Conditions: Human Immunodeficiency Virus
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 27
Healthy volunteers: no

Brief summary:
The purpose of the study is to assess: 1 safety and tolerability of adenovirus serotype 26 (Ad26) prime and Modified Vaccinia Ankara (MVA) boost versus placebo in participants on suppressive antiretroviral therapy (ART) that was initiated during acute Human Immunodeficiency Virus (HIV) infection; 2) Measure the frequency and duration of sustained viremic control after receiving Ad26 prime/MVA boost or placebo, defined as greater than 24 weeks with plasma HIV ribonucleic acid (RNA) lesser than (\<)50 copies/ml after antiretroviral (ARV) analytical treatment interruption (ATI).

Detailed description:

Arms:
arm 1: Participants will receive adenovirus serotype 26-Mosaic -Human Immunodeficiency Virus (Ad26.Mos.HIV) 0.5 milliliter (mL) injection intramuscularly (containing 5 \* 10\^10 viral particles \[vp\]) at Weeks 0 and 12 followed by modified Vaccinia Ankara-Mosaic (MVA mosaic) 0.5 mL injection (containing 10\^8 Plaque-forming unit \[pfu\]) at Week 24 and 48. arm 2: 0.5 mL Sodium Chloride Injection United States Pharmacopeia (USP) 0.9% will be administered by intramuscular (IM) injection.

Interventions: intervention 1: Ad26.Mos.HIV intervention 2: MVA-Mosaic intervention 3: Placebo
Intervention descriptions: intervention 1: Recombinant replication-deficient Ad26 vectored vaccine and consists of 3 Ad26 vectors, one containing a mosaic insert of envelope (Env) sequence, and 2 vectors containing mosaic inserts of Gag and Pol sequences (Ad26.Mos.1.Env + Ad26.Mos1.Gag-Pol + Ad26.Mos2.Gag-Pol). Total dose is 5\*10\^10 viral particle per 0.5 milliliter (mL) injection administered intramuscularly. intervention 2: Recombinant live attenuated MVA virus-vectored vaccine that has been genetically engineered to express 2 mosaic Gag, Pol, and Env sequences (Mosaic 1 and Mosaic 2). Total dose is 10\^8 plaque-forming unit per 0.5 mL injection administered intramuscularly. intervention 3: Participants will receive placebo intramuscularly Weeks 0, 12, 24 and 48.

Protocol document:
Prot_000.pdf: 
Page 1 of 139
Status: Approved, Date: 11 October 2017
Protocol VAC89220HTX1001 Amendment 3
RV405/SEARCH023/IPCAVD007/WRAIR#2217
Ad26.Mos.HIV/MVA-Mosaic
A Combined Phase 1/2a, Exploratory Study of a Therapeutic Vaccine Using an Adenovirus 
Type 26 Vector Prime and Modified Vaccinia Ankara Boost Combination With Mosaic Inserts 
in HIV-1 Infected Adults who Initiated Antiretroviral Treatment During Acute HIV Infection
Sponsor: 
Janssen Vaccines & Prevention B.V.*
Funding Agencies: 
Janssen Vaccines & Prevention B.V.*
Military HIV Research Program (MHRP, through Cooperative Agreement #W81XWH-11-
2-0174)
*Janssen Vaccines & Prevention B.V. (formerly known as Crucell Holland B.V.) is a Janssen pharmaceutical 
company of Johnson & Johnson and is hereafter referred to as the sponsor of the study. The sponsor is identified on 
the Contact Information page that accompanies the protocol.
Protocol Chair:
Jintanat Ananworanich, MD, PhD
Local Principal Investigator: Nittaya Phanuphak, MD, PhD
Sponsor Clinical Leader: 
Frank Tomaka, MD
Local Project Leader: 
, MD, MPH
Status:
Approved
Date:
11 October 2017
EDMS number: EDMS-ERI-90101185, 17.0
GCP Compliance: This study will be conducted in compliance with Good Clinical Practice (GCP), and applicable regulatory 
requirements. 
Confidentiality Statement
The information in this document contains trade secrets and commercial information that are privileged or confidential and may 
not be disclosed unless such disclosure is required by applicable law or regulations. In any event, persons to whom the 
information is disclosed must be informed that the information is privileged or confidential and may not be further disclosed by 
them. These restrictions on disclosure will apply equally to all future information supplied to you that is indicated as privileged or 
confidential.
NCT02919306
PPD


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TABLE OF CONTENTS
TABLE OF CONTENTS ............................................................................................................................... 2
LIST OF TABLES......................................................................................................................................... 5
LIST OF FIGURES ....................................................................................................................................... 6
PROTOCOL AMENDMENTS....................................................................................................................... 7
LIST OF ABBREVIATIONS........................................................................................................................13
SYNOPSIS.................................................................................................................................................. 15
1.
INTRODUCTION................................................................................................................................19
1.1.
Overall Rationale of Study Design................................................................................................. 20
1.1.1.
Rationale for Modified Vaccinia Ankara (MVA) .......................................................................... 22
1.1.2.
Rationale for Adenovirus Type 26 Vector................................................................................... 22
1.1.3.
Rationale for Mosaic Sequence Inserts...................................................................................... 25
1.1.4.
Rationale for Heterologous Vectors............................................................................................25
1.2.
Rationale for ARV ATI.................................................................................................................... 26
1.3.
Description of Ad26.Mos.HIV......................................................................................................... 27
1.4.
Description of MVA-Mosaic Sequence ..........................................................................................27
1.5.
Preclinical Studies..........................................................................................................................27
1.6.
Clinical Studies...............................................................................................................................31
2.
STUDY HYPOTHESIS AND OBJECTIVES...................................................................................... 37
2.1.
Hypothesis ..................................................................................................................................... 37
2.2.
Primary Objectives.........................................................................................................................37
2.3.
Secondary Objectives .................................................................................................................... 37
2.4.
Exploratory Objectives ................................................................................................................... 38
3.
STUDY DESIGN AND POPULATION...............................................................................................38
3.1.
Study Design.................................................................................................................................. 38
3.2.
Subject Selection and Withdrawal ................................................................................................. 41
3.2.1.
Inclusion Criteria.........................................................................................................................41
3.2.2.
Exclusion Criteria........................................................................................................................42
3.2.3.
Prohibitions and Restrictions...................................................................................................... 44
3.2.4.
Withdrawal or Elimination Criteria ..............................................................................................45
3.2.5.
Cohort Criteria for Proceeding to ARV ATI Phase of Study.......................................................45
3.2.6.
Individual Subject Criteria for ARV ATI at Week 60 ................................................................... 45
3.2.6.1.
Inclusion Criteria for ARV ATI.................................................................................................. 45
3.2.6.2.
Exclusion Criteria for ARV ATI ................................................................................................45
3.3.
Treatment Arm Allocation............................................................................................................... 46
3.4.
Method of Blinding and Unblinding ................................................................................................46
4.
TREATMENTS................................................................................................................................... 47
4.1.
Study Treatments...........................................................................................................................47
4.1.1.
Ad26.Mos.HIV Vaccine............................................................................................................... 47
4.1.2.
MVA-Mosaic Vaccine.................................................................................................................. 47
4.1.3.
Placebo....................................................................................................................................... 48
4.1.4.
Vaccine Administration Schedule...............................................................................................48
4.1.5.
Monitoring During ARV ATI ........................................................................................................ 48
4.1.6.
Criteria to Reinitiate ART During ARV ATI................................................................................. 48


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4.1.7.
Management of Subjects After Reinitiating ART........................................................................ 49
4.1.8.
Management of Subjects With Sustained Virologic Control at End of Study.............................49
4.1.9.
Management of Subjects Who Become Pregnant ..................................................................... 50
4.2.
Handling of Study treatments......................................................................................................... 51
4.2.1.
Packaging and Labeling ............................................................................................................. 51
4.2.2.
Shipment and Storage................................................................................................................ 51
4.2.3.
Dose Preparation and Administration.........................................................................................51
4.2.4.
Accountability..............................................................................................................................52
4.3.
Prior and Concomitant Treatment.................................................................................................. 52
4.4.
Investigational Product Compliance...............................................................................................53
5.
CONDUCT OF THE STUDY.............................................................................................................. 53
5.1.
General Aspects.............................................................................................................................53
5.1.1.
Screening.................................................................................................................................... 60
5.1.2.
Randomization and Vaccination 1/Day 1/Week 0 (Stage 1) ...................................................... 61
5.1.3.
Vaccinations 2 – 4 / Weeks 12, 24, 48 (Stage 1) ....................................................................... 62
5.1.4.
Post-Vaccination Follow-Up Visits / After Study Visits Week 0, 12, 24, 48) (Stage 1)............... 63
5.1.5.
Safety and Immunogenicity Visits / Weeks 4, 16, 26, 30, 36, 50 (Stage 1) ...............................63
5.1.6.
Preparation for ARV ATI / Week 58 (Stage 1)............................................................................ 63
5.1.7.
ARV ATI / Week 60 (Stage 1)..................................................................................................... 63
5.1.8.
Monitoring After ARV ATI / Week 61-96 (Stage 2)..................................................................... 63
5.1.9.
Unscheduled Visits ..................................................................................................................... 64
5.1.10.
Neuropsychiatric Examination.................................................................................................... 64
5.1.11.
Optional Procedures................................................................................................................... 64
5.1.12.
Visit Windows .............................................................................................................................65
5.2.
Vaccination Discontinuation........................................................................................................... 66
5.3.
Study Discontinuation .................................................................................................................... 67
5.3.1.
Early Discontinuation or Withdrawal of Subjects........................................................................ 67
5.3.2.
Premature Termination of the study ...........................................................................................68
5.3.2.1.
Study Pausing Rules ............................................................................................................... 68
5.4.
Laboratory Evaluations .................................................................................................................. 70
5.5.
Potential Risks and Benefits .......................................................................................................... 71
5.5.1.
Risks Related to Vaccines.......................................................................................................... 71
5.5.2.
Risk of Myo/Pericarditis .............................................................................................................. 71
5.5.3.
Risks Related to Blood Draws.................................................................................................... 72
5.5.4.
Risk Related to ATI..................................................................................................................... 72
5.5.5.
Unknown Risks...........................................................................................................................72
5.5.6.
Potential Benefits........................................................................................................................72
6.
SAFETY ASSESSMENTS AND REPORTING ................................................................................. 73
6.1.
Definitions ...................................................................................................................................... 73
6.1.1.
Adverse Event Definitions and Classifications ........................................................................... 73
6.1.2.
Rebound in Viral Replication, Study Endpoint not Considered as AE/ADR...............................74
6.1.3.
Surveillance, Reporting, and Documentation of Adverse Events............................................... 74
6.2.
Special Reporting Situations.......................................................................................................... 75
6.2.1.
Reporting Requirements to the Local IRB.................................................................................. 75
6.3.
Procedures..................................................................................................................................... 75
6.3.1.
All Adverse Events......................................................................................................................75
6.3.1.1.
Post-Vaccination Reactions Occurring Immediately After Each Vaccination..........................76
6.3.1.2.
Local Reactions Occurring Within Seven Days After Each Vaccination................................. 77
6.3.1.3.
Systemic Reactions Occurring Within Seven Days Post-Vaccination..................................... 77
6.3.2.
Serious Adverse Events ............................................................................................................. 78
6.3.3.
Reporting requirements to the Local IRB ................................................................................... 79


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6.3.4.
Causality Assessment of Study Vaccines to Adverse Events.................................................... 79
6.3.5.
Severity of Adverse Events ........................................................................................................ 80
6.3.6.
Follow-Up of Ongoing Adverse Events and Assessment of Outcome....................................... 81
6.3.6.1.
Follow-Up of Non-Serious Adverse Events............................................................................. 81
6.3.7.
Treatment of Adverse Events..................................................................................................... 81
6.3.8.
Pregnancy................................................................................................................................... 81
6.4.
Physical Examination..................................................................................................................... 82
6.5.
Routine Safety Laboratory ............................................................................................................. 82
6.6.
Vital Signs ...................................................................................................................................... 82
6.7.
Electrocardiogram..........................................................................................................................83
7.
PRODUCT QUALITY COMPLAINT HANDLING.............................................................................. 83
7.1.
Procedures..................................................................................................................................... 83
7.2.
Contacting Sponsor Regarding Product Quality............................................................................ 84
8.
PROTOCOL SAFETY REVIEW TEAM AND DATA AND SAFETY MONITORING 
COMMITTEE...................................................................................................................................... 84
8.1.
Protocol Safety Review Team........................................................................................................ 84
8.2.
Data and Safety Monitoring Committee.........................................................................................84
9.
DATA EVALUATION AND ANALYSIS.............................................................................................85
9.1.
Analysis Populations......................................................................................................................85
9.1.1.
Safety Population........................................................................................................................85
9.1.2.
Immunogenicity Population ........................................................................................................ 85
9.1.3.
Efficacy Population ..................................................................................................................... 85
9.1.4.
Secondary and Exploratory Analyses.........................................................................................85
9.2.
Primary Endpoints..........................................................................................................................85
9.3.
Secondary and Exploratory Measurements................................................................................... 85
9.3.1.
Assays That Will be Implemented for Immunogenicity:..............................................................86
9.3.1.1.
Humoral Immunogenicity......................................................................................................... 86
9.3.1.2.
Cellular Immunogenicity .......................................................................................................... 86
9.3.1.3.
Other Immunological Evaluations............................................................................................86
9.3.1.4.
Viral Expression and Reservoir...............................................................................................87
9.4.
Sample Size Consideration............................................................................................................ 87
9.5.
Data Analysis ................................................................................................................................. 87
9.5.1.
Primary Objectives and Endpoints .............................................................................................88
9.5.1.1.
Safety....................................................................................................................................... 88
9.5.1.2.
Efficacy .................................................................................................................................... 88
9.5.1.3.
Clinical Outcomes.................................................................................................................... 88
9.5.2.
Analysis Time Points .................................................................................................................. 88
10.
STUDY-SPECIFIC MATERIALS....................................................................................................... 89
11.
ETHICAL AND LEGAL REQUIREMENTS .......................................................................................89
11.1.
General Requirements................................................................................................................... 89
11.2.
Institutional Review Board/Ethics Committee................................................................................ 89
11.3.
Regulatory Authorities.................................................................................................................... 90
11.4.
Subject Information and Informed Consent ................................................................................... 90
11.5.
Data Access and Protection........................................................................................................... 91
11.6.
Data Coding ................................................................................................................................... 91
11.7.
Future use and Storage of Blood Samples.................................................................................... 92
11.8.
Compensation................................................................................................................................92
11.9.
Compliance with NIH Guidelines for Research Involving Products Containing 
Recombinant DNA .........................................................................................................................92


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11.10.
Institutional Biosafety Committee (IBC) .........................................................................................93
12.
QUALITY CONTROL AND QUALITY ASSURANCE....................................................................... 93
12.1.
Monitoring ...................................................................................................................................... 93
12.2.
Audit and Inspections..................................................................................................................... 93
12.3.
Data Quality Assurance ................................................................................................................. 94
13.
ADMINISTRATIVE REQUIREMENTS ..............................................................................................94
13.1.
Protocol Amendments and Protocol Deviations ............................................................................ 94
13.2.
Continuing Review and Close-out Reports.................................................................................... 95
13.3.
Regulatory Documentation ............................................................................................................ 96
13.3.1.
Regulatory Approval/Notification ................................................................................................96
13.3.2.
Required Prestudy Documentation.............................................................................................96
13.4.
Subject Identification, Enrollment, and Screening Logs ................................................................97
13.5.
Source Documentation................................................................................................................... 97
13.6.
Case Report Form Completion ...................................................................................................... 98
13.7.
Archiving ........................................................................................................................................ 98
13.8.
Study Completion/Termination....................................................................................................... 99
13.8.1.
Study Completion .......................................................................................................................99
13.8.2.
Study Termination.......................................................................................................................99
13.9.
Volunteer Registry Database....................................................................................................... 100
13.10.
Use of Information and Publication..............................................................................................100
14.
REFERENCES................................................................................................................................. 102
15.
SIGNATURES.................................................................................................................................. 107
16.
APPENDICES.................................................................................................................................. 110
16.1.
Definitions of HIV-1 Related Disease...........................................................................................110
16.2.
Case Definition for Acute Retroviral Syndrome (ARS) ................................................................112
16.3.
Test of Understanding.................................................................................................................. 113
16.4.
Research Specimen Laboratory Testing,..................................................................................... 114
16.5.
Neurological Examination Form, Abridged Neuropsychological Assessment Test 
Battery, Substance use and Sleep, CDR..................................................................................... 118
17.
ATTACHMENTS..............................................................................................................................129
17.1.
Protocol Team Roster .................................................................................................................. 129
17.2.
Study Personnel Roles and Responsibilities ............................................................................... 134
17.3.
Collaborating Institutions and Investigators ................................................................................. 137
LIST OF TABLES
Table 1:
Study Design: Repeated-Dose GLP Toxicology Study in New Zealand White Rabbits........... 28
Table 2:
Design of a 4-cycle IM Repeated Dose Toxicity Study With Prime-Boost 
Combinations of Ad26.Mos.HIV, MVA-Mosaic, and Clade gp140 DP ..................................... 29
Table 3:
Characteristics of 150 Subjects Enrolled in the RV254/SEARCH010 Acute Infection 
Cohort ....................................................................................................................................... 35
Table 4:
Schedule of Follow-Up and Testing From Screening to After all Vaccinations and 
Prior to ARV ATI (Stage 1) ....................................................................................................... 55
Table 5:
Schedule of Follow-Up and Testing From ARV ATI to the end of the Study (Stage 2)............ 57
Table 6:
AE Notification and Safety Pause/AE Review Rules1...............................................................69


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LIST OF FIGURES
Figure 1:
After 6 Months of HAART, RV254/SEARCH010 Subjects Have Very low Levels of 
HIV DNA in PBMCs, Similar to Levels Seen in Elite Controllers (Ananworanich, 
2012)......................................................................................................................................... 36
Figure 2:
Study Design.............................................................................................................................39


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PROTOCOL AMENDMENTS
Protocol Version 
Issue Date
Original Protocol
7 Jan 2015
Amendment 1
12 Jun 2015
Amendment 2
1 Oct 2015
Amendment 3
This document
Amendment 3 (This document)
The overall reason for the amendment: Reduction in sample size.
Rationale: The sample size has been reduced from 36 to 27 subjects, with 18 subjects in the vaccine 
arm and 9 in the placebo arm instead of the original 24 and 12 subjects, respectively. The decision to 
stop study enrollment prior to the full anticipated recruitment is due to limitations of clinical supplies. 
This is in part due to unexpected delays in approvals as well as challenges in timely recruitment of 
subjects.
SYNOPSIS
3.1. Study Design
9.4. Sample Size Consideration
Rationale: Addition of information on the interim immunology analysis.
9.5.2. Analysis Time Points
Rationale: Clarification that volunteers for this study are participants in the study RV254 and will 
continue to participate in RV254 related activities during the course of the study. Updated wording on 
the inclusion of RV254 related activities as data points in this study.
3.1. Study Design
4.1.7. Management of Subjects After Reinitiating ART
4.1.8. Management of Subjects With Sustained Virologic Control at End of Study
4.1.9. Management of Subjects Who Become Pregnant
Rationale: Extending exceptions to ARV changes as justified by recent clinical practice.
4.3. Prior and Concomitant Treatment
Rationale: Addition of an alternative and more sensitive assay for detection of vector specific vaccine 
responses, reflecting experience from other studies.
3.2.5. Cohort Criteria for Proceeding to ARV ATI Phase of Study
9.5.2. Analysis Time Points
Rationale: The case definition for Acute Retroviral Syndrome has been updated, reflecting experience 
from other studies and consistency with other study protocols including ATI.
16.2. Case Definition for Acute Retroviral Syndrome (ARS)


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Rationale: In alignment with other RV254 protocols, the location for the performance of the optional 
procedures, specifically lymph node biopsy and the leukapheresis, has been updated to include the 
 
 in addition to the 
.
5.1.11. Optional Procedures
Rationale: From 1 June 2016 onwards, the name of Crucell Holland B.V. has been changed to Janssen 
Vaccines & Prevention B.V. Company logo has been updated accordingly.
Title page
SYNOPSIS
1.3. Description of Ad26.Mos.HIV
8.1. Protocol Safety Review Team
15. SIGNATURES
17.1. Protocol Team Roster
Rationale: It is clarified that neuropsychiatric evaluations and brain imaging, including MRI, MRS, and 
DTI are part of required continuous monitoring of the subjects from Study RV254.
5.1.10. Neuropsychiatric Examination
5.1.11. Optional Procedures
16.5. Neurological Examination Form, Abridged Neuropsychological Assessment Test Battery, 
Substance use and Sleep, CDR
Rationale: Changes are made in the fever criteria classification from 37.7ºC to 38.0ºC to align with the 
Division of AIDS (DAIDS) toxicity table.
5.1.2. Randomization and Vaccination 1/Day 1/Week 0 (Stage 1)
5.1.3. Vaccinations 2 – 4 / Weeks 12, 24, 48 (Stage 1)
6.3.1.1. Post-Vaccination Reactions Occurring Immediately After Each Vaccination
6.3.1.3. Systemic Reactions Occurring Within Seven Days Post-Vaccination
Rationale: Minor clarifications.
3.2.5. Cohort Criteria for 
Proceeding to ARV ATI 
Phase of Study and 3.2.6. 
Individual Subject 
Criteria for ARV ATI at 
Week 60
5.1. General Aspects
5.1. General Aspects
5.1. General Aspects and 
5.1.1. Screening
5.1. General Aspects and
5.1.12. Visit Windows
Minor clarification of individual subject criteria for ARV ATI.
Reference to Section 5.1.8 added to footnote 4.
Updated references for total blood volume drawn.
Minor clarification regarding testing for syphilis and hepatitis B.
Visit days and visit windows added.
Rationale: Minor changes.
PPD
PPD


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3.2.6.1. Inclusion Criteria 
for ARV ATI and 3.2.6.2. 
Exclusion Criteria for 
ARV ATI
3.4. Method of Blinding 
and Unblinding
SYNOPSIS, 4.1.1. 
Ad26.Mos.HIV Vaccine, 
5.1.10. Neuropsychiatric 
Examination and 9.3.1.2. 
Cellular Immunogenicity
4.1.9. Management of 
Subjects Who Become 
Pregnant
6.3.2 Serious Adverse 
Events; 5.3.2.1 Study 
Pausing Rules
13.6. Case Report Form 
Completion
16.4 Research Specimen 
Laboratory Testing,
17.1. Protocol Team 
Roster
17.2 Study Personnel Roles 
and Responsibilities
17.3. Collaborating 
Institutions and 
Investigators
Numbering of inclusion/exclusion criteria for ARV ATI.
Specified that blind should not be broken before eDC (electronic data 
capture) database is closed.
Typographical error.
Administrative correction on forms to be used to report cases of pregnancy. 
Administrative change.
Removed statement that all data should be recorded in CRF.
Janssen laboratory added to list.
Updated names and affiliations.
Administrative update.
Updated list of collaborating institutions and investigator.
Amendment 2 (1 Oct 2015)
The overall reason for the amendment: It was requested by the local Institutional Review Board (IRB) 
 and the Ethical Committee (EC) of the Ministry of 
Public Health (MoPH) of Thailand to clarify the language explaining:
that all subjects will return to their usual antiretroviral therapy (ART) after Week 96 (post Stage 2)
that a placebo group is required
that the study consists of 2 separate stages (phases)
the rationale for blood volume requirements
the rationale for ATI and risks related to ATI
the procedures for the Test of Understanding (TOU)
the compensation for the subjects
Rationale: It is clarified that after week 96 (post Stage 2) all study participants will resume ART.
4.1.8 Management of Subjects With Sustained Virologic Control at End of Study
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Rationale: It is clarified that the study will occur in 2 stages (Stage 1/Phase 1: vaccination period; 
Stage 2/Phase 2a: ART interruption [ATI]). It is further clarified that placebo only refers to the vaccine
component of the trial (Stage 1). Subjects in both arms (vaccine, placebo) will receive standard ART:
For HIV treatment during the first 60 weeks of the trial prior to ATI (Stage 1)
If they meet any of the criteria to restart ART listed in Section 4.1.6 (Stage 2)
At the end of the trial at Week 96 (post Stage 2)
SYNOPSIS
2.1 Hypothesis
3.1 Study Design
3.2.5 Cohort Criteria for Proceeding to ARV ATI Phase of Study
4.1.4 Vaccine Administration Schedule
4.1.5 Monitoring During ARV ATI
Table 4
Table 5
5.1.2 Randomization and Vaccination 1/Day 1/Week 0 (Stage 1)
5.1.3 Vaccinations 2 – 4 / Weeks 12, 24, 48 (Stage 1)
5.1.4 Post-Vaccination Follow-Up Visits / After Study Visits Week 0, 12, 24, 48) (Stage 1)
5.1.5 Safety and Immunogenicity Visits / Weeks 4, 16, 26, 30, 36, 50 (Stage 1)
5.1.6 Preparation for ARV ATI / Week 58 (Stage 1)
5.1.7 ARV ATI / Week 60 (Stage 1)
5.1.8 Monitoring After ARV ATI / Week 61-96 (Stage 2)
5.2 Vaccination Discontinuation
8.2 Data and Safety Monitoring Committee
9.1.3 Efficacy Population
Rationale: It is clarified by an additional table (Appendix 16.4) that several types of biological samples 
will (optionally) be collected to address study objectives. Additionally, blood volumes are reduced 
during Stage 2 of the study (see Table 5). Minor textual changes are made conform MoPH guidelines for 
blood handling.
Table 5
5.4 Laboratory Evaluations
11.7 Future use and Storage of Blood Samples
16.4 Research Specimen Laboratory Testing
Rationale: Rationale for ATI and clinical studies (new safety results) are updated. The risk of ATI is 
added in a separate Section 5.5.4.
1.2 Rationale for ARV ATI
1.6 Clinical Studies
5.1 General Aspects
5.5.4 Risk Related to ATI
5.5.5 Unknown Risks
14 References
Rationale: The TOU is explained in more detail (as part of the consent procedure).
11.4 Subject Information and Informed Consent


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Rationale: The compensation for the subjects is explained in more detail.
11.8 Compensation
Rationale: To align language among HIV vaccine protocols, in accordance with FDA, Study Holding 
Rules are adapted to avoid unnecessary pausing of vaccination for DSMC review of certain 
reactogenicity events, common to most vaccines.
5.3.2.1 Study Pausing Rules
Rationale: To align language among HIV vaccine protocols, wording is added to exclusion criterion 22 
to exclude employees of the sponsor or its partners from enrollment.
3.2.2 Exclusion Criteria
Rationale: To allow more flexible scheduling of the visits to administer last dose of study vaccine, 
without expected impact on immunogenicity, the visit window of the last vaccination is extended from 
±1 week to ±2 weeks.
5.1.3 Vaccinations 2 – 4 / Weeks 12, 24, 48 (Stage 1)
Rationale: Minor changes.
List of Abbreviations
1.6 Clinical Studies
3.1 Study Design
3.2.1 Inclusion Criteria
4.1.1 Ad26.Mos.HIV Vaccine
4.1.6 Criteria to Reinitiate ART During ARV ATI
4.2.1 Packaging and Labeling
Table 4
Table 5
5.1.1 Screening
5.1.11 Optional Procedures
5.2 Vaccination Discontinuation
5.5.3 Risks Related to Blood Draws
6.1.2 Rebound in Viral Replication, Study Endpoint not Considered as AE/ADR
9.3.1.1 Humoral Immunogenicity
9.3.1.2 Cellular Immunogenicity
10 Study-Specific Materials
11.2 Institutional Review Board/Ethics Committee
17.1 Protocol Team Roster
17.2 Study Personnel Roles and Responsibilities
Amendment 1 (12 Jun 2015)
This amendment is considered nonsubstantial in that it does not significantly impact the safety or 
physical/mental integrity of subjects, nor the scientific value of the study. 


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The overall reason for the amendment: It was requested by the local Institutional Review Board (IRB) 
 to clarify the language explaining how subjects will be followed 
after the study end and how they may return to their usual antiretroviral therapy (ART), and to align the 
storage time of blood samples in the protocol with the information in the Informed Consent Form (ICF). 
The protocol was updated accordingly. In addition, the Division of AIDS Table for Grading the Severity 
of Adult and Pediatric Adverse Events (DAIDS AE Grading Table) has recently been updated. 
Therefore, in the protocol, the reference to this table was updated.
Rationale: In response to a question from the local IRB, the language explaining how subjects will be 
followed after the study end and how they may return to their usual ART was clarified. It was specified 
that after study completion (96 weeks), subjects will be offered enrollment in study RV412 for 
continued observation, followed by a return to the RV254 parent protocol afterwards. In these follow-up 
studies, all subjects will have access to ART, and those subjects who are not on ART can decide to 
resume their treatment at any time.Thus, subjects enrolled in the RV405 protocol will have access to 
long-term treatment and monitoring.
4.1.7 Management of Subjects After Reinitiating ART
4.1.8 Management of Subjects With Sustained Virologic Control at End of Study
Rationale: In response to a question from the local IRB, the language explaining the storage time of 
blood samples in the protocol was clarified and aligned with the ICF. It was specified that samples can 
be stored for up to 10 years for future testing, unless there would be scientific merit in additional testing 
or storage beyond 10 years. Then, permission for continued storage should be requested from both the 
relevant IRBs and the subjects (if possible). 
11.7 Future use and Storage of Blood Samples
Rationale: The reference to the DAIDS AE Grading Table Version 1.0 dated December 2004 
(Clarification, August 2009) was updated to Version 2.0 dated November 2014. 
6.3.5 Severity of Adverse Events
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LIST OF ABBREVIATIONS
Ad26
adenovirus serotype 26
ADCC
antibody-dependent cell-mediated cytotoxicity
AE
adverse event
AIDS
acquired immunodeficiency syndrome
ALT
alanine aminotranferase
ALVAC-HIV
canary pox Avipox vectored HIV vaccine candidate 
ART
antiretroviral therapy
ARV
Antiretroviral
AST
aspartate aminotransferase
ATI
analytical treatment interruption
BUN
blood urea nitrogen
CBC
complete blood count
CDC
Centers for Disease Control and Prevention
CD4+
a functional subclass of helper T lymphocytes that are necessary for augmentation and 
coordination of innate and adaptive effector responses, humoral and cellular
CD8+
cytotoxic T-Cells that destroy host cells, which have become infected by viruses or 
other intracellular pathogens
CFR
Code of Federal Regulations
CMDR
Chiang Mai Double Recombinant
CSF
cerebrospinal fluid
CTL
cytotoxic T lymphocyte
DAIDS
Division of AIDS
DNA
deoxyribonucleic acid
DoD
Department of Defense (US)
DP
drug product
DSMC
Data and Safety Monitoring Committee
EC
ethical committee
ECG
electrocardiogram 
(e)CRF
(electronic) case report form
eDC
electronic data capture
ELISA
enzyme-linked immunosorbent assay
ELISPOT
enzyme-linked immunospot
Env
Envelope
FDA
US Food and Drug Administration
FIH
first-in-human
GCP
Good Clinical Practice
GGT
gamma-glutamyl transpeptidase
GLP
Good Laboratory Practice
HAART
highly active antiretroviral therapy
HIV and HIV-1
human immunodeficiency virus, type 1
HLA
human leukocyte antigen
HVTN
HIV Vaccine Trials Network
IB 
investigator’s brochure
IBC
Institutional Biosafety Committee
ICF
Informed Consent Form
ICH
International Conference on Harmonization
ICS
intracellular cytokine staining
ID
intradermal 
IFN
Interferon
IM
intramuscular 
IP
investigational product


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IPCAVD
Integrated Preclinical/Clinical AIDS Vaccine Development
IQR
inter-quartile range
IRB
Institutional Review Board
LTR
long terminal repeats
MHRP
(US) Military HIV Research Program
MRI
magnetic resonance imaging
MRS
magnetic resonance spectroscopy
MoPH
Ministry of Public Health
MVA
Modified Vaccinia Ankara
NIH
National Institutes of Health
NNRTI
non-nucleoside reverse transcriptase inhibitor
ORP
Office of Research Protection
PBMC
peripheral blood mononuclear cell
pfu
plaque forming units (equivalent to International Units)
PI
principal investigator
PIR
post-injection reactogenicity
PQC
product quality complaint
PSRT
Protocol Safety Review Team
QVOA
quantitative viral outgrowth assay
RAC
Recombinant DNA Advisory Committee
RBC
red blood cell 
rMVA
recombinant MVA
RNA
ribonucleic acid
SAE
serious adverse event
SIV
simian immunodeficiency virus
SMC
Safety Monitoring Committee
SOE
schedule of evaluations
SUSAR
suspected unexpected serious adverse reaction
TILDA
Tat/Rev Induced Limiting Dilution Assay 
TOU
Test of Understanding
TULDA
Tat/Rev Uninduced Limiting Dilution Assay
UCSF
University of California San Francisco
ULN
upper limits of normal
US
United States
USP
US Pharmacopeia
USAMRMC
US Army Medical Research and Materiel Command
vp
viral particles
WBC
white blood cell 
WHO
World Health Organization
WRAIR
Walter Reed Army Institute of Research


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SYNOPSIS
Protocol Number
Janssen Vaccines & Prevention B.V. protocol VAC89220HTX1001 Amendment 3
Protocol Numbers of Collaborators:
MHRP RV 405
SEARCH 023
IPCAVD 007
A Combined Phase 1/2a, Exploratory Study of a Therapeutic Vaccine Using an Adenovirus 
Type 26 Vector Prime and Modified Vaccinia Ankara Boost Combination With Mosaic Insert s in 
HIV-1 Infected Adults who Initiated Antiretroviral Treatment During Acute HIV Infection
Clinical Phase
Phase 1/2a
Study Vaccine
Vaccines used in this study are adenovirus serotype 26 mosaic human immunodeficiency virus 
(Ad26.Mos.HIV) and Modified Vaccinia Ankara-Mosaic (MVA-Mosaic):
Ad26.Mos.HIV vaccine contains the following drug substances in a 2:1:1 ratio:

Ad26.Mos1.Env: recombinant, replication-incompetent adenovirus serotype 26 expressing a 
mosaic 1 Human Immunodeficiency Virus Type 1 (HIV-1) Envelope (Env) protein

Ad26.Mos1.Gag-Pol: recombinant, replication-incompetent adenovirus serotype 26 expressing 
mosaic 1 HIV-1 Gag and Pol proteins,

Ad26.Mos2.Gag-Pol: recombinant, replication-incompetent adenovirus serotype 26 expressing 
mosaic 2 HIV-1 Gag and Pol proteins.
MVA-Mosaic contains the following vaccine products, supplied in separate vials and administered in a 
1:1 ratio:

MVA-Mosaic1: Modified Vaccinia Ankara virus expressing Mosaic 1 HIV-1 Gag, Pol and Env 
proteins

MVA-Mosaic2: Modified Vaccinia Ankara virus expressing Mosaic 2 HIV-1 Gag, Pol and Env 
proteins
Study Design
This is a combined Phase 1/2a randomized, double-blind, placebo-controlled study to investigate the 
safety, immunogenicity and effect on viremic control after antiretroviral (ARV) analytical treatment 
interruption (ATI) of a vaccine regimen consisting of an Ad26.Mos.HIV prime and an MVA-Mosaic
boost. The study will include subjects who started on antiretroviral therapy (ART) during acute HIV 
infection, who are on a current stable ART and who have achieved absence of viremia (HIV ribonucleic 
acid [RNA] <50 copies/ml) for ≥48 weeks prior to initiation of vaccine/placebo. The study will be 
carried out in two stages (Phases). During the first stage (Phase 1), each subject will receive a total of 


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four vaccinations or four placebo injections. During the second stage of the study (Phase 2a), ATI will 
be started and all ARV drugs will be discontinued in all subjects.
Treatments
Subjects in the vaccine arm will receive Ad26.Mos.HIV at Weeks 0 and 12, and MVA-Mosaic at 
Weeks 24 and 48 (Stage 1). Subjects in the placebo arm will receive placebo at Weeks 0, 12, 24 and 48
(Stage 1). At Week 60, ATI will be started and all ARV drugs will be discontinued at that time in both 
arms (Stage 2). 
Objectives and Hypothesis
Primary Objectives
1.
Determine the safety and tolerability of Ad26 prime/MVA boost versus placebo in subjects on 
suppressive ART that was initiated during acute HIV infection
2.
Measure the frequency and duration of sustained viremic control after receiving Ad26 
prime/MVA boost or placebo, defined as greater than 24 weeks with plasma HIV RNA 
<50 copies/ml after ARV ATI
Secondary Objectives
1.
Determine the immunogenicity of Ad26 prime/MVA boost in subjects on suppressive ART that 
was initiated during acute HIV infection
2.
Characterize biomarkers of HIV reservoir at baseline, after vaccine therapy prior to ARV ATI
(Week 48-60) and after ARV ATI (Week 60-96)
3.
Compare the duration of viremic control (HIV RNA <50 copies/ml) between vaccine and 
placebo recipients who failed to achieve sustained viremic control (undetectable plasma HIV 
RNA [<50 copies/ml] at Week 24 after ARV ATI)
4.
Describe the frequency, magnitude, specificity and functional capacity of humoral and cellular 
immune responses to vaccine and other immunogens
5.
Describe the molecular sequence sieve effects of vaccine therapy on breakthrough rebound 
viremia before and after cessation of therapy 
6.
Describe the peripheral blood mononuclear cells (PBMC) phenotype, pattern of soluble factors 
and immune functional responses before and after ARV ATI in both the vaccine and placebo 
arms and compared to historical untreated acute infection cases in RV217
7.
Describe the clinical outcomes in terms of the frequency, severity, duration and treatment for 
acute retroviral syndrome post ARV ATI.
8.
Evaluate and characterize HIV resistance to ARV drugs in subjects who experience rebound 
viremia after ARV ATI. 
Exploratory Objectives
1.
Analyze the markers of HIV reservoir as predictors of sustained viral suppression
2.
Analyze the markers of HIV specific immune responses as predictors of sustained viral 
suppression.
3.
Analyze the expression profiles and transcriptome analysis of
sorted and/or unsorted
lymphocytes and monocyte/macrophages in the different arms of the study
4.
Compare the mucosal immunity (vaginal and rectal for humoral and gut mucosa for cellular 
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5.
Describe the markers of immune activation in the peripheral blood and various reservoirs
6.
Describe the viral load response to resumption of ART and impact on markers of HIV reservoir 
among those who fail to suppress HIV RNA off therapy.
Hypothesis
This is an exploratory study to collect preliminary data and establish sufficient experience to inform and 
develop subsequent studies to address the following hypothesis: Vaccine therapy administered in the 
form of Ad26 vector with mosaic inserts for gag/pol and env genes of HIV-1 at 0 and 12 weeks
(Stage 1), boosted with the MVA vector with homologous mosaic inserts for gag/pol and env genes at 
24 and 48 weeks (Stage 1), among individuals with fully suppressed HIV will be safe and well-tolerated, 
will result in a measurable immune response, and will result in over 50% of vaccine recipients achieving 
sustained viremic control after ATI (Stage 2).
Study Center
 Thailand
Study duration Planned
The total duration for each subject will be 96 weeks, including 48 weeks for the vaccination period
(Stage 1), 12 weeks between the final vaccination and ARV ATI (Stage 1), and 36 weeks of follow-up
(Stage 2).
Number of Subjects
Up to 27 subjects are planned to be enrolled. Subjects will be randomly assigned in a 2:1 schedule to 
either receive Ad26.Mos.HIV and MVA-Mosaic (total of 18 subjects) or placebo (total of 9 subjects).
Population
HIV-infected adults who started ART during acute infection, who are on a current stable ART for at 
least 4 weeks prior to screening, who are clinically stable while on treatment and who have undetectable 
plasma HIV RNA (<50 copies/ml) for at least 48 weeks. 
Safety Monitoring 
A Protocol Safety Review Team (PSRT) and an independent Data and Safety Monitoring Committee 
(DSMC) will oversee the conduct and safety monitoring of the study. 
Endpoints/Criteria for Evaluation of Safety, Immunogenicity, and Efficacy 
Safety and Tolerability:

For safety and tolerability: adverse events (AEs) during the course of the study and 
reactogenicity for 1 week after each study vaccination.
PPD


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Immunogenicity:

This is an exploratory study; the main criteria for evaluation of immunogenicity will include but
are not limited to the frequency, magnitude, and breadth of epitope recognition by 
enzyme-linked immunospot (ELISPOT), polyfunctinality of T-cell responses, binding antibody 
to various HIV-1 Env regions and neutralization of a variety of HIV strains (tier 1, 2).
Efficacy:

The main endpoint for efficacy is the proportion of subjects with undetectable plasma HIV RNA 
(<50 copies/ml) at 24 weeks after ARV ATI. 
Statistical Methods
The primary outcomes of the study are safety and efficacy of the vaccine. The sample size is within the 
range of subjects recommended in the Code of Federal Regulations (CFR 312.21) for each of the 
products in this investigation and will allow evaluation of epitope enumeration between arms. Placebo 
recipients are included for blinding purposes, safety and efficacy analyses, and will provide additional 
control specimens for immunogenicity assays.
While mild to moderate vaccine reactions (local site and systemic responses) are expected, AEs that 
preclude further dose administration or more serious ones that would limit product development are not 
anticipated. With 18 subjects in the vaccine arm, the observation of 0 such reactions would result in a 
two-sided exact 95% confidence interval with an upper limit of 18. 50%.
Immunogenicity
Blood and genital secretions for the determination of systemic and local cellular and humoral immune 
responses will be collected at the time points as described in the schedule of evaluations. In a subset of 
consenting subjects, sigmoid biopsy, lymph node biopsy and cerebrospinal fluid (CSF) samples will also 
be collected to evaluate immune responses in gastro-intestinal and other reservoirs. Leukapheresis will 
be an optional procedure to collect a larger number of cells for analysis. Brain magnetic resonance 
imaging (MRI), magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI) will also 
be performed as optional procedures.
Reviewing Institutional Review Board (IRB): 
PPD


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1.
INTRODUCTION
HIV Epidemiology
Globally, an estimated 35.3 million people were living with HIV in 2012, an increase from 
previous years as a result of the wider availability of life-saving ART. There were 2.3 million 
new HIV infections globally, showing a 33% decline in the number of new infections from 
3.4 million in 2001. At the same time the number of acquired immunodeficiency syndrome 
(AIDS) deaths is also declining with 1.6 million AIDS deaths in 2012, down from 2.3 million in 
2005. From 1996 to 2012, ART averted 6.6 million AIDS-related deaths worldwide, including 
5.5 million deaths in low- and middle-income countries (UNAIDS Report on the Global AIDS 
Epidemic 2013). The world is within reach of providing ART to 15 million people by 2015. In 
2012, 9.7 million people in low- and middle-income countries received ART, representing 61% 
of all who were eligible under the 2010 World Health Organization (WHO) HIV treatment 
guidelines. However, under the 2013 WHO guidelines, the HIV treatment coverage in low - and 
middle-income countries represented only 34% of the 28.3 million people eligible in 2013. 
Despite its proven success at saving lives, there are significant challenges to initiating and 
maintaining ART for all of those that need it in the world. ART must be taken life-long with 
near perfect adherence in order to be effective. This places extreme pressure and costs on 
international donors and over-taxed health systems in developing countries where HIV 
prevalence rates are highest. ART has both short-term and long-term side effects for users, and 
drug resistance rates rise as more people are on treatment for longer periods of time. Alternative 
or complementary treatments, including a therapeutic vaccine, which could induce a true or 
“functional” cure of HIV infection and lessen or eliminate the need for lifelong ART for HIV 
infected individuals, would therefore be of great benefit. 
HIV Vaccines
A Phase 3 community-based study conducted in Thailand (RV144) provided the first evidence 
that an HIV-1 vaccine could provide protective efficacy against HIV-1 acquisition. The 
prime-boost vaccine regimen consisted of a recombinant canarypox vector, ALVAC-HIV prime 
(vCP1521, expressing gag, protease subtype B (LAI) and env gp120 CRF01_AE with a gp41 
subtype B (LAI) transmembrane anchor) and a bivalent AIDSVAXgp120 B/E MN and 
CRF01_AE (A244) boost. The vaccine regimen was safe and well tolerated (Pitisuttithum 
2011). A modified intent-to-treat analysis showed an estimated 31.2% efficacy after 42 months 
of follow-up (Rerks-Ngarm 2009; Gilbert 2011). Post hoc analysis of the interaction of risk 
status and acquisition efficacy was significant with greater benefit in low-risk individuals (Robb 
2012). Vaccine efficacy appeared to be higher (60%) at 12 months post vaccination than at later 
timepoints, suggesting an early, but nondurable, vaccine effect. There was no effect on early 
post-infection HIV-1 RNA viral load or CD4+ T-cell count. Vaccination did not affect the 


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clinical course of HIV-1 disease after infection, though there was evidence of reduction in 
seminal fluid viral load (Rerks-Ngarm 2013). 
The RV144 study provided a unique opportunity to perform a case control study of correlates of 
risk. Plasma IgG antibody binding to scaffolded gp70 V1V2 Env proteins correlated inversely 
with risk, while Env plasma IgA correlated directly with risk, raising the hypotheses that IgA 
responses against Env and IgG responses directed against V1V2 may be mechanistically 
associated with protection. Neither low levels of V1V2 antibodies nor high levels of 
Env-specific IgA antibodies were associated with higher rates of infection than in the placebo 
group. In vaccinees, low levels of Env-specific IgA antibodies, IgG avidity, antibody-dependent 
cell-mediated cytotoxicity (ADCC), neutralizing antibodies, and Env-specific CD4+ T cells, 
were inversely correlated with risk of infection (Haynes 2012; Karasavvas 2012; Zolla-Pazner 
2013). Two weeks post last vaccination, 97% of RV144 studied plasma samples from vaccine 
recipients contained antibodies to V2 region synthetic peptides, falling to 19% at 48 weeks, 
suggesting that waning vaccine efficacy may be correlated to waning V2 antibody response
(serum IgG). Interestingly, gp70 V1V2 antibodies were lower in HVTN 505 compared to 
RV144. The response to V3 CRF01_AE also inversely correlated with the risk of HIV infection 
in vaccine recipients with lower levels of Env-specific plasma IgA and neutralizing antibodies. 
In Vax003 and Vax004 (no protection), serum IgG responses targeted the same epitopes as in 
RV144 with the exception of an additional C1 reactivity in Vax003 and infrequent V2 reactivity 
in Vax004. These results along with a recent sieve analysis (Rolland 2013) generate the 
hypothesis that IgG to linear epitopes in the V2 and V3 regions of gp120 are part of a complex 
interplay of immune responses that contributed to protection in RV144 (Gottardo 2013). 
1.1.
Overall Rationale of Study Design
Studies of HIV vaccine in HIV-uninfected and infected subjects suggest that a successful HIV 
vaccine program will need to induce immunity against the diverse strains and subtypes 
predominating in the target populations. Improving magnitude, breadth and depth of epitope 
coverage is thought to be a key to development of a successful T-cell based preventive HIV 
vaccine. Published primate data indicate that the number of epitope specific responses induced 
by a vaccine may be an important immune correlate of viral load control in the simian 
immunodeficiency virus (SIV) challenge system (Chen 2001). Strategies to accomplish this 
include using multivalent vaccines containing immunogens from a number of prevalent 
subtypes or using mosaic sequences, proteins assembled from natural sequences by in silico 
recombination, which are optimized for potential T-cell epitopes.
The enhancement of host-mediated clearance of residual virus represents a new additional 
approach to HIV functional cure (Carcelain 2013). Findings of several studies have shown the 
importance of cellular immunity in the control of HIV reservoir size. HIV-1 Gag-specific CD8+ 
T cells isolated from elite controllers, but not from patients given ART, were shown to kill 


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autologous resting CD4+ T cells in which the virus was reactivated with vorinostat. Moreover, 
functional anti-viral CD8 T cells are associated with reduced size of the central memory CD4 T 
cell reservoir in patients controlling their virus without ART. High-avidity multifunctional CD8 
cytotoxic T lymphocytes (CTL) that target vulnerable regions in Gag are especially important in 
limiting virus diversity and reservoirs in individuals infected with HIV who have protective 
human leukocyte antigen (HLA) class I alleles. Therapeutic vaccines could re-stimulate CD8+ 
CTL to prevent or control virus relapses and re-establish latent infection in CD4+ T cells after 
treatment interruptions. A few therapeutic vaccine studies such as the Ad5 HIV-1 gag vaccine 
(ACTG A5197 NCT00080106), and infusions of dendritic cells pulsed with inactivated HIV 
particles have shown transient viral suppression after treatment interruption. Eramune-02 is 
testing whether a deoxyribonucleic acid (DNA) prime, replication defective, recombinant 
adenovirus serotype-5 boost strategy, with the Vaccine Research Center’s polyvalent HIV-Gag, 
Pol, Nef, and Env vaccine can reduce the viral reservoir in patients undergoing an 
ARV-intensification regimen (Katlama 2013). 
Liu et al. (Liu 2009) showed that a heterologous rAd26 prime/rAd5 boost vaccine regimen 
expressing SIV Gag elicited cellular immune responses with augmented magnitude, breadth and 
polyfunctionality as compared with the homologous rAd5 regimen. After SIVmac251 
challenge, monkeys vaccinated with the rAd26/rAd5 regimen showed a 1.4 log reduction of 
peak and a 2.4 log reduction of setpoint viral loads as well as decreased AIDS-related mortality 
as compared with control animals. These data demonstrate that durable partial immune control 
of a pathogenic SIV challenge for more than 500 days can be achieved by a T-cell-based 
vaccine in Mamu-A*01-negative rhesus monkeys in the absence of a homologous Env antigen. 
Recently Barouch et al. (Barouch 2012) have shown that heterologous Ad26/MVA vaccine 
regimens containing Gag, Pol, and Env antigens, can protect against acquisition of infection 
following repetitive, heterologous, intrarectal challenges with a neutralization-resistant virus 
(SIV MAC251) in rhesus monkeys. Moreover, immunological correlates analyses suggested that 
multiple cellular and humoral immune responses correlate with virological control, although the 
actual mechanisms of protection remain to be determined. 
This study will explore a combination prime-boost vaccine regimen. Here we propose to 
evaluate the regimen in HIV-infected individuals who began suppressive ART during very 
early acute HIV infection and therefore have minimal viral reservoir and more intact immune 
systems. While previous vaccine-based immunotherapy strategies failed to control viral load in 
chronically HIV-infected subjects, we hypothesize that vaccine-induced immune responses may 
continue to control viral replication after ART stop in subjects treated very early during acute 
infection. This setting provides an optimal opportunity to demonstrate the concept of vaccine 
therapy to achieve sustained control of HIV without ART (functional cure). 


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1.1.1.
Rationale for Modified Vaccinia Ankara (MVA) 
The use of live attenuated MVA virus recombinants as expression vectors for HIV genes either 
alone or in various prime-boost combinations has been shown to be safe and immunogenic in 
humans (Cebere 2006; Currier 2010; Excler 2013; Jaoko 2008; Peters 2007; Ramanathan 2009; 
Vasan 2010). In particular, MVA-based recombinants have been shown to induce antibodies 
and specific cellular immune responses mediated by CTLs. The Walter Reed Army Institute of 
Research (WRAIR) and National Institutes of Health (NIH) have produced live recombinant
poxvirus vectors, MVA-A/C/E, that are genetically engineered to express Gag, Pol, and Env 
from HIV-1 subtypes A, C, and E. 
1.1.2.
Rationale for Adenovirus Type 26 Vector 
All three of the HIV-1 vaccine efficacy studies utilizing Ad5 and DNA/Ad5 vaccines (HVTN 
502, HVTN 503, HVTN 505) showed no efficacy against HIV-1 infection. In the first two 
studies increased HIV-1 infection was observed in vaccinees as compared with placebo 
subjects. The mechanism for this possible increase in HIV-1 acquisition risk remains unclear, 
but a leading hypothesis involves activation of vector-specific CD4+ T lymphocytes at mucosal 
surfaces following Ad5 vaccination, potentially resulting in increased targets for HIV-1 
infection.
The rationale to continue clinical development of Ad26 vector-based vaccines for HIV-1 is 
based on data showing that: (1) Ad26 is biologically substantially different than Ad5; (2) Ad26-
based vaccines afford superior protective efficacy compared with Ad5-based vaccines against 
stringent SIVmac251 challenges in rhesus monkeys; and (3) Ad26 did not increase the number 
or activation status of total or vector-specific CD4+ T lymphocytes at mucosal surfaces in 
healthy HIV-uninfected humans following vaccination in a randomized, double-blinded, 
placebo-controlled clinical study (IPCAVD003).
Biological Differences Between Ad5 and Ad26
Ad5 seroprevalence is nearly universal with high titers in the developing world, whereas Ad26 
seroprevalence is lower with substantially lower titers in the developing world (Barouch 2011, 
Mast 2010). Baseline Ad5 neutralizing antibodies have been shown to suppress the 
immunogenicity of Ad5-based vaccines in both pre-clinical and clinical studies. International 
seroepidemiology studies have demonstrated a much lower prevalence of antibodies to Ad26, 
Ad35, and Ad48 compared to Ad5 (Barouch 2011, Mast 2010). For example, Ad26 
seroprevalence in adults in South Africa, Kenya, Uganda, and Thailand were 43%-53%, 66%, 
68%, and 55%, respectively, compared to Ad5 seroprevalence of 88%-90%, 91%, 86%, and 
82% in the same cohorts, respectively. The majority (61%-79%) of adults in these regions also 
demonstrated high Ad5 neutralizing antibody titers, whereas only 5%-18% of adults had high 
Ad26 neutralizing antibody titers. 


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Ad5 utilizes the coxsackievirus and adenovirus receptor as its primary cellular receptor, 
whereas Ad26 utilizes CD46 (Li 2012). Ad5 exhibits primarily liver tropism in vivo, whereas 
Ad26 exhibits no liver tropism (Waddington 2008). Moreover, Ad5 and Ad5 immune 
complexes are more stimulatory to human dendritic cells than are Ad26 and Ad26 immune 
complexes (Perreau 2012).
Ad5 and Ad26 also exhibit profoundly different innate immune profiles following vaccination 
of human subjects, as demonstrated by differential gene expression profiles on Days 1, 3, and 7 
after immunization. Certain genes are up- or down-regulated by Ad5 but not Ad26, whereas 
others are up- or down-regulated by Ad26 but not Ad5. Genes associated with general 
inflammation and interferon (IFN) pathways were up-regulated by both Ad5 and Ad26, as 
expected, but this effect was substantially more robust and more durable with Ad5 as compared 
with Ad26. These data demonstrate that Ad5 and Ad26 trigger phenotypically very distinct 
innate inflammatory profiles, and that Ad5 stimulates more robust and more durable common 
inflammatory pathways than does Ad26.
Ad5 and Ad26 vectors also exhibit profoundly different adaptive immune phenotypes. In mice, 
Ad5 vectors elicited high magnitude but dysfunctional and exhausted T cells, characterized by 
low levels of CD127, CD62L, and IFN-gamma, and high levels of PD-1, whereas Ad26 vectors 
elicited polyfunctional T cells characterized by high levels of CD127, CD62L, and IFN-gamma, 
and low levels of PD-1 (Penaloza-MacMaster 2013, Tan, 2013). Moreover, Ad26-induced 
T cells expanded more robustly than did Ad5-induced T cells upon re-exposure to antigen and 
afforded superior protective efficacy in murine challenge models (Penaloza-MacMaster 2013, 
Tan, 2013).
Taken together, these data show that Ad5 and Ad26 are biologically substantially different in 
terms of key aspects of seroepidemiology, basic virology, innate immune profiles, and adaptive 
immune phenotypes.
Improved Protective Efficacy of Ad26 Compared With Ad5 in Rhesus Monkeys
Consistent with the results of the HVTN 505 clinical study, DNA/Ad5 vaccines afforded no 
protective 
efficacy 
against 
stringent, 
repetitive, 
intrarectal 
challenges 
with 
the 
neutralization-resistant virus SIVmac251 in rhesus monkeys (Letvin 2011). In contrast, 
DNA/Ad5 
vaccines 
afforded 
partial 
protective 
efficacy 
against
less 
stringent, 
neutralization-sensitive SIVsmE660 challenges, indicating that the low stringency SIVsmE660 
challenge model was not predictive of the clinical results observed in HVTN 505. In contrast, 
the lack of protection in the high stringency SIVmac251 challenge model was consistent with 
the lack of efficacy observed in HVTN 505. Ad5 only vaccines also did not afford protection 
against SIVmac239 (Casimiro 2005), consistent with the results of the Step study.


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In contrast, Ad26/MVA and Ad26/Ad35 vaccine regimens afforded substantial protective 
efficacy against repetitive, intrarectal challenges with the neutralization-resistant virus 
SIVmac251 in rhesus monkeys (Barouch 2012). Thus, Ad26/MVA vaccines afforded partial 
protection (76-83% reduction in the per-exposure risk of infection) in the stringent SIVmac251 
challenge model in which DNA/Ad5 failed.
No Activation of Mucosal Total or Vector-Specific CD4+ T Lymphocytes Following Ad26 
Vaccination in Humans
The mechanism for the possible increase in HIV-1 acquisition risk in individuals who received 
Ad5-based vaccines remains unclear, but a leading hypothesis involves the potential activation 
of total or vector-specific CD4+ T lymphocytes at mucosal surfaces following Ad5 vaccination, 
which theoretically could result in increased targets for HIV-1 infection. To assess the 
possibility that this effect may occur with Ad26 vectors, we performed a randomized, 
double-blinded, placebo-controlled clinical study (IPCAVD003) to determine whether 
vaccination of healthy human subjects with an Ad26 vector expressing HIV-1 Env would result 
in increased numbers or activation status of total or vector-specific CD4+ T lymphocytes in 
colorectal mucosa. As shown below, 24 subjects were enrolled in this study. Group 1 (N = 16) 
consisted of individuals who were baseline Ad26 seronegative. Group 2 (N = 8) consisted of 
individuals who were baseline Ad26 seropositive. Subjects were randomly allocated to receive 
vaccine or placebo in a 3:1 ratio. All subjects received a single intramuscular (IM) injection of 
vaccine (5x1010 viral particles [vp] Ad26.ENVA.01) or placebo on Day 0. Blood samples, 
colorectal biopsies, and colorectal weck-cel sponges were collected prior to immunization and 
at Weeks 2 and 24 following vaccination. Of 72 planned biopsies, 71 were performed.
HIV-1 Env-specific humoral and cellular immune responses were observed in both peripheral 
blood and colorectal mucosa following Ad26 vaccination (data not shown). Ad26 
vector-specific CD4+ T-lymphocyte responses were substantially lower in colorectal mucosa 
than in peripheral blood and did not appear to increase following vaccination. The degree of 
inflammation in colorectal mucosa was also assessed. Normal histology was observed in 71/71 
biopsies. No increases in CD4, CD8, CD3, or CD25 cells were observed in colorectal mucosa 
following Ad26 vaccination. Moreover, no increases in Ki67 or HLA-DR activation were 
observed in colorectal mucosa following Ad26 vaccination. The activation status of total and 
vector-specific CD4+ T lymphocytes in colorectal mucosa was assessed by flow cytometry. No 
increased Ki67 activation or CCR5 expression was observed on total or vector-specific CD4+ T 
lymphocytes in colorectal mucosa following Ad26 vaccination. These data demonstrate that 
Ad26 vaccination did not result in increased numbers or activation status of total or vector-
specific CD4+ T cells in colorectal mucosa in human subjects.


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1.1.3.
Rationale for Mosaic Sequence Inserts 
Multiple studies have supported the necessity of an HIV vaccine to elicit CD8+ T-cell 
responses in order to control or clear HIV infection (Bhattacharya 2007, Brumme 2008, Borrow 
1997, Betts 2006, Fellay 2007). A global HIV vaccine will have to elicit CD8+ T-cell responses 
to a diverse group of viruses. HLA class I-presented epitopes are usually 9-mers and each HIV 
genome contains thousands of potential T-cell epitopes. The ideal sequence to use in a global 
vaccine would be one that would match all 9-mers in the M group of HIV viruses; however, 
given the diversity of strains, the best natural strain only matches approximately one-quarter of 
the 9-mers in Env and one-half in Pol (Korber 2009). In order to increase the coverage of 
potential T-cell epitopes, mosaic sequences have been created. Mosaic sequences are in 
silico-derived recombinants optimized to match the most potential T-cell epitopes of M-group 
virus. Studies in rhesus monkeys have shown that mosaic sequences increase the breadth and 
depth of immune response compared with consensus or natural sequences (Barouch 2010, 
Santra 2010). 
1.1.4.
Rationale for Heterologous Vectors 
Heterologous, as opposed to homologous viral vectors in a prime boost immunization strategy 
have been shown to increase magnitude, breadth and polyfunctionality of cellular immune 
responses to core protein epitopes. Ratto-Kim et al. characterized prime-boost vaccine regimens 
using heterologous and homologous vector and gene inserts. Heterologous regimens o ffer a 
promising approach that focuses the cell-mediated immune response on the insert and away 
from vector-dominated responses (Ratto-Kim 2012). Ad35-GRIN/ENV (Ad35-GE) vaccine is 
comprised of two vectors containing gene sequences from HIV-1 subtype A gag, rt, int, nef
(Ad35-GRIN) and env (Ad35-ENV). MVA-Chiang Mai Double Recombinant (CMDR), 
MVA-KEA (MVA-K) and MVA-TZC (MVA-T) vaccines contain gag, env and pol genes from 
HIV-1 subtypes CRF01_AE, A and C, respectively. Balb/c mice were immunized with different 
heterologous and homologous vector and insert prime-boost combinations. HIV and vector-
specific immune responses were quantified post-boost vaccination. Gag-specific IFN-gamma 
ELISPOT, intracellular cytokine staining (ICS) (CD107a, IFN-gamma, TNF-a, and IL-2), 
pentamer staining and T-cell phenotyping were used to differentiate responses to inserts and 
vectors. Ad35-GE prime followed by boost with any of the recombinant MVA constructs 
(rMVA) induced CD8+ Gag-specific responses superior to Ad35-GE-Ad35-GE or 
rMVA-rMVA prime-boost combinations. Notably, there was a shift toward insert-focus 
responses using heterologous vector prime-boost regimens. Gag-specific central and effector 
memory T cells were generated more rapidly and in greater numbers in the heterologous 
compared to the homologous prime-boost regimens. These results suggest that heterologous 
prime-boost vaccination regimens enhance immunity by increasing the magnitude, onset and 
multifunctionality of the insert-specific cell-mediated immune response compared to 
homologous vaccination regimens. This approach has also resulted in reduced viral load and 
improved survival in subsequently infected non-human primates. Furthermore, the magnitude 


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of these effects varies by vector. Specifically, the combination of Ad26/Ad5 has been shown to 
be superior to Ad35/Ad5, which is superior to the homologous Ad5/Ad5 (O’Brien 2009). Early 
phase human studies with heterologous vectors are currently in progress (VRC 012, HVTN 
083).
1.2.
Rationale for ARV ATI
ATI is not the standard of care for HIV infection. The Thai National HIV Treatment Guidelines, 
recently revised in 2014, now recommend lifelong ART for all persons living with HIV 
(PLHIV). However, the possibility of safely stopping ART would hold great benefit both for 
patients, who are inconvenienced by having to take medications that require strict adherence 
and have a number of proven short-term and long-term toxicities, and by national health 
programs, which are committed to providing medications to hundreds of thousands or even 
millions of patients for decades to come.
ATI studies are an integral and necessary part of an international research agenda to find a 
“cure” for HIV, or at the minimum a treatment regimen that would provide an “HIV remission” 
in which the body could control HIV infection without the need for daily medication 
(Volberding 2014; Li 2015). Dr. Anthony Fauci, Director of the National Institute of Allergy 
and Infectious Diseases at the United States National Institutes of Health and recipient of the 
2013 Prince Mahidol Award, has written that clinical trials of HIV-cure therapies, including 
ATI trials, are promising and are feasible to implement (Fauci 2014). HIV-cure trials have also 
been deemed ethical to conduct (Sugarman 2013). The Bangkok RV254/SEARCH010 cohort is 
an ideal study population for ATI studies because, as explained in the following paragraph, 
there is evidence that patients who start ART very early in HIV infection may have a lower 
chance of viral rebound after stopping ART.
In contrast to patients begun on ART in the course of chronic HIV infection, many patients who 
begin ART at the time of acute HIV infection demonstrate blunted or delayed rebound viremia 
after ATI (Gianella 2011; Goujard 2012; Hamlyn 2012; Lodi 2012; Saez-Cirion 2013). Several 
studies have shown sustained viremic control after treatment interruption in 5- 16% of patients 
initiated on ART at the time of acute infection (Gianella 2011; Goujard 2012; Grijsen 2012; 
Lodi 2012; Saez-Cirion 2013). In these studies, factors associated with successful viremic 
control included shorter duration from HIV onset to ART initiation, longer duration on ART 
and low PBMC-associated HIV DNA (Williams 2014). In the RV254/SEARCH010 study in 
Thailand, early initiation of ART significantly restricted integration of HIV DNA in PBMCs, 
including central memory CD4+ T cells
(Ananworanich 2012; Ananworanich 2013; 
Ananworanich 2014). Total blood HIV DNA at initiation of therapy predicted reservoir size 
after 24 weeks of aggressive ART, underscoring the importance of early initiation of ART. 
Prior evidence has shown that reservoir distribution in shorter-lived cells is associated with elite 
control status (Buckheit 2013; Fonseca 2011; Shacklett 2011). Patients in RV254/SEARCH010 


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demonstrated this favorable pattern of reservoir distribution. For these reasons, patients who 
have received ART early in acute HIV infection are ideal candidates for the evaluation of novel 
therapies that seek to achieve a functional cure of HIV.
During ARV ATI, subjects will be monitored closely for clinical and laboratory indications to 
resume ART. These criteria are designed to protect the subjects as much as possible from any 
possible clinical, immunological, or virological adverse effects of ATI. Frequent virological 
monitoring after ATI, up to two times per week during the first 6 weeks when rebound viremia 
is most likely to occur, will minimize risk for the subjects. In the event that rebound viremia 
above 1,000 copies/ml does occur, the frequency of monitoring will allow for early re-initiation 
of ART as soon as possible and before clinical or immunological adverse effects would be 
expected. Moreover, the safety of this approach is proven in a recent study of chronically 
infected adults, many of who had low pre-ART CD4+ count. With twice-weekly viral load 
monitoring, rapid resumption of ART and viral suppression was demonstrated. No subjects had 
clinical adverse events due to ATI and CD4+ counts remained stable (Rothenberger 2015).
1.3.
Description of Ad26.Mos.HIV
Ad26.Mos.HIV vaccine was developed as part of the National Institute of Allergy and 
Infectious Diseases-sponsored U19 AI078526 Integrated Preclinical/Clinical AIDS Vaccine 
Development (IPCAVD) program at Beth Israel Deaconess Medical Center and Janssen 
Vaccines & Prevention B.V. It is a recombinant replication-incompetent live Ad26 virus-
vectored vaccine that is comprised of 3 adenoviruses, each of which has been genetically 
engineered to express one of 2 mosaic gag, pol genes or one mosaic env gene, each optimized 
for maximal coverage of potential T-cell epitopes. The Ad26.Mos.HIV vaccine will be used as 
the prime at 0 and 12 weeks. For comprehensive information refer to the most recent version of 
the Ad26.Mos.HIV Investigator's Brochure (IB).
1.4.
Description of MVA-Mosaic Sequence
Modified Vaccinia Ankara Mosaic Sequence (MVA-Mosaic sequence) is a recombinant live 
attenuated modified vaccinia virus vector that has been genetically engineered to express 
2 mosaic Gag, Pol, and Env sequences optimized for maximal coverage of potential T-cell 
epitopes. The MVA-Mosaic vaccine will be used as the booster at Week 24 and 48. For 
comprehensive information refer to the most recent version of the MVA-Mosaic Vaccines IB.
1.5.
Preclinical Studies
Safety of Ad26 and MVA-Vectored Vaccines in Preclinical Studies
The safety of the MVA-vectored vaccines is supported by the comprehensive preclinical studies 
conducted previously with an identical vector and similar inserts. The safety of MVA-HIV 
expressing gag, pol, and env inserts derived from an HIV-1 CRF01_AE isolate (CM235 


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env/CM240 gag/pol), from Chiang Mai, Thailand (referred to as MVA-CMDR) has been 
established based on a repeat-dose toxicity study conducted in rabbits and based on human 
clinical data from study RV158 (Currier 2010).
A repeated-dose Good Laboratory Practice (GLP) toxicity and local tolerance study in 
New Zealand white rabbits with different combinations of Ad26.ENVA.01 and MVA-Mosaic 
or MVA-Natural given by the IM route, including a 14-day reversibility period, was performed 
(Table 1).
Table 1:
Study Design: Repeated-Dose GLP Toxicology Study in New Zealand White Rabbits
Group
Treatment
Dose
Total 
dose 
volume
(mL)
Dose interval 
(Days)
Number of animals
Treatmenta
Necropsy
Total
(M/F)
Terminal 
(M/F)b
Recovery 
(M/F)c
1
Placebo
0
1.0
1, 22, 43, 64, 85, 
106
14/14
4/4
3/3
2
MVA-Natural
1x108 pfu/mL
1.0
1, 22, 43, 64, 85
10/10
5/5
5/5
3
MVA-Mosaic
1x108 pfu/mL
1.0
1, 22, 43, 64, 85
10/10
5/5
5/5
4
Ad26ENVA.01
5x1010 vp/dose
0.5
1, 22, 43
10/10
5/5
5/5
MVA-Natural
1x108 pfu/mL
1.0
64, 85, 106
5
Ad26ENVA.01
5x1010 vp/dose
0.5
1, 22, 43
10/10
5/5
5/5
MVA-Mosaic
1x108 pfu/mL
1.0
64, 85, 106
M/F: male/female; pfu: plaque-forming units
a The skin over the hind legs was shaved free of hair at least 24 hours prior to dose administration.
b Terminal necropsies occurred on Day 87 for Groups 1-3 and on Day 108 for Groups 4 and 5.
c Recovery necropsies occurred on Day 99 for Groups 1-3 and on Day 120 for Groups 4 and 5.
Vaccine-related changes in body temperatures, clinical pathology parameters and pathology 
data were generally resolved and/or were trending toward recovery, were not deemed to be 
adverse, and were considered a normal immunologic/inflammatory response to the vaccine 
administration. None of the effects noted during the study for animals receiving any of the test 
articles were adverse to the health of the animal.
Barouch, et al (Barouch 2010) evaluated cellular immune responses to mosaic HIV-1 vaccines 
in rhesus monkeys immunized with a single dose of recombinant adenovirus serotype 26 
(rAd26) expressing mosaic, consensus, or natural HIV-1 Gag, Pol, and Env sequences. The 
breadth of immune responses was greatest for the mosaic antigens; however there was no 
difference in magnitude of CD4+ and CD8+ T-cell responses between the groups. The depth of 
immune response (defined as the number of simultaneously elicited variant T-cell epitopes for a 
particular region) was greatest with the mosaic antigens. Humoral immune responses after a 
heterologous vector boost with the same insert as initial vaccination were also measured and the 
mosaic Env immunogens elicited non-inferior antibody responses as compared with the 
consensus or natural sequence Env antigens. This study showed that 2-valent mosaic HIV-1 
Gag, Pol, and Env antigens expanded cellular immune breadth and depth in rhesus monkeys as 
compared with consensus or natural sequence antigens without compromising the magnitude of 


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antigen-specific T lymphocyte responses as compared with consensus or natural sequence HIV-
1 antigens in rhesus monkeys.
A 4-cycle IM repeated-dose toxicity study (TOX10873, see IB of Ad26.Mos.HIV) was 
performed with prime-boost combinations of Ad26.Mos.HIV, MVA-Mosaic and Clade C 
gp140 drug product (DP). The objective of this study was to assess potential toxicity and local 
tolerance of the vaccines when administered to New Zealand white rabbits by IM injection once 
every 3 weeks up to 9 weeks (ie, a total of 4 injection days) and to evaluate the reversibility, 
persistence or delayed occurrence of any adverse effects on the vaccines during a 21 -day 
recovery observations period following the last injection. In this study, the full human doses are 
being tested. Details concerning the dose groups and regimen are given in Table 2.
Table 2:
Design of a 4-cycle IM Repeated Dose Toxicity Study With Prime-Boost Combinations of 
Ad26.Mos.HIV, MVA-Mosaic, and Clade gp140 DP
Group
Treatment(s)
Dose Levels
Dose 
Volume
(mL)
Dosing Day/Dosing Sitea
Number of 
Animalsb
1
22
43
64
Caudal
Cranial
Male
Female
1
Placebo (Saline)
0
0.5 or 0.75
ADM 1
ADM 2
ADM 3; 
ADM 4
ADM 5; 
ADM 6
10
10
2
Ad26.Mos.HIVc
5x1010 vp
0.5
ADM 1
ADM 2
ADM 3
ADM 5
10
10
gp140/AdjuPhos
250 mcg/425 mcg
0.5
--
--
ADM 4
ADM 6
3
Ad26.Mos.HIV
5x1010 vp
0.5
ADM 1
ADM 2
--
--
10
10
gp140/AdjuPhos
250 mcg/425 mcg
0.5
--
--
ADM 4
ADM 5
4
Ad26.Mos.HIV
5x1010 vp
0.5
ADM 1
ADM 2
--
--
10
10
MVA Mosaic
1x108 pfu
0.75
--
--
ADM 3
ADM 5
gp140/AdjuPhos
250 mcg/425 mcg
0.5
--
--
ADM 4
ADM 6
5
Ad26.Mos.HIV
5x1010 vp
0.5
ADM 1
ADM 2
--
--
10
10
MVA Mosaic
1x108 pfu
0.75
--
--
ADM 3
ADM 6
6
Ad26.Mos.HIV
5x1010 vp
0.5
ADM 1
ADM 2
ADM 3
ADM 6
10
10
Note: 
Day 1 is the first day of dosing
MVA-Mosaic is a 1:1 combination of MVA-Mosaic 1 and MVA-Mosaic 2
a
Doses were administered to either the left (ADM 1, 3 and 5) and/or right (ADM 2, 4 and 6) thigh (caudal and/or cranial 
muscle) via IM bolus injection.
b
Five animals/sex/group were maintained for a 21-day recovery period. Terminal animals were necropsied on Day 66, while 
recovery animals were necropsied on Day 85. 
c
On Days 43 and 64 control animals received 2 injections, 1 in left and 1 in right thigh, to provide appropriate controls for 
Groups 2 and 4
AdjuPhos = aluminum phosphate = established adjuvant; ADM = administration site; gp140 = Clade C gp140 DP
The different dose regimens were well tolerated when administered once every 3 weeks for up 
to 9 weeks (ie, 4 injections). The initial immune response, which was triggered after 
2 injections with Ad26.Mos.HIV, was increased after the next 2 injections for all tested vaccine 
regimens shortly after the last injection and 3 weeks later. The observed increases in C-reactive 
protein, fibrinogen, globulin and body temperature and the transient decrease in food 
consumption are considered to reflect a normal, non-adverse response to the vaccine 


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administration. Non-adverse test article-related findings were seen in iliac lymph nodes, in the 
spleen, and at the injection sites. Injection-site lesions had recovered after 3 weeks while the 
findings in lymph nodes and spleen (ie, immunogenic response) were still present after 
recovery.
Immunogenicity and Protective Efficacy Against SHIV-SF162P3 Challenge of Ad/MVA and 
Ad/Ad Regimens Expressing HIV-1 Mosaic Gag,Pol and Env Antigens in Rhesus Monkeys
To provide maximal coverage of potential T-cell epitopes, bivalent Mosaic HIV-1 Gag, Pol and 
Env antigens were developed. The goal of this study was to determine the immunogenicity of 
these vectors expressing mosaic antigens and the protective efficacy of these vaccine regimens 
against repetitive, low-dose, intrarectal challenges with the heterologous, neutralization-
resistant virus SHIV-SF162P3 (Barouch 2013). In particular, the study assessed whether the 
synthetic HIV-1 mosaic Env would elicit Env-specific antibody responses and afford 
protection. Adult Rhesus monkeys (N=6/group) were primed at Week 0 with Ad26 or Ad35 
vectors and boosted at Week 12 with Ad26, Ad35 or MVA vectors expressing HIV-1 mosaic 
Gag, Pol and Env immunogens, by the IM route. The dose was 4x1010 vp for Ad26 and Ad35 
vectors and 107 pfu for MVA vectors. Starting at Week 52, all monkeys received 6 low-dose, 
intrarectal challenges with SHIV-SF162P3. All vector regimens induced comparable magnitude
and breadth of cellular immune responses against matched Mos1 and Mos2 peptides as well as 
against global potential T-cell epitope peptides. All vector regimens induced binding antibody 
responses against multiple Env strains from clades A, C, D, F and M (namely UG37, UG92, 
CN54, ZA9, UG21, BR29 and Mosaic 1 respectively). Both the Ad/MVA and Ad/Ad vaccines 
afforded partial protection against acquisition of SHIVSF162P3 infection, corresponding to an 
87-90% reduction of per exposure acquisition risk, although the majority of the vaccinees 
became infected by the end of the challenge series. The vaccinated monkeys also exhibited a 
modest but significant 0.64 log reduction of setpoint viral loads, which was measured 84 days
after infection, as compared with the controls.
Recently, a non-human primate challenge study addressed correlates of protection by 
interrogating virus sequences and vaccine-induced immune responses. Animals were vaccinated 
with DNA and Ad5 expressing either mosaic Gag or mosaic heterologous Env, or heterologous 
Env based on a natural SIVmac239 sequence. Env-elicited immune response is necessary and 
sufficient to provide protection from acquisition from intrarectal SIVsmE660 with a strong 
sieving effect of Env immunization, selecting for minor variants in the challenge swarm. In 
addition, a sequence signature in the SIV Env, possibly shared by HIV, was identified that 
programs the neutralization phenotype of the viruses. There was no association between 
protection from infection and protection from pathogenesis, suggesting that humoral responses 
that effectively block acquisition are not necessarily correlated with cellular responses that 
control pathogenesis. The Env-induced CTL suppressed acute viraemia better than Gag CTL, 


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but suppressed chronic viraemia less effectively, resulting in reduced transmitted-founder 
viruses in breakthrough infections (Roederer 2013).
Taken together, both rhesus monkey studies demonstrated increased breadth and depth of CD8+ 
T-cell responses to mosaic HIV-1 vaccines. This increased immune response may protect 
against infection by diverse viruses and may limit the formation of variant “escape” viruses. 
1.6.
Clinical Studies
Several Phase 1 studies have evaluated Ad26 vectored HIV-1 vaccines under the IPCAVD 
NIH-sponsored program (see Ad26.Mos.HIV IB). IPCAVD001 is a phase 1 randomized, 
double-blind, placebo controlled dose escalation study to evaluate the safety and 
immunogenicity of recombinant Ad26.ENVA.01 in healthy, non-HIV-infected adults. The 
study is now completed (N=60) and overall, the Ad26.ENVA.01 vaccine has been generally 
safe and well tolerated at all doses tested (1x109, 1x1010, 5x1010, and 1x1011 vp) and up to 3 
vaccinations. There were no vaccine-related serious adverse events (SAEs). Minimal 
reactogenicity was seen at the 5x1010 dose. At the 1011 dose, reactogenicity (fatigue, myalgias, 
and chills) was seen 12 hours after the 1st vaccination, which usually resolved within 24 hours. 
These symptoms were not seen after the 2nd vaccination. Regarding immunogenicity, consistent 
Env antibody titers and Env T cell responses were seen following the prime, which increased 
after the 6-month boost and were durable at 1 year in vaccinated subjects. There was a dose 
response effect observed for Ad26 neutralizing antibody responses and EnvA enzyme-linked 
immunosorbent assay (ELISA) responses with 109 vp <1010 vp ~1011 vp. These results were the 
basis for the choice of the selected dose of Ad26.ENVA.01 (5x1010vp) used in the following 
and in the current study.
IPCAVD003 was a phase 1 randomized, double-blind, placebo-controlled study to evaluate the 
safety, mucosal immunogenicity, and innate immune responses of Ad26.ENVA.01 in 
24 healthy, HIV-1 uninfected adults. The vaccinations were generally well tolerated. The 
majority of subjects reported mild or no tenderness at the vaccination site overall (21 subjects or 
88%). Two vaccine recipients noted moderate and 1 vaccine recipient noted severe local 
reactogenicity over the week post vaccination. Four out of 18 vaccine recipients experienced at 
least 1 systemic reactogenicity symptom with a maximal grade of moderate; 5 vaccine 
recipients experienced at least 1 systemic reactogenicity symptom with a maximal grade of 
severe. In the latter group, the severe reactogenicity symptoms largely occurred between Day 0 
to 1 and were typically a combination of headache, chills, joint pain, myalgia, and 
malaise/fatigue. There were no study product related SAEs. Immunology results demonstrated 
that all subjects developed detectable Ad26-specific NAbs by 8 weeks. 16 of the 18 subjects 
who received Ad26.ENVA.01 had detectable peripheral EnvA IgG ELISA titer by 2 weeks with 
persistence in most subjects at 24 weeks. Mucosal EnvA-specific IgG ELISA responses were 
detected 2 weeks post vaccination in 11 of 18 subjects receiving Ad26.EnvA. This response rate 


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increased slightly at 24 weeks to 14 of 17 subjects having a detectable response. EnvA-specific 
CD8+ T-cell response increased post vaccination to 0.25%-0.43% at 2 weeks and 0.18%-0.26% 
at 24 weeks. A similar pattern was seen with EnvA CD4+ T-cell responses. 12 of 18 vaccinated 
subjects had detectable systemic IFN-gamma EnvA-specific ELISPOT responses by Week 2 
and 16 of 18 vaccinated subjects had a detectable response by Week 24. Mucosal EnvA-specific 
IFN-gamma CD8+ T-cell responses were undetectable at baseline in all 3 groups and remained 
undetectable in the placebo group at all time points. In Ad26- subjects, responses were detected
in half of subjects at 2 weeks and 24 weeks with mean responses of 0.07% and 0.13% at 2 and 
24 weeks, respectively. In Ad26+ subjects, similar mean responses of 0.05% and 0.06% at 2 
weeks and 24 weeks were observed. Mucosal EnvA-specific IFN-gamma CD4+ T-cell 
responses were 0%, 0.02% and 0% (Ad26-, Ad26+, and placebo) at baseline; 0.03%, 0.14%, 
and 0% at 2 weeks; and 0.03%, 0.03 and 0% at 24 weeks. There were stable levels of Ki67+ 
cellular activation in total and vector specific CD4+ T lymphocytes following Ad26.ENVA.01 
vaccination. There was no increase in CCR5 expression in total or vector specific CD4+ T 
lymphocytes in PBMCs or colorectal mucosa following vaccination. 
A third Phase 1 randomized, double blind, placebo controlled study (IPCAVD004 – IAVI 
B003) was conducted in the United States (US) and in South and East Africa to evaluate the 
safety and immunogenicity of Ad26.ENVA.01 and Ad35Env HIV vaccines in 217 healthy 
HIV-uninfected adult subjects. There was no pre-screening of Ad26 or Ad35 serology and the 
dose of each vaccine was 5x1010vp. The study assessed different prime boost regimens 
including 3- vs. 6-month intervals, prime boost directionality, and homologous versus 
heterologous vector regimens. The majority of reported local reactogenicity reactions was 
graded as mild or moderate (mild 65.0%; moderate 12.4%) and resolved spontaneously. 
Similarly, the majority of systemic reactogenicity reactions was graded as mild or moderate 
(mild 43.2%; moderate 29.6%) and resolved spontaneously. The majority of AEs were mild 
(75.5%) or moderate (23.2%) in severity. There were three unrelated SAEs. 4 weeks post -
second vaccination T-cell response rate (by IFN-gamma ELISPOT) was 44 -100%. There was 
induction of Ad26 NAbs in the majority of vaccinees 4 weeks after two Ad26.ENVA 
immunizations. EnvA ELISA response rate was 97-100% after the second injection.
A large body of clinical experience was acquired when non-recombinant MVA was used for 
primary vaccination against smallpox by intradermal (ID), subcutaneous, and IM routes in over 
120,000 humans in southern Germany and Turkey. The original idea of using an attenuated 
vector such as MVA was to sequentially immunize first with MVA and then boost with the 
classical vaccinia vaccine to offer protection against smallpox with fewer complications. 
During these extensive field studies no serious side effects were associated with the use of the 
MVA vaccine.
Several rMVA vaccines (e.g. MVA-malaria, MVA-HIV) are currently in Phase 1 and early 
phase 2 international studies. For example, protocol HVTN 055 [A Phase I Trial to Evaluate the 


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Safety and Immunogenicity of rMVA-HIV (rMVA-HIV env/gag+rMVA-HIV tat/rev/nef-RT) 
and rFPV-HIV (rFPV-HIV env/gag+rFPV-HIV tat/rev/nef-RT) Vaccines, Alone or in 
Combination, in Healthy, Vaccinia-Naïve HIV-1 Negative Participants] assessed the safety and 
immunogenicity of the simultaneous administration of two modified vaccinia Ankara (MVA) -
vectored HIV vaccines as priming doses (one containing env/gag and the other containing 
tat/rev/nef-RT), followed by boosting doses of two fowlpox-vectored HIV vaccines containing 
identical inserts. Overall, the vaccines were well tolerated and there were no SAEs. One or 
more AEs were reported by 91% of study subjects; however 97% of these were mild or 
moderate and they were infrequently considered to be definitely/probably (6%) or possibly 
(4%) related to the vaccine. Pain at the injection site was common but was usually mild with 
only one vaccinee reporting severe pain. Six subjects had systemic symptoms; 3 experienced 
them after the fowlpox vaccination and 3 after the MVA vaccination. There was no evidence 
that any subject developed subclinical or overt myo/pericarditis (Keefer 2011). 
Another group based in Germany gave an rMVA-HIV (Bavarian Nordic) to HIV infected 
patients in an effort to ‘boost’ their immune responses (ie, therapeutic vaccination). The patients 
tolerated the injections well and enhanced immune responses were observed (Harrer 2005). 
This study in HIV-infected individuals provided very important data, because the subjects were 
immune suppressed; yet they tolerated the live attenuated MVA vaccine quite well and may 
have benefitted from the immunization as it boosted their immune responses.
A third group, GeoVax, developed an MVA-HIV vaccine to follow a DNA prime. Data on the 
combination of clade B products, JS7 DNA followed by MVA/HIV62 also demonstrate this 
combination to be safe (local and systemic symptoms were mild or moderate) and that T-cell 
responses not detected with available assays from seemingly non-immunogenic priming 
regimens are enhanced by MVA recombinant vectors (Goepfert 2005).
Another MVA construct, MVA-CMDR was tested by the Military HIV Research program 
(MHRP as a single agent in a Phase 1 study (RV158) and administered by IM or ID route in 
HIV-uninfected adults (Currier 2010). A total of 51 healthy, HIV-negative, young adult subjects 
were enrolled in the study and randomized 5:1 to receive vaccine or placebo. Subjects were 
vaccinated with up to 107 pfu of MVA-CMDR by the ID route (0.1 mL into the volar aspect of 
forearm) or with up to 108 pfu by the IM route (1 mL into the deltoid). Three doses of vaccine 
were given on Days 0, 28 and 84. Safety data at all three sites show the vaccine to have an 
excellent safety profile, with the majority of reactogenicity being due to local cutaneous 
reactions in the group administered vaccine by the ID route (Grade 1 erythema, induration, 
pain, and tenderness). A few subjects experienced mild systemic reactions such as fatigu e, 
headaches, myalgias, and arthralgia. One subject had a Grade 2 fever (38.7°C), lasting one day 
after vaccination. No significant laboratory abnormalities were associated with vaccination. No 
subjects showed signs/symptoms of myo/pericarditis (clinical or subclinical) despite intensive 
monitoring with regularly scheduled electrocardiograms (ECGs), cardiac enzymes, and AE


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solicitation of cardiac symptoms and diary cards. Cell-mediated immune responses were 
moderate in magnitude but high in response rate. Immune responses were predominantly HIV 
Env-specific CD4+T cells and were durable for at least 6 months. Binding antibodies against 
gp120 and p24 were detectable in all vaccination groups. The high dose (10 8 pfu) delivered by 
IM route was the most immunogenic.
Preliminary Safety Data From Healthy Adult Subjects
The following candidate vaccine components are currently being studied in 2 first-in-human 
(FIH) studies in HIV-1 uninfected subjects: MVA-Mosaic and Ad26.Mos.HIV.
HIV-V-A002
HIV-V-A002 is a single-center, randomized, parallel-group, placebo-controlled, double-blind 
FIH Phase 1 study to evaluate safety, tolerability, and immunogenicity of MVA-Mosaic, both in 
previously unvaccinated healthy adult subjects and in subjects previously vaccinated with 
Ad26.ENVA.01. 
A prototype of MVA-Mosaic, referred to as MVA-CMDR, has previously been found to be 
immunogenic, safe, and well tolerated.
The independent Safety Monitory Committee (SMC) has performed a safety evaluation 
(unblinded to group level for SMC members only) at Week 16 (4 weeks after all of the 
25 enrolled subjects had received their second vaccination). MVA-Mosaic was well tolerated 
and no safety concerns were identified. Most AEs were of mild and moderate severity. At the 
time of this SMC evaluation, there were no withdrawals or discontinuations. One SAE was 
reported, assessed as not related to study vaccine. The SMC concurred with proceeding with the 
clinical development as planned and with administration of MVA-Mosaic in HIV-V-A004.
HIV-V-A004
HIV-V-A004 is a multi-center, randomized, parallel-group, placebo-controlled, double-blind 
Phase 1/2a study to evaluate safety, tolerability, and immunogenicity of various prime/boost 
regimens containing Ad26.Mos.HIV, MVA-Mosaic, and/or Clade C gp140 (with aluminum 
phosphate adjuvant) components in healthy HIV-uninfected adult subjects. For Ad26.Mos.HIV, 
this is a FIH study.
A prototype of Ad26.Mos.HIV, called Ad26.ENVA.01, has previously been found to be 
immunogenic, safe, and well tolerated.
As pre-specified in the protocol, enrollment was paused when approximately 10% of subjects 
had received their first injection (Ad26.Mos.HIV) and the Protocol Safety Review Team 


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(PSRT) has reviewed blinded safety data of 2 weeks after this first injection to determine if 
enrollment could continue. The PSRT has reviewed blinded 2 week safety follow-up data of 
39 subjects and on 18 Jun 2015 all members agreed that the study could resume enrollment. 
Most AEs were of mild and moderate severity. Three grade 3 AEs were reported that were 
considered related to vaccination: headache, chills, and myalgia. At the time of the PSRT 
review, there were no SAEs, withdrawals, or discontinuations.
Preliminary Data From the Acute HIV Infection Cohort
RV254 has extensive feasibility data for enrolling and retaining subjects, and performing 
specimen collection, processing, storage, international shipping and laboratory studies. 
Screening for acute HIV infection is performed in real-time by pooled nucleic acid testing. 
RV254 has an accompanying ARV protocol in which all subjects are offered immediate ART. 
Enrollment in the cohort is ongoing with an average of 30-50 new enrollees per year. 
Table 3 shows characteristics of the first 150 enrolled subjects in RV254. All except two 
elected to initiate ART, and the mean time from enrollment to ART initiation was 2 days. The 
RV254 follow-up schedule includes visits at 12-week intervals until a maximum of 416 weeks, 
and offers additional optional intensive studies (leukapheresis, lumbar puncture, sigmoid 
biopsy, inguinal lymph node biopsy).
Table 3:
Characteristics of 150 Subjects Enrolled in the 
RV254/SEARCH010 Acute Infection Cohort
Characteristics
Acute HIV Cohort
Age, Median (IQR)
28 (24 - 32)
Male gender
141 (94%)
Infection duration, days (n=123)
   Min-Max
3 - 48
   Median (IQR)
17 (12 - 22)
Fiebig, n(%)
    I
51 (35%)
    II
24 (16%)
    III
59 (40%)
    IV
8 (5%)
    V
6 (4%)
CD4 (cells/mm3), median (IQR)
361 (267 - 496)
HIV RNA (log10cps/mL), median (IQR)
5.7 (5.0 - 6.6)
Treatment arm (n=147)
    HAART
68 (46%)
    megaHAART
79 (54%)
CRF_01AE (n=62)
47 (76%)
R5 tropism (n=39)
38 (97%)
IQR: inter-quartile range
Relevant to HIV functional cure, RV254 has demonstrated limited seeding of the viral reservoir 
in early acute HIV infection with subsequent decrease following ART initiation 


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(Ananworanich 2012). Subjects evaluated during Fiebig I exhibit lower total and undetectable 
integrated HIV DNA compared to those sampled during Fiebig stage III. More importantly, 
after 24 weeks of ART, Fiebig I subjects display an extremely low frequency of infected cells,
comparable to historical controls including elite controllers and much lower than in subjects 
who have initiated ART in chronic infection, despite 4 years of viral suppression. These results 
suggest that very early ART (Fiebig I) may limit the initial seeding of the HIV reservoir (Figure
1).
These preliminary data indicate that the subjects in the RV254 cohort have a lower HIV viral 
reservoir than patients who initiate ART at a later time after HIV infection. This study is based 
on the hypothesis that the vaccine regimen will boost the immune system enough that it will be 
able to control HIV replication in the absence of ART after ATI. 
Figure 1:
After 6 Months of HAART, RV254/SEARCH010 Subjects Have Very low Levels of HIV DNA 
in PBMCs, Similar to Levels Seen in Elite Controllers (Ananworanich, 2012)
We propose to evaluate safety and efficacy of a therapeutic vaccine administered in the form of 
Ad26 vector with mosaic inserts for gag/pol and env genes of HIV-1, boosted with the MVA 
vector with homologous mosaic inserts. These vaccine products have been shown to be 
immunogenic and safe. The vaccine population will be HIV-infected adults who began ART 
during acute HIV infection and have undetectable plasma HIV RNA (<50 copies/ml) for at least 
48 weeks. Patients in this unique cohort have been shown to have very low HIV viral reservoirs 
and therefore have a high chance for maintained viral suppression in the absence of ART 
(functional cure of HIV infection).


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2.
STUDY HYPOTHESIS AND OBJECTIVES
2.1.
Hypothesis
This is an exploratory study to collect preliminary data and establish sufficient experience to 
inform and develop subsequent studies to address the following hypothesis: Vaccine therapy 
administered in the form of Ad26 vector with mosaic inserts for gag/pol and env genes of 
HIV-1 at 0 and 12 weeks (Stage 1), boosted with the MVA vector with homologous mosaic 
inserts for gag/pol and env genes at 24 and 48 weeks (Stage 1), among individuals with fully 
suppressed HIV will be safe and well-tolerated, will result in a measurable immune response, 
and will result in over 50% of vaccine recipients achieving sustained viremic control after ATI
(Stage 2).
2.2.
Primary Objectives
1.
Determine the safety and tolerability of Ad26 prime/MVA boost versus placebo in subjects 
on suppressive ART that was initiated during acute HIV infection
2.
Measure the frequency and duration of sustained viremic control after receiving Ad26 
prime/MVA boost or placebo, defined as greater than 24 weeks with plasma HIV RNA 
<50 copies/ml after ARV ATI
2.3.
Secondary Objectives
1.
Determine the immunogenicity of Ad26 prime/MVA boost in subjects on suppressive ART 
that was initiated during acute HIV infection
2.
Characterize biomarkers of HIV reservoir at baseline, after vaccine therapy prior to ARV 
ATI (Week 48-60) and after ARV ATI (Week 60-96)
3.
Compare the duration of viremic control (HIV RNA <50 copies/ml) between vaccine and 
placebo recipients who failed to achieve sustained viremic control (undetectable plasma 
HIV RNA [<50 copies/ml] at Week 24 after ARV ATI)
4.
Describe the frequency, magnitude, specificity and functional capacity of humoral and 
cellular immune responses to vaccine and other immunogens
5.
Describe the molecular sequence sieve effects of vaccine therapy on breakthrough rebound 
viremia before and after cessation of therapy 
6.
Describe the PBMC phenotype, pattern of soluble factors and immune functional responses 
before and after ARV ATI in both the vaccine and placebo arms and compared to historical 
untreated acute infection cases in RV217
7.
Describe the clinical outcomes in terms of the frequency, severity, duration and treatment 
for acute retroviral syndrome post ARV ATI
8.
Evaluate and characterize HIV resistance to ARV drugs in subjects who experience 
rebound viremia after ARV ATI


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2.4.
Exploratory Objectives
1.
Analyze the markers of HIV reservoir as predictors of sustained viral suppression.
2.
Analyze the markers of HIV specific immune responses as predictors of sustained viral 
suppression.
3.
Analyze the expression profiles and transcriptome analysis of sorted and/or unsorted 
lymphocytes and monocyte/macrophages in the different arms of the study.
4.
Compare the mucosal immunity (vaginal and rectal for humoral and gut mucosa for 
cellular immunogenicity) in vaccine versus placebo arms.
5.
Describe the markers of immune activation in the peripheral blood and various reservoirs.
6.
Describe the viral load response to resumption of ART and impact on markers of HIV 
reservoir among those who fail to suppress HIV RNA off therapy. 
3.
STUDY DESIGN AND POPULATION
3.1.
Study Design
This is a combined Phase 1/2a randomized, double-blind, placebo-controlled study to 
investigate the safety, immunogenicity and effect on HIV viremic control after ARV ATI of a 
vaccine regimen consisting of an Ad26.Mos.HIV prime and an MVA-Mosaic boost. The study 
will include subjects who started on ART during acute HIV infection, who are on a current 
stable ART for at least 4 weeks prior to screening and who have achieved absence of viremia 
(HIV RNA <50 copies/ml) for ≥ 48 weeks prior to initiation of vaccine/placebo.
The study will be conducted at one study site in 
 Thailand 
 
. Volunteers for this study will be participants on RV254. The RV254 cohort 
is largely composed of men who have sex with men and transgender subjects and has a limited 
number of women (Table 3). Participants for this study will continue to participate in other 
RV254 related activities in parallel. This will include regular and unscheduled visits. No 
research blood draws or invasive procedures are scheduled during the RV254 related visits 
performed in parallel with this study. Laboratory tests in RV254 for HIV clinical care, such as 
screening and monitoring for co-infection may be performed. In addition, blood draws for 
clinical care may be performed. The results of the activities/procedures specific for RV254 will 
only be reported if relevant to this study. Data from RV254 will be shared with the current 
study as needed to assess endpoints and objectives. As an example, sequences for the founder 
virus from the RV254 study would be available for analysis within this study.
PPD
PPD


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Figure 2:
Study Design
The study will enroll 27 subjects in a 2:1 ratio of vaccine to placebo (Figure 2). The study will 
occur in two stages. Before the first stage, recruitment, consent and baseline assessments of 
subjects who meet the entry criteria of sustained absence of viremia for at least 48 weeks and of
initiation of ART during acute HIV infection will take place. It is anticipated that enrollment 
will be rapid as the RV254 study has over 100 early-treated patients with viral suppression. The 
first stage is administration of the vaccine or placebo, and observation after vaccination and 
prior to ARV ATI (Phase 1). The second stage will be ARV ATI with frequent assessment of 
viral load for rebound viremia (Phase 2a). In this period, the duration of viral control will be 
determined, and follow-up for those subjects who resume therapy, ie, fail to maintain viremic 
control will be performed. Durability of immunogenicity will be evaluated during the ARV ATI
period.
An independent DSMC will be involved in addition to a PSRT, which will have representatives 
from the sponsor and protocol team.


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Measured outcomes include:
1)
Safety endpoints
2)
HIV specific antibody and cellular immune responses in blood and mucosal 
compartments
3)
Viremic control
4)
Biomarkers for HIV expression and reservoir in blood and mucosal compartments
5)
Markers for immune status, including phenotype and activation markers in blood and 
mucosal compartments
The subjects will receive 4 single injections of the respective vaccine or placebo. Subjects, 
clinical staff, investigators and the sponsor are blinded to treatment allocation. The subjects 
receiving placebo serve as a control for safety, efficacy and immunogenicity assessments. 
Placebo only refers to the vaccine component of the study (Stage 1). Subjects in both the 
vaccine and placebo arms will receive standard ART:

For HIV treatment during the first 60 weeks of the trial prior to ATI (Stage 1)

If they meet any of the criteria to restart ART listed in Section 4.1.6 (Stage 2)

At the end of the trial at Week 96 (post Stage 2)
To assess the safety of the administered vaccines, subjects will be observed for reactogenicity in 
the clinic for a minimum of 30 minutes after each vaccination. Vital signs will be taken and 
qualified study personnel will evaluate for any signs or symptoms of local or systemic 
reactions. The subjects will document local and systemic reactions 6 hours after each 
vaccination and then daily for the next 7 days and investigators will document any reported AEs 
at each visit throughout the study. Rebound in viral replication post ARV ATI (HIV RNA 
>50 copies/ml) will not be considered an AE/SAE as it is a study endpoint. To assess the 
subjects' cellular and humoral immune response, blood samples will be taken during the clinic 
visits. Safety, efficacy and immunogenicity visits will be conducted as outlined in the study 
Schedule of Evaluations (SOE) in Table 4 and Table 5.
The study will last approximately 96-100 weeks per subject, including up to 6 weeks for 
screening, 48 weeks for the vaccination period (Stage 1), 12 weeks between the final 
vaccination and ARV ATI (Stage 1), and 36 weeks of follow up after ARV ATI (Stage 2). End 
of study is defined as the last visit of the last subject undergoing the study.
All subjects in this study are participants in RV254 and will continue to participate in other 
RV254 activities in parallel which include regular and scheduled visits. No research blood 
draws are scheduled during this visits, but blood draws or procedures scheduled for clinical care 


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may be performed. The results of the activities/procedures specific for the RV254 will only be 
reported if relevant to the current study.
In addition, all subjects will be offered enrollment in study RV412 (Safety and Virologic 
Outcomes after Analytic Treatment Interruption in Thai Patients who Initiated Antiretroviral 
Therapy During Early Acute HIV Infection). This requires a separate consent process. RV412 is 
a protocol that will enroll all subjects who complete any clinical study that includes ARV ATI 
at SEARCH. The protocol provides regular clinical, immunological and virogical monitoring 
for at least 48 weeks or until HIV-1 RNA is undetectable on 2 consecutive assessments after 
reinitiating ART. RV412 provides a transition period of more frequent monitoring for safety 
and clinical endpoints before subjects return to routine follow-up visits of every 12 weeks under 
the RV254 study protocol. Data from RV412 will be shared with the current study as needed.
3.2.
Subject Selection and Withdrawal
3.2.1.
Inclusion Criteria
1.
Confirmed HIV-1 infected and started ART during acute infection (Fiebig stages I, II, III or 
IV)
2.
Treatment with current stable ART (no changes to treatment) for at least 4 weeks prior to 
screening.
3.
HIV RNA <50 copies/ml for at least 48 weeks at screening.
a.
One blip of HIV RNA >50 and <200 copies/ml within 48 weeks is acceptable, 
provided that the most recent (before screening) HIV RNA <50 copies/ml.
4.
Each subject must have voluntary signed an informed consent form indicating that he/she 
understands the purpose of and procedures required for the study and is willing to 
participate in the study.
5.
Age 18 to 50 years old at the time of study enrollment.
6.
Laboratory criteria during screening:
a.
Hemoglobin: Women: ≥11 g/dL; Men ≥12.5 g/dL
a.
White cell count: 2,500 to 11,000 cells/mm3
b.
Platelets: 125,000 to 450,000 per mm3
c.
Alanine aminotransferase (ALT)/aspartate aminotransferase (AST) ≤1.5x institutional 
upper limits of normal (ULN)
d.
Creatinine ≤1.5x institutional ULN
e.
CD4 > 400 cells/mm3
f.
Troponin <1 x ULN


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7.
All female subjects of childbearing potential must have a negative serum pregnancy test 
(β-human chorionic gonadotropin) at the screening visit, and a negative urine pregnancy 
test prior to vaccination on Day 11.
8.
A woman must be either:
a. Not of childbearing potential: postmenopausal (>45 years of age with amenorrhea for at 
least 2 years, or any age with amenorrhea for at least 6 months and a serum follicle 
stimulation hormone [FSH] level >40 IU/L); surgically sterile; or
b. Of child-bearing potential and practicing an effective double method of birth control 
(e.g. prescription oral contraceptives, contraceptive injections, intrauterine device, 
contraceptive patch, or vaginal ring, in conjunction with either a female condom or one 
of the methods for male contraception indicated in Inclusion Criterion 9, before entry2
and through 3 months after the last vaccination.
9.
A man who is sexually active with a woman of child-bearing potential and has not had a 
vasectomy, must agree to use an adequate contraception method as deemed appropriate by 
the investigator (e.g. condom, sterilization [after vasectomy, zero sperm must be confirmed 
if procedure occurred less than 1 year ago]) in conjunction with one of the methods for 
female contraception indicated in Inclusion Criterion 8.
10. Available and willing to participate for the duration of the study visits and follow up.
11. Pass the Test of Understanding (TOU) (See Appendix 16.3)
3.2.2.
Exclusion Criteria
1.
A woman who is pregnant, breastfeeding, or planning to become pregnant while enrolled in 
this study, or within 3 months after the last vaccination.
2.
Subject is a man who plans to father a child while enrolled in this study, or within 3 months 
after the last vaccination.
3.
Any clinically significant acute or chronic medical condition that in the opinion of the 
investigator 
would 
preclude 
participation 
(e.g., 
history 
of 
seizure 
disorders, 
bleeding/clotting disorder, autoimmune disease, active malignancy, poorly controlled 
asthma, active tuberculosis or other systemic infections, etc).
4.
Any history of HIV-related illness under Centers for Disease Control and Prevention 
(CDC) category C. 
5.
Major surgery within the 4 weeks prior to study entry or planned major surgery through the 
course of the study.
                                                
1 Note: negative urine pregnancy test also required prior to second, third and fourth vaccinations
2 Verbal assurance should be given that adequate birth control measures have been followed for 28 days prior to 
vaccination.


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6.
History of myocarditis, pericarditis, cardiomyopathy, congestive heart failure with 
permanent sequelae, clinically significant arrhythmia (including any arrhythmia requiring 
medication, treatment, or clinical follow-up).
7.
ECG with cardiology reading showing clinically significant findings, or features that would 
interfere with the assessment of myo/pericarditis, including any of the following:
a.
Conduction disturbance (complete left or complete right bundle branch block or 
nonspecific intraventricular conduction disturbance with QRS ≥120 ms, PR interval 
≥220 ms, any 2nd or 3rd degree AV block, or QTc prolongation [> 450 ms]);
b.
Significant repolarization (ST segment or T wave) abnormality;
c.
Significant atrial or ventricular arrhythmia; frequent atrial or ventricular ectopy (e.g., 
frequent premature atrial contractions, 2 premature ventricular contractions in a row)
d.
ST elevation consistent with ischemia; or evidence of past or evolving myocardial 
infarction.
8.
History of diabetes mellitus type 1 or type 2, including cases controlled with diet alone. 
(Not excluded: history of isolated gestational diabetes.)
9.
Chronic active hepatitis B or active hepatitis C (eg, positive serology with confirmatory 
positive polymerase chain reaction) or active syphilis infection. Active syphilis 
documented by examination or serology unless positive serology is due to past treated 
infection.
10. Thyroidectomy, or thyroid disease requiring medication during the last 12 months.
11. Hypertension:
a.
If a subject has been diagnosed with hypertension during screening or previously,
exclude for hypertension that is not well controlled. Well-controlled hypertension is 
defined as blood pressure consistently ≤ 140 mm Hg systolic and ≤ 90 mm Hg 
diastolic, with or without medication, with only isolated, brief instances of higher 
readings 
b.
If a subject has NOT been diagnosed with hypertension during screening or 
previously, exclude for systolic blood pressure ≥ 150 mm Hg at enrollment or diastolic 
blood pressure ≥ 100 mm Hg at enrollment. 
12. Major psychiatric illness and/or substance abuse problems during the past 12 months that 
in the opinion of the investigator would preclude participation
13. Receipt of any vaccine within 30 days prior to the first vaccination or plans to receive 
within 30 days post-vaccination. In the case of medically indicated vaccines, the 
vaccination should be given at least 2 weeks before or after the first vaccination. However, 
if a vaccine is indicated in a post-exposure setting (eg, rabies or tetanus), it must take 
priority over the study vaccine.3
                                                
3 The same rules will apply for the second, third and fourth vaccinations.


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14. Use of experimental therapeutic agents within 30 days of study entry.
15. Receipt of blood products or immunoglobulin in the past 3 months. 
16. History of anaphylaxis or other serious adverse reactions to vaccines or vaccine products, 
or neomycin or streptomycin or egg products.
17. History of chronic urticaria (recurrent hives).
18. Chronic or recurrent use of medications which modify host immune response, e.g. cancer 
chemotherapeutic agents, parenteral corticosteroids (short course oral steroids given for 
non-chronic conditions not expected to recur is not an exclusion criteria, topical steroid use 
is not an exclusion criteria), etc. but not including ART.
19. Recipient of an HIV vaccine candidate at any time, receipt of a pox-virus vaccine in the 
last 10 years, and receipt of other experimental vaccine(s) within the last 5 years.
a.
Exceptions may be made for vaccines that have subsequently undergone licensure by 
the US Food and Drug Administration (FDA). For potential subjects who have 
received placebo or experimental vaccine (greater than 5 years ago), documentation of 
the identity of the study placebo or vaccine must be provided to the PSRT, who will 
determine eligibility on a case-by-case basis.
20. Subjects who cannot communicate reliably with the Investigator.
21. Subjects who are, in the opinion of the Investigator, unlikely to cooperate with the 
requirements of the study.
22. A study site employee directly supervised by members of the study team, as well as family 
members of the investigator, or an employee of the sponsor or its partner(s).
NOTE: The investigator should ensure that all study enrollment criteria have been met when the 
subject is screened. If a subject’s status changes (including laboratory results or receipt of 
additional medical records) after screening but before the first administration of study vaccine 
such that they now meet an exclusion criterion, they should be excluded from participation in 
the study. 
3.2.3.
Prohibitions and Restrictions
Vaccination with any vaccine within 30 days prior or after any of the study vaccinations is 
disallowed. Medically indicated vaccines, other than those described above, (e.g. influenza, 
tetanus, Hepatitis A, Hepatitis B or rabies) are not exclusionary but should, if possible, be given 
at least two weeks before or two weeks after any study vaccination to avoid potential confusion 
of adverse reactions. However, if a vaccine is indicated in the post-exposure setting (e.g. rabies 
or tetanus), it must take priority over the study vaccine. 


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3.2.4.
Withdrawal or Elimination Criteria
Under certain circumstances, a subject will be terminated from participating in further 
vaccinations or further study visits. These specific experiences are outlined in Section 5.2 and 
Section 5.3, respectively.
A subject who discontinues will not be replaced, unless the subject did not yet receive any study 
vaccine. Replacement of these subjects will only be allowed until the end of the recruitment 
period.
3.2.5.
Cohort Criteria for Proceeding to ARV ATI Phase of Study
ICS or ELISPOT will be performed on cryopreserved PBMC using peptide pools for gag, env 
and pol matching the vaccine inserts. Participants will be considered to have a positive response 
following vaccination if after the third vaccination (2 injections of Ad26 and a single MVA 
injection) there is ≥2-fold increase in magnitude of IFN-gamma producing cells relative to
baseline for any one of the matching peptide pools. If the baseline response is negative, then 
any detectable response is considered a positive response. Placebo recipients will be evaluated 
to maintain blinding. The ARV ATI (Stage 2) will proceed if at least 50% of subjects who 
received the active vaccine have a positive response (as evaluated by an independent, unblinded 
statistician).
Individual subject criteria for proceeding to ARV ATI are provided in Section 3.2.6 below.
3.2.6.
Individual Subject Criteria for ARV ATI at Week 60
Subjects who complete 4 vaccine/placebo injections and remain in the study through Week 58 
will undergo ARV ATI at Week 60 if they meet all of the criteria listed in Section 3.2.6.1. 
Subjects will be excluded from ARV ATI if they meet one of the criteria listed in 3.2.6.2.
3.2.6.1.
Inclusion Criteria for ARV ATI
1. HIV-1 RNA <50 copies/mL for previous 52 weeks (blips <200 copies/ml are allowed, 
provided that the most recent result is <50 copies/ml)
2. Most recent (within 3 months) peripheral blood CD4 count > 400 cells/mm3
3. No CDC Category B or C HIV-related illness within the last 6 months (Appendix 16.1)
3.2.6.2.
Exclusion Criteria for ARV ATI
1. Pregnancy or breast-feeding.
2. New hepatitis B surface antigen positive since enrollment.
3. New hepatitis C antibody positive since enrollment.
4. Serious medical or psychiatric illness that, in the opinion of the site investigator, would 
interfere with the ability to adhere to study requirements.


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5. Active drug or alcohol use or dependence that, in the opinion of the site investigator, would 
interfere with adherence to study requirements.
3.3.
Treatment Arm Allocation
Subjects will be randomly assigned to one of the vaccine and placebo arms based on a 
computer-generated randomization schedule prepared before the study by or under the 
supervision of the sponsor. A list of treatment allocation assignments will be sent to the 
designated pharmacist at the study site.
Distribution and randomization will be 2:1 in the vaccine and placebo arms, respectively. 
A subject who discontinues will not be replaced, unless the subject did not yet receive any study 
vaccine. Replacement of these subjects will only be allowed until the end of the recruitment 
period.
3.4.
Method of Blinding and Unblinding
The subjects, the clinical staff, the Principal Investigator (PI) and the sponsor will be blinded to 
treatment allocation. The pharmacist with primary responsibility for vaccine dispensing will not 
be blinded to the treatment and maintains the randomization code and completes assignments of 
subjects according to the randomization allocation. 
The blind will be broken only if, in the opinion of the site PI, PSRT or DSMC, immediate 
unblinding is necessitated by an acute safety concern in order to make medical management 
decisions. Whenever possible, the sponsor and PSRT should be contacted before breaking the 
blinded emergency code. Emergency unblinding, without prior notification to the sponsor and 
PSRT, should occur only if necessary to make urgent medical management decisions. The 
sponsor's Pharmacovigilance Department (Global Medical Safety) has to be informed of 
unblinding within 1 working day.
Accidental unblinding (e.g., investigator sees the investigational product [IP] administration 
logs) must be reported within 1 working day to the sponsor who then advises on the next 
corrective and preventative steps to be taken. 
Subjects for whom the code was broken will be withdrawn from further vaccination. If less than 
4 study injections were received then they will not receive further injections and will not 
undergo ARV ATI. These subjects will continue follow-up through Week 50, which will be 
considered the final study visit and all investigations scheduled for the exit visit (Week 96) will 
be completed at that time. If unblinding occurs after all 4 study injections were received then 
the subject will continue participation in the study per protocol.


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Routine unblinding of treatment allocations may occur only after all subjects have had their last 
study visit and the eDC database is finalized.
4.
TREATMENTS
4.1.
Study Treatments
Detailed instructions for study treatment management including preparation, storage and 
documentation are provided under separate cover in the IP Management Manual. Additional 
information is also provided in the IBs for Ad26.Mos.HIV and MVA-Mosaic.
4.1.1.
Ad26.Mos.HIV Vaccine
Ad26.Mos.HIV is a trivalent vaccine containing the following three active pharmaceutical 
ingredients pre-mixed in a 1:1:2 vp ratio:

Ad26.Mos1.Gag-Pol = recombinant, replication-incompetent Adenovirus Serotype 26 
expressing mosaic 1 HIV-1 Gag and Pol proteins, manufactured in PER.C6® Cells
(JNJ-55471494-AAA).

Ad26.Mos2.Gag-Pol = recombinant, replication-incompetent Adenovirus Serotype 26 
expressing mosaic 2 HIV-1 Gag and Pol proteins, manufactured in PER.C6 Cells
(JNJ-55471520-AAA).

Ad26.Mos1.Env
= recombinant, replication-incompetent Adenovirus Serotype 26 
expressing a mosaic 1 HIV-1 Env protein, manufactured in PER.C6 Cells
(JNJ-55471468-AAA).
The trivalent Ad26.Mos.HIV vaccine will be manufactured and provided under the 
responsibility of the sponsor. It is formulated as a clear to slightly opalescent solution for IM 
injection. The vaccine will be supplied as a frozen liquid to be thawed prior to use, and will be 
essentially free from particles. The vaccine will be provided in individual dosage vials at a 
concentration of 1 x 1011 vp/mL. Each 2 mL DIN 2R vial contains a volume of approximately 
0.75 mL, with an extractable volume of 0.5 mL for a dose of 5 x 1010 vp. See IP Management 
Manual for further details.  
4.1.2.
MVA-Mosaic Vaccine
Modified Vaccinia Ankara Mosaic Sequence is comprised of the following 2 vaccine products 
supplied in separate vials and administered in a 1:1 ratio:

MVA Mosaic 1 = Modified Vaccinia Ankara virus expressing Mosaic 1 HIV-1 Gag, Pol 
and Env proteins (JNJ-55471533-AAA)

MVA Mosaic 2 = Modified Vaccinia Ankara virus expressing Mosaic 2 HIV-1 Gag, Pol 
and Env proteins (JNJ-55471572-AAA)


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Each vaccine is provided in individual dosage vials at the concentration of 2 x 108 pfu/mL.
Each stoppered and sealed 2 mL glass vial contains a volume of approximately 1.2 mL. They 
are supplied as white to opaque, slightly to moderately cloudy liquids. Each of the MVA 
products is mixed at the site research pharmacy in a 1:1 ratio and a 0.5 mL dose withdrawn to 
deliver a net dose of MVA mosaic-1 and MVA mosaic-2 of 1 x 108 pfu. See IP Management 
Manual for further details. 
4.1.3.
Placebo
Sodium Chloride Injection US Pharmacopeia (USP), 0.9% will be used as the placebo for 
Ad26.Mos.HIV and MVA-Mosaic vaccinations. See the IP Management Manual for further 
details.
4.1.4.
Vaccine Administration Schedule
The schedule of follow up is shown in Table 4 (schedule from screening to completion of 
vaccinations; Stage 1) and Table 5 (schedule from ARV ATI to the end of the study; Stage 2). 
Subjects will receive Ad26.Mos.HIV or placebo at Weeks 0 and 12, and will receive MVA-
Mosaic or placebo at Weeks 24 and 48. In subjects who are on non-nucleoside reverse 
transcriptase 
inhibitor 
(NNRTI)-based 
treatment, 
a 
boosted 
protease 
inhibitor 
(lopinavir/ritonavir or atazanavir/ritonavir or darunavir/ritonavir) will be given instead of the 
NNRTI from Weeks 58 to 60 to reduce the risk for NNRTI resistance. At Week 60, all ARV 
drugs will be discontinued.
4.1.5.
Monitoring During ARV ATI
During ARV ATI (Stage 2), subjects will be monitored frequently for clinical and laboratory 
indications to resume ART (Section 4.1.6). These criteria are designed to protect the subjects 
from any possible clinical, immunological, or virological adverse effects during the ARV ATI. 
Prior investigation has shown that virologic controllers may experience transient low-level 
viremias after ARV ATI and that these reversible low-level viremias do not always indicate 
viral rebound (saez-Cirion 2013). Therefore transient viremia <1,000 copies/mL is not 
considered an indication for resumption of ART. This low level of viremia is also unlikely to 
produce clinical disease or CD4 decline within the time interval of ARV ATI in this study. 
4.1.6.
Criteria to Reinitiate ART During ARV ATI
Subjects will require re-initiation of ART after ATI using the same regimen as their previous 
treatment (before Week 60) for any of the following reasons:
1.
HIV-1 RNA above 1,000 copies/mL on 2 consecutive determinations at least 1 week apart.
2.
CD4+ T-cell counts below 350 cells/mm3 on 2 consecutive determinations at least 2 weeks 
apart.


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3.
CD4+ T-cell count decline of > 50% from Week 60 prior to ATI (as reference/baseline).
4.
Clinical progression to CDC Category B or C disease (see Appendix 16.1).
5.
Diagnosis of Acute Retroviral Syndrome (ARS, see Appendix 16.2).
6.
Pregnancy
7.
Subject requests re-initiation of ART.
4.1.7.
Management of Subjects After Reinitiating ART
If the decision is made to reinitiate ART for any reason (except pregnancy), the subject will 
continue the planned visits as in Table 5. Management of subjects who become pregnant is 
detailed in Section 4.1.9.
If ART is reinitiated due to rebound viremia with HIV-1 RNA >1,000 copies/mL, then HIV 
genotype testing will be performed at the time of ART resumption. The results of genotype 
testing will be available within seven days and will be used to guide ART selection. The final 
regimen will be determined by discussion between the subject and the treating physician. 
Subjects will undergo quantitative HIV-1 RNA testing every two weeks for the first four weeks 
and then every four weeks until the end of the study.
After 96 weeks, the protocol will be completed and all subjects will remain on ART. They will 
be offered enrollment in protocol RV412, which provides frequent and intensive clinical and 
laboratory monitoring, as noted in Section 3.1.  
All subjects who complete enrollment in study RV412 will continue to participate in the RV254 
parent protocol. Therefore, subjects enrolled in this VAC89220HTX1001/RV405 protocol will 
have access to long-term treatment and monitoring.
4.1.8.
Management of Subjects With Sustained Virologic Control at End of 
Study
Subject can request re-initiation of ART at any time during the study, regardless of their 
virologic control.
After 96 weeks, the protocol will be completed and subjects will be asked to resume ART. ART 
will be provided by the study site. They will be offered enrollment in protocol RV412 for 
continued observation. The RV412 protocol will help to ensure the safety of the subjects with
frequent and intensive viral load and safety laboratory monitoring. 
ART regimens are based on the subject’s last known ART regimen or another empiric regimen 
as determined by discussion with the subject’s medical provider. 


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All subjects who complete enrollment in RV412 will continue to participate in the RV254 
parent protocol. Therefore, subjects enrolled in this VAC89220HTX1001/RV405 protocol will 
have access to long-term treatment and monitoring.
4.1.9.
Management of Subjects Who Become Pregnant
Subjects who become pregnant during the study period will complete all procedures scheduled 
for study end at the time the pregnancy is diagnosed. If the subject was scheduled to receive 
additional vaccine or placebo, these injections will not be administered after pregnancy has 
been determined. The visit at which pregnancy is diagnosed or immediately following the 
diagnosis (if diagnosed outside the study) will be considered the subject’s last study visit. 
Pregnant subjects will subsequently be offered enrollment in RV412 for continued observation. 
They will be advised to resume ART as soon as possible in consultation with the study 
investigator, PI and an obstetrician/gynecologist experienced with the management of HIV 
during pregnancy. If the subject completes participation in RV412 before the pregnancy is 
completed, she will subsequently continue her participation in the RV254 parent study. 
Pregnant subjects prescribed ART will also be advised to enroll in the ARV pregnancy registry 
(http://www.apregistry.com/who.htm).
The site PI or designated associate investigator will be responsible for reporting any pregnancy 
to the PSRT and the study sponsor. This information will be reviewed by the PSRT regularly in 
aggregate with other safety data. The PSRT will forward notification as necessary to IRBs, 
study sponsor, and regulatory agencies.
A pregnancy should be followed to term, any premature terminations reported, and the health 
status of the mother and child including date of delivery should be reported to the sponsor after 
delivery. Pregnancy outcomes will be recorded via a standardized form. Information 
documented on this form will include date of last menstrual period, date pregnancy confirmed, 
history of complications during prior pregnancies (such as congenital abnormalities or 
spontaneous abortions), and the outcome of the pregnancy including date of termination or 
delivery, any complications of pregnancy, and the status of the child. A separate form will be 
completed for the delivered child to document date of delivery, gender, weight, presence of any 
congenital abnormalities, APGAR score, HIV status, and any other complication of delivery.
Any abnormality considered related to the study vaccine by the responsible physician during 
pregnancy and delivery, and health of mother and baby must be reported through an expedited 
report.


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4.2.
Handling of Study treatments
4.2.1.
Packaging and Labeling
The Ad26.Mos.HIV vaccine and MVA-Mosaic vaccines will be manufactured and vialed under 
supervision of the sponsor. Sodium Chloride Injection USP, 0.9% will be purchased 
commercially.
All study vaccines will be manufactured and packaged in accordance with Current Good 
Manufacturing Practice. 
Further details for the packaging and labeling for both the study vaccine and placebo can be 
found in the IP Management Manual.
No study vaccine can be repacked or relabeled without prior approval from the sponsor.
4.2.2.
Shipment and Storage
Authorization to ship the IP to the site will be provided in writing by the sponsor, upon 
confirmation that all required critical documents for shipment authorization are completed. The 
IP will be shipped to the site according to the required storage conditions.
Upon receipt of study treatment supplies, the pharmacist must immediately inspect all vials for 
damage, temperature recorded during shipment, packaging and labeling, and inventory 
compared to the packing list. The study drug will be shipped with a continuous temperature 
monitoring device. The condition of the vials and the temperature of the shipment must be 
documented. Any deviations or problems identified must be documented. The sponsor should 
be immediately notified of any temperature excursion and/or damage to vials in shipment 
and/or storage of IP.
Study vials must be stored in a secured location with no access for unauthorized personnel. 
Guidance on storage temperature is provided in the pharmacy manual/study site investigational 
product manual. Storage temperature must be monitored daily and a log of the monitored 
temperature maintained. The study freezer must be equipped with a continuous temperature 
monitor and alarm. Study freezers should be equipped with back-up power systems. Complete 
storage instructions are provided in the IP Management Manual. In the event that study product 
is out of temperature range, all relevant data will be sent to the sponsor to determine if the 
affected study product will be used or replaced. Dosing should be halted until further instruction 
from the sponsor, and during that period the affected IPs must be quarantined. 
4.2.3.
Dose Preparation and Administration
To prepare the MVA-Mosaic vaccines, the vials must be thawed to room temperature and then 
mixed using a high-speed vortex, as per instructions in the IP Management Manual. 


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To prepare the Ad26.Mos.HIV vaccine, each vial must be thawed to room temperature and then 
prepared as per instructions in the IP Management Manual. 
For MVA-Mosaic, a 0.5 mL injection (108 pfu) will be administered IM.
For Ad26.Mos.HIV, a 0.5 mL injection (5 x 1010 vp) will be administered IM.
For placebo, a 0.5 mL injection of normal 0.9% saline will be administered IM.
All injections may be administered in either deltoid. It is not necessary to use the same deltoid 
at each visit. No local or topical anesthetic will be used prior to the injection via any route of 
administration. The study vaccine will be prepared by the site pharmacist and administered by 
the clinic staff. The IP Manual specifies the maximum time allowed between preparation and 
administration.
Site pharmacists will prepare all doses for administration and dispense to the clinic. All other 
staff members and subjects will remain blinded to the treatment administered. In order to 
preserve blinding, the pharmacist will place an overlay on the syringes. 
4.2.4.
Accountability
The site pharmacist will be responsible for maintaining an accurate record of the randomization 
codes, inventory, and accountability of vaccine supplies for this study. At any time, the 
numbers of supplied, used, and remaining vials have to match. It must be possible to reconcile 
delivery records with those of used and unused stocks. Account must be given of any 
discrepancies.
The site pharmacist will also be responsible for ensuring the security of these documents. 
Partially used vials will not be administered to other subjects or used for in vitro experimental 
studies. Any unused portion of entered vials and expired syringes should be disposed of in a 
biohazard container and incinerated or autoclaved. After the study is completed or terminated, 
the site will receive instruction regarding the final disposition of any remaining study products.
4.3.
Prior and Concomitant Treatment
Concomitant medications are recorded from day of initial screening to first vaccination and at 
every study visit. All subjects are HIV-infected and on suppressive ART. Study vaccinations 
will be administered in addition to ART. The ART cannot be modified, with the following 
exceptions: 
Switches within an ARV class will be allowed for well documented tolerability/toxicity 
reasons (linked to an AE or SAE) and after sponsor approval. A switch to a different 


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formulation of the same ARV is allowed. Switches between lamivudine (3TC) and 
emtricitabine (FTC) are allowed.
If subject fails to comply with the ARV regimen, a change that will prevent efficacy and 
tolerability issues will be allowed for well documented reasons and after sponsor 
approval.
At Week 58, NNRTIs will be replaced by protease inhibitors (see Section 5.1.6).
Study subjects can receive medications such as acetaminophen, non-steroidal anti-inflammatory 
drugs, or antihistamines as needed although they must be documented and use of these 
medications as routine prophylaxis prior to study vaccination is discouraged.
4.4.
Investigational Product Compliance
The study products will be administered intramuscularly to subjects in the clinic by trained and 
qualified study personnel. Study site personnel will keep a log of all study treatment 
administered. 
5.
CONDUCT OF THE STUDY
5.1.
General Aspects
Evaluation of the safety of the vaccine regimens will include clinical laboratory tests, physical 
assessments by clinical staff and subject reports on signs and symptoms following vaccinations. 
Additional study visits may be required if in the investigator’s opinion, further clinical or 
laboratory evaluation is needed. In case a grade 3 or grade 4 laboratory abnormality, or any 
laboratory abnormality accompanied by clinically relevant signs or symptoms occurs (from the 
baseline visit onwards), a confirmatory test should be performed within 48 hours after the 
results have become available. After that, laboratory tests will be repeated weekly until values 
are resolved or stable.
Potential adverse reactions will be further evaluated prior to continuing the vaccination 
schedule. Total blood volume drawn from each subject will not exceed 550 mL in any eight-
week period which is considered acceptable based upon the NIH and US FDA guidelines (NIH 
clinical 
center 
guidelines; 
https://www.fda.gov/scienceresearch/specialtopics/runningclinicaltrials/guidancesinformationsh
eetsandnotices/ucm118099.htm). The schedule for evaluations for the study is completely 
detailed in Table 4 and Table 5.
This study is one of the few studies in the world to follow a unique cohort of acutely treated 
HIV infected patients with extremely low HIV-reservoir size. Recent data have shown that HIV 
mainly resides in lymphoid tissues and other tissue compartments. In chronically treated 


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patients, the viruses in tissues are the sources of viral rebound when ART is interrupted 
(Rothenberger 2015). In RV254 study participants who are treated in acute HIV infection, 
remarkably low cell-associated HIV DNA have been observed in the blood and in the gut, as 
well as low HIV RNA in the CSF and genital compartments (Ananworanich 2012, Valcour 
2012). The levels of these markers of HIV reservoir are undetectable in most patients and lower 
than those previously reported (Goujard 2012, Hocqueloux 2010). However, it is not known
whether replication competent HIV is present in the compartments and whether it could be a 
source of viral rebound.  
Moreover, in this study, it will be important to understand HIV-specific immune responses that 
are pre-existing and those that are generated after vaccination. HIV-specific immune responses 
in tissue may be key to controlling viremia following treatment interruption (Barouch 2012;
Hansen 2013). The RV144 study conducted in Thailand study also illustrated the importance of 
HIV V1/V2 antibody in the mucosa in blocking/promoting infection (Haynes 2012). Therefore, 
it is important that we understand the interactions between the virus and the immune system in 
the tissue compartments as well as in the peripheral blood.
The biological samples collected in the study have been selected as the body compartments 
most likely to harbor quiescent HIV virus: CSF from the central nervous system, lymph node 
tissue, and lymphoid tissue in the gut mucosa. Genital secretions are collected because sexual 
contact is the most common way in which HIV is transmitted and the results of this testing has 
important implications for HIV prevention.
Measuring HIV and the immune responses against HIV in the compartments will afford us the 
ability to identify predictors for successes and failures in controlling viremia. This will be a 
major advance for the field in that it will provide measures to help select patients in the future 
who may have the highest probabilities to control viremia without long-term ART (ie, HIV 
remission).  


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Table 4:
Schedule of Follow-Up and Testing From Screening to After all Vaccinations and Prior to ARV ATI (Stage 1)
Visit Number
1
210
3
4
510
6
7
8
9
10
11
1210
13
Visit Week
Screening
Wk 0
Wk 4
Wk 12
Wk 1613
Wk 
2410
Wk 2613 Wk 3013 Wk 3613 Wk 48
Wk 5013
Visit Day and Window
-45 to -3 
1
29 ± 5
85 ± 7
88 ± 2
113 ± 5
169 
± 7 
183 ± 
5
211 ± 
5
253 ± 
5
337 ± 
14
340 ± 2
351 ± 
5
Eligibility criteria, Consent
X
Randomization
X
Ad26 or placebo
X
X
MVA or placebo
X
X
AE and concomitant medications
recording8
X
X
X
X
X
X
X
X
X
X
X
X
X
1-week memory aid distribution
X
X
X
X
Memory aid return and review
X
X
X
X
History, PE, vital signs
X
X
X
X
X
X
X
X
X
X
ECG9
X
X
CBC11
6
6
6
6
6
6
6
CD4 and CD8 
NB
NB
NB
NB
NB
NB
NB
HIV RNA 
NB
NB
NB
NB
NB
NB
NB
Chemistry1,11
10
6
6
4
10
Troponin 
NB
Serum or urine pregnancy test7
NB
4
NB
NB
NB
Hepatitis B surface antigen, HCV, 
VDRL,TPHA12
NB
NB
Binding Ab, Neutralizing Ab, ADCC
6
6
6
6
6
6
(X)4
6
6
6
ELISPOT, ICS, epitope mapping
170
(X)4
(X)4
(X)4
(X)4
170
(X)4
(X)4
(X)4
170
PBMC, plasma storage2,4
25.5
25.5
25.5
51
25.5
25.5
51
25.5
X
Neuropsychiatric examination
X
X
Optional procedures3
(X)
(X)
Soluble immune activation markers 
6
(X)4
6
(X)4
6
(X)4
(X)4
6
(X)4
6
Gene array
8.5
(X)4
8.5
(X)4
8.5
(X)4
(X)4
8.5
(X)4
8.5
X
Cell-associated RNA
8.5
(X)4
8.5
(X)4
8.5
(X)4
(X)4
8.5
(X)4
8.5
Total, integrated HIV DNA, 2 LTR circles 
8.5
(X)4
8.5
(X)4
8.5
(X)4
(X)4
8.5
(X)4
8.5
QVOA5,6
(X)6
(X)6
(X)6
TILDA5
8.5
8.5
8.5
TULDA5
8.5
8.5
8.5
Single copy HIV RNA5
8.5
(X)4
8.5
(X)4
8.5
(X)4
(X)4
8.5
(X)4
(X)4
Tissue phenotyping, RNA/DNA
(X)
(X)
Approx daily vol (ml)
16
243
32
92
9
32
126
176
26
82
73
34
205


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Approx cumulative vol (ml)
16
259
291
383
392
424
550
726
752
834
907
941
1,146
NB: no additional blood required; CBC: complete blood count; S = safety; S + I = safety + immunogenicity; scr = screening; vac = vaccination
1Chemistry including electrolytes, glucose, blood urea nitrogen (BUN), creatinine, Total/direct bilirubin, ALT, AST, gamma-glutamyl transpeptidase (GGT). 
2Blood draw for PBMC and plasma storage, ELISPOT, ICS epitope mapping and flow cytometry will not be performed at visits for which leukapheresis is done.
3Optional procedures include leukapheresis, sigmoid biopsy, inguinal lymph node biopsy, lumbar puncture, MRI, MRS and DTI. 
4Testing may be performed on selected samples of interest depending on cell yields and at the discretion of the protocol chair or PI. Not all assays listed will be done on all 
timepoints.
5QVOA: quantitative viral outgrowth assay; TILDA, Tat/Rev Induced Limiting Dilution Assay; TULDA, Tat/Rev Uninduced Limiting Dilution Assay; and single copy RNA: 
will be performed only if plasma HIV-1 RNA viral load <50 copies/mL. 
6QVOA will only be performed on subjects undergoing leukapheresis or when cells yields are sufficient
7Women of childbearing potential only. Serum test at screening, urine test at all other visits.
8 AEs are to be collected from signing of informed consent onwards until the end of the study.
9 ECGs are to be taken before blood sampling.
10 Subjects will be contacted (methods of contact may vary and may include but are not limited to phone calls, clinic visits, home visits, or email communications) 24 – 72 hours 
after each vaccination (contacts will be recorded as Visits 2a, 4a, 7a, and 11a as appropriate). The subjects will need to come for a clinic visit at Week 12 +3 days and at Week 48 
+3 days for the planned blood sampling. For the gene array testing ideally the visit should take place exactly at +3 days after the vaccination.
11 In case a grade 3 or grade 4 laboratory abnormality, or any laboratory abnormality accompanied by clinically relevant signs or symptoms occurs (from the baseline visit 
onwards), a confirmatory test should be performed within 48 hours after the results have become available. After that, laboratory tests will be repeated weekly until values are 
resolved or stable.
12For hepatitis C: PCR has to be done when Ab are positive and for syphilis: TPHA needs to be done when VDRL is positive.
13Timing of the visit will be determined relative to the actual day of vaccination.


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Table 5:
Schedule of Follow-Up and Testing From ARV ATI to the end of the Study (Stage 2)
Visit Number
14
15
16-2116
2216
23-
26,26a16
2716
2816
29-3116
3216
33-
3516
3616
37-
3916
4016
41-
4316
4416
4516
4616
4716
4816
4916
5016
Visit Week
Wk 58 Wk 60
Wk 64
Wk 67 Wk 68 Wk 69-
71
Wk 
72
Wk 
73-
75
Wk 
76
Wk 
77-
79
Wk 
80
Wk 
81-
83
Wk 
84
Wk 
86
Wk 
88
Wk 
90
Wk 
92
Wk 
94
Wk 96 
Exit 
visit
Study 
End
Visit Day and Window
407 ± 
5
421 ± 7 428 to 
446
449 ±
2
452 to 
467
470 ± 
2
477 ± 
3
484 to 
498 ± 
3
505 
± 3
512 
to 
526 
± 3
533 
± 3
540 
to 
554
± 3
561 
± 3
568 
to 
582 
± 3
589 
± 3
603 
± 5
617 
± 5
631 
± 5
645 
± 5
659 
± 5
673 
±5
Protease inhibitor
replacement for NNRTI1
X
ART interruption
X
ART resumption2
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
HIV genotype2
6
AE and concomitant 
medications recording14
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
History, PE, vital signs
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CBC3,15
6
6
6
6
6
6
6
6
6
6
6
CD4 and CD83 
NB
NB
NB
NB
NB
NB
NB
NB
NB
NB
NB
HIV RNA4 
NB
NB
NB
NB
NB
NB
NB
NB
NB
NB
NB
Qualitative RNA 2x per 
week
1x6
1x4
Qualitative RNA 1x per 
week
1
1x3
1x3
1x3
1x3
1
1
1
Chemistry5,15
4
4
4
4
Urine pregnancy test13  
X
X
X
X
Binding Ab, Neutralizing 
Ab, ADCC6 
6
6
6
6
6
6
6
6
6
6
ELISPOT, ICS, epitope 
mapping6
(X)11
(X)11
(X)11
(X)11
(X)11
(X)11
(X)11
(X)11
(X)11
(X)11
PBMC, plasma storage7,11
51
51
51
51
51
25.5
25.5
25.5
25.5
51
Neuropsychiatric 
examination
X
X
Optional procedures8
(X)
(X)
Soluble immune 
activation markers 
6
(X)11
(X)11
6
(X)11
(X)11
6
(X)11
(X)11
6
Gene array
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
Cell-associated RNA
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
Total, integrated HIV 
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5


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Table 5:
Schedule of Follow-Up and Testing From ARV ATI to the end of the Study (Stage 2)
Visit Number
14
15
16-2116
2216
23-
26,26a16
2716
2816
29-3116
3216
33-
3516
3616
37-
3916
4016
41-
4316
4416
4516
4616
4716
4816
4916
5016
Visit Week
Wk 58 Wk 60
Wk 64
Wk 67 Wk 68 Wk 69-
71
Wk 
72
Wk 
73-
75
Wk 
76
Wk 
77-
79
Wk 
80
Wk 
81-
83
Wk 
84
Wk 
86
Wk 
88
Wk 
90
Wk 
92
Wk 
94
Wk 96 
Exit 
visit
Study 
End
Visit Day and Window
407 ± 
5
421 ± 7 428 to 
446
449 ±
2
452 to 
467
470 ± 
2
477 ± 
3
484 to 
498 ± 
3
505 
± 3
512 
to 
526 
± 3
533 
± 3
540 
to 
554
± 3
561 
± 3
568 
to 
582 
± 3
589 
± 3
603 
± 5
617 
± 5
631 
± 5
645 
± 5
659 
± 5
673 
±5
DNA, 2 LTR circles 
QVOA9,10
(X)10
(X)10
(X)10
(X)10
TILDA9
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
TULDA9
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
Single copy HIV RNA9 
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
(X)11
(X)11
8.5
Tissue12 phenotyping, 
RNA/DNA
(X)
Approx daily vol (ml)
6
124
6
63
4
1
63
3
124
3
63
3
38
3
99
1
38
1
38
1
130
Approx cumulative vol 
(ml)
1,150
1,274
1,28
0
1,343
1,347
1,34
8
1,411
1,41
4
1,538
1,541
1,604
1,607
1,645
1,648
1,747
1,748
1,786
1,787
1,825
1,826
1,956
LTR: long terminal repeats; NB: no additional blood required
1For subjects who are on NNRTI, the NNRTI will be discontinued at Week 58 and replaced with a protease inhibitor for 2 weeks prior to interrupting all drugs at week 60. This is to prevent possible 
NNRTI resistance.  
2ART will be resumed if subjects meet any of the criteria listed in section 4.1.6. HIV genotype will be done before restarting ART. 
3After ARV ATI, CBC and CD4 are performed every 4 weeks. A repeat CD4 will be performed when CD4 is <350 cells/mm3 and ART will be resumed if CD4 is confirmed to be < 350 cells/mm3 
continuously for 2 weeks. 
4 From Week 61 to 66 HIV RNA testing will be done twice per week and then subsequently once per week during Week 67 to 84 and once every 2 weeks during Week 86 to 96. On visits where no 
venous blood draw is scheduled, testing will be performed via qualitative RNA testing from fingerstick. In case of detectable HIV RNA results, refer to Section 5.1.8 for detailed assessment timings.
Upon restarting ART, subjects will undergo quantitative HIV-1 RNA testing every two weeks for the first four weeks and then every four weeks until the end of the study.
5Chemistry including electrolytes, glucose, BUN, creatinine, Total/direct bilirubin, ALT, AST, GGT. 
6In patients who fulfill ART resumption criteria and no immunogenicity testing is planned at the visit that ART will be re-initiated, additional blood will be drawn for immunogenicity tests prior to 
ART start according to the blood draw schedule of Week 60.
7Blood draw for PBMC and plasma storage will not be performed at visits for which leukapheresis is done.
8Optional procedures include leukapheresis, genital mucosal secretions, sigmoid biopsy, inguinal lymph node biopsy, lumbar puncture, MRI, MRS and DTI. For subjects who need to resume ART 
prior to Week 96, optional procedures would be considered prior to ART initiation in replace of the 96-week time point.
9QVOA, TILDA, TULDA; and single copy RNA: will be performed only if plasma HIV-1 RNA viral load <50 copies/mL. 
10QVOA will only be performed on subjects undergoing leukopheresis or when cells yields are sufficient
11Testing may be performed on selected samples of interest depending on cell yields and at the discretion of the protocol chair or PI. Not all assays listed will be done on all timepoints.
12Tissues/samples are obtained through optional procedures that include sigmoid biopsy, inguinal lymph node biopsy and lumbar puncture (CSF).
13Women of childbearing potential only
14AEs are to be collected from signing of informed consent onwards until the end of the study.


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15 In case a grade 3 or grade 4 laboratory abnormality, or any laboratory abnormality accompanied by clinically relevant signs or symptoms occurs (from the baseline visit onwards), a confirmatory test 
should be performed within 48 hours after the results have become available. After that, laboratory tests will be repeated weekly until values are resolved or stable.
16Timing of the visit will be determined relative to the ARV ATI at Visit 15. Visits from week 61 to 67 are to be scheduled twice a week at least 2 days apart.


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5.1.1.
Screening
A qualified study team member will inform each participant of the study, provide him/her wi th 
any study details and a copy of the informed consent document to aid him/her in his/her choice 
for voluntary participation. Study subjects will receive a briefing from the PI or designee during 
which the study will be explained and participation requirements outlined. The briefing will be 
followed by an opportunity for questions. Study staff will then review the consent form with 
potential subjects and answer any additional questions.
After the informed consent has been signed, the following evaluations will be performed to 
determine eligibility requirements as specified in the inclusion and exclusion criteria: 

Physical examination including vital sign measurement

Medical history

Review of concomitant medications

Review of inclusion/exclusion criteria

Subject must pass the TOU

Blood sampling for CBC with differential, CD4, HIV RNA, blood chemistries, troponin,
hepatitis B and C serologies, and syphilis serology. Details on hepatitis B and syphilis 
testing are indicated in Table 4

ECG with interpretation by cardiologist

Serum pregnancy testing (women of childbearing potential
only)/pregnancy
and 
contraceptive counseling
General eligibility for this clinical study will be dependent on results of laboratory tests and the 
medical assessment. Counseling related to the potential risks of becoming pregnant during this 
study will be provided. Study subjects who qualify for inclusion based on the history, physical, 
and laboratory results will be contacted and scheduled for enrollment and initial vaccination 
(Visit 2, Week 0). 
Subjects with laboratory values or vital signs not meeting eligibility criteria on the initial 
screening visit may be re-assessed for eligibility at a repeat screening visit. Laboratory tests may 
only be repeated if either a laboratory error or a transient condition at screening is suspected. The 
laboratory tests can only be repeated once. Physiologic parameters, such as vital signs, may be 
repeated no more than 3 times if in the judgment of the investigator the abnormal result is the 
result of an acute, short-term rapidly reversible condition. Ineligible subjects will be referred for 
care as clinically indicated.


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5.1.2.
Randomization and Vaccination 1/Day 1/Week 0 (Stage 1)
After confirmation of meeting the eligibility criteria, eligible subjects will be randomly assigned 
as described in Section 3.3.
Day 1 is defined as the day of the first vaccination and the day of study enrollment. If clinical 
assessment on Day 1 suggests significant changes may have occurred since the Screening Visit, 
then the physical examination, hematology tests, and blood chemistries will be repeated as 
needed and the subjects rescheduled for Day 1 assessment. Pregnancy test results for women of 
reproductive potential must be obtained on each vaccination day prior to the vaccination. Eligible 
subjects with acute illnesses or who are febrile (temperature ≥38.0 oC non-axillary) will be 
rescheduled for Day 1 vaccination.
Day 1 evaluations, performed prior to the first vaccination, are used as the baseline for 
subsequent safety assessments. Evaluations to be performed on Day 1, Visit 2 can be found in 
the SOE (Table 4). 
Following each injection, subjects will be observed for reactogenicity in the clinic for a 
minimum of 30 minutes. Vital signs will be taken before vaccination and at least 30 minutes 
post-vaccination.
Qualified study personnel will then evaluate the subjects for any signs or symptoms of local or 
systemic reactions. The injection sites will be examined for local reactions at approximately 
30 minutes post-vaccination. If erythema or induration is present, the diameters will be measured 
and the largest diameter recorded. Findings will be entered on appropriate source documents and 
eCRF.
For life-threatening allergic reactions that occur immediately post vaccination, the clinic has a 
protocol for treatment and immediate transfer to a nearby emergency room for stabilization.
Via a memory aid, subjects will be asked to record occurrences of erythema and induration 
(measured using the ruler supplied), and pain/tenderness, itching, swelling, or warmth at the 
injection sites on the day of vaccination and then daily for 7 days, for a total of 8 daily records. 
The investigator or designee should discuss information from the memory aids, document 
relevant information in the clinic chart (source document) and complete the relevant parts of the 
eCRF. Subjects will also be asked to record and report presence of the following AEs, in addition 
to other spontaneously reported AEs:
o
fever (from temperature)
o
fatigue
o
headache
o
myalgia


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o
arthralgia
o
chills
o
nausea
o
vomiting
o
rashes
o
general itching
5.1.3.
Vaccinations 2 – 4 / Weeks 12, 24, 48 (Stage 1)
Subjects will return to the clinic for the assessments as shown in Table 4. The allowable window 
period for vaccinations 2 – 3 is one week before or after the scheduled dose. For vaccination 4, 
the window is 2 weeks before or after the scheduled dose. 
If after all assessments (history, physical examination, vital signs, and pregnancy testing) have 
been performed the subject still qualifies (ie, subject does not have an acute illness or 
temperature ≥38.0ºC (oral or tympanic) and does not meet any vaccination discontinuation 
criteria (Section 5.2), the next scheduled vaccination will be given.
Following each vaccine administration, subjects will be observed for reactogenicity in the clinic 
for a minimum of 30 minutes. Vital signs will be taken at least 30 minutes post-vaccination. 
Qualified study personnel will then evaluate the subjects for any signs or symptoms of local or 
systemic reactions. The injection sites will be examined for local reactions at approximately 
30 minutes post-vaccination. If erythema or induration is present, the diameters will be measured 
and the largest diameter recorded. Findings will be entered on appropriate source documents and 
eCRF.
For life-threatening allergic reactions that occur immediately post vaccination, the clinic has a 
protocol for treatment and immediate transfer to an emergency room for stabilization.
Subjects will be provided a memory aid, thermometer, and ruler to measure and record local 
solicited AEs. Subjects will also record unsolicited and solicited systemic AEs in the memory aid 
6 hours post vaccination and then daily for the next 7 days. The memory aid will act as a 
memory tool for the subject but is not considered a source document. For a list of solicited AEs, 
See Section 5.1.2.
All AEs will be recorded from signing of informed consent form onwards until the end of the 
study.


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5.1.4.
Post-Vaccination Follow-Up Visits / After Study Visits Week 0, 12, 24, 
48) (Stage 1)
Subjects will be contacted (methods of contact may vary and may include but are not limited to 
phone calls, clinic visits, home visits, or email communications) 24 – 72 hours after each 
vaccination. During this time, subjects should report to the clinic staff any concomitant 
medication and any AEs they may have experienced. Subjects will be reminded to fill out their 
memory aids provided to them. At the follow-up visit after Week 12, sampling for gene array 
will also be performed. At the follow-up visit after Week 48, sampling for gene array and 
PBMC, plasma storage will also be performed.
5.1.5.
Safety and Immunogenicity Visits / Weeks 4, 16, 26, 30, 36, 50
(Stage 1)
Subjects will return to the clinic for safety and immunogenicity assessments as detailed in 
Table 4. At Week 16 only, a 12-lead ECG will be performed.
Subjects will be instructed to contact the investigator immediately should he/she manifest any 
signs or symptoms that are perceived as serious or if hospitalization of the subject occurs.
All AEs will be recorded from signing of informed consent form onwards until the end of the 
study.
5.1.6.
Preparation for ARV ATI / Week 58 (Stage 1)
All subjects will have a physical examination and laboratory screening at 58 weeks in 
preparation for ARV ATI. At this visit clinical, immunological, and virological criteria for ARV 
ATI will be assessed as listed in Section 3.2.5 and 3.2.6. Subjects on ART containing an NNRTI 
drug will have the NNRTI replaced by a protease inhibitor drug at this visit in order to prevent 
the possibility of NNRTI drug resistance developing after ARV ATI.
5.1.7.
ARV ATI / Week 60 (Stage 1)
At Week 60 criteria for ARV ATI will be reviewed, including all clinical, immunological, and 
virological criteria listed in Section 3.2.5. Pregnancy is an exclusion criterion for ARV ATI and 
female subjects of childbearing potential will have a urine pregnancy test at this visit. If the 
subject continues to meet criteria for ATI, then all ARV drugs will be stopped at this visit. 
5.1.8.
Monitoring After ARV ATI / Week 61-96 (Stage 2)
Subjects will be followed at least weekly for the first 24 weeks after ARV ATI (Week 61 to 84) 
and then every 2 weeks for the remainder of the study through Week 96 (exit visit). Each 
monitoring visit will include a physical examination as well as laboratory testing at outlined in
Table 5. 


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Specific laboratory monitoring includes:
HIV-1 RNA: From Week 61 to 66 HIV RNA testing will be done twice per week and then 
subsequently once per week from Week 67 to 84 and once every two weeks from Week 86 
to 96. Testing is done via quantitative RNA testing on visits where venous blood draw is 
scheduled or, on all visits where no venous blood draw is scheduled, via qualitative RNA 
testing from fingerstick. If HIV RNA is detectable in the qualitative RNA test, subjects will 
be asked to return to clinic immediately for a confirmatory HIV RNA via a venous blood 
draw. If HIV RNA is confirmed detectable, HIV RNA will be repeated every 3 days by 
quantitative RNA until the subject restarts ART or until HIV RNA becomes undetectable.
CD4 count will be performed every 4 weeks after ARV ATI. Any result <350 cells/mm3 or 
<50% compared to baseline at Week 60 will be repeated within 2 weeks. 
Other tests: urine pregnancy for female subjects of childbearing potential and routine 
chemistries (Table 5) will performed every 12 weeks after ARV ATI. 
5.1.9.
Unscheduled Visits
If a subject is seen for any other visits other than the visits outlined in Table 4 and Table 5 of the 
protocol, the visit will be captured as an unscheduled visit. The reason for the visit will be 
documented in the eCRF along with the procedures completed during this visit. All protocol 
related data resulting from this visit will be captured in the clinical database (ie, safety laboratory 
results).
5.1.10.
Neuropsychiatric Examination
Detailed neurological examination, including neuropsychological testing, will be performed at 
Week 0, 50, and at end of study. Details of the neuropsychological testing can be found in 
Section 16.5. The tests are used for the continuous monitoring in Study RV254. The results of 
these tests will be analyzed as part of the RV254 parent protocol. They are not part of this study 
endpoints and will only be reported if relevant to this study.
The crossed out positions from the questionnaire are to be used at baseline, and will not be 
included at the future visits.
An abbreviated NPZ-4 assessment will be conducted at Week 72 and will include:

Abridged Neuropsychological Assessment Test Battery

SUBSTANCE USE AND SLEEP
5.1.11.
Optional Procedures
A subset of subjects may additionally consent for optional procedures, including leukapheresis, 
genital mucosal secretions, sigmoid biopsy, inguinal lymph node biopsy, lumbar puncture, MRI, 


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MRS and DTI. Detailed procedures will be outlined in specific site Standard Operational 
Procedures. Sampling will occur at baseline before vaccination and at Week 50, Week 72 and 
Week 96. Due to scheduling logistics, optional procedures may be performed up to one week 
before or after scheduled time points.
Lumbar puncture will be performed by one of the trained research physicians using an atraumatic 
(Sprotte®) needle to decrease risk of headache using an SOP established at SEARCH.
Sigmoidoscopy will be performed by qualified personnel at 
Lymph node biopsy and leukapheresis will be performed by qualified personnel at either 
All brain imaging will be performed at 
. MRI and MRS are non-invasive 
methods to detect brain pathology (MRI) and to determine the in-vivo concentration of brain 
metabolites (MRS). Gadolinium will not be used for research purposes. DTI is a sensitive 
non-invasive magnetic resonance technique to analyze the 3-dimensional diffusion of water 
within brain tissue. The diffusion of water within the brain is highly dependent on the underlying 
microarchitecture of the surrounding tissue, which is affected by both normal physiological 
processes (such as aging) as well as local neuropathological disease processes, such as those seen 
with HIV infection. Several cell types are distinguishable with commonly measured metabolites, 
including neurons (N-acetyl-aspartate) and glial or inflammatory cells (myoinositol and choline), 
often measured as ratios to brain creatine (Talos, 2006). HIV infection is associated with 
increased myoinositol/creatine and choline/creatine ratios, indicating direct effects on microglial 
cell concentrations (Chong 1993, Tarasow 2003). 
For female subjects of childbearing potential, pregnancy tests will be performed and pregnant 
women will be excluded from all of the optional procedures.
Brain imaging procedures (MRI, MRS and DTI) and tissue biopsies are part of the procedures in
the RV254 cohort. The results of these tests will be analyzed as part of the RV254 parent 
protocol. They are not part of this study endpoints and will only be reported if relevant to this 
study.
5.1.12.
Visit Windows
The maximum screening period is from Day -45 to Day -3.
For the study visits, following windows will be allowed as indicated:

Visit 3: Day 29 ± 5 days

Visit 4: Day 85 ± 7 days (second vaccination)
PPD
PPD
PPD


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
Visit 5*: Visit 4 + 3 days (Day 88) ± 2 days (for the gene array testing ideally the visit 
should take place exactly at +3 days after the vaccination)

Visit 6*: Visit 4 + 28 days (Day 113) ± 5 days

Visit 7: Day 169 ± 7 days (third vaccination)

Visit 8*: Visit 7 + 14 days (Day 183) ± 5 days

Visit 9*: Visit 7 + 28 days (Day 211) ± 5 days

Visit 10*: Visit 7 + 84 days (Day 253) ± 5 days

Visit 11: Day 337 ± 14 days (fourth vaccination)

Visit 12*: Visit 11 + 3 days (Day 340) ± 2 days (for the gene array testing ideally the visit 
should take place exactly at +3 days after the vaccination)

Visit 13*: Visit 11 + 14 days (Day 351) ± 5 days

Visit 14: Day 407 ± 5 days

Visit 15: Day 421 ± 7 days (start of ATI)

Visit 16-21: Days 428-446: visits are to be scheduled twice a week at least 2 days apart; all 
visits counted relative to start of ATI at Visit 15

Visit 22: Days 449 ± 2, counted relative to start of ATI at Visit 15

Visit 23-26, 26a: Days 452-467: visits are to be scheduled twice a week at least 2 days apart; 
all visits counted relative to start of ATI at Visit 15

Visit 27: Days 470 ± 2, counted relative to start of ATI at Visit 15

Visit 28-44: Days 477 - 589 ± 3, all visits counted relative to start of ATI at Visit 15

Visit 45-50: Days 603-673 ± 5 days, all visits counted relative to start of ATI at Visit 15
*If a subject is not vaccinated on the given day of vaccination, the timings of the 
post-vaccination visits (see Table 4) will be determined relative to the actual day of vaccination.
If a vaccination visit window is missed due to a study pause (see Section 5.3.2.1), vaccination 
will be assessed on a case-by-case basis upon discussion between investigator and sponsor.
5.2.
Vaccination Discontinuation
Under certain circumstances, a subject will be terminated from participating in further 
vaccinations. These specific experiences include:

Investigator’s clinical judgment is that it is in the best interest of the subject 

Pregnancy

Type 1 hypersensitivity associated with vaccination


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
Grade 3 or 4 AE that is vaccine related, except for grade 3 solicited local and systemic 
reactions.

Serious concomitant illness that is not expected to resolve prior to the next scheduled 
vaccination (according to the investigator's judgement)

Treatment 
with 
chronic 
systemic 
glucocorticoids 
(e.g. 
prednisone) 
or 
other 
immunomodulators other than non-steroidal anti-inflammatory drugs. Inhaled, topical, and 
short course oral steroid use is not exclusionary

Need for concomitant vaccine that requires discontinuation (see Section 3.2.3)

Missing any study vaccination by more than 3 weeks from the scheduled vaccination day

Participant’s request
Subjects who are discontinued from additional study vaccinations will continue to be followed 
according to the schedule to further evaluate safety and monitor adverse experiences through 
Week 50 (Stage 1), unless consent is withdrawn. The investigator will consider the reason for 
discontinuation of vaccination and the condition of the subject in determining if the blood 
collection plan should be altered and, in cases requiring a reduction of blood collection, priority 
is given to laboratory safety over immunogenicity or other study objectives. Subjects who 
receive less than 4 study injections will not undergo ARV ATI (Stage 2). All study procedures 
scheduled for study end (Week 96) will be performed at Week 50, which will be the final study 
visit for those subjects.
5.3.
Study Discontinuation
A subject will be discontinued from the study entirely if:

Repeated failure to comply with protocol requirements

Decision by the study sponsor or the PI to stop or cancel the study

Decision by local regulatory authorities and/or IRBs to stop or cancel the study

Subject’s request
5.3.1.
Early Discontinuation or Withdrawal of Subjects
Each subject has the right to withdraw from the study at any time for any reason without 
affecting the right to treatment by the investigator. The investigator should make an attempt to 
contact subjects who did not return for scheduled visits or follow-up. Although the subject is not 
obliged to give reason(s) for withdrawing prematurely, the investigator should make a reasonable 
effort to ascertain the reason(s) while fully respecting the subject’s rights.


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Subjects who wish to withdraw consent from participation in the study will be offered a single 
exit visit for safety follow-up prior to formal withdrawal of consent. They have the right to 
refuse. 
For those subjects who are unable to continue participation in the study, but who do not 
withdraw consent, an exit visit will be conducted (refer to SOE for list of procedures to be 
conducted).
The investigator also has the right to withdraw a subject, e.g. because of worsening health status, 
intercurrent illness, AEs, or pregnancy (for pregnancy follow-up see safety Section 4.1.9). The 
sponsor reserves the right to request the withdrawal of a subject due to protocol violations, or 
administrative or other reasons.
Any unnecessary withdrawal should be avoided. Should a subject be withdrawn, all efforts 
should be made to complete and report the observations as thoroughly as possible. Whenever a 
subject is withdrawn from the study, independent of the reason, a final evaluation must be 
completed for that subject and the major reason for which the subject was withdrawn must be 
stated. All documentation concerning the subject must be as complete as possible. 
5.3.2.
Premature Termination of the study
The sponsor reserves the right to discontinue the study for safety, ethical, or administrative 
reasons. Should the study be discontinued, no further vaccinations will be administered but 
subjects who are ongoing at the time of discontinuation will be followed through the remainder 
of their follow up visits.
5.3.2.1.
Study Pausing Rules
If a dose of vaccine is considered, by PSRT review, to raise significant safety concerns, all 
enrollment and vaccinations will be suspended until recommendations are issued. The AEs that 
may lead to a safety pause or prompt PSRT AE review are summarized below in Table 6. These 
study holding rules apply to AEs/SAEs occurring up to 4 weeks after the last vaccination.


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Table 6:
AE Notification and Safety Pause/AE Review Rules1
(S)AE and 
Relationship2
Severity
Site Principal Investigator Action
PSRT/DSMC Action
SAE, related
Any 
grade
Phone Study Responsible Physician or designee 
AND fax or email SAE form to Global Medical 
Safety Office, immediately and within 24 h
Immediate vaccination pause 
for PSRT review of safety 
data
SAE, not 
related
Grade 5
Phone Study Responsible Physician or designee 
AND fax or email SAE form to Global Medical 
Safety Office, immediately and within 24 h
PSRT review and 
consideration of pause
AE, related
Grade 3 
or
Grade 4
Phone Study Responsible Physician immediately 
and within 24 h
PSRT review and 
consideration of pause
≥3 subjects with 
a similar related 
AE3
Grade 3 
or
Grade 4
Not applicable
Immediate vaccination pause 
for DSMC review of safety 
data
The telephone number of the Study Responsible Physician (and designee) is in the Contact Information 
page(s). The Study Responsible Physician (or designee) is responsible for the immediate notification of 
PSRT/DSMC members and coordination of a PSRT/DSMC meeting.
1
Applicable for AEs/SAEs occurring up to 4 weeks after the last vaccination. For a Grade 3/4 laboratory-related 
AE, the test must be repeated at least once, within 48 hours of the site becoming aware of the abnormal value. 
PSRT evaluation for consideration of a pause will proceed without waiting for repeat testing. Conduct of 
DSMC review will require a confirmation of the laboratory test within 48 hours.
2
Related: very likely, probably, or possibly related to the study vaccine; not related: doubtful or not related to 
the study vaccine. 
3
Applicable for the following related AEs:
- All Grade 4 AEs (regardless of duration)
- Grade 3 unsolicited AEs (regardless of duration)
- Grade 3 solicited AEs (only if persisting for longer than 72 hours)
After each DSMC review of a similar AE, the DSMC will indicate the conditions under which they require 
further notification and/or review of the subsequent similar AEs.
Vaccinations for an individual subject may be suspended for safety concerns other than those 
described in the table, at the discretion of the investigator if he/she feels the subject’s safety may 
be threatened. The investigator may ask for a PSRT meeting to be held for any single event or 
combination of multiple events which, in his/her professional opinion, jeopardize the safety of 
the subjects or the reliability of the data.
Vaccinations for the study may be suspended for safety concerns other than those described in 
the table, or before pause rules are met, pending DSMC review, if, in the judgment of the PSRT, 
subject safety may be threatened.
For events in the table above, the investigator notifies the sponsor’s study responsible physician 
(or designee) immediately, and in all cases within 24 hours at the latest after the site observes, or 
is notified of, the AE, and the study responsible physician (or contacted sponsor’s representative) 
then notifies the PSRT immediately. If the case(s) is (are) deemed to fulfill the potential holding 


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rules, as specified in Table 6, the PSRT will convene within one business day to review these 
AEs. The PSRT will review and determine disposition, including whether the DSMC needs to 
review the event(s).
If a study pause is triggered by the PSRT, all enrollment and vaccinations will be held until 
review by the PSRT or DSMC is complete. Resumption of enrollment and study treatment may 
be determined by the PSRT or DSMC (in consultation with the FDA, if required) following a 
cumulative review of the available safety data as outlined in the charter. The clinical sites will be 
allowed to resume activities upon receipt of a written notification from the sponsor. As needed, 
the appropriate regulatory authorities will be informed in writing of the decision by the PSRT 
and/or DSMC to resume or discontinue study activities. The site is responsible for notifying their 
IRB according to local standards and regulations. The sponsor is responsible for notifying the 
FDA. 
5.4.
Laboratory Evaluations
Sample handling will be performed according to site-specific SOPs and MoPH guidelines.
The total number of blood draws and total volume of blood that will be collected from each 
subject is presented in Table 4 and Table 5. Time points for optional procedures are also 
presented in the tables. Laboratory testing performed for research purposes will be sent to 
laboratories in Thailand and abroad following the table in Appendix 16.4. Changes to this 
appendix will not constitute a protocol amendment but will be adapted through the lab manual.
In case a grade 3 or grade 4 laboratory abnormality, or any laboratory abnormality accompanied 
by clinically relevant signs or symptoms occurs (from the baseline visit onwards), a confirmatory 
test should be performed within 48 hours after the results have become available. After that, 
laboratory tests will be repeated weekly until values are resolved or stable.
All biological samples must be collected in the appropriate manner. The investigator will ensure 
that the personnel and laboratory under his/her supervision comply with these requirements. 
Further details on shipment, handling, and storage of the samples are provided in the Specimen 
Handling Guidelines.
Any residual samples will be stored at the sponsor designated laboratories in Thailand (includes 
) for a
maximum of 5 years. Any future analyses conducted on the samples will maintain subject 
anonymity. Subjects will have to provide their approval for long-term storage of their biological 
specimens. Subjects will have the right to opt out of having their biological specimens stored 
once all analyses described in the Informed Consent Form are completed. Opting out of this 
procedure does not impact the subject’s ability to participate in the study. 
PPD


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If it is necessary to store biological samples beyond 5 years, consent will be obtained from the 
subjects, the appropriate IRBs, and the Thai Ministry of Public Health (MoPH) Ethics 
Committee. If the investigators are unable to contact subjects, then permission for continued 
storage may be requested from the IRB without subject consent. This information will be 
provided to subjects in the consent forms that they sign at study enrollment.
5.5.
Potential Risks and Benefits
5.5.1.
Risks Related to Vaccines
Subjects may exhibit general signs and symptoms associated with administration of a vaccine or 
placebo vaccination, including fever, chills, rash, aches and pains, nausea, headache, dizziness 
and fatigue. These side effects will be monitored, but are generally short term and do not require 
treatment. 
Subjects may have an allergic reaction to the vaccination. An allergic reaction may cause a rash, 
hives or even difficulty breathing. Severe reactions are rare. To minimize this risk, those with 
allergies to or have had an allergic reaction to vaccines, vaccine products, neomycin, 
streptomycin, eggs, or egg products, will not be allowed to participate in this study. Subjects will 
be observed at the study site for 30 minutes after injection of the study products to monitor the 
development of any acute reactions. Vital signs will be taken before vaccination and at least 30 
minutes post-vaccination. Medications shall be available in the clinic to treat serious allergic 
reactions. 
The effect of this vaccine on a fetus or nursing baby is unknown, so female subjects of child 
bearing potential will be required to agree to use birth control for sexual intercourse beginning 
prior to the first vaccination and continuing at least 3 months after the final vaccine/placebo 
vaccination. Women who are pregnant or nursing will be excluded from the study. 
5.5.2.
Risk of Myo/Pericarditis
The MVA vaccine used in this study is related to the vaccine to prevent smallpox. A very small 
number of people who received the smallpox vaccine developed myocarditis or pericarditis. The 
number of people who had these problems was very small (96 people out of 666,712 who 
received the vaccine). Investigators do not expect these side effects from the MVA vaccine 
because it is an attenuated virus and it cannot replicate. Myocarditis has NOT been reported with 
previous MVA use and has not been seen in Phase 2/3 clinical studies cited in this protocol. 
Subjects will be actively screened to exclude pre-existing cardiac concerns and monitored for 
cardiac complications based on clinical symptoms.


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5.5.3.
Risks Related to Blood Draws
Blood drawing may cause pain, bruising, and, rarely, infection at the site where the blood is 
taken.
5.5.4.
Risk Related to ATI
As this is an exploratory study, all of the risks of ATI may not yet be known. However, the 
frequent monitoring of clinical, immunological, and virological status is designed to minimize 
possible risk to subjects from rebound viremia. If rebound viremia occurs, ART will be restarted 
when HIV viral load rises >1,000 copies/ml, a level and duration of viremia that is expected to 
have minimal, if any, impact on the subject’s CD4+ count and would be expected to have no 
clinical manifestations or symptoms to the subject. Moreover, the safety of this approach is 
proven in a recent study of chronically infected adults, many of who had low pre-ART CD4+
count. With twice-weekly viral load monitoring, rapid resumption of ART and viral suppression 
was demonstrated. No one had clinical adverse events due to ATI and the CD4+ count remained 
stable (Rothenberger 2015).
If subjects develop viremia, there is a risk of transmitting HIV to sexual partners. All subjects 
will be notified of this risk and to use condoms for any sexual contact during participation in the 
study.
There is a risk that subjects could develop HIV resistant virus after ATI. Changing all subjects to 
a PI-based ART regimen is designed to minimize this risk. If HIV resistance develops it could 
potentially lead to a higher risk of treatment failure or death during future ART. To detect and 
address any possible HIV resistance, an HIV genotype test will be done on all patients with
rebound viremia before restarting ART. The patient’s ART regimen will be adjusted to 
compensate for any resistance discovered. 
5.5.5.
Unknown Risks
There may be other serious risks that are not known.
Subjects may believe that this vaccine provides therapeutic benefit for HIV infection. They will 
receive extensive counseling throughout the study to address this potential problem. It is not 
known if the study vaccines increase or decrease the chance of HIV viral rebound after ARV 
ATI.
5.5.6.
Potential Benefits
There is no direct benefit to the subject for participation in this clinical study. Although study 
subjects may benefit from clinical testing and physical examination, they may receive no direct 
benefit from participation. Others may benefit from knowledge gained in this study that may aid 
in the development of future HIV treatment methods. 


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As explained in Section 5.5.5, it is not known if the study vaccines increase or decrease the 
chance of HIV viral rebound after ARV ATI.
6.
SAFETY ASSESSMENTS AND REPORTING
6.1.
Definitions
6.1.1.
Adverse Event Definitions and Classifications
Adverse Events
An AE is any untoward medical occurrence in a clinical study subject administered a medicinal 
(investigational or non investigational) product. An AE does not necessarily have a causal 
relationship with the treatment. An AE can therefore be any unfavorable and unintended sign 
(including an abnormal finding), symptom, or disease temporally associated with the use of a 
medicinal (investigational or non investigational) product, whether or not related to that 
medicinal (investigational or non investigational) product. (Definition per International 
Conference on Harmonisation [ICH])
This includes any occurrence that is new in onset or aggravated in severity or frequency from the 
baseline condition, or abnormal results of diagnostic procedures, including laboratory test 
abnormalities.
Note: The sponsor collects AEs starting with the signing of the ICF to the final visit (see 
Section 6.3.1).
Serious Adverse Events
An SAE based on ICH and EU Guidelines on Pharmacovigilance for Medicinal Products for 
Human Use is any untoward medical occurrence that at any dose:

Results in death

Is life-threatening (The subject was at risk of death at the time of the event. It does not refer 
to an event that hypothetically might have caused death if it were more severe.)

Requires inpatient hospitalization or prolongation of existing hospitalization

Results in persistent or significant disability/incapacity

Is a congenital anomaly/birth defect

Is Medically Important*
*Medical and scientific judgment should be exercised in deciding whether expedited reporting is 
also appropriate in other situations, such as important medical events that may not be 
immediately life threatening or result in death or hospitalization but may jeopardize the subject 


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or may require intervention to prevent one of the other outcomes listed in the definition above. 
These should usually be considered serious.
A suspected transmission of any infectious agent via a medicinal product is always considered as 
an important medical event, ie, an SAE.
If a serious and unexpected AE occurs for which there is evidence suggesting a causal 
relationship between the study vaccine and the event (e.g. death from anaphylaxis), the event 
will be reported as a suspected unexpected serious adverse reaction (SUSAR) by the sponsor to 
Health Authorities and by the investigator to the IRB according to regulatory and local 
requirements.
Adverse Reactions
An adverse reaction is an adverse event judged to be related to study vaccine. An AE is 
considered associated with study vaccine if the attribution is possibly, probably, or very likely 
related by the definitions listed in Section 6.3.4.
Unlisted (Unexpected) Adverse Event/Reference Safety Information
An AE is considered unlisted if the nature or severity is not consistent with the applicable 
product reference safety information. For Ad26.Mos.HIV and MVA-Mosaic, the expectedness of 
an AE will be determined by whether or not it is listed in the respective IBs.
6.1.2.
Rebound in Viral Replication, Study Endpoint not Considered as 
AE/ADR
Rebound in viral replication post ARV ATI (HIV RNA>50 copies/ml) will not be considered an 
AE/SAE as it is a study primary efficacy endpoint which is captured in the clinical study 
database and will be analysed as such.
Rebound in viral replication post ARV ATI (HIV RNA>50 copies/ml) will not be reported as 
individual expedited safety reports to IRB/ethical committees (ECs) or to Health Authorities 
during the course of the study. Such occurrences will be analysed according to treatment group 
allocation and descriptive results made available to IRB/ECs and to Health Authorities with the 
final analyses of the study results in the clinical study report. 
6.1.3.
Surveillance, Reporting, and Documentation of Adverse Events
The recording of AEs is an essential part of study documentation. The investigator is responsible 
for documenting all AEs as follows:
At each visit, all AEs, either observed by the investigator or one of his/her clinical collaborators 
or reported by the subjects spontaneously or in response to a direct question will be evaluated by 
the investigator or designee. All AEs will be recorded on eCRFs. Rebound in viral replication 


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post ARV ATI (HIV RNA>50 copies/ml) will not be considered an AE/SAE (See 
Section 6.1.2.).
As a consistent method to find out about AEs, the investigator should use a non-leading question, 
such as:
‘Have you felt different since receiving the study vaccine or since the previous visit?’
The subjects will be instructed to contact the investigator immediately should they experience 
any signs or symptoms they perceive as serious during the period extending from the first 
study-specific procedure up to and including 6 months after the last administration of the study 
agent.
6.2.
Special Reporting Situations
Safety events of interest of a sponsor study vaccine that may require expedited reporting and/or 
safety evaluation include, but are not limited to:

Overdose of a sponsor study vaccine

Suspected abuse/misuse of a sponsor study vaccine

Inadvertent or accidental exposure to a sponsor study vaccine

Medication error involving a sponsor product (with or without subject/patient exposure to 
the sponsor study vaccine, e.g. name confusion)
Special reporting situations should be recorded in the eCRF. Any special reporting situation that 
meets the criteria of an SAE should be recorded on the SAE page of the eCRF.
6.2.1.
Reporting Requirements to the Local IRB
The site PI will be responsible for providing all Safety Reports and reporting all study pauses, 
social impact, and major deviations to the local regulatory authority, such as a local IRB, and any 
country-specific regulatory agencies, in a timely manner according to the institution’s guidelines.
6.3.
Procedures
6.3.1.
All Adverse Events
All AEs and special reporting situations, whether serious or non-serious, will be reported and 
recorded on the eCRFs by the study site from the time a signed and dated ICF is obtained until 
completion of the subject’s last study-related procedure (ie, Week 96) and may include 
additional contact for follow-up of safety. eCRFs will be completed by the research staff as soon 
as possible after availability of the data.


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Rebound in viral replication post ARV ATI (HIV RNA>50 copies/ml) will not be considered an 
AE/SAE as it is a study endpoint and will be documented in the specific eCRF pages.
All SAEs must be reported to the sponsor during the entire study period using the SAE Form. 
SUSARs are reported even after the study is over, if the sponsor, DSMC or investigator becomes 
aware of them. The sponsor will evaluate any safety information that is spontaneously reported 
by the investigator beyond the time frame specified in the protocol.
The investigator will monitor and analyze study data including all AE and laboratory data as they 
become available and will make determinations regarding the severity of the adverse experiences 
and their relation to study vaccine. AEs will be deemed either related to study vaccine or not 
related to study vaccine, according to Section 6.3.4. 
Post-injection reactogenicity (PIR) includes solicited AEs related to study vaccine and therefore 
must be reported as such. The investigator or designee must review both PIR and other AE data 
to insure prompt and complete identification of all events that require expedited reporting as 
SAEs, invoke study pausing rules or are other serious and unexpected events.
All AEs, regardless of seriousness, severity, or presumed relationship to study vaccine, must be 
recorded using medical terminology in the source document and the eCRF. Whenever possible, 
diagnoses should be given when signs and symptoms are due to a common etiology (e.g. cough, 
runny nose, sneezing, sore throat, and head congestion should be reported as “upper respiratory 
infection”). Investigators must record in the eCRF their opinion concerning the relationship of 
the AE to study vaccine. All measures required for AE management must be recorded in the 
source document and reported according to sponsor instructions.
The sponsor assumes responsibility for appropriate reporting of AEs to the regulatory authorities. 
The sponsor will also report to the investigator (and the head of the investigational institute 
where required) all SUSARs. The investigator (or sponsor where required) must report SUSARs
to the appropriate EC/IRB that approved the protocol unless otherwise required and documented 
by the EC/IRB. A SUSAR will be reported to regulatory authorities unblinded. Participating 
investigators and IRB will receive a blinded SUSAR summary, unless otherwise specified. 
Subjects will be provided with a wallet card referring to 24-hour contact numbers that can be 
used by subjects and medical site staff.
6.3.1.1.
Post-Vaccination Reactions Occurring Immediately After Each 
Vaccination
As described previously, following each vaccine administration, subjects will be observed for 
reactogenicity in the clinic for a minimum of 30 minutes. Vital signs will be taken and qualified 
study personnel will evaluate for any signs or symptoms of local or systemic reactions.


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Possible local reactions that should be recorded include erythema and induration (measured 
using the ruler included with the memory aid that will subsequently be provided to the subject),
pain/tenderness, or warmth at the injection site. In addition, systemic symptoms to be noted 
include fever (≥38.0ºC, non-axillary), fatigue, headache, myalgia, arthralgia, chills, nausea, 
vomiting, rashes, and itching. For life-threatening allergic reactions that occur immediately post 
vaccination, site-specific procedures are in place for handling such emergencies.
6.3.1.2.
Local Reactions Occurring Within Seven Days After Each Vaccination
Memory Aid Completion
Subjects will be provided with a ruler to measure erythema, induration or other observable 
reactions.
The subject will be asked to note occurrences of erythema and induration (measured using the 
ruler included with the memory aid), and pain/tenderness, itching, swelling, or warmth at the 
injection sites daily for 8 days including the day of vaccination. These occurrences should be 
recorded on the memory aid provided to serve as a reminder to the subject for the next clinic visit 
(the memory aids are not considered source documents). 
6.3.1.3.
Systemic Reactions Occurring Within Seven Days Post-Vaccination
Memory Aid Completion
Subjects will be instructed on how to record daily temperature using a thermometer provided for 
home use. Subjects should record oral temperature 6 hours post vaccination, and then daily for 
the next 7 days. 
Subjects will also be instructed on how to record and grade daily symptoms on the memory aid
for 8 days post-vaccination (including the day of vaccination).
The subject will also be asked to record the following occurrences 6 hours post vaccination and 
then daily for 7 days on the memory aid:

temperature (fever ≥38.0°C, non-axillary)

fatigue

headache

myalgia

arthralgia

chills

nausea

vomiting


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
rashes 

itching (general and local)
Post-Vaccination Contact
Between 24 and 72 hours after each vaccination, the investigator or designee will contact the 
subjects to obtain local and systemic reaction data and to assess clinical status as described in 
Section 5.1.4. 
Instructions to Subjects Regarding Unusual or Severe Signs or Symptoms
Subjects will be instructed to call the specified study personnel immediately (information 
included on the memory aid and/or appointment and/or emergency card) if any unusual or severe 
sign or symptom appears after vaccination. Subjects with unusual, moderate, or severe sign 
and/or symptoms post vaccination will be asked to return to the clinic for evaluation. They will 
be followed up clinically until resolution of symptoms. Contacts will be arranged as needed. 
Subjects will be instructed to inform treating medical personnel of their participation in the study 
and provide them with the contact information on the emergency card provided.
6.3.2.
Serious Adverse Events
All SAEs occurring during the study must be reported to the appropriate sponsor contact person 
by study-site personnel within 24 hours of their knowledge of the event.
Information regarding SAEs will be transmitted to the sponsor using the SAE Form, which must 
be completed and signed by a physician from the study site, and transmitted to the sponsor 
within 24 hours. The initial and follow-up reports of a SAE should be made by facsimile (fax)
and/or email.
All SAEs that have not resolved by the end of the study, or that have not resolved upon 
discontinuation of the subject’s participation in the study, must be followed until any of the 
following occurs:

The event resolves

The event stabilizes

The event returns to baseline, if a baseline value/status is available

The event can be attributed to agents other than the study vaccine or to factors unrelated to 
study conduct

It becomes unlikely that any additional information can be obtained (subject or health care 
practitioner refusal to provide additional information, lost to follow up after demonstration 
of due diligence with follow-up efforts)


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Study Protocol 
Amendment 3 
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Status: Approved, Date: 11 October 2017

Any event requiring hospitalization (or prolongation of hospitalization) that occurs during 
the course of a subject’s participation in a study must be reported as an SAE, except 
hospitalizations for the following:

Hospitalizations not intended to treat an acute illness or AE (e.g. social reasons such as 
pending placement in long-term care facility)

Surgery or procedure planned before entry into the study (must be documented in the 
eCRF). Note: Hospitalizations that were planned before the signing of the ICF, and where 
the underlying condition for which the hospitalization was planned has not worsened, will 
not be considered SAEs. Any AE that results in a prolongation of the originally planned 
hospitalization is to be reported as a new SAE.
The cause of death of a subject during the entire study period, whether or not the event is 
expected or associated with the study vaccine, is considered an SAE and must be reported.
6.3.3.
Reporting requirements to the Local IRB
The site PI will be responsible for providing all Safety Reports and reporting all SAEs, study 
pauses, social impact, and major deviations to the local regulatory authorities, such as a local 
IRB, and any country-specific IRBs, in a timely manner according to the institution’s guidelines 
and to local regulations. 
6.3.4.
Causality Assessment of Study Vaccines to Adverse Events
Every effort should be made by the investigator to explain any AE and assess its potential causal 
relationship, ie, to administration of the study vaccine or to alternative causes (e.g. natural 
history of the underlying diseases, concomitant therapy). This applies to all AEs, whether serious 
or non-serious.
The investigator will use the following guidelines to assess the causal relationship of an AE to 
study vaccine:
Not related:
An AE that is not related to the use of study vaccine.
Doubtful:
An AE for which an alternative explanation is more likely, e.g. 
concomitant drug(s), concomitant disease(s), or the relationship in time 
suggests that a causal relationship is unlikely.
Possible:
An AE that might be due to the use of study vaccine. An alternative 
explanation, e.g. concomitant drug(s), concomitant disease(s), is 
inconclusive. The relationship in time is reasonable; therefore, the causal 
relationship cannot be excluded.


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Status: Approved, Date: 11 October 2017
Probable:
An AE that might be due to the use of study vaccine. The relationship in 
time is suggestive (e.g. confirmed by dechallenge). An alternative 
explanation is less likely, e.g. concomitant drug(s), concomitant 
disease(s).
Very likely:
An AE that is listed as a possible adverse reaction and cannot be 
reasonably explained by an alternative explanation, e.g. concomitant 
drug(s), concomitant disease(s). The relationship in time is very 
suggestive (e.g. it is confirmed by dechallenge and rechallenge).
An AE is considered associated with study vaccine if the attribution is possibly, probably, or 
very likely related. 
6.3.5.
Severity of Adverse Events
All recorded AEs will be coded for severity using the Division of AIDS (DAIDS) Table for 
Grading the Severity of Adult and Pediatric Adverse Events (DAIDS AE Grading Table), 
Version 
2.0 
dated 
November
2014
found 
on 
the 
website 
http://rsc.tech-
res.com/Document/safetyandpharmacovigilance/DAIDS_AE_GRADING_TABLE_v2_NOV201
4.pdf.
For AEs not identified in the grading table the following guidelines will be applied: 
Mild
Grade 1
Symptoms causing no or minimal interference with usual 
social & functional activities
Moderate
Grade 2
Symptoms causing greater than minimal interference with 
usual social & functional activities
Severe
Grade 3
Symptoms causing inability to perform usual social & 
functional activities
Potentially
Life-Threatening
Grade 4
Symptoms causing inability to perform basic self-care 
functions OR medical or operative intervention indicated to 
prevent permanent impairment, persistent disability or 
death
Fatal
Grade 5
For any AE where the outcome is death, the severity of the 
AE is classified as Grade 5
The clinical research team will ascertain accurate recording of all AEs during the study. AE 
eCRFs will be completed by the research staff as soon as possible after availability of the data. 
The clinical investigators will monitor and analyze study data including all AE and laboratory
data as they become available and will make determinations regarding the severity of the adverse 
experiences and their relation to study product.


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PIR are solicited AEs and related to study and therefore must be reported as such. The PI or 
designee must review both PIR and other AE data to insure prompt and complete identification 
of all events that require expedited reporting as SAEs, study pause rules or other serious and 
unexpected events. 
The clinical research team will follow all recorded AEs to resolution or study completion, 
whichever occurs first. 
6.3.6.
Follow-Up of Ongoing Adverse Events and Assessment of Outcome
6.3.6.1.
Follow-Up of Non-Serious Adverse Events
Non-serious AEs already documented in the eCRF at a previous assessment and designated as 
‘ongoing’ should be reviewed at subsequent visits. If the event has resolved, the documentation 
in the eCRF should be completed. If the frequency or severity of a non-serious AE changes 
significantly, a new record of the AE has to be started. If the AE becomes serious, the procedures 
for reporting of SAEs have to be followed (Section 6.3.2).
Ongoing non-serious AEs will be monitored until the Week 96 visit.
Outcome will be assessed as:
1
Recovered or Resolved
2
Recovering or Resolving
3
Not Recovered or Not Resolved 
4
Recovered or Resolved with Sequelae
5
Fatal
6
Unknown
Abnormal laboratory values will be followed up until they have returned to normal, stabilized, or 
a satisfactory explanation has been provided.
6.3.7.
Treatment of Adverse Events
Treatment of any AE is at the sole discretion of the investigator and according to current 
available best treatment. The applied measures should be recorded in the eCRF.
6.3.8.
Pregnancy
All initial reports of pregnancy must be reported to the sponsor by the study-site personnel 
within 24 hours of their knowledge of the event using the appropriate pregnancy notification 
form. Abnormal pregnancy outcomes (e.g. spontaneous abortion, stillbirth, and congenital 
anomaly) are considered SAEs and must be reported using the SAE Form.


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Status: Approved, Date: 11 October 2017
Any subject who becomes pregnant during the study will complete all procedures scheduled for 
study end at the time the pregnancy is diagnosed. If the subject was scheduled to receive 
additional vaccine or placebo, these injections will not be administered after pregnancy has been 
determined. Pregnant subjects will be offered enrollment in RV412 for continued observation 
(see Section 4.1.9).
Because the effect of the study vaccine on sperm is unknown, pregnancies in partners of male 
subjects included in the study will be reported by the study-site personnel within 24 hours of 
their knowledge of the event using the appropriate consent and notification forms.
Follow-up information regarding the outcome of the pregnancy and any postnatal sequelae in the 
infant will be required to be sent to the sponsor.
6.4.
Physical Examination
Physical examinations will be performed by the investigator or designated medically trained 
clinician. The time points of these examinations are specified in the SOE (Table 4 and Table 5). 
Any abnormalities or changes in severity noted during the review of body systems should be 
documented in the source document and recorded on the eCRF.
A new, clinically significant finding (in the opinion of the investigator) not noted at screening 
must be captured as an AE. In addition, resolution of any abnormal findings during the study will 
be noted in the source document and in the eCRF.
6.5.
Routine Safety Laboratory
Samples for routine safety laboratory parameters will be collected at the time points specified in 
Table 4 and Table 5. 
The following routine safety laboratory parameters will be determined:

Serum chemistry: electrolytes, glucose, BUN, creatinine, total/direct bilirubin, ALT, AST, 
GGT

Hematology safety parameters: hemoglobin, hematocrit, white blood cells (WBCs), WBC 
count, WBC differentiation, red blood cell (RBC) count, platelet count
All abnormal laboratory values will be reported as AEs if considered clinically significant. 
Changes in grade toxicity will be reported in aggregate analysis. 
6.6.
Vital Signs
Vital signs measurements will be performed at each clinical visit that includes a medical history 
and physical examination (according to time points provided in the schedule of evaluations).


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The following measurements will be performed: 

Heart rate (bpm), systolic blood pressure (mmHg) and diastolic blood pressure (mmHg) will 
be measured (sitting position). A confirmatory vital signs measurement can be performed if 
inconsistent with a prior measurement.

Body temperature (oral or tympanic) 

Respiratory rate (breaths per minute)
If any clinically significant changes in vital signs are noted, they must be reported as AEs and 
followed to resolution, or until reaching a clinically stable endpoint. 
6.7.
Electrocardiogram
Supine 12-lead ECGs will be performed at screening and at Week 16, and only repeated if 
symptoms warrant.
For 30 minutes prior to the ECG, subjects should refrain from meals, hot or cold beverages, 
strenuous exercise, and should remain in a room with a comfortable temperature. Each ECG 
should be obtained after the subject has been at rest for at least 5 minutes. All ECG interpretation 
will be provided by a cardiologist at a referral hospital along with cardiac consultation, if needed.
ECGs are to be taken before blood sampling. 
7.
PRODUCT QUALITY COMPLAINT HANDLING
A product quality complaint (PQC) is defined as any suspicion of a product defect related to 
manufacturing, labeling, or packaging, ie, any dissatisfaction relative to the identity, quality, 
durability, or reliability of a product, including its labeling or package integrity. A PQC may 
have an impact on the safety and efficacy of the product. Timely, accurate, and complete 
reporting and analysis of PQC information from studies are crucial for the protection of subjects, 
investigators, and the sponsor, and are mandated by regulatory agencies worldwide. The sponsor 
has established procedures in conformity with regulatory requirements worldwide to ensure 
appropriate reporting of PQC information; all studies conducted by the sponsor or its affiliates 
will be conducted in accordance with those procedures.
7.1.
Procedures
All initial PQCs must be reported to the sponsor by the study-site personnel within 24 hours after 
being made aware of the event.
If the defect is combined with an SAE, the study-site personnel must report the PQC to the 
sponsor according to the SAE reporting timelines. A sample of the suspected product should be 
maintained for further investigation if requested by the sponsor.


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7.2.
Contacting Sponsor Regarding Product Quality
The names (and corresponding telephone numbers) of the individuals who should be contacted 
regarding product quality issues are listed on the Contact Information page(s), which will be 
provided as a separate document.
8.
PROTOCOL SAFETY REVIEW TEAM AND DATA AND SAFETY 
MONITORING COMMITTEE
8.1.
Protocol Safety Review Team 
The PSRT will review all AEs on a regular and expedited basis as needed. In addition, the PSRT 
will review aggregate safety data reports on a regular basis. This team includes the following: 
Study Chair, Site PI, the Janssen Vaccines & Prevention B.V. Clinical Leader and study 
responsible physician and global medical safety officer. Additional members could include 
associate investigators, and senior clinical research nursing staff. A quorum is established with 
the Study Chair, the site PI, the study responsible physician. The PSRT will decide by consensus 
whether AEs should also be reviewed by the DSMC.
8.2.
Data and Safety Monitoring Committee
An independent DSMC will be set up. The DSMC will evaluate safety, tolerability, and efficacy
data from each cohort on an ongoing basis. The DSMC may review an individual SAE or it may 
choose to review AEs, SAEs, and laboratory and vital sign data. The DSMC may unblind any 
amount of safety information needed to conduct their assessment. The conclusions of the DSMC 
will be communicated to the investigators and the IRB/Ethics Committees and the national 
regulatory authorities in a blinded manner as appropriate. The sponsor agrees to abide by the 
decision of the DSMC and any directives issued by the national regulatory authorities, the IRBs 
or Ethics Committees. 
The committee will comprise of five members including one infectious diseases specialist, one 
HIV clinician, one HIV virologist, one biostatistician and one community representative. The 
committee will review data from the study at the following time points: 1) the first 12 enrolled 
subjects have completed 2 injections (Week 12) (Stage 1), and 2) the first 12 enrolled subjects 
complete Week 72 (12 weeks after ARV ATI) (Stage 2). All SAEs will be reviewed by the 
committee Additional meetings may be called at the discretion of the chair of the DSMC. 
In addition, the Community Advisory Board at the 
 will 
review and follow the progress of this study.
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9.
DATA EVALUATION AND ANALYSIS
9.1.
Analysis Populations
9.1.1.
Safety Population
All subjects who received at least 1 injection of Ad26.Mos.HIV or MVA-Mosaic, or placebo, 
and for whom any post-dose data is available will be included in the safety population.
9.1.2.
Immunogenicity Population
The immunogenicity population will consist of all subjects who received at least 1 dose of 
Ad26.Mos.HIV or MVA-Mosaic and who have at least 1 measured post-dose blood sample
collected.
9.1.3.
Efficacy Population
The primary efficacy population will consist of all subjects who undergo ARV ATI at Week 60
and interrupt at least one dose of ART (Stage 2), regardless of the time or outcome of treatment 
interruption. Additionally, as a sensitivity analysis, efficacy will be analyzed in the ITT 
population consisting of all subjects who received at least one injection (with subjects not 
undergoing ARV ATI counted as efficacy failures).
9.1.4.
Secondary and Exploratory Analyses
These analyses are exploratory and descriptive. The number of subjects included in each analysis 
will depend on the number of specimens available and the requirements for each specific testing 
protocol.
9.2. Primary Endpoints
1.
Safety: Vaccine related - grade 3 or greater reactogenicity and AE that are product related or 
probably product related
2.
Efficacy: HIV RNA <50 copies/ml at 24 weeks after ARV ATI. 
9.3. Secondary and Exploratory Measurements
This is an exploratory study. Therefore, the research assays will be disparate and exploratory in 
nature. Both cellular and humoral responses will be measured together with biomarkers and 
transcriptome analysis. These multiple assays will allow correlations that may reveal critical 
information on therapeutic vaccination and treatment interruption. Assays for the evaluation of 
immunogenicity will include but are not limited to:
1.
Frequency, magnitude and breadth of epitope recognition by ELISPOT, polyfunctinality of 
T cell responses, binding antibody to Env regions and neutralization of a variety of HIV-1
strains (tier 1, 2).


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Status: Approved, Date: 11 October 2017
2.
Cell-associated HIV RNA in total CD4+ T cells, and in the memory CD4 subsets
3.
Cell-associated HIV DNA (total, integrated and 2 LTR circles) in total CD4+ T cells, and 
the memory CD4 subsets
4.
Viral outgrowth and inducible HIV RNA in total CD4+ T cells, and the memory CD4 
subsets
5.
Single copy HIV RNA in samples with HIV RNA <50 copies/ml pre and post ARV ATI
6.
Frequency, severity and duration of acute retroviral syndrome following ARV ATI
7.
HIV specific immune responses in genital mucosal compartments and all immunogenicity 
assays.
8.
Analysis of the expression profiles and transcriptome analysis of sorted and/or unsorted 
lymphocytes and monocyte/macrophages in the different arms of the study.
9.3.1.
Assays That Will be Implemented for Immunogenicity:
Evaluation of immunogenicity is performed as described in the schedule of evaluations. Tissue 
samples will be evaluated in a similar fashion at Weeks 0, 50, 72 and 96 (or prior to ART 
resumption) in subjects who are willing to undergo optional invasive procedures.
9.3.1.1.
Humoral Immunogenicity
1.
Binding antibody to immunogen inserts and a panel of HIV envelope proteins representing 
the circulating HIV-1 strains. 
2.
Neutralizing antibody using a representative panel of tier 1 and tier 2 HIV strains with 
emphasis on the strain infecting the subjects and, if available, the pseudovirus expressing the 
subjects transmitted founder virus
3.
Functional antibody assays (ADCC as an example) using a representative panel of HIV 
strains with emphasis on the strain infecting the subjects and, if available the pseudovirus 
expressing the subjects transmitted founder virus.
9.3.1.2.
Cellular Immunogenicity
1.
IFN-gamma ELISPOT assays of PBMC using peptide pools representing the immunogen 
inserts and other HIV specific reagents available.
2.
Phenotypic and functional characterization of CD4 and CD8 T cells, B cells, 
monocyte/macrophages by flow cytometry, transcriptome analysis and T-cell receptor 
analysis.
9.3.1.3.
Other Immunological Evaluations
Evaluation of biomarkers and markers for viral expression and reservoir will be performed on 
stored samples, particularly those from the same time points as the immunogenicity evaluation. 


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Soluble markers of immune activation will be measured with high throughput immunoassay 
platforms (Luminex or similar platform).
9.3.1.4.
Viral Expression and Reservoir
These evaluations will be done on total CD4+ T cells, and tissue (if available) and sorted 
populations of interest. The assays that will be used will include but are not limited to:
1.
Cell-associated HIV viral RNA 
2.
Total, integrated HIV DNA and 2 LTR 
3.
Viral out-growth assay 
4.
TILDA
9.4.
Sample Size Consideration
This is an exploratory analysis of a unique group of early-treated subjects with extremely low 
HIV reservoir size and preserved immunity. The sample size of 18 in the vaccine arm and 9 in 
the placebo arm is based on feasibility in regards to number of subjects. The sample size is 
within the range of subjects recommended in the Code of Federal Regulations (CFR 312.21) for 
each of the products in this investigation and will allow evaluation of epitope enumeration 
between arms. It is estimated that the majority of subjects will proceed to ARV ATI. The rate of 
functional cure is unknown; but we expect that the proportion of subjects achieving drug-free 
remission of HIV will be similar or higher than the VISCONTI study (15%), because our study 
subjects have ART initiated earlier and they have exhibited lower HIV DNA. There will not be 
an adequate statistical power to test differences in post-treatment viremic controllers between 
arms. Placebo recipients are included for blinding, efficacy and safety purposes and will provide 
additional control specimens for immunogenicity assays. The study findings will be important in 
informing the effect of Ad26/MVA on HIV reservoir/immunity and functional cure. 
While mild to moderate vaccine reactions (local site and systemic responses) are expected, AEs 
that preclude further dose administration or more serious ones that would limit product 
development are not anticipated. With 18 subjects in the vaccine arm, the observation of 0 such 
reactions would result in a two-sided exact 95% confidence interval with an upper limit of 
18.50%.
9.5.
Data Analysis
Subject baseline characteristics will be summarized by study arm. Continuous variables will be 
expressed as median and IQR. Categorical variables will be presented as number and percentage. 
Proportion of subjects with AEs and subjects with HIV RNA <50 copies/mL post ARV ATI
between study arms will be compared using Fisher's exact test. Differences in the other outcomes 


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between treatment arms will be assessed using the non-parametric Mann Whitney U test. All 
hypothesis tests will be two-sided. Statistical significance will be taken at a level of 5 percent.
9.5.1.
Primary Objectives and Endpoints 
9.5.1.1.
Safety 
History and physical examination and standard laboratory assessments following each 
vaccination for safety and tolerability as follows:
1.
AEs during the course of the study
2.
Vaccine reactogenicity rates for individual and combined symptom subgroups for 1 week 
after each vaccination using a memory aid
3.
Laboratory evaluation will be performed every 12 weeks during the vaccine series, and 
2 weeks after the fourth vaccination (CBC only). During ARV ATI, serum chemistry will be 
performed every 12 weeks and hematology every 4 weeks
a.
Serum chemistry: electrolytes, BUN, creatinine, glucose, total/direct bilirubin, ALT, 
AST and GGT
b.
Hematology safety parameters: hemoglobin, hematocrit, WBCs, WBC count, WBC 
differentiation, RBC count, platelet count
9.5.1.2.
Efficacy
The proportion of patients with HIV RNA <50 copies/mL at all timepoints after ARV ATI as 
well as a 95% confidence interval will be reported for each treatment arm. 
9.5.1.3.
Clinical Outcomes
Clinical outcomes will be described in the two treatment arms, including:
1.
Change in CD4 count during ARV ATI.
2.
Time to reinitiating ART.
3.
Frequency, severity and duration of acute retroviral syndrome. 
4.
Frequency and characterization of HIV drug resistance in subjects that experience viral 
rebound during ARV ATI.
9.5.2.
Analysis Time Points
The final analysis will be performed once all subjects have completed their final study visit or 
discontinued earlier. Interim analyses may be performed prior to the final analysis. These 
analyses would occur in a group-unblinded manner, but no subject-level unblinding would occur.
ICS or ELISPOT will be performed on baseline and Week 26 samples and completed prior to the 
Week 60 visit to determine whether the study can proceed to Stage 2 (see Section 3.2.5).


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10.
STUDY-SPECIFIC MATERIALS
The investigator will be provided with the following supplies:

IBs for Ad26.Mos.HIV and MVA-Mosaic.

Pharmacy manual/site investigational product procedures manual

Laboratory manual

Randomization instructions

eCRF completion guideline

Master ICF

Memory aid

TOU

Rulers, thermometers

Subject wallet cards, including:

study number;

statement, in the local language, that the subject is participating in a clinical study;

investigator's name and 24-hour contact telephone number;

local sponsor's name and 24-hour contact telephone number (for medical staff only);

subject number;

information about who should be contacted in case of emergency.

Recruitment tools, as applicable
11.
ETHICAL AND LEGAL REQUIREMENTS
11.1.
General Requirements
The study will be performed according to this Study Protocol and in compliance with the 
Declaration of Helsinki, the guidelines of the International Conference on Harmonization (ICH) 
GCP and the respective local legal requirements including the following: US Code of Federal 
Regulations 45CFR Pt 46; 21CFR Pt 50, 21CFR Pt 56 and 21CFR Pt 312.
11.2. Institutional Review Board/Ethics Committee
Before the start of the study, the investigator will submit the Study Protocol, Subject 
Information, Informed Consent Form, Information on compensation for study-related injuries, 
and other study-related documents as required by applicable laws and regulations to the 
responsible IRBs
 
PPD


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for written approval.
The investigator will inform the IRB according to applicable laws and regulations about 
Amendments to the Study Protocol including, but not limited to, any new information that 
require an ethical reconsideration of the Study Protocol.
In addition to the written approval of the IRB the investigator should obtain a statement from the 
IRB confirming that the institution is composed and organized according to and adheres to GCP 
and applicable regulations.
Unless otherwise instructed by the IRB(s) or local regulation the investigator must submit to the 
IRBs defined above:

All subsequent Amendments to the Study Protocol, changes to the Informed Consent Form 
or revisions of other documents originally submitted for review

New or revised subject recruiting materials approved by the sponsor, if applicable

All subsequent changes of logistical or administrative aspects in submitted protocol 
documents (for information)

Serious and/or unexpected AEs occurring during the study, where required

New information that may affect adversely the safety of the subjects or the conduct of the 
study

New edition(s) of the IB and amendments/addenda

Annual update and/or request for re-approval, where required 

Date of study completion, where required

Close out Report
11.3. Regulatory Authorities
Before initiating the study, the sponsor will submit any required application to the regulatory 
authorities and obtain approval according to applicable laws and regulations. The sponsor will 
also inform the regulatory authorities about Amendments to the Study Protocol including, but not 
limited to, any new information that require an ethical reconsideration of the Study Protocol.
11.4. Subject Information and Informed Consent
The study informed consent from is included in a separate document file. It describes the IPs to 
be used and all aspects involved in protocol participation. The investigator will provide a copy of 
the approved informed consent to the subject and a signed copy will be maintained in the 
subject’s record file. Before a subject’s participation in the study, the investigators will obtain 
written informed consent from the subject, after adequate explanation of the aims, methods, 
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anticipated benefits, and potential hazards of the study and before any protocol-specific 
procedures or study medications are administered.
Consent will be obtained in the SEARCH clinic at the 
 
. In order to ensure that participation is voluntary and to demonstrate that refusal to 
participate does not have any effect on subject’s clinical care in the main protocol 
SEARCH010/RV254, consent will be obtained by a research staff who does not provide clinical 
care to that subject in the main protocol. Subjects will also be informed that the participation is 
voluntary and that subjects have the right to withdraw at any time without giving the reasons and 
without any disadvantages for their subsequent care. Subjects will confirm their consent in 
writing before study start and any study-specific procedure.
As part of the consent procedure, potential subjects will have to pass the TOU that includes 
10 questions (Appendix 16.3). The TOU will be given after the study staff have explained the 
study procedures, the subjects has read the information sheets and consent forms and asked any 
question, and before the subject signs the consent forms. Seven out of 10 correct responses will 
be considered a passing score, and questions 1, 2, and 5 must be answered correctly. Subjects 
may take the test up to 3 times to achieve a passing score.
11.5. Data Access and Protection
Confidentiality is extended to cover testing of biological samples in addition to the clinical 
information relating to participating subjects. The study protocol, documentation, data, and all 
other information generated will be held in strict confidence. No information concerning the 
study or the data will be released to any unauthorized third party without prior written approval 
of the sponsor.
The monitors, auditors, the IRB, and the regulatory authorities will be granted direct access to 
the subject’s original medical records for verification of clinical study procedures and/or data, or 
quality assurance reviews and audits without violating the confidentiality of the subject, to the 
extent permitted by the applicable law and regulations. By signing the written Informed Consent 
Form, subjects will authorize such access. Subjects should be informed about the purpose of the 
planned computer data processing and the publication of the data.
Disclosure of the subject’s identity will occur only in case of emergency to avoid health risks.
11.6. Data Coding
All subjects consented will be assigned an 8-digit Subject identification. The first four digits 
represent the study code, followed by a 4-digit number that will be assigned sequentially.
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11.7.
Future use and Storage of Blood Samples 
Each study subject will be asked to voluntarily consent for their blood samples to be stored for 
other research studies that may be done after this study is completed. In case the subject is 
unwilling to have their blood samples stored for future use, they can consent to participate in this 
study only, without having their blood samples stored for future testing. In this case, their blood 
samples will be destroyed after all the tests specified for this study have been concluded.
All samples for which consent has been obtained and for which additional material is available 
after study specified testing is complete will be stored for up to 5 years for future testing. All 
applicable approvals will be sought before any such samples are used for analysis not specified 
in the protocol or a protocol amendment approved by the IRB. In the event that the study team 
decides that there is scientific merit in additional testing or storage beyond 5 years, then 
permission for continued storage will be requested from the subjects, the relevant IRBs, and the 
Thai MoPH Ethics Committee. If subjects are not able to be contacted, then continued storage 
can occur with sole permission of the IRBs.
11.8.
Compensation 
Subjects will be compensated for time and inconvenience in accordance with the usual standards 
and legal obligations for compensation followed at the study site. Compensation amounts for 
study visits and optional procedures are similar to the main protocol SEARCH010/RV254 and 
will be paid in cash at the time of the visit or procedure. Applicable guidelines by the local IRB
for compensation of research subjects will be followed.
11.9.
Compliance with NIH Guidelines for Research Involving Products 
Containing Recombinant DNA
Because this study is evaluating products containing recombinant DNA, per NIH Guidelines for 
Research Involving Recombinant DNA Molecules, the study must be submitted to the site 
Institutional Biosafety Committee (IBC) and must be approved before participants are enrolled at 
each respective institution. Investigators at each site are responsible for obtaining IBC approval 
and periodic review of the research per NIH guidelines section IV-B07-b-(6) and 
section IV-B-2-b. IBC review and approval must be documented by the investigator and 
submitted as part of protocol registration for this study.
The NIH guidelines also require that human gene transfer studies conducted at or sponsored by 
institutions that receive NIH funds must be submitted to the NIH Office of Biotechnology 
Activities (OBA) for review by the Recombinant DNA Advisory Committee (RAC). The NIH 
guidelines create exceptions to RAC review for vaccines. Although the exception may apply 
because the study procedure involves electroporation of the DNA, the Protocol team will submit 
the application with the study concept proposal for RAC review and response.


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11.10. Institutional Biosafety Committee (IBC)
The 
 IBC is responsible for reviewing all research studies involving biological agents as 
described in the Biological Safety Manual. The 
 IBC reviews research, safety 
procedures, and protocols with the objective of safeguarding the public health and occupational 
health and providing environmental safeguards for the surrounding community. The 
 
IBC reviews, approves, and oversees projects in accordance with the responsibilities defined in 
the NIH Guidelines for Research Involving Recombinant DNA Molecules. The PI/Laboratory 
Director is responsible for submitting any procedures that fall under the NIH Guidelines and for 
completing the IBC Registration Form for Recombinant DNA prior to research initiation and 
annually thereafter. The form provides Investigators with an efficient means of notifying the
 IBC regarding NIH compliance and assists investigators when determining their 
compliance requirements.
12.
QUALITY CONTROL AND QUALITY ASSURANCE
12.1.
Monitoring
The sponsor will monitor this study in accordance with 21 CFR 312. Monitoring visits to the 
clinic will be made regularly to ensure that the study is carried out according to this Study 
Protocol and in compliance with GCP and applicable legal requirements. 
Source documents will be reviewed for verification of consistency with eCRF data. The 
investigator guarantees direct access to source documents for monitoring purposes. Source data 
verification will be performed in accordance with data protection regulations and guidelines. All 
information reviewed will be handled according to these rules and regulations.
The monitor will review each subject’s data as outlined in the study specific Clinical Monitoring 
Plan.
12.2.
Audit and Inspections
Representatives of the sponsor’s clinical quality assurance department may visit the study site at 
any time during or after completion of the study to conduct an audit of the study in compliance 
with regulatory guidelines and company policy. These audits will require access to all study 
records, including source documents, for inspection and comparison with the eCRFs. Subject 
privacy must, however, be respected. The investigator and study-site personnel are responsible 
for being present and available for consultation during routinely scheduled study-site audit visits 
conducted by the sponsor or its designees. 
Similar auditing procedures may also be conducted by agents of any regulatory body, either as 
part of a national GCP compliance program or to review the results of this study in support of a 
PPD
PPD
PPD
PPD


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regulatory submission. The investigator should immediately notify the sponsor if he or she has 
been contacted by a regulatory agency concerning an upcoming inspection.
12.3. Data Quality Assurance
All eCRF data will be entered into a validated, 21CFR Part 11 compliant, computerized clinical 
data management system. 
The site is required to have a plan in place for assuring the quality of the research being 
conducted.
Steps to be taken to ensure the accuracy and reliability of data include the selection of qualified 
investigators and appropriate study sites, review of protocol procedures with the investigator and 
study-site personnel before the study, and periodic monitoring visits by the sponsor. Written 
instructions will be provided for collection, handling, storage, and shipment of samples. 
Guidelines for eCRF completion will be provided and reviewed with study-site personnel before 
the start of the study. The sponsor will review eCRFs for accuracy and completeness during 
on-site monitoring visits and after transmission to the sponsor; any discrepancies will be resolved 
with the investigator or designee, as appropriate. After upload of the data into the study database
they will be verified for accuracy and consistency with the data sources.
13.
ADMINISTRATIVE REQUIREMENTS
13.1.
Protocol Amendments and Protocol Deviations
Protocol Amendments
Protocol amendments are planned changes to a protocol that are documented in a controlled
manner through numbered amendments by the sponsor.
Neither the investigator nor the sponsor will modify this protocol without a formal amendment 
by the sponsor. All protocol amendments must be issued by the sponsor, and signed and dated by 
the investigator. Protocol amendments must not be implemented without prior IRB approval, or 
when the relevant competent authority has raised any grounds for non-acceptance, except when 
necessary to eliminate immediate hazards to the subjects, in which case the amendment must be 
promptly submitted to the IRB and relevant competent authority. Documentation of amendment 
approval by the investigator and IRB must be provided to the sponsor. When the change(s) 
involves only logistic or administrative aspects of the study, the IRB only needs to be notified.
During the course of the study, in situations where a departure from the protocol is unavoidable, 
the investigator or other physician in attendance will contact the appropriate sponsor 
representative (see Contact Information page(s) provided separately). Except in emergency 
situations, this contact should be made before implementing any departure from the protocol. In 


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all cases, contact with the sponsor must be made as soon as possible to discuss the situation and 
agree on an appropriate course of action. The data recorded in the eCRF and source documents 
will reflect any departure from the protocol, and the source documents will describe this 
departure and the circumstances requiring it.
Protocol Deviations
Protocol deviations are unplanned excursions from the protocol that occur during the conduct of 
the study. Protocol deviations may (major protocol deviations) or may not (minor protocol 
deviations) affect the safety of the subject or the integrity of clinical study data. 
Protocol deviations to inclusion/exclusion criteria may be identified post hoc during routine site 
monitoring. In such cases, the Study Responsible Physician must decide on a case-by-case basis 
the appropriate action to take.
Other protocol deviations can include non-clinically significant variations determined by good 
medical judgment not to affect the integrity of the study or safety of the subject. Also included 
are occasional insignificant variations to timings of investigational drug/study-related procedures 
or visit windows (which may be foreseen given the circumstances of an individual subject), or 
procedures that do not affect the integrity of the study or the safety of the subject, or 
non-clinically significant test results in the opinion of the investigator which pose no clinical 
safety concerns. Both the investigator and the sponsor should discuss these deviations prior to or 
during their occurrence. A record of the outcome (e.g., Note to File) must be retained in the Trial 
Master File and Trial Center File.
Any protocol deviations identified during routine monitoring are documented in Monitoring 
Visit Reports in accordance with the appropriate Sponsor procedures. Documentation of protocol 
deviations on a study basis, reporting of protocol deviations to local health authorities and IRBs, 
and documentation in the Clinical Study Report must be compliant with the appropriate Sponsor 
procedures.
13.2.
Continuing Review and Close-out Reports
Continuing review reports will be submitted at intervals designated (and at least annually) by the 
IRB of record in accordance with 32 CFR 219. The continuing review report should be submitted 
to the 
IRBs for review and approval on no 
less than an annual basis. A final study report will be submitted to each IRB for review and 
approval upon completion of the study.
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13.3.
Regulatory Documentation
13.3.1.
Regulatory Approval/Notification
This protocol and any amendment(s) must be submitted to the appropriate regulatory authorities 
in each respective country, if applicable. A study may not be initiated until all local regulatory 
requirements are met.
13.3.2.
Required Prestudy Documentation
The following documents must be provided to the sponsor before shipment of study drug to the 
study site:

Protocol and amendment(s), if any, signed and dated by the PI

A copy of the dated and signed (or sealed, where appropriate per local regulations), written 
IRB approval of the protocol, amendments, ICF, any recruiting materials, and if applicable, 
subject compensation programs. This approval must clearly identify the specific protocol by 
title and number and must be signed (or sealed, where appropriate per local regulations) by 
the chairman or authorized designee.

Name and address of the IRB, including a current list of the IRB members and their 
function, with a statement that it is organized and operates according to GCP and the 
applicable laws and regulations. If accompanied by a letter of explanation, or equivalent, 
from the IRB, a general statement may be substituted for this list. If an investigator or a 
member of the study-site personnel is a member of the IRB, documentation must be 
obtained to state that this person did not participate in the deliberations or in the 
vote/opinion of the study.

Regulatory authority approval or notification, if applicable

Documentation of investigator qualifications (eg, curriculum vitae)

Completed investigator financial disclosure form from the PI, where required

Signed and dated clinical trial agreement, which includes the financial agreement

Any other documentation required by local regulations
The following documents must be provided to the sponsor before enrollment of the first subject:

Completed investigator financial disclosure forms from all subinvestigators, where required

Documentation of subinvestigator qualifications (eg, curriculum vitae)

Name and address of any local laboratory conducting tests for the study, and a dated copy of 
current laboratory normal ranges for these tests, if applicable

Local 
laboratory 
documentation 
demonstrating 
competence 
and 
test 
reliability 
(eg, accreditation/license), if applicable


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13.4.
Subject Identification, Enrollment, and Screening Logs
The investigator agrees to complete a subject identification and enrollment log to permit easy 
identification of each subject during and after the study. This document will be reviewed by the 
sponsor study-site contact for completeness.
The subject identification and enrollment log will be treated as confidential and will be filed by 
the investigator in the study file. To ensure subject confidentiality, no copy will be made. All 
reports and communications relating to the study will identify subjects by subject identification 
and date of birth. In cases where the subject is not randomized into the study, the date seen and 
date of birth will be used.
The investigator must also complete a subject screening log, which reports on all subjects who 
were seen to determine eligibility for inclusion in the study.
13.5.
Source Documentation
At a minimum, source documentation must be available for the following to confirm data 
collected in the eCRF: subject identification, eligibility, and study identification; study 
discussion and date of signed informed consent; dates of visits; results of safety and efficacy 
parameters as required by the protocol; record of all AEs and follow-up of AEs; concomitant 
medication; drug receipt/dispensing/return records; study drug administration information; and 
date of study completion and reason for early discontinuation of study drug or withdrawal from 
the study, if applicable. 
In addition, the author of an entry in the source documents should be identifiable.
At a minimum, the type and level of detail of source data available for a subject should be 
consistent with that commonly recorded at the study site as a basis for standard medical care. 
Specific details required as source data for the study will be reviewed with the investigator 
before the study and will be described in the monitoring guidelines (or other equivalent 
document).
Source documents are original documents, data, and records from which the subject’s data are 
obtained. These include but are not limited to hospital records, clinical and office charts, 
laboratory and pharmacy records, microfiches, radiographs, and correspondence. 
The investigator and staff are responsible for ensuring maintenance of a comprehensive and 
centralized filing system of all study-related (essential) documentation, suitable for inspection at 
any time by representatives from the sponsor, the DoD, DAIDS, FDA, and/or applicable 
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All eCRF entries have to be verifiable by the source data in the subject file. This does not apply 
to eCRF entries that are defined as source data. 
13.6.
Case Report Form Completion
CRFs are provided for each subject in electronic format. 
EDC will be used for this study. The study data will be transcribed by study-site personnel from 
the source documents onto an electronic CRF, and transmitted in a secure manner to the sponsor 
within the timeframe agreed upon between the sponsor and the study site. The electronic file will 
be considered to be the CRF.
Data must be entered into eCRFs in English. Study site personnel must complete the eCRF as 
soon as possible after a subject visit, and the forms should be available for review at the next 
scheduled monitoring visit.
The investigator must verify that all data entries in the eCRFs are accurate and correct. All eCRF 
entries, corrections, and alterations must be made by the investigator or other authorized 
study-site personnel. 
If corrections to an eCRF are needed after the initial entry into the eCRF, this can be done in 
3 different ways:

Study site personnel can make corrections in the eDC tool at their own initiative or as a 
response to an auto query (generated by the eDC tool).

Study site personnel can make corrections in the eDC tool in response of a query from Study 
site manager.

Study site personnel can make corrections in the eDC tool in response of a query from 
clinical data manager. 
13.7.
Archiving
The investigator is responsible for the archiving of the investigator’s file, the subject’s file, and 
the source data according to national and international legal requirements. 
Any records related to the conduct of the study may not be destroyed without written 
authorization by the sponsor. 
In compliance with the ICH/GCP guidelines, the investigator/institution will maintain all eCRFs 
and all source documents that support the data collected from each subject, as well as all study 
documents as specified in ICH/GCP Section 8, Essential Documents for the Conduct of a 
Clinical Trial, and all study documents as specified by the applicable regulatory requirement(s). 


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The investigator/institution will take measures to prevent accidental or premature destruction of 
these documents.
Essential documents must be retained until at least 2 years after the last approval of a marketing 
application in an ICH region and until there are no pending or contemplated marketing 
applications in an ICH region or until at least 2 years have elapsed since the formal 
discontinuation of clinical development of the IP. These documents will be retained for a longer 
period if required by the applicable regulatory requirements or by an agreement with the sponsor. 
It is the responsibility of the sponsor to inform the investigator/institution as to when these 
documents no longer need to be retained.
If the responsible investigator retires, relocates, or for other reasons withdraws from the 
responsibility of keeping the study records, custody must be transferred to a person who will 
accept the responsibility. The sponsor must be notified in writing of the name and address of the 
new custodian. Under no circumstance shall the investigator relocate or dispose of any study 
documents before having obtained written approval from the sponsor.
If it becomes necessary for the sponsor or the appropriate regulatory authority to review any 
documentation relating to this study, the investigator/institution must permit access to such 
reports.
13.8.
Study Completion/Termination
13.8.1.
Study Completion
The study is considered completed with the last visit for the last subject participating in the 
study. The final data from the study site will be sent to the sponsor (or designee) after completion 
of the final subject visit at that study site, in the time frame specified in the Clinical Trial 
Agreement.
13.8.2.
Study Termination
The sponsor reserves the right to close the study site or terminate the study at any time for any 
reason at the sole discretion of the sponsor. Study sites will be closed upon study completion. A 
study site is considered closed when all required documents and study supplies have been 
collected and a study-site closure visit has been performed.
The investigator may initiate study-site closure at any time, provided there is reasonable cause 
and sufficient notice is given in advance of the intended termination.
Reasons for the early closure of a study site by the sponsor or investigator may include but are 
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
Failure of the investigator to comply with the protocol, the requirements of the IRB or local 
health authorities, the sponsor’s procedures, or GCP guidelines

Inadequate recruitment of subjects by the investigator

Discontinuation of further study vaccine development
13.9.
Volunteer Registry Database
The USAMRMC has a policy to collect information from every volunteer who participates in 
this study in a “Volunteer Registry Database”. There are two objectives of this data collection.
First, to answer questions about the volunteers involved in research sponsored by the
USAMRMC, and second, to ensure that volunteers are informed of risks or new information.
Such information includes

Name, Surname

Identification Number

Date of Birth

Contact information

Study title, date of study participation, and reason for withdrawal if applicable

Severe adverse reactions including unexpected events that occurred due to vaccination 
during research participation

Details of the research product received
This Volunteer Enrollment Database will be kept in Thailand for 75 years under the auspices of 
the Armed Forces Research Institute of Medical Sciences.
13.10. Use of Information and Publication
All information, including but not limited to information regarding Ad26.Mos.HIV and 
MVA-Mosaic or the sponsor’s operations (eg, patent application, formulas, manufacturing 
processes, basic scientific data, prior clinical data, formulation information) supplied by the 
sponsor to the investigator and not previously published are considered confidential. The 
investigator agrees to maintain this information in confidence and use this information only to 
accomplish this study, and will not use it for other purposes without the sponsor’s prior written 
consent.
The investigator understands that the information developed in the study will be used by the 
sponsor in connection with the continued development of Ad26.Mos.HIV and MVA-Mosaic, and 
thus may be disclosed as required to other clinical investigators or regulatory agencies. 
The results of the study will be reported in a Clinical Study Report generated by the sponsor in 
conjunction with the investigator. Results of analyses performed after the Clinical Study Report 


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has been issued will be in a separate report and will not require a revision of the Clinical Study 
Report. Study subject identifiers will not be used in publication of results.
Registration of Clinical Studies and Disclosure of Results
The sponsor will register and/or disclose the existence of and the results of clinical studies as 
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14.
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Li H, Rhee EG, et al. (2012) Adenovirus serotype 26 utilizes CD46 as a primary cellular receptor and 
only transiently activates T lymphocytes following vaccination of rhesus monkeys. J Virol. 86:10862-5.
Li JZ, Smith DM, Mellors JW. (2015) The need for treatment interruption studies and biomarker 
identification in the search for an HIV cure. AIDS. 29(12):1429-32.
Liu J, O-Brien K, Lynch D, et al. (2009) Immune control of an SIV challenge by a T-cell-based vaccine 
in rhesus monkeys. Nature. 457:87-91.
Lodi S, Meyer L, Kelleher AD, et al. (2012) Immunovirologic control 24 months after interruption of 
antiretroviral therapy initiated close to HIV seroconversion. Archives of internal medicine. 172(16):1252-
5.
Mast TC, Kierstead L, et al. (2010) International epidemiology of human pre-existing adenovirus (Ad) 
type-5, type-6, type-26 and type-36 neutralizing antibodies: correlates of high Ad5 titers and implications 
for potential HIV vaccine trials. Vaccine. 28:950-7.
NIH clinical center guidelines: “Guidelines For Blood Drawn For Research Purposes In The Clinical 
Center” (M95-9). NIH Policy Manual. 3014 -
NIH Human Research Protection Program. 
https://policymanual.nih.gov/3014. Accessed 06/07/2017.
O'Brien KL, Liu J, et al. (2009) Adenovirus-specific immunity after immunization with an Ad5 HIV-1 
vaccine candidate in humans. Nat Med. Aug;15(8):873-5. Epub 2009 Jul 20.
Penaloza-MacMaster P, Provine NM, et al. (2013) Alternative serotype adenovirus vaccine vectors elicit 
memory T cells with enhanced anamnestic capacity compared to Ad5 vectors. J Virol. 87:1373-84.
Perreau M, Welles HC, et al. (2012) The number of Toll-like receptor 9-agonist motifs in the adenovirus 
genome correlates with induction of dendritic cell maturation by adenovirus immune complexes. J Virol. 
86:6279-8.


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Peters BS, Jaoko W, Vardas E, et al. (2007) Studies of a prophylactic HIV-1 vaccine candidate based on 
modified vaccinia virus Ankara (MVA) with and without DNA priming: effects of dosage and route on 
safety and immunogenicity. Vaccine. 25(11):2120-7. 
Pitisuttithum P, Rerks-Ngarm S, Bussaratid V, et al. (2011) Safety and reactogenicity of canarypox 
ALVAC-HIV-1 (vCP1521) and HIV-1 gp120 AIDSVAX B/E vaccination in an efficacy trial in Thailand. 
PLoS one. 6: e27837.
Ramanathan VD, Kumar M, Mahalingam J, et al. (2009) A phase 1 study to evaluate the safety and 
immunogenicity of a recombinant HIV type 1 subtype C-modified vaccinia Ankara virus vaccine 
candidate in Indian volunteers. AIDS Res Hum Retroviruses. 25:1107-1116. 
Ratto-Kim S, Currier JR, Cox JH, et al. (2012) Heterologous Prime-Boost Regimens Using rAd35 and 
rMVA Vectors Elicit Stronger Cellular Immune Responses to HIV Proteins Than Homologous Regimens. 
PLoS one. 7(9): e45840.
Rerks-Ngarm S, Pitisuttithum P, Nitayaphan S, et al. (2009) Vaccination with ALVAC and AIDSVAX to 
Prevent HIV-1 Infection in Thailand. N Engl J Med. 361:1-12.
Rerks-Ngarm S, Paris RM, Chunsutthiwat S, et al. (2013) Extended evaluation of the virologic, 
immunologic, and clinical course of volunteers who acquired HIV-1 infection in a phase III vaccine trial 
of ALVAC-HIV and AIDSVAX(R) B/E. J Infect Dis. 207:1195-1205.
Robb ML, Rerks-Ngarm S, Nitayaphan S, et al. (2012) Risk behaviour and time as covariates for efficacy 
of the HIV vaccine regimen ALVAC-HIV (vCP1521) and AIDSVAX B/E: a post-hoc analysis of the 
Thai phase 3 efficacy trial RV144. Lancet Infect Dis. 12:531-537.
Roederer M, Keele BF, Schmidt SD, et al. (2013) Immunological and virological mechanisms of vaccine-
mediated protection against SIV and HIV. Nature. 
Rolland M, Edlefsen PT, Gottardo R, et al. (2013) Genetic and immunological evidence for a role of Env-
V3 antibodies in the RV144 trial. AIDS Res Human Retroviruses. 29:A-168 [P03-73 LB].
Rothenberger MK. (2015) Large number of rebounding/founder HIV variants emerge from multifocal 
infection in lymphatic tissues after treatment interruption.Proc Natl Acad Sci USA. 112(10):E1126-1134.
Saez-Cirion A, Bacchus C, Hocqueloux L, et al. (2013) Post-treatment HIV-1 controllers with a long-term 
virological remission after the interruption of early initiated antiretroviral therapy ANRS VISCONTI 
Study. PLoS pathogens. 9(3):e1003211.
Santra S, Liao HX, et al. (2010) Mosaic vaccines elicit CD8+ T lymphocyte responses that confer 
enhanced immune coverage of diverse HIV strains in monkeys. Nat Med. 16(3):324-8.
Shacklett BL, Ferre AL. (2011) Mucosal immunity in HIV controllers: the right place at the right time. 
Current opinion in HIV and AIDS. 6(3):202-7.
Sugarman J. (2013) HIV Cure Research: Expanding the Ethical Considerations. Annals of Internal 
Medicine. 159(7):490-491.
Talos IF, Mian AZ, Zou KH, et al. (2006) Magnetic resonance and the human brain: anatomy, function 
and metabolism. Cell Mol Life Sci. 63:1106-1124.
Tan WG, Jin HT, et al. (2013) Comparative analysis of simian immunodeficiency virus gag-specific 
effector and memory CD8+ T cells induced by different adenovirus vectors. J Virol. 87:1359-72.


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Tarasow E, Wiercinska-Drapalo A, Kubas B, et al. (2003) Cerebral MR spectroscopy in neurologically 
asymptomatic HIV-infected patients. Acta Radiol. 44:206-212.
UNAIDS/WHO. Report on the global HIV/AIDS epidemic. 2013
US Food and Drug Administration. Conditions for IRB Use of Expedited Review. 
https://www.fda.gov/scienceresearch/specialtopics/runningclinicaltrials/guidancesinformationshe
etsandnotices/ucm118099.htm. Accessed 06/07/2017.
Valcour V, Chalermchai T, Sailasuta N, et al. (2012) Central nervous system viral invasion and 
inflammation during acute HIV infection. J Infect Dis. 206(2):275-282.
Vasan S, Schlesinger SJ, Chen Z, et al. (2010) Phase 1 safety and immunogenicity evaluation of 
ADMVA, a multigenic, modified vaccinia Ankara-HIV-1 B'/C candidate vaccine. PLoS one. 5:e8816.
Volberding P. (2014) HIV cure: the target is clearer but not yet close. Annals of Internal Medicine. 
160:505-6.
Waddington SN, McVey JH, et al. (2008) Adenovirus serotype 5 hexon mediates liver gene transfer. 
Cell.132:397-409.
Williams JP, Hurst J, Stöhr W, et al. (2014) HIV-1 DNA predicts disease progression and post-treatment 
virological control. Elife. 3:e03821.
Zolla-Pazner S, Decamp AC, Cardozo T, et al. (2013) Analysis of V2 Antibody Responses Induced in 
Vaccinees in the ALVAC/AIDSVAX HIV-1 Vaccine Efficacy Trial. PLoS one. 8:e53629.


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15.
SIGNATURES
SPONSOR: Janssen Vaccines & Prevention B.V.; Archimedesweg 4, 2333 CN Leiden, The 
Netherlands
This Study Protocol was subject to critical review and has been approved.
Medical Lead
[electronic signature appended at the 
end of the protocol]
Frank Tomaka, MD (Medical Lead) 
Janssen Vaccines & Prevention B.V.
Date


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PROTOCOL CHAIRS
This Study Protocol has been approved.
Protocol Chair
Jintanat Ananworanich, MD, PhD
US Military HIV Research 
Program (MHRP)
Date
PI
Nittaya Phanuphak, MD, PhD
 
Date
PPD


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INVESTIGATORS:
I have reviewed this Study Protocol, including Appendices. I will conduct the clinical study as 
described and will adhere to ICH/GCP and all the ethical and regulatory requirements stated. 
I have read and understood the contents of the Investigator's Brochure.
I will provide copies of the protocol and all pertinent information to all individuals responsible to 
me who assist in the conduct of this study. I will discuss this material with them to ensure that 
they are fully informed regarding the study vaccine, the conduct of the study, and the obligations 
of confidentiality.
Principal Investigator
Signature
Date


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16.
APPENDICES
16.1.
Definitions of HIV-1 Related Disease
(CDC Classification, 1993: modified for Thailand with addition of Penicilliosis to Category C) 
Category B:  Symptomatic Conditions in HIV-Infected Patients
o
Bacillary angiomatosis
o
Candidiasis, oropharyngeal (thrush)
o
Candidiasis, vulvovaginal; persistent, frequent, or poorly responsive to therapy
o
Cervical dysplasia (moderate or severe)/cervical carcinoma in situ
o
Constitutional symptoms, such as fever (38.5 °C) or diarrhea lasting >1 month
o
Hairy leukoplakia, oral
o
Herpes zoster (shingles), involving at least two distinct episodes or more than one 
dermatome
o
Idiopathic thrombocytopenic purpura
o
Listeriosis
o
Pelvic inflammatory disease, particularly if complicated by tubo-ovarian abscess
o
Peripheral neuropathy
Category C:  AIDS-Defining Diagnoses
o
Candidiasis of bronchi, trachea, or lungs
o
Candidiasis, esophageal
o
Cervical cancer, invasive
o
Coccidioidomycosis, disseminated or extrapulmonary
o
Cryptococcosis, extrapulmonary
o
Cryptosporidiosis, chronic intestinal (>1 month duration)
o
Cytomegalovirus disease (other than liver, spleen or nodes)
o
Cytomegalovirus retinitis (with loss of vision)
o
Encephalopathy, HIV-related
o
Herpes simplex: chronic ulcer(s) (>1 month duration); or bronchitis, pneumonitis or 
esophagitis
o
Histoplasmosis, disseminated or extrapulmonary
o
HIV encephalopathy (dementia)
o
Isosporiasis, chronic intestinal (>1 month duration)
o
Kaposi’s sarcoma
o
Lymphoma, Burkitt’s (or equivalent term)
o
Lymphoma, immunoblastic (or equivalent term)
o
Lymphoma, primary, of brain
o
Mycobacterium avium complex or M kansasii, disseminated or extrapulmonary
o
Mycobacterium, other species or unidentified species, disseminated or extrapulmonary
o
Penicilliosis (addition to CDC classification for Thailand)
o
Pneumocystis carinii pneumonia
o
Pneumonia, recurrent
o
Progressive multifocal leukoencephalopathy
o
Salmonella septicemia, recurrent


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o
Toxoplasmosis of brain
o
Wasting syndrome


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16.2.
Case Definition for Acute Retroviral Syndrome (ARS)
Case definition must include all of the following:

Occurs within 30 days after documented rise in plasma HIV-1 RNA of >1 log10 or 
to >1,000 copies/mL

Occurs during ATI or within 7 days of restarting ARV medication after ATI

Documented fever (temperature >38.5°C)

At least two major criteria

At least two minor criteria

No other cause for signs and symptoms identified
Major criteria (must be present for >7 days):

Pharyngitis

Fatigue

Morbilliform rash

Myalgia/arthralgia

Lymphadenopathy

Subjective fever as reported by subject (may be intermittent)
Minor criteria (must be present for >7 days):

Headache

Nausea/vomiting

Diarrhea

Mucocutaneous ulceration (oral, genital, and/or anorectal)

Meningismus/aseptic meningitis

Night sweats

Thrombocytopenia (platelets <150,000)

Leukopenia (WBC <4,000)

Malaise

Abdominal pain

Weight loss (loss of >5% of body weight)


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16.3.
Test of Understanding
Please read each question and answer whether the statement is True or False.
True
  
False
  
1.    The vaccines you will receive in this study have been proven to cure HIV.
True
  
False
  
2.    You will need to come to the clinic for about 50 scheduled visits over the 
next 2 years.
True
  
False
  
3.    The vaccines in this study can give you side effects such as redness, bruising, 
and pain at the injection site..
True
  
False
  
4.    One purpose of this study is to determine if these vaccines are safe to 
administer to humans.
True
  
False
  
5.    Participants in this study will need to avoid engaging in activities that may 
expose others to HIV infection.
True
  
False
  
6.    You may take other experimental (test) vaccines while you are taking part in 
this study.
True
  
False
  
7.    You may withdraw from the study at any time if you choose or your 
participation may be stopped if the study team decides it is in your best 
interest.
True
  
False
  
8.    Women participating in this study are permitted to become pregnant during 
the study.
True
  
False
  
9.    A participant in this study may donate blood during the study.
True
 
False
 
10.  During the study staff members may contact you by telephone or email or 
text message.


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16.4.
Research Specimen Laboratory Testing4,5
Co-Investigating 
Site
Specimen 
type
Planned Usage
Specimen Disposition
Overseeing IRB
Sub-project/
Research Questions
Post-testing 
plan
Data Usage
Blood
Storage
Storage
RV254 study team 
N/A
CSF
Storage
Storage
RV254 study team 
Gut tissue
Storage
Storage
RV254 study team 
Mucosal 
secretions
Storage
Storage
RV254 study team 
Lymph node 
tissue
Storage
Storage
RV254 study team 
 
Lymph node 
tissue
Characterization of virological 
and immunological events in 
lymphoid tissue
Storage
Local 
coinvestigating site 
and RV254 study 
team
Investigation of viral 
reservoir and 
immunologic markers 
and fibrosis
Blood
HIV-RNA, 
genotyping/sequencing, 
immunologic studies of T-cell, B-
cell and antibody responses to 
HIV and host genetic studies to 
evaluate potential genes 
associated with HIV infection or 
diseases progression
Specimen 
discarded by 
site
RV254 study team 
Immunologic/virologic 
characterization 
CSF
HIV-RNA, immune response and 
immunohistopathology
RV254 study team 
Characterize viremia and 
immunologic responses 
in the CNS compartment 
Gut tissue
immune response and 
immunohistopathology
RV254 study team 
Characterization of T 
cell responses in gut 
tissues 
Mucosal 
HIV-RNA, immune response and 
RV254 study team 
HIV viremia in the 
                                                
4 Any changes to this table will not consistute a protocol amendment but will be adapted through the lab manual.
5 Updated 11 Oct 2017
PPD
PPD
PPD
PPD
PPD
PPD


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secretions
immunohistopathology
genital compartment 
Lymph node 
tissue
Immunologic and virologic 
studies
RV254 study team 
Investigation of viral 
reservoir.
 
Blood
Performance of novel biomarkers
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
 
Characterization of 
neuropathology 
CSF
Performance of novel CSF 
biomarkers
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Blood
Inflammatory and coagulation 
and tissue fibrosis makers such as 
CP, sCD14, D-dimer and 
hyaluronic acid, HIV viral load 
by high sensitivity assay, 
immunophenotyping
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Characterization of 
novel biomarkers 
CSF
Inflammatory and immunologic 
markers
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Gut tissue
Immunohistochemistry, in situ 
hybridization, RNA and DNA 
scope
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Characterization of CD4 
T cells and associated 
factors 
Mucosal 
secretions
Inflammatory and immunologic 
markers
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Investigation of viral 
burden and immunologic 
markers
Lymph node 
tissue
Immunohistochemistry, markers 
of immune activation and 
fibrosis, in situ hybridization, 
RNA and DNA scope
Investigation of systemic 
inflammation and change to the 
global T-cell receptor (TCR) and 
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Investigation of viral 
reservoir.
Characterization of 
lymphoid 
immuno(patho)logical 
events 
PP
D
PPD
PP
D
PPD


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B-cell receptor (BCR) repertoires
 
Blood
Investigation of viral reservoir 
size and immunologic responses
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
 
Investigate reservoir size 
and immunologic 
responses in particular, T 
and B cell function 
CSF
Investigation of cellular response
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Gut tissue
Investigation of viral reservoir 
size
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
 
Blood
Investigation of monocytes, 
macrophages and innate 
responses 
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Characterization of the 
inflammatory profile of 
monocytes 
CSF
Investigation of monocytes, 
macrophages and innate 
responses
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Gut tissue
Investigation of monocytes, 
macrophages and innate 
responses
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
Lymph node 
tissue
Investigation of monocytes, 
macrophages and innate 
responses
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
 
Lymph node 
tissue
Characterization of virological 
and immunological events in 
lymphoid tissue, 
immunohistochemistry, in situ 
hybridization, RNA and DNA 
scope
Specimen 
discarded by 
site
Local 
coinvestigating site 
and RV254 study 
team
 
Investigation of viral 
reservoir and 
immunologic markers 
and fibrosis
 
Blood
Trofile (CCR5 tropism) testing
Specimen 
discarded by 
RV254 study team
N/A
None
PPD
PPD
PPD
PPD
PPD
PPD
PPD


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site
Janssen Vaccines & 
Prevention B.V.
Blood
Evaluation of immunogenicity
Storage
RV254 study team
N/A
N/A
PPD


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16.5.
Neurological Examination Form, Abridged Neuropsychological 
Assessment Test Battery, Substance use and Sleep, CDR
SEARCH010                                   PID:  ___ ___ ___ ___          Date: ___ ___/___ ___/___ ___
Visit #: ___ ____                              PTT Int. ____ ____                            Month / day / year
MACRONEUROLOGICAL EXAM 2.0 (REVISED 1/2016)
Place an “X” if ALL questions and neurological tests reveal normal results (“0”)
Default to “0” for questions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 17, 18, 19, 20, 22, 23, 24, 25, 28b/c, 29, 30, 
31, 32, 35, 36, 37
COGNITIVE FUNCTION
1.  Concentration/Speed of thought      
0
Normal mental ability
1
Notes some diminished capacity for concentration or need for longer time to accomplish normal tasks but 
able to manage nearly all of daily affairs
2
Definitely loses track of conversation or task, takes more than twice as long to complete some tasks, needs 
help to manage more difficult tasks of daily life (e.g. financial records) or complex work activities
3
Marked slowing of mental processes, perseveration or loss of train of thought, needs help to manage almost 
all daily affairs requiring cognition
4
Only rudimentary cognition
5
Cannot evaluate
6
Not elicited
2.  Reading (or TV)                                                                                                                      
0
Normal
1
Notes mild increase in effort or occasionally loses place in either reading or TV
2
Reduced reading or TV because of slowness, clearly more laborious, or requires repeated efforts, but can 
follow news or similar subjects
3
Reading or TV markedly reduced because of difficulty attending or following plot but occasionally reads or 
watches with reduced attention or comprehension
4
Unable to read or watch TV at all because cannot make sense of subject, etc.
8
Cannot evaluate
9
Not elicited  
3. Memory                                                                                                                               
0
Normal
1
Mildly more forgetful than usual, will occasionally miss appointments
2
Definite memory difficulty, needs to keeps lists, forgets events of day or activities in mid task
3
Frankly confused at times about events, places, persons
4
Persistently disoriented to time, frequently to place or person
8
Cannot evaluate
9
Not elicited
4.   Speech 
0
Normal
1
Mild hesitancy, extra deliberation of occasional use of inappropriate words
2
Definite slowness or difficulty finding words of use of wrong or inappropriate words
3
Reduced speech output or frequent speech errors


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4
Speech limited to a few phrases at best
8
Cannot evaluate
9
Not elicited
MOTOR FUNCTION
5. Gait                                                                                                                                    
0
Normal
1
Mild unsteadiness, imbalance, slowness or weakness, but fully ambulatory without assistance
2
Unsteady or weak, requiring cane or some prop
3
Requires bilateral (e.g. walker or human) prop
4
Unable to walk
8
Cannot evaluate
9
Not elicited
6. Upper Extremity Coordination                                                                                                             
0
Normal dexterity
1
Mild slowing or clumsiness of hands without functional impairment
2
Moderate clumsiness of hands, altering handwriting or slowing regular hand activities (eating, etc.)
3
Marked clumsiness resulting in difficulty performing ADL (e.g. difficulty feeding without manual help of 
others)
4
Unable to perform hand-related ADL because of incoordination
8
Cannot evaluate
9
Not elicited
7. Involuntary Movement (Includes Tremor, Myoclonus & Chorea)                                                                   
0
None or normal mild tremor
1
Noticeable new or increase in adventitious movements, but does not interfere with ADL
2
New or increased adventitious movements impair handwriting or other manual tasks, but still can function 
independently for most ADL
3
Marked adventitious movements interfering with most ADL
4
Severe adventitious movements necessitate nearly full-time care
8
Cannot evaluate
9
Not elicited
BEHAVIOR/MOOD
8.  Mood     
0                 Normal spirits, no depression
1               Depressed about illness but does not intrude on function
3               Severe depression, requires suicide precautions
8               Cannot evaluate
9             Not elicited
9. Social Apathy/Withdrawal
0                Normal socialization
1                Diminished interest but continues with normal activities
2                Activities reduced because of lack of interest or initiative but maintains most contacts
3                Marked loss of interest in friends and normal social activities, does not seek contact
4                Little or no meaningful socialization
8                Cannot evaluate
9                Not elicited
10. Emotional Lability/Agitation                                                                                               


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0                Normal
1                Irritable, more easily agitated or excited, but does not alter social ability
2                Clearly labile or hyperactive so that interactions are altered, with occasional inappropriate social
                     interaction
3                Nearly constant hyperactivity or hypomania interfering with social interactions
4               Frank delirium or psychosis
MENTAL STATUS and AFFECT                                                              
11. Response slowing
0
Normal or hyperactive
1
Mild slowing of response time 
2
Moderate slowing of response time
3
Severely slowed, nearly fully or mute
8
Cannot evaluate
9
Not elicited
SENSATION
12. Neuropathy Symptoms                                                                                                    
0
None
1
Mild distal paresthesia or pain not requiring more than occasional non-narcotic analgesics and not 
interfering with Activities of Daily Living           
2
Moderate distal paresthesia/pain requiring daily non-narcotic
3 
Severe distal paresthesia or pain requiring daily narcotic analgesic and substantially interfering with 
Activities of Daily Living
8
Cannot evaluate
9
Not elicited
     
SEIZURES
13.  Seizures
00
None
01 
Single partial or generalized seizure
02 
Repeated (<3/wk) partial or generalized seizures
03
Frequent (>4/wk) partial or generalized seizures
08
Cannot evaluate
09 
Not elicited
OCULAR MOTILITY
14. Smooth Pursuits (Have the participant follow finger a minimum of 2 times horizontally from side to side and 2 
times vertically from up to down)
0
Normal
1
Mildly abnormal (mild cogwheeling)
2
Moderately - severely abnormal
8
Cannot evaluate
9
Not elicited
15. Saccades   [Place finger in participant’s lateral field and ask participant to look at finger (without snapping 
finger) then your nose, alternating.  Repeat for opposite lateral direction, as well as vertically (up and downward)]
00
Normal


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01
Mildly abnormal (mild cogwheeling)
02 
Moderately - severely abnormal
08
Cannot evaluate
09
Not elicited
STRENGTH
Strength score reflects worst major muscle group or groups. Scores of 0-3 refer to weakness related to ADC only. 
Test by asking patient to do a deep knee bend, walk on toes and heels, and hop on each foot.
16. Right Leg 
0
Normal
1
Mild weakness (4/5)
2
Moderate weakness (3/5)
3
Severe weakness (2/5 or worse)
4
Other _______________________________________
8
Cannot evaluate
9
Not elicited
16a.  Is weakness related to neuropathy?
0
No weakness found
1
No, neuropathy not present
2
No, neuropathy present, but not thought to be etiology of weakness
3
Yes, neuropathy somewhat affecting strength or measure of strength
4
Yes, neuropathy responsible for majority of finding.
8
Cannot evaluate
9
Not elicited
17. Left Leg   
0
Normal
1
Mild weakness (4/5)
2
Moderate weakness (3/5)
3
Severe weakness (2/5 or worse)
4
Other _______________________________________
8
Cannot evaluate
9
Not elicited
COORDINATION     
18. Gait Coordination (Test by asking subject to walk quickly down hall, turn rapidly and return) 
0
Normal gait
1
Mild impairment (evident only on rapid turns or tandem)
2
Moderate impairment (clear difficulty of unassisted gait)
3
Severe impairment (walking only with assistance)
4
Non-ambulatory
5
Weakness precludes assessment of gait coordination
8
Cannot evaluate
9
Not elicited
19. Limb Coordination (Test by opposition of the first and second fingers, rapid wrist rotation, and rapid foot 
tapping)
0
Normal
1
Mild slowness or clumsiness (compared to examiner, the movement is slightly slower, fatigues or breaks 
down earlier)
2
Moderate slowness or clumsiness


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3
Weakness precludes assessment of gait coordination
8
Cannot evaluate
9
Not elicited
REFLEXE
20. Deep Tendon Reflexes
0
Normal
          
5
Increased on one side
1
Increased in legs only
          
6
Other ____________________________
2
Generalized increase
          
7
Depressed or absent ankle jerks only(o/w normal)
3
General decrease (most notably distally)          
8
Cannot evaluate
4
Increased proximally and depressed distally     
9
Not tested
21. Jaw Jerk
00 
Absent 
01
Present
02
Unusually brisk 
08
Cannot evaluate
09         Not elicited
NEUROPATHY
22. Severity of Sensory Loss
0
Normal 
1
Mild impairment (e.g. increased threshold to position, vibration, or light touch in great toes)
2
Moderate impairment (e.g. decrease in position or vibration, pin or cold to ankles)
3
Severe impairment (e.g. decrease or loss of sensation to knees or wrists or comparable level)
8
Cannot evaluate
9
Not elicited
23. Type of Neuropathy
0
None
1
Predominantly distal sensory
2 
Predominantly motor
3 
Mixed distal
4 
Mononeuritis or localized radiculopathy
5
Mononeuritis multiplex or polyradicutopathy
8
Cannot assess
9
Not elicited
MOTOR UPDRS EXAMINATION 
24. Speech
0 
Normal
1 
Slight loss of expression, diction and/or volume
2 
Monotone, slurred but understandable
3
Marked impairment, difficult to understand
4 
Unintelligible
8 
Cannot assess
9 
Not elicited
25. Facial Expression                  
                                                                                 


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0  
Normal
1  
Slight hypomimia, could be normal "Poker Face"
2 
Slight but definitely abnormal diminution of facial expression
3
Moderate hypomimia; lips parted some of the time
4 
Masked or fixed face with severe or complete loss of facial expression; lips parted 1/4 inch or more
8
Cannot assess
9
Not elicited
26a-e. Tremor at Rest (see scoring box)
26a. 
Face, lips, and chin
26b. 
Arm, right
26c. 
Arm, left 
26d. 
Leg, right
26e.        Leg left
27a-b. Action or Postural 
Tremor
a.   RIGHT ARM
b.   LEFT ARM
28a-e. Rigidity (Judged on passive movement of major joints with subject relaxed in sitting position. Cogwheeling to 
be ignored.  Performed with and without distraction; see scoring box)
28a. 
Neck, 
28b. 
Arm, right
28c. 
Arm, left  
28d. 
Leg, right  
28e. 
Leg, left
29a-b. Finger Taps (Subject taps thumb with index finger in rapid succession with widest amplitude possible, each 
hand separately).  Please record patient’s handedness: 
right
left
a.   RIGHT FINGERS
b.   
LEFT 
FINGERS
00
Absent
01
Slight and infrequently present
02
Mild in amplitude and present most of the time
03
Moderate in amplitude and present most of the time
04
Marked in amplitude and present most of the time
08
Cannot assess
09          Not elicited
00
Absent
01
Slight; present with action
02 
Moderate in amplitude present with action only
03 
Moderate in amplitude with posture holding as well as action
04 
Marked in amplitude; interferes with feeding 
08 
Cannot assess
0
Absent
1
Slight and infrequently present
2
Mild and present most of the time
3
Moderate and present most of the time
4
Marked and present most of the time
8
Cannot assess
9          Not elicited
0 
Normal (15 or more/5 sec)
1
Mild slowing and/or reduction in amplitude (11-14/5 sec)
2 
Moderately impaired. Definite and early fatiguing. May have occasional arrest in 
movement (7-10/5 sec)
3 
Severely impaired. Frequent hesitation in initiating movements or arrests in ongoing 
movements (3-6/5 sec)
4 
Can barely perform the task (0-2/5 sec) 
8 
Cannot assess
9
Not elicited


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30a-b. Hand Movements   [Subject opens and closes hands in rapid succession with widest amplitude possible (full 
digit extension/flexion), each hand separately]
a.   RIGHT HAND
b.   LEFT HAND
31a-b. Rapid Alternating Movements of Hands/Arms (Pronation-supination of hands, vertically or horizontally, 
with as large an amplitude as possible, both hands simultaneously)
a.   RIGHT ARM
b.   LEFT ARM
32a-b. Leg Agility
(with knees bent, 
subject taps heel on 
ground in rapid 
succession, picking 
up entire leg. Amplitude should be about 3 inches.)
a.   RIGHT LEG
b.   LEFT LEG
33. Arising from chair (subject attempts to arise from a straight back wood or metal chair, with arms folded across 
chest)
00 
Normal
01 
Slow; or may need more than one attempt
02
Pushes self up from arms of seat
03 
Tends to fall back and may have to try more than one time, but can get up without help
04
Unable to arise without help 
08 
Cannot assess
09
Not elicited
34. Posture
00 
Normal
01 
Not quite erect, slightly stooped posture; could be normal for older person
02
Moderately stooped posture, definitely abnormal; can be leaning slightly to one side
03 
Severely stooped posture with kyphosis; can be leaning moderately to one side
04
Marked flexion with extreme abnormality of posture 
08 
Cannot assess
09
Not elicited
0 
Normal                                                                          
1
Mild slowing and/or reduction in amplitude 
2
Moderately impaired. Definite and early fatiguing. May have occasional arrest in 
movement
3
Severely impaired. Frequent hesitation in initiating movements
4
Can barely perform the task 
8 
Cannot assess 
9
Not elicited
0
Normal                                                                          
1
Mild slowing and/or reduction in amplitude 
2
Moderately impaired. Definite and early fatiguing. May have occasional arrest in 
movement
3
Severely impaired. Frequent hesitation in initiating movements
4
Can barely perform the task 
8
Cannot assess 
9 
Not elicited
0 
Normal       
1
Mild slowing and/or reduction in amplitude
2
Moderately impaired. Definite and early fatiguing. May have occasional arrest in movement
3 
Severely impaired. Frequent hesitation in initiating movements or arrests
4
Can barely perform the task
8 
Cannot assess
9
Not elicited


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35. Gait
0 
Normal
1 
Walks slowly, may shuffle with short steps
2
Walks with difficulty, but requires little or no assistance
3 
Severe disturbance of gait, requiring assistance
4
Cannot walk at all even with assistance 
5 
Spastic
6
Circumductive
8
Cannot assess
9
Not elicited
36. Postural Stability (Response to sudden posterior displacement produced by pull on shoulders while subject is  
                                      erect, with eyes open and feet slightly apart. Subject is prepared.)
0 
Normal (2 or less steps)
1 
Retropulsion, but recovers unaided
2
Absence of postural response; would fall if not caught by examiner
3 
Very unstable, tends to lose balance spontaneously
4
Unable to stand without assistance 
8
Cannot assess 
9
Not elicited
37. Body Bradykinesias and Hypokinesia
(Combining slowness, hesitance, decreased arm swing, small amplitude and poverty of movement in general)
0 
Normal 
1   
Minimal slowness, giving movement a deliberate character; could be normal for some persons. Possibly 
reduced amplitude
2 
Mild degree of slowness and poverty of movement which is definitely abnormal
3   
Moderate slowness, poverty or small amplitude of movement
4  
Marked slowness, poverty or small amplitude of movement 
8   
Cannot assess
9
Not elicited
Examiner Signoff: ________________________________


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Abridged Neuropsychological Assessment Test Battery
A.1
COLOR TRAILS I
Time to complete: _______________ seconds 
A.2
COLOR TRAILS II
Time to complete: _______________ seconds
B.
GROOVED PEGBOARD TEST
Non-dominant Hand Time
_________________
Non-dominant Hand Number of Drops
_________________
C.
TRAIL-MAKING TEST A
Time to complete: _______________ seconds
Error: _______________
D.        FLANKER TEST
Completed?                         
Yes                               
No
Abridged Neuropsychological Assessment Reporting Form
1. Color Trails 1 and 2 
00 
Reliable, standardized test administration
01
Questionable test results (specify reason)
02 
Invalid test results (specify reason)
Examiner Comments and 
Observations:_____________________________________________
2. Grooved Pegboard
00 
Reliable, standardized test administration
01
Questionable test results (specify reason)
02 
Invalid test results (specify reason)
Examiner Comments and 
Observations:_____________________________________________
3. Trail-making Test A
00 
Reliable, standardized test administration
01
Questionable test results (specify reason)
02 
Invalid test results (specify reason)
Examiner Comments and 
Observations:_______________________________________


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SUBSTANCE USE AND SLEEP
1. Do you use recreational drugs? (may include drugs made of local herbs/plants) 
01 
NEVER 
02
YES, TODAY OR YESTERDAY  
03
YES, THIS WEEK BUT NOT TODAY OR YESTERDAY  
04
YES, MORE THAN ONE WEEK AGO
05
ONLY IN THE PAST, MORE THAN ONE YEAR AGO 
08
REFUSE TO ANSWER  
09
DON’T KNOW 
2. If you have used recreational drugs today or yesterday, please specify which 
ones you used (may choose more than one). 
01
MARIJUANA/CANNABIS/BHANG  
02
HASHISH   
03
CRACK   
04
COCAINE 
05
METHAMPHETAMINES   
06
ECSTASY   
07
KHAT   
08
HEROIN   
09
GLUE/PETROL SNIFFING   
10
LSD   
11
POPPERS   
99
OTHER, SPECIFY: _____________________________________
3. Do you drink alcohol? 
01
NEVER   
02
YES, TODAY OR YESTERDAY   
03
YES, THIS WEEK, BUT NOT TODAY OR YESTERDAY   
04
YES, MORE THAN ONE WEEK AGO   
05
ONLY IN THE PAST, MORE THAN ONE YEAR AGO   
08
REFUSE TO ANSWER   
09
DON’T KNOW   
4. If you have consumed alcohol today or yesterday, how many drinks have you 
had in this time period?
01
1-2 DRINKS   
02
3-4 DRINKS   
03
5-6 DRINKS   
04
7 OR MORE DRINKS   
08
REFUSED TO ANSWER   
09
DON’T KNOW   
  5. How many hours sleep did you get in the last 24 hours?  
  6. How tired do you feel right now? 
(Rate from 1= Not tired, 5= moderately tired to 10= Very tired) 


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Clinical Dementia Rating (CDR) Scale
None
0
Questionable
0.5
Mild
1
Moderate
2
Severe
3
Memory
No memory loss or 
slight inconsistent 
forgetfulness
Consistent slight 
forgetfulness; partial 
recollection of events; 
“benign” forgetfulness
Moderate memory loss; 
more marked for recent 
events; defect interferes 
with everyday activities
Severe memory loss; only 
highly learned material 
retained; new material 
rapidly lost
Severe memory loss; 
only fragments 
remain
Orientation
Fully oriented
Fully oriented expect for 
slight difficulty with time 
relationships
Moderate difficulty with time 
relationships; oriented for 
place at examination; may 
have geographic 
disorientation elsewhere
Severe difficulty with time 
relationships; usually 
disoriented to time & often 
to place
Oriented to person 
only
Judgment & 
Problem 
Solving
Solves everyday 
problems, handles 
business & financial 
affairs well: judgment 
good in relation to past 
performance
Slight impairment in 
solving problems, 
similarities, & differences
Moderate difficulty in 
handling problems, 
similarities, 
&differences;social 
judgment usually 
maintained
Severely impaired in 
handling problems, 
similarities, & differences; 
social judgment usually 
impaired
Unable to make 
judgments or solve 
problems
Community 
Affairs
Independent function at 
usual level in job, 
shopping, volunteer & 
social groups
Slight impairment in these 
activities
Unable to function 
independently at these 
activities although may still 
be engaged in some; 
appears normal to casual 
inspection
No pretense of independent 
function outside home; 
appears well enough to be 
taken to function outside a 
family home
No pretense of 
independent function 
outside home; 
appears too ill to be 
taken to functions 
outside a family 
home
Home and 
Hobbies
Life at home, hobbies, 
& intellectual interests 
well maintained
Life at home, hobbies, & 
intellectual interests 
slightly impaired
Mild but definite impairment 
of function at home; more 
difficult chores abandoned; 
more complicated hobbies 
& interests abandoned
Only simple chores 
preserved; very restricted 
interests, poorly maintained
No significant 
function in home
Personal Care
Fully capable of self-care
Needs prompting
Requires assistance in 
dressing, hygiene, keeping 
of personal effects
Requires much help 
with personal care; 
frequent incontinence


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17.
ATTACHMENTS
17.1.
Protocol Team Roster6
Protocol Chair:
Jintanat Ananworanich MD, PhD 
 USA
   
Principal Investigator:
Nittaya Phanuphak, MD, PhD 
Thailand
 
Sponsor Clinical Leader
Frank Tomaka, MD
Clinical Leader, 
Janssen Vaccines & Prevention B.V.
Project Leader
 Thailand
 
Clinical Trials Specialist (Thailand)
                                                
6 Changes to the roster will not constitute a protocol amendment.
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 Thailand
Thailand
Statistical Leader
, MSc
Janssen Pharmaceutica NV
BELGIUM
Data Manager
 PhD
Janssen Pharmaceutica NV
BELGIUM
Clinical Virologist
BELGIUM
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Head biomarkers viral vaccines
The Netherlands
Clinical Immunologist
The Netherlands
Global Trial Leader
Malaysia
Study Pharmacist
 Thailand
Laboratory coordinator
 USA
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Investigators
 
hailand
 Thailand
 
 Thailand
Associate Investigators
hailand
 
 Thailand 
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 Thailand
 Thailand
 Thailand
 20817, USA
 
 USA
 
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USA
Consultant
 Thailand 
 
 
Laboratory Investigator
 Thailand
 
17.2.
Study Personnel Roles and Responsibilities
Protocol Chair: Responsible for overall direction and the leadership for the study. The protocol 
chair acts as liaison between the PI with the vaccine developers, sponsors, and funders, and 
supports the site PI with clinical study guidance, protocol preparation, protocol review and 
regulatory approvals.
Principal Investigator: Provides overall project management and the analysis and reporting of 
the study data. The PI is responsible for local IRB submission and approval, study conduct, and 
reporting all unanticipated problems and AEs to the protocol team and the IRB. The PI will act 
as the qualified physician responsible for all study site related medical decisions and will take all 
necessary precautions to ensure that the study obtains the proper clearance for all publication and 
abstracts, and maintain a study regulatory file as instructed by the study sponsor.
Sponsor Clinical Leader: Is a qualified research physician who provides technical assistance 
and supervision on the study design, protocol development and study implementation. The 
sponsor clinical leader is the main liaison between the sponsor, the PI, and the complementing 
team.
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Project Leader: Coordinates implementation of the study at the Thailand clinic site, ensures that 
all study procedures are conducted in accordance with the study protocol, provides training and
supervision to study clinic staff, and supervises quality assurance activities. The project leader 
will assist the PI and protocol chair with local and international IRB submissions and reporting 
requirements, including the reporting of problems and AEs to the protocol team and study 
sponsors. 
Clinical Trials Specialist (Thailand): Is based at the study site in Thailand and under the 
supervision of the PI coordinates all management and administrative issues for the study, 
including local IRB submission and other regulatory affairs. 
Clinical Operation Group, Clinical Research Division: Is based at Department of 
Retrovirology, 
 and under the supervision of the protocol chair and PI coordinates all 
management and administrative issues in the USA for the study, including US DoD submissions 
and other regulatory affairs.
Statistician: Will receive coded data from the data manager and use those data to perform all 
investigational data analyses in collaboration with MHRP and the study team. 
Data Manager: Responsible for overall data management and providing a final data transfer 
following final database closure, including, but not limited to, the data dictionary, formats, 
codes, and any accompanying memorandums. The media format and transfer specifications will 
be agreed upon with the Sponsor.
Virologist: Is an expert on HIV virology who will provide technical assistance to the project on 
study design, protocol development, laboratory evaluations, and data analysis and reporting.
Laboratory Coordinator: A qualified laboratory researcher who will lead the development and 
implementation of all laboratory analyses conducted as part of the protocol. The laboratory 
coordinator coordinates receiving, storing and distributing of biological specimens from the 
study. The laboratory coordinator will not have any access to any subjects’ identifiers or private 
information.
Site Investigators: Conduct the participant study visits, and assist with AE assessment and 
reporting. Site investigators will report any findings and study status directly to the study PI and 
will also coordinate with the PI in the planning, design, and execution of the study.
Senior Investigators: Are experienced clinical researchers in Thailand who will provide 
technical assistance to the PI on study design and implementation. 
Associate Investigators: Assist the PI and senior investigators in performing specific study tasks 
and/or procedures under their supervision.
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Laboratory Investigators: Laboratory investigators supervise clinical and research laboratory 
activities for the study. Laboratory investigators will not have contact with study subjects or 
identifiers.
Study Pharmacist: Qualified pharmacist with responsibility for overseeing the import, 
preparation, blinding, distribution, inventory, and accountability of vaccine, communication with 
sponsor / MHRP regarding pharmacy related issues, training and consultation with the study site 
pharmacy personnel.
Clinical Staff: Can be physicians, nurses, or any specifically trained study personnel to perform 
clinical duties at the study site, including physical examination, counseling, and vaccine 
administration.
Department of Defense (DoD) Research Monitor: The role and qualifications of the DoD
research monitor are identified as follows:
The DoD research monitor is a member of the PSRT of the study. In the context as a member of 
the PSRT, the DoD research monitor is required to review all SAEs and unanticipated problems 
involving risk to volunteers or others, social harms, and all volunteer deaths associated with the 
protocol and provide an unbiased written report of the event. At a minimum, the DoD research 
monitor should comment on the outcomes of the event or problem, and in the case of a SAE, 
social harms, or death, comment on the relationship to participation in the study. The DoD
research monitor should also indicate whether he/she concurs with the details of the report 
provided by the site principal investigator.
The medical monitor will also in the context of the PSRT:
1. Discuss research progress, consult on individual cases, or evaluate suspected adverse reaction 
reports on behalf of the 
/IRB.
2. May at the direction of the 
 IRB, be involved in oversight functions (e.g., 
observe recruitment, enrollment procedures, and the consent process for individuals, groups or 
units; oversee study interventions and interactions; review monitoring plans and UPIRTSO 
reports; and oversee data matching, data collection, and analysis).
3. Promptly report discrepancies or problems to the 
 IRB.
4. Have the authority, in the context of his PSRT membership, to be involved in all decision 
making regarding stopping a research study in progress, removingindividual subjects from a 
study, and taking whatever steps are necessary to protect the safety and well-being of research 
subjects until the IRB can assess the research monitor's report
Consultants: Protocol consultants are responsible for providing input for the study design, 
protocol development, and serve as technical advisors and subject matter experts for study 
execution.
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17.3.
Collaborating Institutions and Investigators
Vaccine Research Center 
AIDS and Cancer Virus Program
 
 
 
 
                                                
7 These collaborating institutions will receive coded samples and/or data.
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Status: Approved, Date: 11 October 2017
 
SWEDEN 
 
National Institutes of Health7
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SIGNATURES 
 
 
 
 
Signed by  
Date  
 
Justification  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Frank Tomaka
13Oct2017, 20:39:38 PM, UTC
Document Approval

SAP_001.pdf: 
1 
Approved, Date: 17 October 2018  
 
Janssen Vaccines & Prevention B.V.* 
 
 
Statistical Analysis Plan 
 
A Combined Phase 1/2a, Exploratory Study of a Therapeutic Vaccine Using an Adenovirus 
Type 26 Vector Prime and Modified Vaccinia Ankara Boost Combination With Mosaic 
Inserts in HIV-1 Infected Adults who Initiated Antiretroviral Treatment During Acute 
HIV Infection 
 
Protocol VAC89220HTX1001; Phase 1/2a 
 
 Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Janssen Vaccines & Prevention B.V. (formerly known as Crucell Holland B.V.) is a Janssen 
pharmaceutical company of Johnson & Johnson and is hereafter referred to as the sponsor of the study.  
Status: 
Approved   
Date: 
17 October 2018 
Prepared by: 
Janssen Vaccines & Prevention B.V.   
Document No.: EDMS-ERI-166858471 
 
Compliance:  The study described in this report was performed according to the principles of Good Clinical Practice (GCP).  
Confidentiality Statement 
The information in this document contains trade secrets and commercial information that are privileged or confidential and may 
not be disclosed unless such disclosure is required by applicable law or regulations. In any event, persons to whom the 
information is disclosed must be informed that the information is privileged or confidential and may not be further disclosed by 
them. These restrictions on disclosure will apply equally to all future information supplied to you that is indicated as privileged or 
confidential. 
NCT02919306


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TABLE OF CONTENTS 
TABLE OF CONTENTS ............................................................................................................................... 2 
AMENDMENT HISTORY .............................................................................................................................. 4 
1. 
INTRODUCTION.................................................................................................................................. 4 
1.1. 
Trial Objectives ................................................................................................................................ 4 
1.2. 
Trial Design ...................................................................................................................................... 4 
1.3. 
Statistical Hypotheses for Trial Objectives ....................................................................................... 5 
1.4. 
Sample Size Justification ................................................................................................................. 5 
1.5. 
Randomization and Blinding ............................................................................................................ 5 
1.6. 
Changes to planned analyses .......................................................................................................... 5 
2. 
GENERAL ANALYSIS DEFINITIONS ................................................................................................ 5 
2.1. 
Study phases ................................................................................................................................... 5 
2.1.1. 
Phase definitions .......................................................................................................................... 6 
2.1.1.1. 
Stage 1 ...................................................................................................................................... 6 
2.1.1.2. 
Stage 2 ...................................................................................................................................... 7 
2.2. 
Pooling Algorithm for Analysis Centers ............................................................................................ 8 
2.3. 
Analysis Sets .................................................................................................................................... 8 
2.3.1. 
Full Analysis Set (FAS) ................................................................................................................. 8 
2.3.2. 
Immunogenicity Analysis Set ........................................................................................................ 8 
2.3.2.1. 
Immunogenicity Population (IP)................................................................................................. 8 
2.3.3. 
Efficacy Analysis Set .................................................................................................................... 8 
2.3.3.1. 
Efficacy Population (EP) ............................................................................................................ 8 
2.3.3.2. 
Per Protocol Efficacy Population (PPE) .................................................................................... 9 
2.4. 
Definition of Subgroups .................................................................................................................... 9 
3. 
INTERIM ANALYSIS AND DATA MONITORING COMMITTEE REVIEW ......................................... 9 
4. 
SUBJECT INFORMATION .................................................................................................................. 9 
4.1. 
Demographics and Baseline Characteristics ................................................................................... 9 
4.2. 
Disease Baseline Characteristics .................................................................................................. 10 
4.3. 
Disposition Information................................................................................................................... 10 
4.4. 
Treatment Compliance ................................................................................................................... 10 
4.5. 
Protocol Deviations ........................................................................................................................ 10 
4.6. 
Concomitant Medications ............................................................................................................... 10 
5. 
SAFETY ............................................................................................................................................. 11 
5.1. 
Adverse Events (AE) ...................................................................................................................... 11 
5.1.1. 
Definitions ................................................................................................................................... 11 
5.1.2. 
Analysis of Adverse Events ........................................................................................................ 12 
5.1.3. 
Transforming On-site Assessments and Diaries of Solicited Adverse Events into an 
Analysis Format ............................................................................ Error! Bookmark not defined. 
5.1.4. 
Phase allocation of Unsolicited Adverse Events ........................................................................ 12 
5.1.4. 
Missing Data ............................................................................................................................... 14 
5.2. 
Laboratory, ECG and Vital Signs ................................................................................................... 14 
6. 
IMMUNOGENICITY ANALYSIS ........................................................................................................ 15 
6.1. 
Parameters ..................................................................................................................................... 15 
6.2. 
Handling of Missing and/or Unquantifiable Immune Response Data ............................................ 16 
6.3. 
Immune Response Analysis .......................................................................................................... 16 
6.3.1. 
Immunogenicity against the insert: ............................................................................................. 16 
6.3.1.1. 
Humoral assays ....................................................................................................................... 16 
6.3.1.2. 
Cellular assays ........................................................................................................................ 17 
6.3.2. 
Immunogenicity against the vector: ............................................................................................ 17 


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7. 
EFFICACY ANALYSIS ...................................................................................................................... 23 
7.1. 
Immunogenicity Assays ................................................................... Error! Bookmark not defined. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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AMENDMENT HISTORY 
SAP Version  
Approval Date 
Original SAP 
17 July 2018 
Final SAP 
17 Oct 2018 
The statistical analysis plan (SAP) from the interim analysis for cohort criteria for proceeding to 
ARV ATI phase is available in ERIS (EDMS-ERI-150930582, V1.0). 
The statistical analysis plan (SAP) from the final analysis is available in ERIS (EDMS-ERI-
150930582, V2.0). 
 
1. 
INTRODUCTION 
This SAP is applicable to the final analysis defined in the VAC89220HTX1001 protocol (EDMS-
ERI-90101185, V 17.0). The final analysis will be performed once all subjects have completed 
their final study visit or discontinued earlier. Interim analyses may be performed prior to the final 
analysis. These interim analyses would occur in a group-unblinded manner, but no subject-level 
unblinding would occur. The final analysis will be unblinded on subject-level. 
Data from subject 37 will be excluded (EC decision: “Subject to be removed from the 
project.  Subject’s data shall not be used for analysis. This is due to the fact that the subject is not 
eligible to participate since the beginning of the trial). This subject will be excluded from the 
safety, efficacy and immunogenicity analysis.  
1.1. 
Trial Objectives 
See CTP, Section 2.1. 
1.2. 
Trial Design 
See CTP, Section 3.1. 


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1.3. 
Statistical Hypotheses for Trial Objectives 
See CTP, Section 2.2. 
1.4. 
Sample Size Justification 
See CTP, Section 9.4. 
1.5. 
Randomization and Blinding 
See CTP, Section 3.3 & 3.4. 
1.6. 
Changes to planned analyses 
NAP 
2. 
GENERAL ANALYSIS DEFINITIONS 
2.1. 
Study phases 
A baseline (or reference) value will be defined as the value of the last available assessment prior 
to the first vaccination on Day 1. 


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The safety analysis will present all results by phase. Listings will be shown per phase and time 
point. 
Immunogenicity results will be presented per scheduled time point as appropriate.  
Study day or relative day is defined as follows: 
Study Day = visit date - date of Day 1 + 1; if visit date≥ date of Day 1 (date of first vaccination). 
Study Day = visit date – date of Day 1; if visit date < date of Day 1 (date of first vaccination). 
 
2.1.1. 
Phase definitions 
 
For the immunogenicity analyses no phases will be constructed.  
The periods/phases will be used primarily for safety and concomitant medication allocation.  The 
post-dose periods and stage 2 (and the regimen phase) are considered active periods/phase, the 
screening and follow-up phases are considered non-active phases. 
For descriptive statistics over time, assessments (regardless of the investigated parameter) will be 
allocated to an analysis visit based on the visit number as captured in the database. 
 
 
 
2.1.1.1. 
Stage 1 
 
Phase 
Phase 
number 
Period 
 
Period 
numbe
r 
Interval 
From 
To 
Screening 
1 
 
00:00 of the date of 
signing the 
informed consent 
forma 
One minute prior to Dose 1 on Day 1   
Regimen 
2 
Post-
Dose 1 
1 
Date and time of 
Dose 1 (Day  1) 
Minimum of:  
a) Maximum (28 days after first vaccination at 
23.59, scheduled visit 4 weeks after first 
vaccination at 23:59) 
b)  23:59 at the date of last contact (for early 
discontinuation) 
Follow-Up 
1 
3 
 
1 minute after end 
of Post-Dose 1 
period 
Minimum of:  
a) One minute prior to date and time of the next 
vaccination 


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Phase 
Phase 
number 
Period 
 
Period 
numbe
r 
Interval 
From 
To 
b) 23:59 at the date of last contact (for early 
discontinuation) 
Regimen 
2 
Post-
Dose 2 
2 
Date and time of 
Dose 2  
Minimum of:  
a) Maximum (28 days after second vaccination at 
23.59, scheduled visit 4 weeks after second 
vaccination at 23:59) 
b) 23:59 at the date of last contact (for early 
discontinuation) 
Follow-Up 
2 
4 
 
1 minute after end 
of Post-Dose 2  
period 
Minimum of:  
a) One minute prior to date and time of the next 
vaccination 
b) 23:59 at the date of last contact (for early 
discontinuation) 
Regimen 
2 
Post-
Dose 3 
3 
Minimum of Date 
and Time Dose 3  
Minimum of:  
a) Maximum (28 days after third vaccination at 
23.59, scheduled visit 4 weeks after third 
vaccination at 23:59) 
b) 23:59 at the date of last contact (for early 
discontinuation) 
Follow-Up 
3 
5 
 
1 minute after end 
of Post-Dose 3  
period 
Minimum of:  
a) One minute prior to date and time of the next 
vaccination 
b) 23:59 at the date of database cut-offc in case of 
interim 
c) 23:59 at the date of last contact (for early 
discontinuation) 
Regimen 
2 
Post-
Dose 4 
4 
Minimum of Date 
and Time Dose 4  
Minimum of:  
a) Maximum (28 days after fourth vaccination at 
23.59, scheduled visit 2 weeks after fourth 
vaccination at 23:59) 
b) 23:59 at the date of database cut-offc in case of 
interim 
c) 23:59 at the date of last contact (for early 
discontinuation) 
Follow-Up 
4 
6 
 
1 minute after end 
of Post-Dose 4  
period 
Minimum of:  
a) 23:59 of the date prior to ART interruption 
b) 23:59 at the date of last contact (for early 
discontinuation) 
NOTE:  
a  The start time of screening phase is 00:00. In case an earlier date is available (e.g. for lab or vital signs then use the very first date in order to 
include all data) 
b In case a dose is not administered, the observations end up in the previous Follow-Up phase 
c For safety interim analyses a cut-off date will be identified: all data included in the analysis will go up to the cut-off date, any data referring to a 
later time point will not be included; ongoing events will be included and duration will be up to the cut-off date. 
 
 
 
2.1.1.2. 
Stage 2 
 


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Phase 
Phase 
number 
Period 
 
Period 
number 
Interval 
From 
To 
Stage 2 
7 
ATI 
5 
00:00 of the date of ART 
interruption 
Minimum of:  
a) Last available visit for completers (no 
ART resumption) 
b) One minute prior to ART resumption. If 
no time available: 23:59 of the date prior to 
ART resumption 
Stage 2 
7 
ART 
resumption 
6 
Date time of ART 
resumption. If no time 
available: 00:00 of the date 
of ART resumption 
Minimum of:  
a) Last available visit for completers 
b) 23:59 at the date of last contact (for early 
discontinuation) 
2.2. 
Pooling Algorithm for Analysis Centers 
NAP 
2.3. 
Analysis Sets 
Vaccination assignment will follow the as treated principle. 
2.3.1. 
Full Analysis Set (FAS) 
The full analysis set will include all randomized subjects with at least one vaccine administration 
documented. Subject 37 as indicated above will be excluded from this analysis set. 
2.3.2. 
Immunogenicity Analysis Set 
2.3.2.1. 
Immunogenicity Population (IP) 
All immunogenicity analyses will be performed on subjects that were randomized and received at 
least 3 vaccinations. For the unblinded final analysis only: vaccinations should be given within the 
protocol-specified windows +/- 7 days for whom immunogenicity data are available excluding 
subjects with major protocol deviations expecting to impact the immunogenicity outcomes. 
 
In addition, all samples obtained after missed doses will be excluded from the analysis (eg if a 
subject in group 1 received their first 3 vaccinations, but not the last, samples pertaining to the first 
3 vaccinations can still be used, but not for the 4th vaccination. Similarly, if a subject received their 
first two and their last vaccination but missed the third vaccination then samples up and till the 
second vaccination will be used, later samples will be excluded). 
 
2.3.3. 
Efficacy Analysis Set 
2.3.3.1. 
Efficacy Population (EP) 
The primary efficacy population will consist of all subjects who undergo ARV ATI at Week 60 
(Stage 2), regardless of the time or outcome of treatment interruption. Subject 37 as indicated 
above will be excluded from this analysis set. 


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2.3.3.2. 
Per Protocol Efficacy Population (PPE) 
The per protocol efficacy population will include all randomized and fully vaccinated subjects for 
whom efficacy data concerning endpoint measures are available excluding subjects with major 
protocol deviations expecting to impact the efficacy outcomes. 
If there is no difference in subjects between the EP and PPE population, all the outputs will state 
the analysis is done on the EP population. 
2.4. 
Definition of Subgroups 
Three subgroups based on their time to rebound will be made. The cut-off values are to be chosen 
such that 33.3% of all vaccinated subjects from the active arm fall in the first subgroup (1st tertile), 
33.3% fall in the second subgroup (2nd tertile), 33.3% fall in the third subgroup (3rd tertile). These 
groups will be shown in a different color in the graphs. Analogously, tertiles subgroups will be 
computed in the placebo arm. Here the cut-off values are to be chosen such that 33.3% of all 
vaccinated subjects from the placebo arm fall in the first subgroup (1st tertile), 33.3% fall in the 
second subgroup (2nd tertile), 33.3% fall in the third subgroup (3rd tertile). 
3. 
INTERIM ANALYSIS AND DATA MONITORING COMMITTEE REVIEW 
The final analysis will be performed once all subjects have completed their final study visit or 
discontinued earlier. Interim analyses may be performed prior to the final analysis. These analyses 
would occur in a group-unblinded manner, but no subject-level unblinding would occur. One 
interim analysis is performed on W26 ICS data to evaluate the subjects entering in ATI. One 
interim analysis is performed on immunogenicity data only up to W50. The final analysis will 
evaluate safety, immunogenicity and efficacy. 
4. 
SUBJECT INFORMATION 
Subject information will be analyzed based on the FA analysis set unless otherwise specified.  
Continuous variables will be summarized using the following statistics, as appropriate: number of 
observations, arithmetic mean (mean), 95% confidence interval (CI) for the mean, standard 
deviation (SD), standard error (SE), median, quartiles (Q1 and Q3), minimum and maximum. The 
minimum and maximum will be presented to the same number of decimal places as the original 
data. The mean and median will be rounded to one more decimal place than the original data, while 
the SD, SE and 95% CI to two more decimal places. 
4.1. 
Demographics and Baseline Characteristics 
The following demographic and baseline characteristics will be summarized, where available. 
• Sexa (Female/Male)     
 
 
• Age (years) 
• Race 
• Ethnicity 
                                                 
 
a At screening 


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• Region 
• Country 
• Height (cm) 
• Weight (kg) 
• BMI (kg/m2), calculated from the recording of baseline height and weight 
 
4.2. 
 Disease Baseline Characteristics 
The following disease baseline characteristics will be summarized, where available: 
• HIV RNA 
• Date of HIV diagnosis 
• ART history 
• Fiebig Stage (I, II, III or IV) 
• CD4 Cell Count 
• CD8 Cell Count 
• Hepatitis B/C at screening 
• VDRL at screening 
• TPHA at screening 
4.3. 
Disposition Information 
The number and percentage of subjects screened, subjects in the FAS, IP, EP, PPE and subjects 
vaccinated and not randomized, subjects randomized and not vaccinated and discontinued subjects 
(study discontinuation and vaccination discontinuation) with the reason of discontinuation will be 
tabulated per vaccine group and overall. 
Also the number of subjects and percentage per phase will be tabulated. 
4.4. 
Treatment Compliance 
The number of missed vaccinations will be tabulated. 
4.5. 
Protocol Deviations 
Major protocol deviations will be summarized.  
4.6. 
Concomitant Medications 
The analysis of concomitant medications will be using the WHO drug dictionary as provided in 
the clinical database. 
Based on their start and stop date, concomitant therapies will be reported in each period during 
which they were applied. 
If a concomitant therapy record misses components of its start and/or stop dates (Day and/or month 
and/or year):  


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- 
In case of partial start or stop dates, the concomitant therapy records will be allocated to 
periods using the available partial information, without imputations. If, for example, only month 
and year are available, these will be compared to the month and the year of the periods, and the 
concomitant therapy record will be allocated to the period(s) where these date parts match.   
- 
In case of a completely missing start date, the concomitant therapy will be considered as 
having started before the trial.  
- 
In case of a completely missing end date, the concomitant therapy will be considered as 
ongoing at the end of the trial/cut-off date. 
Concomitant therapies will be tabulated. There will be special attention to Acetaminophen, 
NSAIDS or Antihistamines to identify medication that can mask local or systemic solicited events 
in the 8 days after the vaccinations and to Glucocorticosteroids during the whole study duration 
for possible influence on immunogenicity results. Glucocorticosteroids will be flagged in the 
overall CM listing.  
 
ART will be listed before and after ATI separately for each subject. 
 
 
5. 
SAFETY 
 
Safety analyses will be performed on the FAS. Continuous variables will be summarized using the 
following statistics, as appropriate: number of observations, arithmetic mean (mean), 95% CI for 
the mean (if applicable, large studies), standard deviation (SD), standard error (SE), median, 
quartiles (Q1 and Q3), minimum and maximum. Frequencies and percentages (one decimal place) 
will be generated for categorical variables. No formal comparisons between groups will be 
provided.  
Safety data will be analysed by vaccine regimens as designed per protocol (per regimen analysis). 
In case of multiple vaccinations, data will be presented by phase as well as over the entire regimen. 
For stage 2 safety data will be shown by phase and period, where applicable (no solicited adverse 
events). Denominator for the percentages is the number of subjects in the considered population 
and phase for a certain regimen (incidence per 100 subjects/phase).  
 
5.1. 
Adverse Events (AE) 
 
5.1.1. 
Definitions 
Solicited AEs shown in the tables are extracted from the diary pages/ on-site assessment pages of 
the CRF (systemic solicited events: fever, fatigue, headache, myalgia, arthralgia, chills, nausea, 
vomiting, rash, general itching and local solicited events: erythema, warmth, itching, swelling, 


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induration and pain/tenderness). For unsolicited AEs, only the AEs within the 28-day period 
following each vaccination will be presented in the safety tables except for SAEs, which will be 
captured and tabulated in the outputs covering the whole study period. All other collected 
unsolicited adverse events will be presented through listings. For stage 2, AEs will be shown by 
phase and period, where applicable. 
Solicited local AEs will be by definition considered as related to the study vaccine. Solicited events 
are always allocated to the respective Post Dose period. 
The severity of the AEs will be classified as grade 1 to 4, using the grades in SDTM. Solicited 
events that are graded less than grade 1, are not considered as AE. In case no grades are available 
the grading of the solicited events will be based on the Division of AIDS (DAIDS) Table for 
Grading the Severity of Adult and Pediatric Adverse Events (DAIDS AE Grading Table), Version 
2.1 
dated 
July 
2017 
found 
on 
the 
website: 
http://rsc.tech-res.com/docs/default-
source/safety/daidsgradingcorrecetedv21.pdf?sfvrsn=6. 
Solicited events which meet the AE criteria are reported in the AE domain. Separated analysis 
datasets should be created for unsolicited AEs and solicited AEs. 
 
5.1.2. 
Analysis of Adverse Events 
Number and percentage of subjects with at least one particular AE (unsolicited/solicited) will be 
tabulated. Unsolicited AEs will be summarized by System Organ Class and Preferred Term. 
Solicited AEs will be summarized by class (local, systemic) and preferred term. 
For solicited AEs following tables will be provided: summary, by worst severity grade, grade 3, 
related (systemic only), time to onset (in days) and duration (in days) for most frequent events and 
body temperature. Note: Duration is defined as number of days from the start of the event until 
resolution of the event. The time to first onset is defined as (date of first onset – reference date + 
1). The reference date is the start date of the vaccination period. 
For unsolicited AEs following tables will be provided: summary table (including SAE, fatal 
outcome, and discontinuation), all events, most frequent, at least grade 3, permanent stop of 
vaccine, related and SAE. 
 
Listings and/or subject narratives will be provided as appropriate, for those subjects who die, 
discontinue study vaccinations due to an AE, or experience a related grade 3 or 4 event or SAE. 
 
5.1.3. 
Phase allocation of Unsolicited Adverse Events 
 
Step 1: Allocation of events to the periods: 


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Adverse events in the SDTM database are allocated to periods based on their start date/time. If the 
start date/time of an event falls between (or on) the start and stop date/time of a period, the AE is 
attributed to that period (treatment-emergent principle). 
- In case of partial start or stop dates (i.e. time and/or day and/or month and/or year missing), the 
events are allocated to the periods using the available partial information on start and end date; no 
imputation will be done. If, for instance, the AE start date only month and year are available, these 
data are compared to the month and year information of the periods. This rule may lead to 
multiplication of the event as a consequence of its assignment to multiple periods. 
- In case of a completely missing end date, the date is imputed by the cut-off date of the analysis 
for subjects still ongoing in the study, and by the end date of the last period for subjects who 
discontinued or completed the trial. 
Step 2: Combination of events: 
Overlapping/consecutive events are defined as events of the same subject with the same preferred 
term which have at least 1 day overlap or for which the start date of an event is 1 day after the end 
date of the preceding event. Overlapping/consecutive events may be combined into one AE or not, 
according to the following rules: 
1) If overlapping/consecutive events start in one of the following periods - Screening or post dose 
extension (i.e. non-active periods) - followed by an AE in - post-dose period (active period) - they 
are allocated to their respective periods and are considered as separate events. 
2) In case overlapping/consecutive events start within a single period, they are considered as one 
and the same AE. The individual events which contribute to this AE are retained as individual 
records in the ADaM database but are assigned the same onset, period, and total duration. All 
related attributes to the AE/phase/period should also be consistent with the new event. 
3) In case overlapping/consecutive events start in both an active period followed by a non active 
period, they are allocated to the active period only and are considered as one and the same AE. 
The individual events which contribute to this AE are retained as individual records in the ADaM 
database but are assigned the same onset, treatment period, and total duration. All related attributes 
to the AE/phase/period should also be consistent with the new event. 
4) In case an active period is followed by another active period, and the overlapping/consecutive 
events start in both periods, they are allocated to their respective period and are considered as 
separate AEs. The same rule applies for 2 non-active periods.  
Remarks: 
1. Events can only be combined into one and the same AE if their start and stop dates are known. 
2. In case the completely missing end date is imputed (for period allocation), this date is also 
considered as a complete date. 


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3. Time is not considered when determining overlap of events. 
 
5.1.4. 
Missing Data 
Missing data will not be imputed. Subjects who do not report an event will be considered as 
subjects without an event. The analysis of the solicited AEs will include only documented safety 
data (i.e. in case severity is missing it is not considered an event). 
 
5.2. 
Laboratory, ECG and Vital Signs 
For laboratory safety parameters, vital signs and ECG (if applicable) only abnormalities emerging 
after vaccination will be tabulated by worst abnormality grade using the table in attachment 1. QT 
interval abnormalities will be defined only for corrected QT values (QTcF or QTcB) if available 
(no recalculation from QT values will be performed). Abnormalities on the changes from 
reference, if available, for QT interval will be categorized in ‘<30ms’, ‘[30; 60]ms’ and ‘>60ms’. 
Increases of less than 30ms will not be considered as an abnormal increase.  
A listing of subjects with fever according to the DAIDS grading table will be provided. 
An abnormality (toxicity grades according to the DAIDS (website: http://rsc.tech-
res.com/docs/default-source/safety/daidsgradingcorrecetedv21.pdf?sfvrsn=6) 
or 
abnormality 
based on normal ranges) will be considered as emerging in a particular period if it is worse than 
the baseline value. If the baseline is missing, the abnormality is always considered emerging. A 
shift from ‘abnormally low’ at baseline to ‘abnormally high’ post baseline (or vice versa) is also 
emerging. In case a laboratory test result is censored (no numeric value is available, but only a 
verbatim term) then a numeric value will be imputed by a value exceeding the cut-off value with 
one unit. (<x: subtract 1 unit from x, >x: add 1 unit to x; <3.45 is imputed with 3.44). 
In case no toxicity grades are defined for a test, the abnormalities (above/below normal range) will 
be used. In determining toxicity grades, the following rules are applied: 
-worst grades/abnormalities are determined over the whole observational period for each trial 
period separately, including all post-baseline measurements of that period. 
- The abnormalities ‘abnormally low’ and ‘abnormally high’ are considered equally important, i.e. 
if a subject has as well an abnormally low as an abnormally high value post-baseline, both 
abnormalities are shown in the tables. (This means that the sum of the percentages can by more 
than 100%) 
- Note: as the grading scale for some parameters in the grading table has some gaps (zones where 
no toxicity grade definition exists), laboratory results falling in these zones will be allocated to the 
adjacent worst-case grade. 
- If a lab value falls within the grading as specified in the grading table but also within the local 
lab normal limits, the value is considered as normal.  


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For the grades, no distinction will be made between test results of samples obtained under fasting 
and under non-fasting conditions: in case limits under fasting and non-fasting conditions differ, 
the limits of the conditions (fasting/non-fasting) of scheduled visits as planned in the CTP will 
always be used, also for samples obtained under a different condition (e.g. samples of withdrawal 
visits). 
 
6. 
IMMUNOGENICITY ANALYSIS 
The analysis of immunogenicity will be done on the IP set. Also, in addition to the general graphs, 
graphs with the following specifications will be made. Three subgroups based on their time to 
rebound will be made. The cut-off values are to be chosen such that 33.3% of all vaccinated 
subjects from the active arm fall in the first subgroup (1st tertile), 33.3% fall in the second 
subgroup (2nd tertile), 33.3% fall in the third subgroup (3rd tertile). These groups will be shown 
in a different color in the graphs. Analogously, tertiles subgroups will be conducted in the placebo 
arm. Here the cut-off values are to be chosen such that 33.3% of all vaccinated subjects from the 
placebo arm fall in the first subgroup (1st tertile), 33.3% fall in the second subgroup (2nd tertile), 
33.3% fall in the third subgroup (3rd tertile). Subjects who are still undetectable on the W84 visit 
for immunogenicity will be shown with a different symbol. 
The timing of the rebound is defined as follows: 
A) Using the first time of multiple consecutive determinations of HIV-1 RNA above 1000 
copies/ml at least 1 week apart for the real rebounders. 
B) Using the last determination of HIV-1 RNA above 1000 copies/ml before restart of ART 
for the others. 
 
In addition, subgroups will be created based on AD26 baseline seropositivity. 
6.1. 
Parameters 
The following humoral and cellular immune responses are measured by immunogenicity against 
the insert. 
Humoral response 
- Env ELISA IgG-t gp140 (Clade A (92UG037.1); Clade B (1990a); Clade C (Con C); Clade C (C97ZA.012);  
Mos1)  
- ELISA IgG 1-4 (Clade C (C97ZA.012) IgG1; Clade C (C97ZA.012) IgG2; Clade C (C97ZA.012) IgG3;  
Clade C (C97ZA.012) IgG4) 
- ADCC  
Cellular response 
- ELISpot 
- ELISpot breadth 
- ICS 
The following humoral responses are measured by immunogenicity against the vector: 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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- Ad 26 VNA 
 
6.2. 
Handling of Missing and/or Unquantifiable Immune Response Data  
Missing immune response data will not be imputed.   
Values below the lower limit of quantification (LLOQ) or limit of detection (LOD) will be handled 
as follows:  
- 
Calculation of geomean and median:  
o values<LLOQ are imputed=LLOQ/2.  
- 
Calculation of fold increases from baseline:  
o values <LLOQ are imputed with LLOQ.  
- 
Note: for ADCP the LOD will be considered and the same rules apply using LOD as 
reference. 
                                
Values above the upper limit of quantification (ULOQ) will be handled as follows:  
- 
Calculation of geomean and median:  
o Values>ULOQ are imputed=ULOQ.  
- 
Calculation of fold increases from baseline:  
o Values >ULOQ are imputed with ULOQ.  
                                
6.3. 
Immune Response Analysis 
No formal hypothesis on immunogenicity will be tested. 
6.3.1. 
Immunogenicity against the insert: 
6.3.1.1. 
Humoral assays 
For the humoral assays following results will be calculated and tabulated: N, geometric mean§ 
and corresponding 95% CI, fold increases from baseline and responder rates. §calculate the mean and 
corresponding 95%CI of the log10 transformed values, back-transform this mean [i.e. 10^mean] and CI [i.e. 10^CI]. 
For all the humoral assays the immune response values will be log10-transformed before any 
further handling. The log10-transformed values will be used throughout the analysis. In the graphs, 
original values will be displayed on the log10 scale.   
Graphical presentations will be provided displaying dots for the subject values and including the 
N, geometric mean and the percentage of responders. Baseline values will be summarized by 
pooling the groups and will be displayed on the left of each graph. In the graphs the actual values 
will be shown and the LLOQ cut-off will be visualized as a horizontal dotted line together with 
the reference ‘LLOQ’. The values below LLOQ will be visualized with the value imputed as 
described in section 6.4 (column LLOQ or LOD).  


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Statistical Analysis Plan VAC89220HTX1001  
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In general a sample is considered positive, if the value is above the assay LLOQ. A response will 
be defined if the sample interpretation is negative at baseline and positive post-baseline, or, if 
sample interpretation is positive at both time points and there is a >x-fold increase from baseline 
(x-fold increase on the original scale). The definition of responders is defined in section 6.4. for 
each assay.  
6.3.1.2. 
Cellular assays 
For cellular assays following results will be calculated: N, median, quartile range, and min-max 
range of the actual values together with responders will be tabulated and graphically presented.  
Tables with the corresponding descriptive statistics will be provided. 
Actual values are shown as plots with dots for subject values, and the corresponding median and 
interquartile range per time point for each assay. At the bottom the N, median value and percentage 
of responders will be shown. For Elispot, subject values post-baseline will be defined as responder 
when the value is above threshold and threshold is defined as the 95th percentile of all baseline 
data calculated per peptide pool (reference A004 data). The definition of responders is defined in 
section 6.4. 
For the graphs, original values will be displayed on the log10 scale. In the graphs the actual values 
will be shown and the P95 cut-off will be visualized as a horizontal dotted line together with the 
reference ‘P95=value for each peptide pool. The values below LLOQ will be visualized with the 
value imputed as described in section 6.2.  
6.3.2. 
Immunogenicity against the vector: 
For VNA following statistics will be calculated: N, geometric mean (see above for the calculation) 
and corresponding 95% CI of the actual values. 
Actual values are tabulated and shown as dot plots with dots for subject values, and the 
corresponding geometric mean and 95% CI at baseline.


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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6.4. 
Immunogenicity Assays 
 
The details for each immune response assay are listed in following table 
 
Assay 
Test  
Lab 
Time 
Points 
LLOQ 
LOD 
Treshold ULOQ 
Unit 
Responder definition 
(R) 
Fold increase (FI) 
calculation 
Env ELISA  
IgG-t gp140 
Clade A (92UG037.1) 
Clade B (1990a) 
Clade C (Con C) 
Clade C (C97ZA.012) 
Mos1 
Janssen 
Vaccines 
&Prevention 
Wk 0, 4, 16, 
26, 50, 60, 
84, 96 
 
625 
156.25 
625 
156.25 
78.125 
0 
EU/ml 
1) if baseline <LLOQ, 
R>LLOQ  
2) if baseline 
>=LLOQ, R=3-fold 
increase from baseline  
1) if baseline 
>LLOQ, FI=Value 
post-baseline/Value 
wk0 
2) if 
baseline<LLOQ, 
FI=value post-
baseline/LLOQ 
Env ELISA 
IgG1-4 
Clade C (C97ZA.012) IgG1 
Clade C (C97ZA.012) IgG2 
Clade C (C97ZA.012) IgG3 
Clade C (C97ZA.012) IgG4  
BIDMC 
Wk0 
Wk 50  
12.3 
28.7 
12.4 
13.2 
4 
EC50 
ADCC 
HIV ENV gp140 Mos1 Fun Ab 
Schuetz 
(MHRP) 
Wk0 
Wk 50 
  
NAP 
 
NAP 
% 
 
 
Ad26 VNA 
Ad26 Vector Neutralization 
Janssen 
Vaccines& 
Prevention 
Wk0 
17 
NAP 
IC90 
>LLOQ 
NAP 
ELISpot 
 
 
 
 
 
 
HIV IFNg ENV pep pool (Mos1) 
HIV IFNg Pol pep pool (Mos1) 
HIV IFNg Gag pep pool (Mos1) 
HIV IFNg ENV pep pool (Mos2) 
HIV IFNg Pol pep pool (Mos2) 
HIV IFNg Gag pep pool (Mos2) 
Env peptide pool PTE (Env peptide pool 1 PTE, 
Env peptide pool 2 PTE, Env peptide pool 3 
PTE) 
HIV IFNg ENV1 pep pool (Clinical PTE) 
HIV IFNg ENV2 pep pool (Clinical PTE) 
HIV IFNg ENV3 pep pool (Clinical PTE) 
HIV IFNg Pol pep pool (Clinical PTE) 
 
HIV IFNg Gag pep pool (Clinical PTE) 
HIV IFNg ENV1 pep subpool (Mos1) 
HIV IFNg ENV2 pep subpool (Mos1) 
HIV IFNg ENV3 pep subpool (Mos1) 
HIV IFNg ENV4 pep subpool (Mos1) 
HIV IFNg ENV5 pep subpool (Mos1) 
HIV IFNg ENV6 pep subpool (Mos1) 
BIDMC 
Wk0 
Wk26 
Wk 50 
  
 
 
 
 
 
 
 
 
 
55 
55 
55 
 
 
55 
0 
73a 
112 
87 
70 
63 
55 
100 
105 
181 
SFC/10
6 
PBMC   
1) if 
baseline<threshold, 
R>threshold. 2) if 
baseline >= threshold, 
R=3-fold increase 
from baseline. 
(reference A004) 
 
 
 
 
 
 
 
 For the subpools: 
 Responder: >55 
 
 
NAP 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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HIV IFNg ENV7 pep subpool (Mos1) 
HIV IFNg ENV8 pep subpool (Mos1) 
HIV IFNg ENV9 pep subpool (Mos1) 
HIV IFNg ENV10 pep subpool (Mos1) 
HIV IFNg ENV11 pep subpool (Mos1) 
HIV IFNg ENV12 pep subpool (Mos1) 
HIV IFNg ENV13 pep subpool (Mos1) 
HIV IFNg ENV14 pep subpool (Mos1) 
HIV IFNg ENV15 pep subpool (Mos1) 
HIV IFNg ENV16 pep subpool (Mos1) 
HIV IFNg ENV17 pep subpool (Mos1) 
HIV IFNg Gag1 pep subpool (Mos1) 
HIV IFNg Gag2 pep subpool (Mos1) 
HIV IFNg Gag3 pep subpool (Mos1) 
HIV IFNg Gag4 pep subpool (Mos1) 
HIV IFNg Gag5 pep subpool (Mos1) 
HIV IFNg Gag6 pep subpool (Mos1) 
HIV IFNg Gag7 pep subpool (Mos1) 
HIV IFNg Gag8 pep subpool (Mos1) 
HIV IFNg Gag9 pep subpool (Mos1) 
HIV IFNg Gag10 pep subpool (Mos1) 
HIV IFNg Gag11 pep subpool (Mos1) 
HIV IFNg Gag12 pep subpool (Mos1) 
HIV IFNg Pol1 pep subpool (Mos1) 
HIV IFNg Pol2 pep subpool (Mos1) 
HIV IFNg Pol3 pep subpool (Mos1) 
HIV IFNg Pol4 pep subpool (Mos1) 
HIV IFNg Pol5 pep subpool (Mos1) 
HIV IFNg Pol6 pep subpool (Mos1) 
HIV IFNg Pol7 pep subpool (Mos1) 
HIV IFNg Pol8 pep subpool (Mos1) 
HIV IFNg Pol9 pep subpool (Mos1) 
HIV IFNg Pol10 pep subpool (Mos1) 
HIV IFNg Pol11 pep subpool (Mos1) 
HIV IFNg Pol12 pep subpool (Mos1) 
HIV IFNg Pol13 pep subpool (Mos1) 
HIV IFNg Pol14 pep subpool (Mos1) 
HIV IFNg Pol15 pep subpool (Mos1) 
HIV IFNg Pol16 pep subpool (Mos1) 
HIV IFNg Pol17 pep subpool (Mos1) 
HIV IFNg Pol18 pep subpool (Mos1) 
HIV IFNg Pol19 pep subpool (Mos1) 
HIV IFNg Pol20 pep subpool (Mos1) 
HIV IFNg Pol21 pep subpool (Mos1) 
HIV IFNg ENV1 pep subpool (PTE) 
HIV IFNg ENV2 pep subpool (PTE) 
HIV IFNg ENV3 pep subpool (PTE) 
HIV IFNg ENV4 pep subpool (PTE) 
HIV IFNg ENV5 pep subpool (PTE) 
HIV IFNg ENV6 pep subpool (PTE) 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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HIV IFNg ENV7 pep subpool (PTE) 
HIV IFNg ENV8 pep subpool (PTE) 
HIV IFNg ENV9 pep subpool (PTE) 
HIV IFNg ENV10 pep subpool (PTE) 
HIV IFNg ENV11 pep subpool (PTE) 
HIV IFNg ENV12 pep subpool (PTE) 
HIV IFNg ENV13 pep subpool (PTE) 
HIV IFNg ENV14 pep subpool (PTE) 
HIV IFNg ENV15 pep subpool (PTE) 
HIV IFNg ENV16 pep subpool (PTE) 
HIV IFNg ENV17 pep subpool (PTE) 
HIV IFNg Gag1 pep subpool (PTE) 
HIV IFNg Gag2 pep subpool (PTE) 
HIV IFNg Gag3 pep subpool (PTE) 
HIV IFNg Gag4 pep subpool (PTE) 
HIV IFNg Gag5 pep subpool (PTE) 
HIV IFNg Gag6 pep subpool (PTE) 
HIV IFNg Gag7 pep subpool (PTE) 
HIV IFNg Gag8 pep subpool (PTE) 
HIV IFNg Gag9 pep subpool (PTE) 
HIV IFNg Gag10 pep subpool (PTE) 
HIV IFNg Gag11 pep subpool (PTE) 
HIV IFNg Gag12 pep subpool (PTE) 
HIV IFNg Pol1 pep subpool (PTE) 
HIV IFNg Pol2 pep subpool (PTE) 
HIV IFNg Pol3 pep subpool (PTE) 
HIV IFNg Pol4 pep subpool (PTE) 
HIV IFNg Pol5 pep subpool (PTE) 
HIV IFNg Pol6 pep subpool (PTE) 
HIV IFNg Pol7 pep subpool (PTE) 
HIV IFNg Pol8 pep subpool (PTE) 
HIV IFNg Pol9 pep subpool (PTE) 
HIV IFNg Pol10 pep subpool (PTE) 
HIV IFNg Pol11 pep subpool (PTE) 
HIV IFNg Pol12 pep subpool (PTE) 
HIV IFNg Pol13 pep subpool (PTE) 
HIV IFNg Pol14 pep subpool (PTE) 
HIV IFNg Pol15 pep subpool (PTE) 
HIV IFNg Pol16 pep subpool (PTE) 
HIV IFNg Pol17 pep subpool (PTE) 
HIV IFNg Pol18 pep subpool (PTE) 
HIV IFNg Pol19 pep subpool (PTE) 
HIV IFNg Pol20 pep subpool (PTE) 
HIV IFNg Pol21 pep subpool (PTE) 
HIV IFNg ENV1 pep subpool (Mos2) 
HIV IFNg ENV2 pep subpool (Mos2) 
HIV IFNg ENV3 pep subpool (Mos2) 
HIV IFNg ENV4 pep subpool (Mos2) 
HIV IFNg ENV5 pep subpool (Mos2) 
HIV IFNg ENV6 pep subpool (Mos2) 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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HIV IFNg ENV7 pep subpool (Mos2) 
HIV IFNg ENV8 pep subpool (Mos2) 
HIV IFNg ENV9 pep subpool (Mos2) 
HIV IFNg ENV10 pep subpool (Mos2) 
HIV IFNg ENV11 pep subpool (Mos2) 
HIV IFNg ENV12 pep subpool (Mos2) 
HIV IFNg ENV13 pep subpool (Mos2) 
HIV IFNg ENV14 pep subpool (Mos2) 
HIV IFNg ENV15 pep subpool (Mos2) 
HIV IFNg ENV16 pep subpool (Mos2) 
HIV IFNg ENV17 pep subpool (Mos2) 
HIV IFNg Gag1 pep subpool (Mos2) 
HIV IFNg Gag2 pep subpool (Mos2) 
HIV IFNg Gag3 pep subpool (Mos2) 
HIV IFNg Gag4 pep subpool (Mos2) 
HIV IFNg Gag5 pep subpool (Mos2) 
HIV IFNg Gag6 pep subpool (Mos2) 
HIV IFNg Gag7 pep subpool (Mos2) 
HIV IFNg Gag8 pep subpool (Mos2) 
HIV IFNg Gag9 pep subpool (Mos2) 
HIV IFNg Gag10 pep subpool (Mos2) 
HIV IFNg Gag11 pep subpool (Mos2) 
HIV IFNg Gag12 pep subpool (Mos2) 
HIV IFNg Pol1 pep subpool (Mos2) 
HIV IFNg Pol2 pep subpool (Mos2) 
HIV IFNg Pol3 pep subpool (Mos2) 
HIV IFNg Pol4 pep subpool (Mos2) 
HIV IFNg Pol5 pep subpool (Mos2) 
HIV IFNg Pol6 pep subpool (Mos2) 
HIV IFNg Pol7 pep subpool (Mos2) 
HIV IFNg Pol8 pep subpool (Mos2) 
HIV IFNg Pol9 pep subpool (Mos2) 
HIV IFNg Pol10 pep subpool (Mos2) 
HIV IFNg Pol11 pep subpool (Mos2) 
HIV IFNg Pol12 pep subpool (Mos2) 
HIV IFNg Pol13 pep subpool (Mos2) 
HIV IFNg Pol14 pep subpool (Mos2) 
HIV IFNg Pol15 pep subpool (Mos2) 
HIV IFNg Pol16 pep subpool (Mos2) 
HIV IFNg Pol17 pep subpool (Mos2) 
HIV IFNg Pol18 pep subpool (Mos2) 
HIV IFNg Pol19 pep subpool (Mos2) 
HIV IFNg Pol20 pep subpool (Mos2) 
HIV IFNg Pol21 pep subpool (Mos2) 
ICS 
HIV ENV1 pep pool (Mos1) 
HIV ENV2 pep pool (Mos1) 
HIV Pol1 pep pool (Mos1) 
HIV Pol2 pep pool (Mos1) 
HIV Gag pep pool (Mos1) 
 
Schuetz 
(MHRP) 
W0 
W26 
W50 
W60 
W84 
W96 
CD4 for 
IFNg %:.03 
CD8 for 
IFNg %:.09 
 
 
% of 
CD4+/CD8+ 
T cells 
expressing 
IFNg 
Only for IFNg: 
1)Baseline <LLOQ: if 
post baseline values 
≥LLOQ  
 
NAP 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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CD4+INFg+ 
CD8+INFg+ 
CD4+IL2+ 
CD8+IL2+ 
CD4+TNFa+ 
CD8+TNFa+ 
 
 
CD4+IL2-IFNg-TNFa+ 
CD4+IL2-IFNg+TNFa- 
CD4+IL2-IFNg+TNFa+ 
CD4+IL2+IFNg-TNFa- 
CD4+IL2+IFNg-TNFa+ 
CD4+IL2+IFNg+TNFa- 
CD4+IL2+IFNg+TNFa+ 
CD8+IL2-IFNg-TNFa+ 
CD8+IL2-IFNg+TNFa- 
CD8+IL2-IFNg+TNFa+ 
CD8+IL2+IFNg-TNFa- 
CD8+IL2+IFNg-TNFa+ 
CD8+IL2+IFNg+TNFa- 
CD8+IL2+IFNg+TNFa+ 
CD4+CD154+ 
CD4+CD154+INFg+ 
CD4+CD154+IL2+ 
CD4+CD154+TNFa+ 
CD8+CD107a+ 
CD8+CD107a+INFg+ 
CD8+CD107a+IL2+ 
CD8+CD107a+TNFa+ 
CD8+CD107a+GrzB+ 
CD4+CD154+GrzB+ 
CD4+GrzB+ 
CD4+IL4+ 
CD4+IL4+IL2+ 
CD4+IL4+INFg+ 
CD4+IL4+IL2-INFg+ 
CD4+IL4+IL2+INFg- 
2)Baseline >=LLOQ : 
if post baseline values 
are 2 times the 
baseline value 
 
 
a:Threshold for ELISpot test is based on the 95 percentile from the baseline values of about 350 subjects on that test in the HIV-V-A004 study 


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Statistical Analysis Plan VAC89220HTX1001  
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7. 
EFFICACY ANALYSIS 
 
The frequency and duration of sustained viremic control (> 24 weeks of plasma HIV RNA <50 
copies/ml) after ATI will be tabulated. In addition this will be repeated for HIV RNA < 1000 
copies/ml).  
A tabulation will be provided of the restart of ART. Median (min, max, quartiles) time to ART 
restart will also be tabulated. 
A Kaplan-Meier plot of the time to ART restart after ATI together with a log-rank test will be 
provided comparing the placebo and the vaccine group. 
Individual subject profiles of plasma HIV RNA copies/ml will be graphically shown together with 
relevant data (eg start of ART reinitiating and immunogenicity assay results (titer, %), biomarkers 
of HIV reservoir (HIV total DNA), acute retroviral syndrome, CD4/CD8 cell counts) aligning 
timepoints.  
For the total HIV DNA the frequency of subjects that are undetectable at each timepoint will be 
tabulated and graphically shown. 
If corresponding data is available the following analyses will be performed: 
A tabulation will be made on any viral resistance that occurred after ATI. 
A listing with subjects experiencing acute retroviral syndrome post ARV ATI will be 
provided. 
The molecular sequence sieve effects of vaccine therapy on breakthrough rebound viremia 
before and after cessation of therapy will be evaluated. 
 
The peripheral blood mononuclear cells (PBMC) phenotype, pattern of soluble factors and 
immune functional responses before and after ARV ATI in both the vaccine and placebo 
arms and compared to historical untreated acute infection cases in RV217 will be 
evaluated. 
 


Ad26.Mos.HIV/MVA-Mosaic  
 
 
 
Statistical Analysis Plan VAC89220HTX1001  
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7.1. 
Efficacy Assays  
 
LBTESTCD 
LBTEST 
LBDESCR 
LBCAT 
LBSPEC 
LBMETHOD 
LBSCAT 
LBORRESU 
LBLLOQ 
LBLOD 
TESTTYPE 
LBNAM 
POST PROCESSING 
COMMENTS 
HIVDNA 
HIV DNA 
PBMC HIV 
DNA 
CHEMISTRY 
PERIPH
ERAL 
BLOOD 
MONO
NUCLE
AR 
CELL 
QUANTIT
ATIVE 
REVERSE 
TRANSCR
IPTASE 
POLYMER
ASE 
CHAIN 
REACTIO
N 
 
copies/10E6 
cells 
x 
x 
CONTIN
UOUS 
x 
 
HIVVLD 
HIV Viral Load 
Quantitative 
HIV RNA  
CHEMISTRY 
 
COBAS 
TAQMAN  
 
Copies/mL 
 
 
CONTIN
UOUS