranosyl Lipid Adjuvant 

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HBsAg 
 
hepatitis B surface antigen 
HCV 
 
hepatitis C virus 
HIPAA 
 
Health Insurance Portability and Accountability Act 
HIV 
 
human immunodeficiency virus 
HVTN 
 
HIV Vaccine Trials Network 
IAVI 
 
International AIDS Vaccine Initiative 
IB 
 
Investigator’s Brochure 
ICF 
 
informed consent form 
ICH 
 
International Council on Harmonisation of Technical 
Requirements for Pharmaceuticals for Human Use 
ICS 
 
intracellular cytokine staining 
IDRI 
 
Infectious Disease Research Institute 
IFN-γ 
 
interferon gamma 
IgG 
 
immunoglobulin G 
IgM 
 
immunoglobulin M 
IL 
 
interleukin 
IM 
 
intramuscular 
IND 
 
Investigational New Drug 
IP 
 
intraperitoneal 
IRB 
 
Institutional Review Board 
IUD 
 
intrauterine device 
KI 
 
knock-in 
KYNU 
 
kynureninase 
mAb 
 
monoclonal antibody 
MAR 
 
missing at random 
M-B 
 
Magnitude–Breadth 
MCAR 
 
missing completely at random 
MedDRA  
Medical Dictionary for Regulatory Activities 
MMR 
 
measles, mumps, and rubella 
MOP 
 
Manual of Operations 
MPER 
 
membrane proximal external region 
MPLA 
 
monophosphoryl lipid A 
nAb 
 
neutralizing antibody 
NAEPP 
 
National Asthma Education and Prevention Program 
NHP 
 
nonhuman primate 
NIAID 
 
National Institute of Allergy and Infectious Diseases (US NIH) 
NIH 
 
US National Institutes of Health 
OHRP 
 
US Office for Human Research Protections 
OPV 
 
oral polio vaccine 

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PAB 
 
DAIDS Pharmaceutical Affairs Branch 
PBMC 
 
peripheral blood mononuclear cell 
PCA 
 
principal component analysis 
PCR 
 
polymerase chain reaction 
PI 
 
Principal Investigator 
PSRT 
 
Protocol Safety Review Team 
PTE 
 
prothrombin time 
pTfh 
 
Peripheral T follicular helper cells (pTfh) 
PTID 
 
participant identification number 
PubID 
 
de-identified publication identification number 
RAB 
 
DAIDS Regulatory Affairs Branch 
RE 
 
Regulatory Entity 
RSC 
 
DAIDS Regulatory Support Center 
SAE 
 
serious adverse event 
SAP 
 
Statistical Analysis Plan 
SCHARP  
Statistical Center for HIV/AIDS Research and Prevention 
SDMC 
 
statistical and data management center 
SHIV 
 
simian/human immunodeficiency virus 
SMB 
 
Safety Monitoring Board 
SPT 
 
DAIDS Safety and Pharmacovigilance Team 
SSP 
 
Study Specific Procedures 
SUSAR 
 
Suspected, Unexpected Serious Adverse Reactions 
TB 
 
tuberculosis 
Tfh 
 
follicular helper T-cell 
TNF 
 
tumor necrosis factor 
UA 
 
unmutated ancestor 
UW-VSL  
University of Washington Virology Specialty Laboratory 
VH 
 
heavy chain variable region  
VISP 
 
Vaccine induced seropositivity 
VL 
 
light chain variable region 
VRC 
 
Vaccine Research Center (NIAID) 
 

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38. James G, Witten D, Hastie T, Tibshirani R. An introduction to statistical 
learning with applications in R. New York: R. Springer; 2013 2013. 
39. Seaman M, Janes H, Hawkins N, randpre 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. 
40. 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. 
 

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Appendix A 
Sample informed consent form 
Title: A phase 1 clinical trial to evaluate the safety and immunogenicity of an 
HIV-1 gp41 MPER-656 liposome vaccine in healthy, HIV-uninfected adult 
participants 
HVTN protocol number: HVTN 133 
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. 
About the study 
The HIV Vaccine Trials Network (HVTN) and [Insert site name]  are doing a 
study to test an HIV vaccine. HIV is the virus that causes AIDS. 
About 24 people will take part in this study at multiple sites. The researcher in 
charge of this study at this clinic is [Insert name of site PI]. The US National 
Institutes of Health (NIH) is paying for the study. 
1. We are doing this study to answer several questions. 
• Is the study vaccine safe to give to people? 
• Are people able to take the study vaccine without becoming too 
uncomfortable? 
• How do people’s immune systems respond to the study vaccine? (Your 
immune system protects you from disease.) 
• Does the vaccine have different effects at different doses? 
2. The study vaccine cannot give you HIV. 
The study vaccine is not made from actual HIV. It is impossible for the study 
vaccine to give you HIV. Also, it cannot cause you to give HIV to someone else. 

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3. We do not know if the study vaccine will decrease, increase, or not change your 
risk of getting HIV if you 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. 
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. We can tell you about the differences. 
We do not know whether the vaccine in this study will affect your risk of getting 
HIV from another person. The risk could be higher, lower, or unchanged. It’s very 
important to avoid exposure to HIV during and after the study. We will tell you 
how to avoid HIV. 
4. This study vaccine is experimental. 
The study vaccine is called MPER-656 liposomes. From here on out, we will just 
call it the study vaccine. 
The study vaccine has not been given to people before. It is an experimental HIV 
vaccine. That means we do not know if the study vaccine will be safe to use in 
people, or if it will work to prevent HIV infection.  
The study vaccine has been tested in animals and 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 used only in research studies. 
The study vaccine was developed by Duke University, working with the 
Infectious Disease Research Institute (IDRI). This study vaccine has man-made, 
short pieces of a protein called a peptide. This peptide looks like part of a protein 
found in HIV. The peptide in this study vaccine is combined with a tiny bit of fat 
called a liposome. The liposome helps keep the peptide in a shape that might 
make it easier for your immune system to respond to it. If you want to know more 
about the study vaccine, ask the study staff.  
The study vaccine is mixed with an adjuvant. An adjuvant is a substance added to 
the vaccine to help the immune system respond better. The adjuvant in this study 
is called aluminum hydroxide or alum. Aluminum has been used in vaccines for 
more than 60 years. 

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General risks of vaccines: 
All vaccines can cause fever, chills, rash, aches and pains, nausea, headache, 
dizziness, 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 you 
have 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. 
After getting the study vaccine, you could make antibodies that recognize parts of 
cells in humans. These are called autoantibodies. In animal studies, these 
autoantibodies did not show effects on the health of the animals. 
Liposomes have been used for decades in cancer vaccines, animal vaccines, and 
drug treatments. When injected, liposomes can cause symptoms like other 
vaccines. The most common are redness and swelling where you got the injection 
and fever. 
The adjuvant, alum, is the most widely used vaccine adjuvant. It has been used in 
licensed vaccines given to hundreds of millions of people all over the world. 
People can have the same kinds of side effects from the adjuvant as they do with 
vaccines, such as pain where they got the injection, muscle aches, or a fever. 
Joining the study 
5. It is completely up to you whether or not to join the study. 
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 leave it 
after you have joined, your other care at this clinic and the benefits or rights you 
would normally have will not be affected. 
If you join this study, you may not be allowed to join other HIV vaccine or HIV 
prevention studies now or in the future. You cannot be in this study while you are 

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in another study where you get a study product. Being in more than one study 
may not be safe. 
Also during the study, you should not donate blood or tissue. 
If you choose not to join this study, you may be able to join another study. 
Site: Remove item 6 if you use a separate screening consent that covers these 
procedures. 
6. If you want to join the study, we will screen you to see if you are eligible. 
Screening involves a physical exam, HIV test and health history. A physical exam 
may include, but is not limited to: 
• Checking your weight, temperature and blood pressure 
• Looking in your mouth and throat 
• Listening to your heart and lungs 
• Feeling your abdomen (stomach and liver) 
We will also do blood and urine tests, which will tell us about some aspects of 
your health, such as how healthy your kidneys, liver, and immune system are. We 
will also do tests to see if you currently have autoantibodies. We will test for 
syphilis, hepatitis B, and hepatitis C. We will ask you about medications you are 
taking. We will ask you about behaviors that might put you at risk for getting 
HIV. If you were assigned female sex at birth, we will test you for pregnancy.  
We will review the screening results with you. The screening results may show 
you are not eligible to join the study, even if you want to.  
Site: adapt the following section so it is applicable to the care available at your site 
7. If we find that you have a health problem during screening or during the 
study, we will tell you about the care that we can give here for free. 
For the care that we cannot give, we will explain how we will help you get care 
elsewhere. For health problems that are unrelated to the study, we will not pay for 
care. 
8. If you were assigned female sex at birth and could become pregnant, you must 
agree to use birth control to join this study. 
Site: If you want to include Appendix B, Approved birth control methods (for sample 
informed consent form), in this consent form, paste it below and delete paragraph 
below. 

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You should not become pregnant during the study because we do not know how 
the study vaccine could affect the developing baby. You must agree to use 
effective birth control from at least 21 days before your first injection through the 
last required protocol clinic visit. We will talk to you about effective birth control 
methods. They are listed on a handout that we will give to you. 
Being in the study 
If you meet the study requirements and want to join, here is what will happen: 
9. You will come to the clinic for scheduled visits about [#] times over about a 
year and a half. 
Site: Insert number of visits and range of visit lengths. (There is site-specific variation 
in screening protocols and in the number of possible follow-up visits between 
protocol-mandated visits.) 
Visits can last from [#] to [#] hours. 
You may have to come for more visits if you have a lab or health issue. 
We will contact you 6 months after your last scheduled visit to the clinic to check 
on your health. (For example, we will ask you if you had any side effects that you 
think may be related to the study vaccine or any of the study procedures.) 
10. We will give you [Site: Insert compensation] for each study visit you complete. 
This amount is to cover the costs of [Site: Insert text] 
Site: Insert any costs to participants (eg, birth control costs for female participants 
who could become pregnant). 
US sites: Include the following paragraph. You can remove the box around the text. 
Payments you receive for being in the study may be taxable. We may need to ask 
you for your Social Security number for tax reasons. 
You do not have to pay anything to be in this study. 
11. We will give you either the study vaccine or a placebo. 
Not everyone in this study will get the study vaccine. Some people will get a 
placebo, a substance that does not contain vaccine. We will compare the results 
from people who got the placebo with results from people who got the study 
vaccine. In this study, the placebo is sterile salt water. 
You have a 5-in-6 chance of getting the study vaccine. Site: Modify the 
randomization metaphor in the next sentence as appropriate to your local culture. 

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Whether you get the study vaccine or the placebo is completely random, like 
flipping a coin. 
We have no say in whether you get the study vaccine or the placebo. We will not 
know which one you are 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 you got the study vaccine or the placebo. This could be several years. 
But, if you have a serious medical problem and need to know what you got before 
the end of the study, we can tell you. 
12. We will give you the study products on a schedule. 
You will be in one of two groups. You will get two injections, one into each of 
your upper arms at four visits during the study. Group 1 will be enrolled first, and 
then Group 2 will be enrolled. 
 
 
Injection Schedule 
 
Group 
First 
injection 
2 months  
6 months  
12 months  
5 people 
1 
Low dose 
(Study vaccine 
at 0.5 mg/mL 
with 500 mcg 
alum) 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
1 person 
 
Placebo 
Placebo 
Placebo 
Placebo 
15 people 
2 
Higher dose 
(Study vaccine 
at 2 mg/mL 
with 500 mcg 
alum) 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
MPER with 
alum 
adjuvant 
3 people 
 
Placebo 
Placebo 
Placebo 
Placebo 
Total 24 people (20 study vaccine recipients and 4 placebo recipients) 
You will have to wait in the clinic for about a half 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 you are feeling and if you 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 

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of each injection, we will also need to be in contact with you to ask how you are 
doing. Contact the clinic staff if you have any issues or concerns after getting an 
injection. If you have a problem, we will continue to check on you until it goes 
away. 
13. In addition to giving you the study products, we will: 
• Do regular HIV testing, as well as counseling on your results and on how to 
avoid getting HIV 
• Do physical exams 
• Do pregnancy tests if you were assigned female sex at birth 
• Ask questions about your health, including medications you may be taking 
• Ask questions about any personal problems or benefits you may have from 
being in the study 
• Take urine and blood samples. 
When we take blood, the amount will depend on the lab tests we need to do. It 
will be some amount between 10 mL and 340 mL (1 tablespoon to a little less 
than 1 and a half cups). Your 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 D, Table of procedures (for informed consent form) in this 
section or distribute it as a separate sheet if it is helpful to your study participants. 
You are not required to do either. 
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 your health, we will tell you. 
14. We will counsel you about protecting yourself from HIV. 
We will ask you personal questions about your HIV risk factors such as sexual 
behavior, alcohol, and drug use. We will talk with you about ways to keep your 
risk of getting HIV low. 

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

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

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compare the genomes of many people, looking for common patterns of genes that 
could help them understand diseases. The researchers may put the information 
from the genome-wide studies into a protected database so that other researchers 
can access it, but your name and other personal information will not be included. 
Usually, no one would be able to look at your genome and link it to you as a 
person. However, if another database exists that also has information on your 
genome and your name, someone might be able to compare the databases and 
identify you. If others found out, it could lead to discrimination or other problems. 
The risk of this is very small. There may be other unknown risks. 
Who will have access to my information in studies using my extra samples? 
People who may see your information are: 
• Researchers who use your extra samples and information for other research 
• Government agencies that fund or monitor the research using your extra 
samples and information 
• The researcher’s Institutional Review Board or Ethics Committee 
• Any regulatory agency that reviews clinical trials 
• The people who work with the researcher 
All of these people will do their best to protect your information. The results of 
any new studies that use your extra samples and information may be published. 
No publication will use your name or identify you personally. 
17. We will do our best to protect your private information. 
US sites: Check HIPAA authorization for conflicts with this section. 
Your study records and samples will be kept in a secure location. We will label all 
of your samples and most of your records with a code number, not your name or 
other personal information. However, it is possible to identify you, if necessary. 
We will not share your name with the lab that does the tests on your samples, or 
with anyone else who does not need to know your name. 
Site: Any change to the following boxed text requires approval from HVTN 
Regulatory Affairs. You can remove the box around the text.  
We do need to share your name with the HVTN in case you need proof in the 
future that you participated in an HIV vaccine study. The HVTN will keep your 
name in a secure file with these items:  
• The name of your study 

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• Your age or date of birth 
• Your study ID number 
• What study product(s) you received 
There are no HIV test results kept in this file. The HVTN will not share any 
information that could identify you without your agreement. The HVTN will 
remove your name from the file if you do not want it there. 
Clinic staff will have access to your study records. Your records may also be 
reviewed by groups who watch over this study to see that we are protecting your 
rights, keeping you safe, and following the study plan. These groups include: 
• The US National Institutes of Health and its study monitors, 
• The US Food and Drug Administration, 
• Any regulatory agency that reviews clinical trials, 
• [Insert name of local IRB/EC] , 
• [Insert name of local and/or national regulatory authority as appropriate], 
• The Division of AIDS and people who work for them, 
• The HVTN and people who work for them, 
• Infectious Disease Research Institute, 
• Duke University, 
• The Safety Monitoring Board and 
• The US Office for Human Research Protections. 
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:  
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. 

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• [Item 1] 
• [Item 2] 
• [Item 3] 
US sites: Include the following boxed text. You can remove the box around the text. 
We have a Certificate of Confidentiality from the US government, to help protect 
your privacy. With the certificate, we do not have to release information about 
you to someone who is not connected to the study, such as the courts or police. 
Sometimes we can’t use the certificate. Since the US government funds this 
research, we cannot withhold information from it. Also, you can still release 
information about yourself and your study participation to others. 
The results of this study may be published. No publication will use your name or 
identify you personally. 
We may share information from the study with other researchers. We will not 
share your name or information that can identify you. 
A description of this clinical trial will be available on 
http://www.ClinicalTrials.gov. This 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. 
18. There are several reasons why we may stop your injections. We may stop them 
even if you want to stay in the study and even if you were scheduled for more 
injections. 
We will stop your injections if you become pregnant. We will encourage you to 
stay in the study if you choose. We will discuss your study options with you. If 
you leave the study while you are still pregnant, we will contact you after your 
due date to ask some questions about your pregnancy and delivery. 
We will stop your injections if you get HIV. We will also take fewer samples, and 
we will help you get care and support. We will encourage you to stay in the study 
for up to 18 weeks after your last injection if you choose. We will discuss your 
study options with you. We will counsel you about having HIV and about telling 
your partner(s). Site: Modify the following sentence as appropriate. We will not 
provide or pay for any of your HIV care directly. 
If we stop your injections, we may ask you to stay in the study to complete other 
study procedures. 
19. We may take you out of the study at any time. 
We may take you out of the study if: 

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• you do not follow instructions, 
• we think that staying in the study might harm you, 
• you enroll in a different research study where you get another study product, 
or 
• the study is stopped for any reason. 
Other Risks 
20. 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 you got the injection. Taking blood can cause a low blood cell 
count (anemia), making you feel tired. 
Personal problems/discrimination/testing HIV antibody positive: 
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, or assume that you have HIV or at high risk and treat 
you unfairly as a result. Rarely, a person has lost a job because the study took too 
much time away from work, or because their employer thought they had HIV. 
The body makes antibodies to fight or prevent infection. Most vaccines cause the 
body to make antibodies as a way of preventing infection. Your body may make 
antibodies to HIV because you received an HIV study vaccine. The study vaccine 
may cause you to test positive on some types of HIV antibody tests, even if you 
do not have HIV. This is called vaccine-induced seropositivity (VISP). VISP 
means that after you get the study vaccine, a routine HIV test done outside this 
clinic may say you have HIV, even if you don’t. For this reason, you should plan 
to get HIV tests only at this clinic during the study. Our tests can tell the 
difference between true HIV infection and a positive result that is caused by the 
study vaccine. 
If you have a positive test result caused by the study vaccine at any time, we can 
arrange free HIV testing for as long as you need it. If this happens, we do not 
know how long you will test positive due to the study vaccine. If you receive a 

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positive HIV test result and we determine it is because you have HIV, we will 
refer you for follow-up care. 
It is unlikely, but you could test negative at the end of the study and positive some 
time later, even though you don’t have HIV. This could happen if different 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 you have 
HIV even if you do not, you could face discrimination and other problems. For 
example, in some countries, you could be denied medical or dental care, 
employment, insurance, a visa, or entry into the military. If you do have a positive 
HIV antibody test caused by the study vaccine, you will not be able to donate 
blood or organs. Your family and friends may treat you differently. We will give 
you a brochure that tells you more about testing HIV positive because of an HIV 
vaccine, and how you can avoid some of these problems. 
If you become pregnant during or after the study and have VISP, we don't know if 
the antibodies could be passed to your baby. We know that this happens with 
other vaccines, like tetanus vaccine. These antibodies from the mother are not a 
danger to the baby, and they go away over time. For most babies antibodies from 
the mother last for about six months. 
You should always tell the delivery staff if you have VISP. However, you may 
still be tested for HIV using the antibody test when you deliver your baby. If your 
test is positive and the delivery staff believes you have HIV, your baby may be 
started on antiretroviral treatment when it is not needed. If this happens, we can 
arrange for you and the baby to have a test that can tell the difference between 
true HIV infection and a VISP result. If you or the baby continue to have VISP, 
we can arrange this testing for free for as long as it is needed. 
Embarrassment/anxiety: 
You may feel embarrassed when we ask about your HIV risks, such as having sex 
and using drugs. Also, waiting for your HIV test results or other health test results 
could make you feel anxious. You could feel worried if your test results show that 
you have HIV. If you feel embarrassed or anxious, please tell us and we will try to 
help you. 
Risks of disclosure of your personal information: 
We will take several steps to protect your personal information. Although the risk 
is very low, it is possible that your personal information could be given to 
someone who should not have it. If that happened, you could face discrimination, 
stress, and embarrassment. We can tell you more about how we will protect your 
personal information if you would like it. 
Risks of genetic testing: 

HVTN 133 Version 1.0 / March 13, 2019 
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It is unlikely, but the genetic tests done on your samples could show you may be 
at risk for certain diseases. If others found out, it could lead to discrimination or 
other problems. However, it is almost impossible for you or others to know your 
test results from the genetic testing. The results are not part of your study records 
and are not given to you. 
US Sites, include the following paragraph In the very unlikely event that your 
genetic information becomes linked to your name, a federal law called the 
Genetic Information Nondiscrimination Act (GINA) helps protect you. GINA 
keeps health insurance companies and employers from seeing results of genetic 
testing when deciding about giving you health insurance or offering you work. 
GINA does not help or protect you against discrimination by companies that sell 
life, disability or long-term care insurance. 
Unknown risks: 
We do not know if the study vaccine will increase, decrease, or not change your 
risk of getting HIV if exposed. If you get HIV, we do not know how the study 
vaccine might affect your HIV infection or how long it takes to develop AIDS. 
We do not know if getting this study vaccine will affect how you respond to any 
future approved HIV vaccine. Currently, no HIV vaccine has been approved for 
use. 
We do not know how the study vaccine will affect a pregnant participant or a 
developing baby. 
Benefits 
21. The study may not benefit you. 
We do not expect the study vaccine to benefit you in any way. However, being in 
the study might still help you in some ways. The counseling that you get as part of 
the study may help you avoid getting HIV. The lab tests and physical exams that 
you get while in this study might detect health problems you don’t yet know 
about. 
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. 
Your rights and responsibilities 
22. If you join the study, you have rights and responsibilities. 
You 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. 

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Leaving the study 
23. Tell us if you decide to leave the study. 
You are free to leave the study at any time and for any reason. Your care at this 
clinic and your legal rights will not be affected, but it is important for you to let us 
know. 
We will ask you to come back to the clinic one last time for a physical exam, and 
we may ask to take some blood and urine samples. We will also ask about any 
personal problems or benefits you have experienced from being in the study. We 
believe these steps are important to protecting your health, but it is up to you 
whether to complete them. 
Injuries 
Sites: 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. 
24. If you get sick or injured during the study, contact us immediately. 
Your health is important to us. (Sites: adjust the following 2 sentences if 
applicable to the care available at your site) We will tell you about the care that 
we can give here. For the care that we cannot provide, we will explain how we 
will help you get care elsewhere. 
If you become sick or injured in this study, there is a process to decide if it is 
related to the [study product(s)]  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. 
The study product provider may agree to pay medical costs for study-related 
injuries that are determined to be caused by the study product. 
The HVTN has limited funds to pay medical costs that it determines are 
reasonable. (Sites: insert locale- appropriate medical insurance language in the 
following sentence) If the injury is not study related, then you and your health 
insurance will be responsible for treatment costs. 
Some injuries are not physical. For example, you might be harmed emotionally by 
being in an HIV vaccine study. Or you might lose wages because you cannot go 
to work. However, there are no funds to pay for these kinds of injuries, even if 
they are study related. 

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You may disagree with the decision about whether your injury is study related. If 
you wish, the HVTN will ask independent experts to review the decision. You 
always have the right to use the court system if you are not satisfied. 
Questions  
25. If you have questions or problems at any time during your participation in this 
study, use the following important contacts. 
If you have questions about this study, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you have any symptoms that you think may be related to this study, contact 
[name or title and telephone number of the investigator or other study staff]. 
This study has been reviewed and approved by a committee called the 
[name of local IRB/EC]. If you have questions about your rights as a research 
participant, or problems or concerns about how you are being treated in this study, 
contact [name or title and telephone number of person on IRB/EC] , at the 
committee. 
Your permissions and signature  
Site: Delete this section if using a separate consent for use of samples and 
information in other studies 
26. In Section 15 of this form, we told you about possible other uses of your extra 
samples and information, outside this study. Please choose only one of the 
options below and write your initials or make your mark in the box next to it. 
Whatever you choose, the HVTN keeps track of your decision about how your 
samples and information can be used. You can change your mind after signing 
this form. 
 

HVTN 133 Version 1.0 / March 13, 2019 
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I allow my extra samples and information to be used for other studies related to HIV, 
vaccines, the immune system, and other diseases. This may include genetic testing and 
keeping my cells growing over time.  
OR 
 
 
I agree to the option above and also to allow my extra samples and information to be 
used in genome wide studies. 
OR 
 
 
I do not allow my extra samples to be used in any other studies. This includes not 
allowing genetic testing, growing more of my cells, or genome wide studies.  
 
27. If you agree to join this study, you will need to sign or make your mark below. 
Before you sign or make your mark on this consent form, make sure of the 
following: 
• You have read this consent form, or someone has read it to you. 
• You feel that you understand what the study is about and what will happen to 
you if you join. You understand what the possible risks and benefits are. 
• You have had your questions answered and know that you can ask more. 
• You agree to join this study. 
You will not be giving up any of your rights by signing this consent form. 
 
 
 
 
 
 
 
Participant’s name (print) 
 
Participant’s signature or mark 
 
Date 
 
Time 
 
 
 
 
 
 
 
Clinic staff conducting consent 
discussion (print) 
 
Clinic staff signature 
 
Date 
 
Time 
For participants who are unable to read or write, a witness should complete the 
signature block below: 
 
 
 
 
 
 
 
Witness’s name (print) 
 
Witness’s signature  
 
Date 
 
Time 
*Witness is impartial and was present for the entire discussion of this consent form. 
 
 

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Appendix B 
Approved birth control methods for persons 
assigned female sex at birth (for sample informed 
consent form) 
Site: Any change to the following boxed text requires approval from HVTN 
Regulatory Affairs at vtn.core.reg@hvtn.org. You can remove the box around the text. 
You should not become pregnant during the study because we do not know how 
the study vaccine could affect the developing baby. 
You must agree to use effective birth control from at least 21 days before your 
first injection through the last required protocol clinic visit.  
Effective birth control means using any of the following methods every time you 
have sex: 
• Birth control drugs that prevent pregnancy—given by pills, shots, patches, 
vaginal rings, or inserts under the skin; 
• Male or female condoms, with or without a cream or gel that kills sperm; 
• Diaphragm or cervical cap with a cream or gel that kills sperm; 
• Intrauterine device (IUD); or 
• Any other contraceptive method approved by the researchers. 
You do not have to use birth control if: 
• You are only having sex with a partner or partners who have had a vasectomy. 
(We will ask you some questions to confirm that the vasectomy was 
successful.); 
• You have reached menopause, with no menstrual periods for one year; 
• You have had a hysterectomy (your uterus removed); 
• You have had your ovaries removed; 
• You have a tubal ligation (your “tubes tied”) or confirmed successful 
placement of a product that blocks the fallopian tubes; 
• You are having sex only with a partner(s) assigned female sex at birth; 
• You only have oral sex; or, 
• You are sexually abstinent (no sex at all). 
Remember: If you are having sex, male and female condoms are the only birth 
control methods that also provide protection against HIV and other sexually 
transmitted infections. 
If you join the study, we will test you for pregnancy at some visits, including 
before each study injection. 

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Appendix C 
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 an 
HIV-1 gp41 MPER liposome vaccine in healthy, HIV-uninfected adult 
participants 
HVTN protocol number: HVTN 133 
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 extra samples to be 
used in other studies if you agree to this. You will mark your decision at the end 
of this form. If you have any questions, please ask. 
1. Do I have to agree? 
No. You are free to say yes or no, or to change your mind after you sign this form. 
At your request, we will destroy all extra samples that we have. Your decision 
will not affect your being in this study or have any negative consequences here. 
2. Where are the samples stored? 
Extra samples are stored in a secure central place called a repository. [Site: 
choose one of the following two sentences. African sites should choose the 
sentence referencing the repository in South Africa. All other sites should choose 
the sentence referencing the repository in the United States.] Your samples will 
be stored in the HVTN repository in South Africa. Your samples will be stored in 
the HVTN repository in the United States. 
3. How long will the samples be stored? 
There is no limit on how long your extra samples will be stored. [Site: Revise the 
previous sentence to insert limits if your regulatory authority imposes them.] 
4. Will I be paid for the use of my samples? 
No. Also, a researcher may make a new scientific discovery or product based on 
the use of your samples. If this happens, there is no plan to share any money with 
you. The researcher is not likely to ever know who you are. 
5. Will I benefit from allowing my samples to be used in other studies? 
Probably not. Results from these other studies are not given to you, this clinic, or 
your doctor. They are not part of your medical record. The studies are only being 
done for research purposes. 

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6. Will the HVTN sell my samples and information? 
No, but the HVTN may share your samples with HVTN or other researchers. 
Once we share your samples and information, we may not be able to get them 
back. 
7. How do other researchers get my samples and information? 
When a researcher wants to use your samples and information, their research plan 
must be approved by the HVTN. Also, the researcher’s institutional review board 
(IRB) or ethics committee (EC) will review their plan. [Site: If review by your 
institution’s IRB/EC/RE is also required, insert a sentence stating this.] IRBs/ECs 
protect the rights and well-being of people in research. If the research plan is 
approved, the HVTN will send your samples to the researcher’s location. 
8. What information is shared with HVTN or other researchers?  
The samples and information will be labeled with a code number. Your name will 
not be part of the information. However, some information that we share may be 
personal, such as your race, ethnicity, gender, health information from the study, 
and HIV status. We may share information about the study product you received 
and how your body responded to the study product. 
9. What kind of studies might be done with my extra samples and information? 
The studies will be related to HIV, vaccines, the immune system and other 
diseases. 
Researchers may also do genetic testing on your samples. 
In some cases, researchers may take cells from your samples and grow more of 
them over time, so that they can continue to do research with them. 
If you agree, your samples could also be used for genome wide studies. In these 
studies, researchers will look at all of your genes (your genome). The researchers 
compare the genomes of many people, looking for common patterns of genes that 
could help them understand diseases. The researchers may put the information 
from the genome-wide studies into a protected database so that other researchers 
can access it, but your name and other personal information will not be included. 
Usually, no one would be able to look at your genome and link it to you as a 
person. However, if another database exists that also has information on your 
genome and your name, someone might be able to compare the databases and 
identify you. If others found out, it could lead to discrimination or other problems. 
The risk of this is very small. There may be other unknown risks. 

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10. What are the risks of genetic testing? 
It is unlikely, but the genetic tests done on your samples could show you may be 
at risk for certain diseases. If others found out, it could lead to discrimination or 
other problems. However, it is almost impossible for you or others to know your 
test results from the genetic testing. The results are not part of your study records 
and are not given to you. 
US Sites, include the following paragraph  
In the very unlikely event that your genetic information becomes linked to your 
name, a federal law called the Genetic Information Nondiscrimination Act 
(GINA) helps protect you. GINA keeps health insurance companies and 
employers from seeing results of genetic testing when deciding about giving you 
health insurance or offering you work. GINA does not help or protect you against 
discrimination by companies that sell life, disability or long-term care insurance. 
11. Who will have access to my information in studies using my extra samples? 
People who may see your information are: 
• Researchers who use your extra samples and information for other research 
• Government agencies that fund or monitor the research using your extra 
samples and information 
• The researcher’s Institutional Review Board or Ethics Committee 
• Any regulatory agency that reviews clinical trials 
• The people who work with the researcher 
All of these people will do their best to protect your information. The results of 
any new studies that use your extra samples and information may be published. 
No publication will use your name or identify you personally. 
Questions  
12. If you have questions or problems about allowing your samples and 
information to be used in other studies, use the following important contacts. 
If you have questions about the use of your samples or information or if you want 
to change your mind about their use, contact 
[name or title and telephone number of the investigator or other study staff]. 
If you think you may have been harmed because of studies using your samples or 
information, contact 
[name or title and telephone number of the investigator or other study staff]. 

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

HVTN 133 Version 1.0 / March 13, 2019 
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Appendix D 
Table of procedures (for sample informed consent form) 
 
 
 
Time after first injection visit 
Procedure 
Screening 
visit(s) 
First 
injection 
visit 
2 
weeks 
2 
months 
2½ 
months 
6 
months 
6½ 
months 
9 
months 
12 
months 
 
12¼ 
months 
 
12½ 
months 
18 
months 
Injection 
 
√ 
 
√ 
 
√ 
 
 
√ 
 
 
 
Medical history 
√ 
 
 
 
 
 
 
 
 
 
 
 
Complete physical 
√ 
 
 
 
 
 
 
 
 
 
 
√ 
Brief physical 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
Urine test 
√ 
 
√ 
 
 
 
 
 
 
 
√ 
 
Blood drawn 
√ 
√ 
√ 
 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
Pregnancy test (participants 
assigned female sex at birth)* 
√ 
√ 
 
√ 
 
√ 
 
 
√ 
 
√ 
 
HIV testing and pretest 
counseling 
√ 
 
 
 
√ 
√ 
 
√ 
√ 
 
 
√ 
Risk reduction counseling 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
Interview/questionnaire 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
√ 
 
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 you received.  
* Persons who had a hysterectomy (removal of the uterus) or removal of both ovaries (verified by medical records), are not required to have a pregnancy test. 

HVTN 133 Version 1.0 / March 13, 2019 
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Appendix E 
Laboratory procedures 
Visit:
1
2
3
4
5
6
7
8
9
10
11
12
Day:
D0
D14
D56
D70
D168
D182
D273
D364
D371
D378
D546
Month:
M0
M0.5
M2
M2.5
M6
M6.5
M9
M12
M12.25
M12.5
M18
VAC1
VAC2
VAC3
VAC4
Procedure
Ship to1
Tube 4
Tube size 
(vol. 
capacity)4
MPER 
or 
Placebo
MPER 
or 
Placebo
MPER 
or 
Placebo
MPER 
or 
Placebo
Total
BLOOD COLLECTION
Screening or diagnostic assays
Screening HIV test
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
5
HBsAg/anti-HCV
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
5
Syphilis
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
5
Anticardiolipin antibodies (ACA), IgG                 
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
5
Antinuclear antibodies (ANA)
Local lab
Local lab
SST
5mL
5
—
—
—
—
—
—
—
—
—
—
—
5
HIV diagnostics9
UW-VSL
UW-VSL
EDTA
10mL
—
—
—
—
10
10
—
10
10
—
—
20
60
Safety labs10
CBC/Diff/Platelets
Local lab
Local lab
EDTA
5mL
5
—
5
—
5
—
5
—
—
—
5
—
25
Chemistry panel5
Local lab
Local lab
SST
5mL
5
—
5
—
5
—
5
—
—
—
5
—
25
Autoantibody assays
Antinuclear antibodies (ANA)
CSR
Non-HVTN Labs 
at Duke-DHVI 
SST
8.5mL
—
y
y
—
y
—
y
—
—
—
y
y
0.0
Anticardiolipin antibodies (ACA), IgG                 
CSR
Non-HVTN Labs 
at Duke-DHVI 
SST
8.5mL
—
y
y
—
y
—
y
—
—
—
y
y
0.0
Host genetics7
CSR
HVTN Labs
ACD
8.5mL
—
17
—
—
—
—
—
—
—
—
—
—
17
Immunogenicity assays6
Humoral assays
HIV-1 binding Ab
CSR
HVTN Labs
SST
8.5mL
—
8.5
8.5
—
8.5
—
8.5
—
—
—
8.5
8.5
51
Lipid binding Ab
CSR
HVTN Labs
SST
8.5mL
—
y
y
—
y
—
y
—
—
—
y
y
0
Ab avidity
CSR
HVTN Labs
SST
8.5mL
—
y
y
—
y
—
y
—
—
—
y
y
0
Neutralizing Ab
CSR
HVTN Labs
SST
8.5mL
—
8.5
8.5
—
8.5
—
8.5
—
—
—
8.5
8.5
51
Cellular assays
HIV-specific ICS
CSR
HVTN Labs
ACD
8.5mL
—
—
—
—
42.5
—
42.5
—
—
—
42.5
42.5
170
Phenotyping (pTfh)
CSR
HVTN Labs
ACD
8.5mL
—
42.5
—
—
—
—
—
—
42.5
42.5
—
42.5
170
Ag-specific B-cell phenotyping
CSR
HVTN Labs
ACD
8.5mL
—
42.5
42.5
—
42.5
—
42.5
—
—
42.5
42.5
42.5
297.5
B-cell lineage
CSR
HVTN Labs
ACD
8.5mL
—
34
34
—
34
—
34
—
—
34
34
34
238
Specimen storage
PBMC
CSR
ACD
8.5mL
—
51
51
—
85
—
85
—
—
25.5
85
85
467.5
Serum
CSR
SST
8.5mL
—
17
17
—
17
—
17
—
8.5
—
17
17
110.5
Visit total
35
221.0
172
0
258.0
10
248.0
10
61
145
248.0
301
1707.5
56-Day total
35
256.0
427.5
427.5
429.5
10
258.0
10
61
206
454
301
URINE COLLECTION10
Urine dipstick11
Local lab
Local lab
X
—
X
—
—
—
—
—
—
—
X
—
Pregnancy test 8
Local lab
Local lab
X
X
—
X
—
X
—
—
X
—
X
—
Screening 
visit3
Assay Location 2
 
 
1 CSR = Central Specimen Repository; UW-VSL = University of Washington Virology Specialty Laboratory (Seattle, Washington, USA). 
2 HVTN Laboratories include Fred Hutchinson Cancer Research Center (Seattle, Washington, USA); Duke University Medical Center (Durham, North Carolina, USA). Non-HVTN laboratories: 
Duke Human Vaccine Institute (Durham, North Carolina, USA) 

HVTN 133 Version 1.0 / March 13, 2019 
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3 Screening may occur over the course of several contacts/visits up to and including day 0 prior to vaccination. 
4 Local labs may assign appropriate alternative tube types for locally performed tests. 
5 Chemistry panels are defined in Section 9.2 (pre-enrollment) and Section 9.4 (postenrollment). 
6 Immunogenicity assays will be performed at M0 (for binding Ab assay), M6.5, and M12.5. Depending on the initial results, assays for humoral and cellular responses may be performed on samples 
collected from participants at other timepoints. Immunogenicity assays at M6.5 may begin as samples become available, and are not contingent on data from the primary immunogenicity timepoint 
at M12.5. 
7 Genotyping may be performed on enrolled participants using cryopreserved PBMC collected at baseline, initially in participants who demonstrate vaccine-induced T-cell responses at 
postvaccination timepoints. 
8 For a participant who was born female (ie, assigned female sex at birth), pregnancy test must be performed on urine or blood specimens within 24 hours of vaccination with negative results received 
prior to vaccination. Persons who have undergone total hysterectomy or bilateral oophorectomy (verified by medical records), are not required to undergo pregnancy testing. 
9 At an early termination visit for a withdrawn or terminated participant who is not HIV-infected (see Section 9.11), blood should be drawn for HIV diagnostic testing, as shown for visit 12 above. If a 
participant has a confirmed diagnosis of HIV infection, do not collect blood for HIV diagnostic testing (see Section 9.13). 
10 For participants with confirmed diagnosis of HIV infection, only specimens required for protocol-specified safety laboratory tests, urinalysis and pregnancy tests will be collected. 
11 And microscopy if needed. 
y = SST collected for serum storage will also cover specimen needs for autoantibody assays, Ab avidity, and lipid binding Ab assays; no separate blood draw is needed. 
 
 

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Appendix F 
Procedures at HVTN CRS 
Visit: 
011  
02 
03 
04 
05 
06 
07 
08 
09 
10 
11 
12 
13 
Post 
Day: 
 
D0 
D14 
D56 
D70 
D168 
D182 
D273 
D364 
D371 
D378 
D546 
D728 
 
Month: 
 
M0 
M0.5 
M2 
M2.5 
M6 
M6.5 
M9 
M12 
M12.25 
M12.5 
M18 
M24 
 
Procedure 
Scr. 
VAC1 
 
VAC2 
 
VAC3 
 
 
VAC4 
 
 
 
AESI 
 
Study procedures2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Signed screening consent (if used) 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Assessment of understanding 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Signed protocol consent 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Medical history  
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Complete physical exam 
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
— 
— 
Abbreviated physical exam 
— 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
— 
— 
Risk reduction counseling3 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
— 
Contraception status assessment4 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
— 
Behavioral risk assessment5 
X 
— 
— 
— 
— 
X 
— 
— 
X 
— 
— 
X 
— 
— 
Confirm eligibility, obtain 
demographics, randomize  
X 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
Social impact assessment 
— 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
— 
— 
Social impact assessment questionnaire 
— 
— 
— 
X 
— 
X 
— 
— 
— 
— 
— 
X 
— 
— 
Outside testing and belief questionnaire 
— 
— 
— 
— 
— 
X 
— 
— 
— 
— 
— 
X 
— 
— 
Concomitant medications 
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 
— 
— 
HIV infection assessment6  
X 
— 
— 
— 
X 
X 
— 
X 
X 
— 
— 
X 
— 
— 
Confirm HIV test results provided to 
participant 
— 
X 
— 
— 
— 
X 
X 
— 
X 
X 
— 
X 
— 
— 
Specimen collection  
(see Appendix E)7 
X 
X 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
Vaccination procedures 5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Vaccination8 
— 
X 
— 
X 
— 
X 
— 
— 
X 
— 
— 
— 
— 
— 
Reactogenicity assessments9 
— 
X 
— 
X 
— 
X 
— 
— 
X 
— 
— 
— 
— 
— 
AESI health contact10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
AESI assessment 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 
— 
Poststudy 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Unblind participant 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
— 
X 

HVTN 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 116 of 121 
 
 
1 Screening may occur over the course of several contacts/visits up to and including day 0 prior to vaccination. 
2 For specimen collection requirements, see Appendix E. 
3 Includes transmission risk reduction counseling for HIV-infected participants. Conduct after BRA questionnaire if both occur at same visit. 
4  Contraception status assessment is required only for participants who were assigned female sex at birth and are capable of becoming pregnant. Persons who are confirmed 
pregnant, pregnancy testing is not required, unless clinically indicated. 
5 Not applicable to HIV-infected participants. Conduct before Risk Reduction Counseling if both occur at same visit. 
6 Includes pretest counseling and HIV testing. A subsequent follow-up contact is conducted to provide post-test counseling and to report results to participant. If a participant has a 
confirmed diagnosis of HIV infection, do not perform HIV infection assessment. 
7 For participants with a confirmed diagnosis of HIV infection, specimens listed under “Safety labs” in Appendix E, urinalysis, and urine pregnancy tests will be collected per the 
protocol schedule. 
8 Blood draws required at vaccination visits must be performed prior to vaccination; however, it is not necessary to have results prior to vaccination, except for results of a urine or 
serum pregnancy test, if indicated. 
9 Reactogenicity assessments performed daily for at least 7 days postvaccination (see Section 9.9). 
10 Clinic visits are not required unless participant indicates symptoms that require further assessment. See Appendix H. 
                                            

HVTN 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 117 of 121 
Appendix G 
HVTN low risk guidelines for the US 
 

HVTN 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 118 of 121 
 
 

HVTN 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 119 of 121 
Appendix H 
Adverse events of special interest 
Adverse events of special interest (AESIs) for this protocol include but are not limited to 
unexpected induction of autoimmune or auto-inflammatory diseases; representative examples 
of AESIs are listed below. 
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, myeloradiculomyelitis) 
• 
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 
• 
Myasthenia gravis, including Eaton-
Lambert syndrome 
• 
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 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 120 of 121 
Appendix I 
HVTN 133 Visit Windows 
Visit 
Number 
Visit Type 
Lower 
Allowable 
Window 
Lower 
Target Day 
Target 
Day 
Upper 
Target Day 
Upper 
Allowable 
Window 
01.0 
Screening 
-56 
- 
 
- 
 
02.0 
Enrollment 
 
- 
0 
- 
 
03.0 
2 Weeks Post-
Vaccination 
 
-4 
14 
+4 
+7 
04.0 
Vaccination 2 
 
-7 
56 
+9 
+14 
05.0 
2 Weeks Post-
Vaccination 
 
-4 
70 
+4 
+7 
06.0 
Vaccination 3 
-28 
-14 
168 
+14 
+28 
07.0 
2 Weeks Post-
Vaccination 
 
-4 
182 
+4 
+7 
08.0 
Follow-up 
-28 
-14 
273 
+14 
+28 
09.0 
Vaccination 4  
-28 
-14 
364 
+14 
+28 
10.0 
1 Week Post-
Vaccination 
-2 
-1 
371 
+1 
+2 
11.0 
2 Weeks Post-
Vaccination 
 
-3 
378 
+4 
+7 
12.0 
Follow-up 
-28 
-14 
546 
+14 
+28 
13.0 
6 Month Health 
Contact 
-28 
-14 
728 
+14 
+28 
1. Target dates are relative to Day 0 (Enrollment), with the exception of post-vaccination visits 
3.0, 5.0, 7.0, 10.0, 11.0 which are relative to the prior vaccination visit. 
2. See Section 9.10. 

HVTN 133 Version 1.0 / March 13, 2019 
HVTN133_v1.0_FINAL / Page 121 of 121 
Appendix J 
Protocol Signature Page 
 
A phase 1 clinical trial to evaluate the safety and immunogenicity of an HIV-1 
gp41 MPER-656 liposome vaccine in healthy, HIV-uninfected adult participants 
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 133 
DAIDS Protocol Version: Version 1.0 
Protocol Date: March 13, 2019