Long-term Immunogenicity of the HIV gp120-NefTat/AS01B Vaccine (GSK SB732461)
Phase: PHASE1
Conditions: Infection, Human Immunodeficiency Virus, HIV Infections
Sponsor: GlaxoSmithKline

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Clinical Study Protocol
Sponsor:
GlaxoSmithKline Biologicals SA
Rue de l’Institut, 89
B-1330 Rixensart - Belgium
Primary Study 
vaccine/product and 
number

GSK Biologicals candidate HIV prophylactic 
vaccine (gp120/ NefTat) adjuvanted to AS01B 
(GSK SB732461)
Other Study 
vaccine/product

Not applicable
eTrack study number and 
Abbreviated Title
201606 (PRO-HIV-013 EXT:002)
Date of protocol
Final Version 1: 12 July 2017
Title
Long-term immunogenicity of the HIV gp120-
NefTat/AS01B vaccine (GSK SB732461)
Detailed Title
A Phase I, open-label study to evaluate the long-term 
immunogenicity of the gp120-NefTat/AS01B vaccine 
administered in HIV-1 uninfected subjects
Co-ordinating author
, Scientific Writer (4Clinics Belgium for 
GSK Biologicals)
Contributing authors

, Clinical Development 
Research Lead

 Project Statistician

, Study Delivery Lead

, Local Delivery Lead

 CLS Clinical Read-Out Team Leader

 & 
, Clinical Safety 
Representatives

, Senior Oversight Data Manager

, Clinical Lab Study Management

, Clinical Regulatory Affairs 
Representative

, Vaccine Development 
Leader

, Director, Biostat & Stat Prog

, Clinical and Epidemiology 
Project Lead
GSK Biologicals’ Protocol DS v 15.0
Copyright 2017 the GlaxoSmithKline group of companies. All rights reserved. Unauthorised 
copying or use of this information is prohibited.
 
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Protocol Sponsor Signatory Approval
eTrack study number and 
Abbreviated Title
201606 (PRO-HIV-013 EXT:002)
Date of protocol
Final Version 1: 12 July 2017
Detailed Title 
A Phase I, open-label study to evaluate the long-term 
immunogenicity of the gp120-NefTat/AS01B vaccine 
administered in HIV-1 uninfected subjects
Sponsor signatory
François Roman, Clinical and Epidemiology Project 
Lead
Signature
Date
 
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Protocol Investigator Agreement
I agree:

To conduct the study in compliance with this protocol, any future protocol 
amendments or protocol administrative changes, with the terms of the clinical 
trial agreement and with any other study conduct procedures and/or study 
conduct documents provided by GlaxoSmithKline (GSK) Biologicals.

To assume responsibility for the proper conduct of the study at this site.

That I am aware of, and will comply with, ‘Good Clinical Practice’ (GCP) and 
all applicable regulatory requirements. 

To ensure that all persons assisting me with the study are adequately informed 
about the GSK Biologicals’ study vaccine/product and other study-related 
duties and functions as described in the protocol.

To acquire the reference ranges for laboratory tests performed locally and, if 
required by local regulations, obtain the laboratory’s current certification or 
Quality Assurance procedure manual.

To ensure that no clinical samples (including serum samples) are retained onsite 
or elsewhere without the approval of GSK Biologicals and the express written 
informed consent of the subject and/or the subject’s legally acceptable 
representative. 

To perform no other biological assays on the clinical samples except those 
described in the protocol or its amendment(s). 

To co-operate with a representative of GSK Biologicals in the monitoring 
process of the study and in resolution of queries about the data.

That I have been informed that certain regulatory authorities require the 
sponsor to obtain and supply, as necessary, details about the investigator’s 
ownership interest in the sponsor or the investigational vaccine(s)/product(s), 
and more generally about his/her financial ties with the sponsor. GSK 
Biologicals will use and disclose the information solely for the purpose of 
complying with regulatory requirements.
Hence I:

Agree to supply GSK Biologicals with any necessary information regarding 
ownership interest and financial ties (including those of my spouse and 
dependent children).

Agree to promptly update this information if any relevant changes occur during 
the course of the study and for one year following completion of the study. 

Agree that GSK Biologicals may disclose any information it has about such 
ownership interests and financial ties to regulatory authorities.

Agree to provide GSK Biologicals with an updated Curriculum Vitae and other 
documents required by regulatory agencies for this study.
 
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eTrack study number and 
Abbreviated Title
201606 (PRO-HIV-013 EXT:002)
Date of protocol
Final Version 1: 12 July 2017
Detailed Title
A Phase I, open-label study to evaluate the long-term 
immunogenicity of the gp120-NefTat/AS01B vaccine 
administered in HIV-1 uninfected subjects
Investigator name
Prof. Dr. I. Leroux-Roels, University of Ghent, 
Belgium
Signature
Date
 
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Sponsor Information
1.
Sponsor
GlaxoSmithKline Biologicals
Rue de l’Institut, 89
B-1330 Rixensart – Belgium
2.
Sponsor Medical Expert for the Study
Refer to the local study contact information document.
3.
Sponsor Study Monitor
Refer to the local study contact information document.
4.
Sponsor Study Contact for Reporting of a Serious Adverse Event
GSK Biologicals Central Back-up Study Contact for Reporting SAEs: refer to 
protocol Section 8.3.2.
 
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SYNOPSIS
Detailed Title
A Phase I, open-label study to evaluate the long-term 
immunogenicity of the gp120-NefTat/AS01B vaccine 
administered in HIV-1 uninfected subjects
Rationale for the 
study and study 
design
Study PRO HIV-002 was a phase I/II, double-blind, 
randomized controlled study conducted in HIV uninfected 
volunteers, to compare the safety and immunogenicity of the 
gp120-NefTat (20 µg/20 µg) vaccine, adjuvanted with either 
AS02A, AS02V or AS01B. The vaccine was administered in a 
0, 1, 3 and 6 months schedule and subjects were followed up 
to 18 months after the last vaccination. The study was 
conducted from February 2003 until February 2005.
The study vaccines were overall well tolerated and induced 
strong and long-lasting HIV-specific CD4+ T cell responses, 
with strongest responses in the gp120-NefTat/AS01B group. 
Strong binding antibody responses were detected against 
gp120, Nef and Tat in all vaccine groups that persisted up to 
18 months after the last vaccination.
The objective of the present study is to evaluate the long-term 
persistence of binding antibody responses against V1V2 and 
gp120 in subjects who were vaccinated with the gp120-
NefTat/AS01B vaccine candidate. Other immune parameters 
like the HIV-specific CD4+ T cell and CD8+ T cell responses
will also be evaluated.
Objectives
Primary

To assess the persistence of immune responses
approximately 14 years after administration of the gp120-
NefTat/AS01B vaccine candidate, in terms of anti-V1V2 
binding antibody response by Binding Antibody 
Multiplex Assay (BAMA) versus historical time points in 
study PRO-HIV-002.
Secondary

To assess the persistence of immune responses 
approximately 14 years after administration of the gp120-
NefTat/AS01B vaccine candidate, in terms of HIV-1 
specific CD4+ T cell and CD8+ T cell responses versus 
historical time points in study PRO-HIV-002.

To assess the persistence of immune responses
approximately 14 years after administration of the gp120-
NefTat/AS01B vaccine candidate, in terms of anti-gp120 
antibody response by BAMA versus historical time points 
 
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in study PRO-HIV-002.
Study design

Experimental design: Phase I, open-label, descriptive
mono-centric, and uncontrolled study with one single 
group. This study will consist of a single sample 
collection visit.

Duration of the study: 1 day (single study visit) for each 
subject

Epoch 001: Prospective data collection at single 
study visit that will occur approximately 14 years 
post vaccination

Primary completion Date (PCD): Year 14

End of Study (EoS): Last reading results released for all 
subjects from Year 14 single study visit

Study group:
Synopsis Table 1
Study groups and epochs foreseen in the study
Study Groups
Number of subjects
Age (Min – Max)
Epochs
Epoch 001 
Group 1
Approximately 30
30 years– 65 years
x

Control: uncontrolled

Vaccination schedule: Not applicable

Treatment allocation: Not applicable 

Blinding: Not applicable
Synopsis Table 2
Blinding of study epochs
Study Epochs
Study Groups
Blinding
Epoch 001
Group 1
Open

Sampling schedule:

Blood samples will be collected from all subjects 
during the single study visit to assess humoral 
immunity and cell-mediated immunity (CMI). 

Retrospective serum samples from subjects in study 
PRO HIV-002 at Day (D)0, D182, and D672 time 
points will be re-tested for humoral immunity using 
exploratory humoral assays. 

Retrospective PBMC samples from subjects in study 
PRO HIV-002 at D0, D98, and D672 time points will 
be re-tested for CMI responses using intracellular 
cytokine staining (ICS) assays.
 
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
Type of study: extension of protocol PRO HIV-002 
(732461)

Data collection: Electronic Case Report Form (eCRF)

Safety monitoring: Not applicable
Number of subjects
Planned target number of subjects is approximately 30 
subjects who received the gp120-NefTat/AS01B vaccine in 
GSK Biologicals-sponsored PRO HIV-002 study
Endpoints
Primary

Seropositivity status and anti-V1V2 (IgG, IgG1, IgG2, 
IgG3, and IgG4) binding antibody concentration as 
measured by BAMA at Y14, and at historical time points 
of study PRO HIV-002 (D0, D182, and D672).
Testing of samples for PRO HIV-002 time points will depend 
on availability of samples.
Secondary

Magnitude, responder status, cytokine co-expression 
profile of HIV-1 specific CD4+ T cell and CD8+ T cell 
responses as assessed by ICS assay at Y14, and at 
historical time points of study PRO HIV-002 (D0, D98, 
and D672).

Seropositivity status and anti-gp120 (IgG, IgG1, IgG2, 
IgG3, and IgG4) binding antibody concentration as 
measured by BAMA at Y14, and at historical time points 
of study PRO HIV-002 (D0, D182, and D672).
Testing of samples for PRO HIV-002 time points will depend 
on availability of samples.
Additional investigations, such as additional CMI parameters, 
memory B cell phenotypes and antibody function may be 
performed based on study results and availability of samples.
 
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TABLE OF CONTENTS
PAGE
SPONSOR INFORMATION ............................................................................................5
SYNOPSIS......................................................................................................................6
LIST OF ABBREVIATIONS...........................................................................................15
GLOSSARY OF TERMS ...............................................................................................16
1.
INTRODUCTION....................................................................................................18
1.1.
Rationale for the study and study design ....................................................18
1.2.
Benefit : Risk Assessment ..........................................................................19
2.
OBJECTIVES.........................................................................................................19
2.1.
Primary objective ........................................................................................19
2.2.
Secondary objectives..................................................................................19
3.
STUDY DESIGN OVERVIEW ................................................................................20
4.
STUDY COHORT...................................................................................................21
4.1.
Number of subjects/centres ........................................................................21
4.2.
Inclusion criteria for enrolment....................................................................22
4.3.
Exclusion criteria for enrolment...................................................................22
5.
CONDUCT OF THE STUDY ..................................................................................23
5.1.
Regulatory and ethical considerations, including the informed 
consent process..........................................................................................23
5.2.
Subject identification and randomisation.....................................................24
5.2.1.
Subject identification....................................................................24
5.2.2.
Randomisation of treatment.........................................................24
5.3.
Method of blinding ......................................................................................24
5.4.
General study aspects ................................................................................24
5.5.
Outline of study procedures ........................................................................24
5.6.
Detailed description of study procedures ....................................................25
5.6.1.
Informed consent.........................................................................25
5.6.2.
Check inclusion and exclusion criteria .........................................25
5.6.3.
Collect demographic data ............................................................25
5.6.4.
Medical and vaccination history ...................................................25
5.6.5.
Blood sampling ............................................................................25
5.6.6.
Study Vaccine/product administration..........................................25
5.6.7.
Recording of SAEs ......................................................................26
5.6.8.
Study conclusion..........................................................................26
5.7.
Biological sample handling and analysis.....................................................26
5.7.1.
Use of specified study materials ..................................................27
5.7.2.
Biological samples.......................................................................27
5.7.3.
Laboratory assays .......................................................................27
5.7.4.
Biological samples evaluation......................................................29
Humoral immunity.......................................................29
5.7.4.1.
Cellular immunity........................................................30
5.7.4.2.
 
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Molecular biology........................................................30
5.7.4.3.
5.7.5.
Immunological correlates of protection.........................................30
6.
STUDY VACCINE/PRODUCT AND ADMINISTRATION ........................................30
7.
HEALTH ECONOMICS ..........................................................................................30
8.
SAFETY.................................................................................................................30
8.1.
Safety definitions ........................................................................................31
8.1.1.
Definition of an adverse event......................................................31
8.1.2.
Definition of a serious adverse event ...........................................31
8.1.3.
Clinical laboratory parameters and other abnormal 
assessments qualifying as adverse events or serious 
adverse events ............................................................................32
8.2.
Detecting and recording serious adverse events.........................................33
8.2.1.
Time period for detecting and recording serious adverse 
events..........................................................................................33
8.2.2.
Post-Study adverse events and serious adverse events..............33
8.2.3.
Evaluation of adverse events and serious adverse events...........33
Active questioning to detect adverse events 
8.2.3.1.
and serious adverse events........................................33
Assessment of adverse events...................................33
8.2.3.2.
8.2.3.2.1.
Assessment of intensity..........................33
8.2.3.2.2.
Assessment of causality .........................34
Assessment of outcomes............................................35
8.2.3.3.
8.3.
Reporting of serious adverse events...........................................................35
8.3.1.
Prompt reporting of serious adverse events to GSK 
Biologicals ...................................................................................35
8.3.2.
Contact information for reporting serious adverse events ............35
8.3.3.
Completion and transmission of SAE reports to GSK 
Biologicals ...................................................................................36
Back-up system in case the electronic reporting 
8.3.3.1.
system does not work.................................................36
8.3.4.
Updating of SAE information after removal of write access 
to the subject’s eCRF...................................................................36
8.3.5.
Regulatory reporting requirements for serious adverse 
events..........................................................................................36
8.4.
Follow-up of serious adverse events...........................................................37
8.4.1.
Follow-up during the study...........................................................37
8.4.2.
Follow-up after the subject is discharged from the study..............37
8.5.
Treatment of adverse events ......................................................................37
8.6.
Subject card................................................................................................37
9.
SUBJECT COMPLETION AND WITHDRAWAL.....................................................38
10. STATISTICAL METHODS......................................................................................38
10.1.
Primary endpoint.........................................................................................38
10.2.
Secondary endpoints ..................................................................................38
10.3.
Determination of sample size......................................................................38
10.4.
Cohorts for Analyses...................................................................................39
10.4.1.
Total vaccinated cohort................................................................39
10.4.2.
Per-protocol cohort for analysis of immunogenicity ......................39
 
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10.5.
Derived and transformed data.....................................................................39
10.6.
Analysis of demographics ...........................................................................40
10.7.
Analysis of immunogenicity.........................................................................40
10.8.
Analysis of safety........................................................................................41
10.9.
Interpretation of analyses............................................................................41
10.10. Conduct of analyses ...................................................................................41
10.10.1. Sequence of analyses..................................................................41
10.10.2. Statistical considerations for interim analyses..............................42
11. ADMINISTRATIVE MATTERS ...............................................................................42
11.1.
Electronic Case Report Form instructions...................................................42
11.2.
Study Monitoring by GSK Biologicals..........................................................42
11.3.
Record retention .........................................................................................43
11.4.
Quality assurance.......................................................................................43
11.5.
Posting of information on publicly available clinical trial registers and 
publication policy ........................................................................................44
11.6.
Provision of study results to investigators ...................................................44
11.7.
Data Sharing...............................................................................................44
12. COUNTRY SPECIFIC REQUIREMENTS...............................................................44
13. REFERENCES.......................................................................................................45
 
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LIST OF TABLES
PAGE
Table 1
Study group and epoch foreseen in the study ........................................21
Table 2
Blinding of study epochs ........................................................................21
Table 3
List of study procedures .........................................................................24
Table 4
Biological samples .................................................................................27
Table 5
Humoral immunity (Antibody determination)...........................................28
Table 6
Cell-mediated Immunity (CMI)................................................................28
Table 7
Humoral immunity read-outs ..................................................................29
Table 8
Cellular immunity read-outs....................................................................30
Table 9
Molecular biology read-outs ...................................................................30
Table 10
Timeframes for submitting serious adverse event to GSK 
Biologicals..............................................................................................35
Table 11
GSK Biologicals’ laboratories .................................................................48
Table 12
Outsourced laboratories.........................................................................48
 
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LIST OF FIGURES
PAGE
Figure 1
Schematic representation of the study design........................................20
 
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LIST OF APPENDICES
PAGE
APPENDIX A LABORATORY ASSAYS .......................................................................46
APPENDIX B CLINICAL LABORATORIES ..................................................................48
 
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LIST OF ABBREVIATIONS
AE
Adverse Event
AS01B
Adjuvant System 01B
BAMA
Binding Antibody Multiplex Assay
CD
Cluster of Differentiation
CFC
Cell Flow Cytometry (= ICS)
CI
Confidence Interval
CLS
Clinical Laboratory Sciences
CMI
Cell-Mediated Immunity
eCRF
electronic Case Report Form
EoS
End of Study
GCP
Good Clinical Practice
GM(C)
Geometric mean (concentration)
gp
Envelope glycoprotein
GSK
GlaxoSmithKline
HIV
Human immunodeficiency virus
ICF
Informed Consent Form
ICH
International Conference on Harmonisation
ICS
Intracellular Cytokine Staining (= CFC)
IEC
Independent Ethics Committee
LTFU
Long-Term Follow-Up
LOQ
Limit Of Quantification
MPL
3-O-desacyl-4’-monophosphoryl lipid A
PBMC
Peripheral Blood Mononuclear Cell
PCD
Primary Completion Date
PP
Per-protocol
QS21
Quillaja saponaria Molina, fraction 21
RT
Reverse Transcriptase
SAE
Serious Adverse Event
SDV
Source Document Verification
SPM
Study Procedures Manual
 
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GLOSSARY OF TERMS
Adverse event:
Any untoward medical occurrence in a patient or clinical 
investigation subject, temporally associated with the use 
of a medicinal product, whether or not considered related 
to the medicinal product. 
An adverse event (AE) can therefore be any unfavourable 
and unintended sign (including an abnormal laboratory 
finding), symptom, or disease (new or exacerbated) 
temporally associated with the use of a medicinal product. 
For marketed medicinal products, this also includes 
failure to produce expected benefits (i.e. lack of efficacy), 
abuse or misuse.
Eligible:
Qualified for enrolment into the study based upon strict 
adherence to inclusion/exclusion criteria.
End of Study 
(Synonym of End of 
Trial)
For studies without collection of human biological 
samples or imaging data EoS is the Last Subject Last 
Visit (LSLV). 
For studies with collection of human biological samples 
or imaging data, EoS is defined as the date of the last 
testing/reading released of the Human Biological 
Samples or imaging data, related to primary and 
secondary endpoints. EoS must be achieved no later than 
8 months after LSLV
Epoch:
An epoch is a set of consecutive time points or a single 
time point from a single protocol. Epochs are defined to 
support a main purpose which either to draw conclusions 
on subject participation or to draw a complete conclusion 
to define or precise the targeted label of the product. 
Supporting means that data collected at the time points 
included in an epoch must be sufficient to fulfil the 
purpose of the epoch.
eTrack:
GSK’s tracking tool for clinical trials.
Evaluable:
Meeting all eligibility criteria, complying with the 
procedures defined in the protocol, and, therefore, 
included in the per-protocol analysis (see Section 10.4 for 
details on criteria for evaluability).
Immunological correlate 
of protection:
The defined immune response above which there is a high 
likelihood of protection in the absence of any host factors 
that might increase susceptibility to the infectious agent.
 
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Investigational 
vaccine/product:
(Synonym of 
Investigational Medicinal 
Product)
A pharmaceutical form of an active ingredient being 
tested in a clinical trial, including a product with a 
marketing authorisation when used in a way different 
from the approved form, or when used for an unapproved 
indication, or when used to gain further information about 
an approved use. 
Primary completion 
date:
The date that the final subject was examined or received 
an intervention for the purpose of final collection of data 
for all primary outcomes, whether the clinical trial was 
concluded according to the pre-specified protocol or was 
terminated.
Protocol amendment:
The International Conference on Harmonisation (ICH) 
defines a protocol amendment as: ‘A written description 
of a change(s) to or formal clarification of a protocol.’ 
GSK Biologicals further details this to include a change 
to an approved protocol that affects the safety of subjects,
scope of the investigation, study design, or scientific 
integrity of the study.
Protocol administrative 
change:
A protocol administrative change addresses changes to 
only logistical or administrative aspects of the study. 
Study vaccine/product
Any investigational vaccine/product being tested and/or 
any authorized use of a vaccine/ product /placebo as a 
reference or administered concomitantly, in a clinical trial 
that evaluates the use of an investigational 
vaccine/product.
Subject:
Term used throughout the protocol to denote an 
individual who has been contacted in order to participate 
or participates in the clinical study, either as a recipient of 
the vaccine(s)/product(s) or as a control.
Subject number:
A unique number identifying a subject, assigned to each 
subject consenting to participate in the study.
Treatment:
Term used throughout the clinical study to denote a set of 
investigational product(s) or marketed product(s) or 
placebo intended to be administered to a subject.
 
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1.
INTRODUCTION
1.1.
Rationale for the study and study design
The RV144 trial provided the first evidence of protective efficacy against HIV-1 
acquisition after vaccination in adults [Rerks-Ngarm, 2009]. This protective effect peaked 
at more than 60% one year after first vaccination but rapidly declined thereafter, 
suggesting the importance of persistent antibody responses to sustain protection over time 
[Robb, 2012]. An immune-correlates analysis of the RV144 trial identified vaccine-
induced anti-V1V2 IgG responses as being inversely correlated with the rate of HIV-1 
infection [Haynes, 2012]. 
Study PRO HIV-002 was a phase I/II, double-blind, randomized controlled study 
conducted in HIV uninfected volunteers, to compare the safety and immunogenicity of 
the gp120-NefTat (20 µg/20 µg) vaccine, adjuvanted with either AS02A, AS02V or 
AS01B. The vaccine was administered in a 0, 1, 3 and 6 months schedule and subjects 
were followed up to 18 months after the last vaccination. The study was conducted from 
February 2003 until February 2005 [Leroux-Roels, 2010].
The study vaccines were overall well tolerated and induced strong and long-lasting HIV-
specific CD4+ T cell responses, with strongest responses in the gp120-NefTat/AS01B
group. Strong binding antibody responses were detected against gp120, Nef and Tat in all 
vaccine groups that persisted up to 18 months after the last vaccination [Leroux-Roels, 
2010].
A post-hoc analysis was performed on PRO HIV-002 samples from 30 subjects who were 
administered the gp120-NefTat/AS01B vaccine candidate in PRO HIV-002 to determine 
the level of anti-V1V2 IgG responses. An ample and persistent anti-V1V2 response was 
observed after vaccination, with 100% of response rate two weeks after the third and 
fourth vaccination and 87% of response rate at 18 months after the last vaccine dose. 
These observations and the potential correlate of protection associated with anti-V1V2 
response as described in RV144 study [Yates, 2011; Yates, 2014] support further 
assessment of long-term persistence of anti-V1V2 response.
GSK contributes to the public-private pox-protein partnership (P5) which is currently 
assessing the safety, immunogenicity and clinical efficacy of a prime-boost regimen 
aimed at preventing HIV transmission (http://www.hvtn.org/en.html). The booster 
component of the candidate vaccine used in this program consists of two gp120 clade C 
proteins administered with an adjuvant. In order to inform the selection of the adjuvant in 
future studies, data on long-term persistence of immunity after vaccination with 
gp120/AS01 would prove useful. The present study was designed to address this question 
using a cohort of volunteers vaccinated several years ago with a candidate vaccine 
containing gp120 and AS01.
The objective of the present study is to evaluate the long-term persistence of binding 
antibody responses against V1V2 and gp120 in subjects who were vaccinated with the 
gp120-NefTat/AS01B vaccine candidate. Other immune parameters like the HIV-specific 
CD4+ T cell and CD8+ T cell responses will also be evaluated.
 
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1.2.
Benefit : Risk Assessment
The following section outlines the risk assessment and mitigation strategy for this study 
protocol:
Important 
Potential/Identified 
Risk
Data/Rationale for Risk
Mitigation Strategy
Study Procedures
Risk from blood 
sampling
Blood sampling-associated 
risk of discomfort, syncope, 
dizziness, infection at the 
site during or after 
venepuncture.
Blood samples will be obtained by a 
trained professional and medical 
assistance will be available.
The potential risk of feeling faint, or 
experiencing mild local pain, bruising, 
irritation or redness at the site where 
blood was taken, will be mentioned in the 
ICF. The amount of blood to be taken for 
sampling will not be harmful to the 
subject’s health.
2.
OBJECTIVES
2.1.
Primary objective

To assess the persistence of immune responses approximately 14 years after 
administration of the gp120-NefTat/AS01B vaccine candidate, in terms of anti-V1V2 
binding antibody response by Binding Antibody Multiplex Assay (BAMA) versus 
historical time points in study PRO-HIV-002.
Refer to Section 10.1 for the definition of the primary endpoint.
2.2.
Secondary objectives

To assess the persistence of immune responses approximately 14 years after 
administration of the gp120-NefTat/AS01B vaccine candidate, in terms of HIV-1 
specific CD4+ T cell and CD8+ T cell responses versus historical time points in 
study PRO-HIV-002.

To assess the persistence of immune responses approximately 14 years after 
administration of the gp120-NefTat/AS01B vaccine candidate, in terms of anti-
gp120 antibody response by BAMA versus historical time points in study PRO-HIV-
002.
Based on the outcome of the primary responses, other exploratory investigations may be 
performed on stored samples (sera and peripheral blood mononuclear cells [PBMCs]) to 
better characterize the effect of vaccination on other immunologic parameters, such as 
 
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antibody function, memory B cells, or other read-outs that might be available at the time 
of the analysis.
Refer to Section 10.2 for the definition of the secondary endpoint(s).
3.
STUDY DESIGN OVERVIEW
Figure 1
Schematic representation of the study design
Protocol waivers or exemptions are not allowed unless necessary for the 
management of immediate safety concerns. Therefore, adherence to the study design 
requirements, including those specified in the outline of study procedures 
(Section 5.5), are essential and required for study conduct. 

Experimental design: Phase I, open-label, descriptive, mono-centric, and 
uncontrolled study with one single group. This study will consist of a single sample 
collection visit.

Duration of the study: 1 day (single study visit) for each subject

Epoch 001: Prospective data collection at single study visit that will occur 
approximately 14 years post vaccination 

Primary completion Date (PCD): Year 14
Refer to glossary of terms for the definition of PCD.

End of Study (EoS): Last reading results released for all subjects from Year 14 single 
study visit
Refer to glossary of terms for the definition of EoS.

Study group:
 
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Table 1
Study group and epoch foreseen in the study
Study Groups
Number of 
subjects
Age (Min - Max)
Epochs
Epoch 001
Group 1
Approximately 
30
30 years - 65 years
x

Control: uncontrolled

Vaccination schedule: Not applicable

Treatment allocation: Not applicable

Blinding: Not applicable
Table 2
Blinding of study epochs
Study Epochs
Study Groups
Blinding
Epoch 001
Group 1
Open

Sampling schedule:

Blood samples will be collected from all subjects during the single study visit to 
assess humoral immunity and cell-mediated immunity (CMI). 

Retrospective serum samples from subjects in study PRO HIV-002 at Day (D)0, 
D182, and D672 time points will be re-tested for humoral immunity using 
exploratory humoral assays. 

Retrospective PBMC samples from subjects in study PRO HIV-002 at D0, D98, 
and D672 time points will be re-tested for CMI responses using intracellular 
cytokine staining (ICS) assays.

Type of study: extension of protocol PRO HIV-002 (732461)

Data collection: Electronic Case Report Form (eCRF)

Safety monitoring: Not applicable
4.
STUDY COHORT
4.1.
Number of subjects/centres
Planned target number of subjects is approximately 30 subjects who received the gp120-
NefTat/AS01B vaccine in GSK Biologicals-sponsored PRO HIV-002 study.
Overview of the recruitment plan
This study will occur in only one medical centre. The recruitment is estimated to be 
completed in approximately 5 weeks.
 
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4.2.
Inclusion criteria for enrolment
Deviations from inclusion criteria are not allowed because they can potentially 
jeopardize the scientific integrity, regulatory acceptability of the study or subject 
safety. Therefore, adherence to the criteria as specified in the protocol is essential.
All subjects must satisfy ALL the following criteria at study entry:

Subjects who, in the opinion of the investigator, can and will comply with the 
requirements of the protocol.

Written informed consent obtained from the subject prior to performing any 
study specific procedure.

A subject who has received at least three doses of the gp120-NefTat/AS01B vaccine 
candidate in GSK Biologicals-sponsored PRO HIV-002 study.
4.3.
Exclusion criteria for enrolment
Deviations from exclusion criteria are not allowed because they can potentially 
jeopardize the scientific integrity, regulatory acceptability of the study or subject 
safety. Therefore, adherence to the criteria as specified in the protocol is essential.
The following criteria should be checked at the time of study entry. If ANY 
exclusion criterion applies, the subject must not be included in the study:

Use of any investigational or non-registered product (drug or vaccine) during 30 days 
prior to study enrolment.

History of HIV-1 or HIV-2 infection.

Participation to another clinical trial of an investigational HIV vaccine between study 
PRO HIV-002 and the present study.

Chronic administration (defined as more than 14 days in total) of 
immunosuppressants or other immune-modifying drugs during the period starting 
one month preceding this study. For corticosteroids, this will mean prednisone 
≥20 mg/day (or equivalent). Inhaled and topical steroids are allowed.

Administration of cytotoxic medication within six months preceding this study.

History of daily, long-term (more than one year) immunosuppressive medication 
between study PRO HIV-002 and the present study.

Administration of immunoglobulins and/or any blood products during the period 
starting 3 months before enrolment.

Any confirmed or suspected immunosuppressive or immunodeficient condition other 
than HIV disease, based on medical history and physical examination (no laboratory 
testing required) between study PRO HIV-002 and the present study.

Past administration of an investigational vaccine containing AS01 other than the 
gp120-NefTat/AS01B vaccine administered in PRO HIV-002 study.
 
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5.
CONDUCT OF THE STUDY
5.1.
Regulatory and ethical considerations, including the 
informed consent process
The study will be conducted in accordance with all applicable regulatory 
requirements.
The study will be conducted in accordance with the ICH Guideline for GCP, all 
applicable subject privacy requirements and the guiding principles of the 
Declaration of Helsinki. 
GSK will obtain favourable opinion/approval to conduct the study from the 
appropriate regulatory agency, in accordance with applicable regulatory 
requirements, prior to a site initiating the study in that country.
Conduct of the study includes, but is not limited to, the following:

Institutional Review Board (IRB)/Independent Ethics Committee (IEC) review 
and favourable opinion/approval of study protocol and any subsequent 
amendments.

Subject informed consent.

Investigator reporting requirements as stated in the protocol.
GSK will provide full details of the above procedures to the investigator, either 
verbally, in writing, or both.
Freely given and written informed consent must be obtained from each subject 
prior to participation in the study. 
GSK Biologicals will prepare a model Informed Consent Form (ICF) which will 
embody the ICH GCP and GSK Biologicals required elements. While it is strongly 
recommended that this model ICF is to be followed as closely as possible, the 
informed consent requirements given in this document are not intended to pre-empt 
any local regulations which require additional information to be disclosed for 
informed consent to be legally effective. Clinical judgement, local regulations and 
requirements should guide the final structure and content of the local version of the 
ICF. 
The investigator has the final responsibility for the final presentation of the ICF 
respecting the mandatory requirements of local regulations. The ICF generated by 
the investigator with the assistance of the sponsor’s representative must be 
acceptable to GSK Biologicals and be approved (along with the protocol, and any 
other necessary documentation) by the IRB/IEC.
 
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5.2.
Subject identification and randomisation
5.2.1.
Subject identification
Subjects will be assigned the identification number they received in PRO HIV-002 study.
5.2.2.
Randomisation of treatment
Not applicable.
5.3.
Method of blinding 
Not applicable.
5.4.
General study aspects
Supplementary study conduct information not mandated to be present in this 
protocol is provided in the accompanying Study Procedures Manual (SPM). The 
SPM provides the investigator and the site personnel with administrative and 
detailed technical information that does not impact the safety of the subjects.
5.5.
Outline of study procedures
Table 3
List of study procedures
Epoch
Epoch 001
Type of contact
Single 
Study Visit
Timepoint (s)
Year 14
Informed consent
●
Check inclusion/exclusion criteria
●
Collect demographic data
●
Record medical history
●
Record vaccination history
●
Laboratory Assays
Blood sampling for HIV test (9 mL)
●
Blood sampling for antibody determination and exploratory characterisation (9 mL)
●
Blood sampling for CMI responses and exploratory characterisation (27 mL)
●
Safety Assessments
Recording of SAEs related to study participation or to a concurrent GSK medication/product
●
Study conclusion
●
● is used to indicate a study procedure that requires documentation in the individual eCRF.
CMI: cell-mediated immunity; SAE: serious adverse event. 
All study procedures will occur during the Single Study Visit.
 
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5.6.
Detailed description of study procedures
5.6.1.
Informed consent
The signed informed consent of the subject must be obtained before study 
participation. Refer to Section 5.1 for the requirements on how to obtain informed 
consent.
5.6.2.
Check inclusion and exclusion criteria
Check all inclusion and exclusion criteria as described in Sections 4.2 and 4.3 before 
enrolment. 
5.6.3.
Collect demographic data
Record demographic data such as sex and race in the subject’s eCRF.
5.6.4.
Medical and vaccination history
Obtain the subject’s medical and vaccination history by interview and/or review of 
the subject’s medical records and record in the eCRF:

any pre-existing conditions or signs and/or symptoms present in a subject within 
30 days before the start of the study, 

past receipt of any vaccine administered within 30 days before the start of the study.

past receipt of any experimental HIV vaccine (other than in PRO HIV-002 study).
5.6.5.
Blood sampling
Blood samples will be taken during the Single Study Visit (as specified in Section 5.5
List of Study Procedures) for the assessment of. 

HIV test,

antibody determination,

CMI responses, and 

exploratory characterisation.
An overall volume of 45 mL will be collected during the entire study period.
Refer to the Module on Biospecimen Management in the SPM for general handling of 
blood samples.
5.6.6.
Study Vaccine/product administration
Not applicable. 
 
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5.6.7.
Recording of SAEs

Refer to Section 8.2 for procedures for the investigator to record SAEs. Refer to 
Section 8.3 for guidelines and how to report SAE to GSK Biologicals.

The subjects will be instructed to contact the investigator immediately should 
the subjects manifest any signs or symptoms they perceive as serious. 
5.6.8.
Study conclusion
The investigator will:

review data collected to ensure accuracy and completeness

complete the Study Conclusion screen in the eCRF.
5.7.
Biological sample handling and analysis
Please refer to the SPM for details on biospecimen management (handling, storage and 
shipment).
Samples will not be labelled with information that directly identifies the subject but will 
be coded with the identification number for the subject (subject number).

Collected samples will be used for protocol mandated research and purposes 
related to the improvement, development and quality assurance of the 
laboratory tests described in this protocol. This may include the management of 
the quality of these tests, the maintenance or improvement of these tests, the 
development of new test methods, as well as making sure that new tests are 
comparable to previous methods and work reliably.

It is also possible that future findings may make it desirable to use the samples 
acquired in this study for future research, not described in this protocol. 
Therefore, all subjects will be asked to give a specific consent to allow GSK or a 
contracted partner to use the samples for future research. Future research will 
be subject to the laws and regulations in Belgium and will only be performed 
once an independent Ethics Committee or Review Board has approved this 
research.
Information on further investigations and their rationale can be obtained from GSK 
Biologicals. 
Any sample testing will be done in line with the consent of the individual subject.
Refer also to the Investigator Agreement, where it is noted that the investigator 
cannot perform any other biological assays except those described in the protocol or 
its amendment(s). 
If additional testing is performed, the marker priority ranking given in Section 5.7.4
may be changed.
 
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Collected samples will be stored for a maximum of 20 years (counting from when 
the last subject performed the last study visit), unless local rules, regulations or 
guidelines require different timeframes or different procedures, which will then be 
in line with the subject consent. These extra requirements need to be communicated 
formally to and discussed and agreed with GSK Biologicals.
5.7.1.
Use of specified study materials
When materials are provided by GSK Biologicals, it is MANDATORY that all 
clinical samples (including serum samples) be collected and stored exclusively using 
those materials in the appropriate manner. The use of other materials could result 
in the exclusion of the subject from the per-protocol analysis (See Section 10.4 for 
the definition of cohorts to be analysed). The investigator must ensure that his/her 
personnel and the laboratory(ies) under his/her supervision comply with this 
requirement. However, when GSK Biologicals does not provide material for 
collecting and storing clinical samples, appropriate materials from the investigator’s 
site must be used. Refer to the Module on Clinical Trial Supplies in the SPM.
5.7.2.
Biological samples
Table 4
Biological samples
Sample type
Quantity
Unit
Timepoint
Blood sample for HIV test
9
ml
Single Study Visit
Blood sample for serology
9
ml
Single Study Visit
Blood sample for CMI
27
ml
Single Study Visit
5.7.3.
Laboratory assays
Please refer to APPENDIX A for a detailed description of the assays performed in 
the study. Please refer to APPENDIX B for the address of the clinical laboratories 
used for sample analysis.
Test for the determination of HIV infection (to exclude subjects with HIV infection) will 
be performed at the Investigator’s laboratory using standardized and validated 
procedures. The diagnostic algorithm used in UZ Gent is: 4th generation screening test 
that detects antibodies and p24 antigen (VIDAS DUO). In case of reactivity, an Inno Lia 
confirmation test (Fujirebio) is performed. This test will allow to distinguish between a 
real seropositivity and a vaccine-induced seropositivity (if any). The result will be 
encoded in eCRF as a qualitative result (HIV infected: yes/no) and raw results will be 
uploaded into the study database.
 
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Serological assays (Table 5) for the determination of antibodies against gp120 and V1V2 
to check the subjects’ response to the vaccine they received in study PRO HIV-002 will 
be performed at Duke Human Vaccine Institute using standardized procedures.
Table 5
Humoral immunity (Antibody determination)
System
Component
Method Kit / Manufacturer
Unit
Cut-off *
Laboratory ***
SER 
Human 
Immunodeficiency 
Virus Ab
Anti-gp120** 
(IgG, IgG1, IgG2, 
IgG3, IgG4, and 
Total IgG Avidity)
BAMA
In-house
MFI
95th percentile 
of the MFI-
background-
Neg from all 
baseline†
Duke Human Vaccine Institute
SER 
Human 
Immunodeficiency 
Virus Ab
Anti-V1V2** (IgG, 
IgG1, IgG2, IgG3, 
IgG4, and Total 
IgG Avidity)
BAMA
In-house
MFI
95th percentile 
of the MFI-
background-
Neg from all 
baseline†
Duke Human Vaccine Institute
SER: serum; BAMA: Binding Antibody Multiplex Assay; MFI: mean fluorescence intensity
† Baseline is defined as Day 0 in PRO HIV-002 study.
* Assay cut-off and unit might be subject to change during the course of the study (e.g., in case of requalification, 
revalidation or standardization). In this case, this will be documented in the clinical report.
** Performed on antigen corresponding to the vaccine strain (other HIV strains or HIV clades).
*** Refer to APPENDIX B for the laboratory addresses.
ICS (CFC) assays (Table 6) on HIV-1 specific CD4+ T cells and CD8+ T cells to check 
the subjects’ response to the vaccine they received in study PRO HIV-002 will be 
performed at CEVAC using standardized procedures.
Table 6
Cell-mediated Immunity (CMI)
System
Component Family
Challenge
Method
Laboratory *
PBMC
Core CFC read-out specific CD4+ T cells and CD8+ T 
cells
Cells CD4.CD40L(+)+Interleukin-2(-)+Tumor Necrosis 
Factor alpha(-)+Interferon gamma(-)
gp120
CFC
CEVAC
PBMC: peripheral blood mononuclear cells; CFC: cell flow cytometry
* Refer to APPENDIX B for the laboratory addresses.
 
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Other assays may be performed on sample repositories taken for this purpose or leftover 
samples, with the aim to explore tertiary objectives of the study. The research may 
include, but is not limited to:

Serological responses to Nef, Tat, or other HIV antigens.

Evaluation of antibody quality by Surface Plasmon Resonance (SPR), Bio-Layer 
Interferometry (BLI) or other methods.

Evaluation of antibody function by antibody profiling and systems serology.

Characterisation of the impact of vaccination on possible new virological markers.

Host transcriptome signature: evaluation of mRNA and/or microRNA signatures by 
microarray and/or RNA sequencing.

Translational research using next generation technologies.
Additional exploratory testing on the vaccine and and/or the disease under study 
may be performed within the framework of the study if deemed necessary for 
accurate interpretation of the data or should such assay(s) become available at 
GSK. These assays may not be represented in the objectives/endpoints of the study 
protocol.
The GSK Biologicals’ clinical laboratories have established a Quality System 
supported by procedures. The activities of GSK Biologicals’ clinical laboratories are 
audited regularly for quality assessment by an internal (sponsor-dependent) but 
laboratory-independent Quality Department.
5.7.4.
Biological samples evaluation
Humoral immunity
5.7.4.1.
Table 7
Humoral immunity read-outs
Blood sampling time point
No. subjects
Component
Components 
priority rank
Type of contact 
and time point
Sampling time point
Single Study Visit
+
Historical PRO HIV-
002 time points
Y14 (PIV)
+
PRO HIV-002 D0 (PRE), 
D182 (PIV), and D672 (PIV)
Approximately 
30 *
Anti-V1V2 antibodies (BAMA)
1
Anti-gp120 antibodies (BAMA)
2
Total IgG Avidity
3
* to be determined based on sample availability.
In case of insufficient blood sample volume to perform assays for all antibodies, the 
samples will be analysed according to priority ranking provided in Table 7. 
 
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Cellular immunity
5.7.4.2.
Table 8
Cellular immunity read-outs
Blood sampling time point
No. subjects
Component
Type of contact and time 
point
Sampling time point
Single Study Visit
+
Historical PRO HIV-002
time points
Y14 (PIV)
+
PRO HIV-002 D0 (PRE), D98 (PIII), 
and D672 (PIV)
Approximately 
30 *
HIV-1 specific CD4+ T cells and 
CD8+ T cells
* to be determined based on sample availability.
Molecular biology
5.7.4.3.
Table 9
Molecular biology read-outs
Blood sampling time point
No. subjects
Component
Type of contact and time 
point
Sampling time 
point
Single Study Visit
Y14 (PIV)
Approximately
30 *
Human Immunodeficiency virus 1 RNA
(QPCR )
* to be determined based on sample availability.
5.7.5.
Immunological correlates of protection
No generally accepted immunological correlate of protection has been demonstrated so 
far for the antigens used in the candidate vaccine/ product.
6.
STUDY VACCINE/PRODUCT AND ADMINISTRATION
Not applicable.
7.
HEALTH ECONOMICS
Not applicable. 
8.
SAFETY
The investigator or site staff is/are responsible for the detection, documentation and 
reporting of events meeting the criteria and definition of an adverse event (AE) or 
serious adverse event (SAE) as provided in this protocol. 
Each subject will be instructed to contact the investigator immediately should 
they/the subject manifest any signs or symptoms they perceive as serious.
 
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8.1.
Safety definitions
8.1.1.
Definition of an adverse event
An AE is any untoward medical occurrence in a clinical investigation subject, 
temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.
An AE can therefore be any unfavourable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product. For marketed medicinal products, 
this also includes failure to produce expected benefits (i.e. lack of efficacy), abuse or 
misuse. 
Examples of an AE include:

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

Pre- or post-treatment events that occur as a result of protocol-mandated 
procedures (i.e. invasive procedures, modification of subject’s previous 
therapeutic regimen).
Examples of an AE DO NOT include:

Medical or surgical procedures (e.g. endoscopy, appendectomy); the condition that 
leads to the procedure is an AE/SAE.

Situations where an untoward medical occurrence did not occur (e.g. social and/or 
convenience admission to a hospital, admission for routine examination).

Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen. 

Pre-existing conditions or signs and/or symptoms present in a subject prior to
the study vaccination. These events will be recorded in the medical history 
section of the eCRF.
8.1.2.
Definition of a serious adverse event 
A SAE is any untoward medical occurrence that:
a.
Results in death,
b.
Is life-threatening,
Note: The term ‘life-threatening’ in the definition of ‘serious’ refers to an event 
in which the subject was at risk of death at the time of the event. It does not 
refer to an event, which hypothetically might have caused death, had it been 
more severe.
 
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c.
Requires hospitalisation or prolongation of existing hospitalisation,
Note: In general, hospitalisation signifies that the subject has been admitted at 
the hospital or emergency ward for observation and/or treatment that would 
not have been appropriate in the physician’s office or in an out-patient setting. 
Complications that occur during hospitalisation are also considered AEs. If a 
complication prolongs hospitalisation or fulfils any other serious criteria, the 
event will also be considered serious. When in doubt as to whether 
‘hospitalisation’ occurred or was necessary, the AE should be considered 
serious.
Hospitalisation for elective treatment of a pre-existing condition (known or 
diagnosed prior to informed consent signature) that did not worsen from 
baseline is NOT considered an AE.
d.
Results in disability/incapacity, OR
Note: The term disability means a substantial disruption of a person’s ability to 
conduct normal life functions. This definition is not intended to include 
experiences of relatively minor medical significance such as uncomplicated 
headache, nausea, vomiting, diarrhoea, influenza like illness, and accidental 
trauma (e.g. sprained ankle) which may interfere or prevent everyday life 
functions but do not constitute a substantial disruption.
e.
Is a congenital anomaly/birth defect in the offspring of a study subject.
Medical or scientific judgement should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalisation but may jeopardise 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition. These should also be considered 
serious. Examples of such events are invasive or malignant cancers, intensive 
treatment in an emergency room or at home for allergic bronchospasm, blood 
dyscrasias or convulsions that do not result in hospitalisation.
8.1.3.
Clinical laboratory parameters and other abnormal assessments 
qualifying as adverse events or serious adverse events 
In absence of diagnosis, abnormal laboratory findings (e.g. clinical chemistry, 
haematology, urinalysis) or other abnormal assessments (e.g., severe fluctuations in 
blood pressure) that are judged by the investigator to be clinically significant will be 
recorded as SAE if they meet the definition of an SAE (refer to Section 8.1.1). 
Clinically significant abnormal laboratory findings or other abnormal assessments 
that are present at baseline and significantly worsen following the start of the study 
will also be reported as SAEs. However, clinically significant abnormal laboratory 
findings or other abnormal assessments that are associated with the disease being studied, 
unless judged by the investigator as more severe than expected for the subject’s 
condition, or that are present or detected at the start of the study and do not worsen, will 
not be reported as AEs or SAEs.
 
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The investigator will exercise his or her medical and scientific judgement in deciding 
whether an abnormal laboratory finding or other abnormal assessment is clinically 
significant.
8.2.
Detecting and recording serious adverse events
8.2.1.
Time period for detecting and recording serious adverse events
SAEs related to study participation or to a concurrent GSK medication/product will be 
collected and recorded from the time the subject consents to participate in the study 
(i.e., ICF signature) until the subject is discharged from the study.
8.2.2.
Post-Study adverse events and serious adverse events
A post-study AE/SAE is defined as any event that occurs outside of the AE/SAE 
reporting period. Investigators are not obligated to actively seek AEs or SAEs in 
former study participants. However, if the investigator learns of any SAE at any 
time after a subject has been discharged from the study, and he/she considers the 
event reasonably related to the study procedure, the investigator will promptly notify 
the Study Contact for Reporting SAEs.
8.2.3.
Evaluation of adverse events and serious adverse events
Active questioning to detect adverse events and serious adverse 
8.2.3.1.
events
When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g. hospital progress notes, laboratory and diagnostics reports) 
relative to the event. The investigator will then record all relevant information 
regarding an AE/SAE in the eCRF. The investigator is not allowed to send 
photocopies of the subject’s medical records to GSK Biologicals instead of 
appropriately completing the eCRF. However, there may be instances when copies 
of medical records for certain cases are requested by GSK Biologicals. In this 
instance, all subject identifiers will be blinded on the copies of the medical records 
prior to submission to GSK Biologicals.
The investigator will attempt to establish a diagnosis pertaining to the event based 
on signs, symptoms, and/or other clinical information. In such cases, the diagnosis 
should be documented as the AE/SAE and not the individual signs/symptoms.
Assessment of adverse events
8.2.3.2.
8.2.3.2.1.
Assessment of intensity
The investigator will assess the maximum intensity that occurred over the duration 
of the event for all unsolicited AEs (including SAEs) recorded during the study. The 
assessment will be based on the investigator’s clinical judgement.
 
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The intensity should be assigned to one of the following categories:
1 (mild)
=
An AE which is easily tolerated by the subject, causing 
minimal discomfort and not interfering with everyday 
activities.
2 (moderate)
=
An AE which is sufficiently discomforting to interfere with 
normal everyday activities.
3 (severe)
=
An AE which prevents normal, everyday activities 
An AE that is assessed as Grade 3 (severe) should not be confused with a SAE. 
Grade 3 is a category used for rating the intensity of an event; and both AEs and 
SAEs can be assessed as Grade 3. An event is defined as ‘serious’ when it meets 
one of the pre-defined outcomes as described in Section 8.1.1.
8.2.3.2.2.
Assessment of causality
The investigator is obligated to assess the relationship between study procedure and 
the occurrence of each AE/SAE using clinical judgement.
There may be situations when a SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK Biologicals. However, it is very 
important that the investigator always makes an assessment of causality for every 
event prior to submission of the Expedited Adverse Events Report to GSK 
Biologicals. The investigator may change his/her opinion of causality in light of 
follow-up information and update the SAE information accordingly. The causality 
assessment is one of the criteria used when determining regulatory reporting 
requirements.
If an event meets the criteria to be determined as ‘serious’ (see Section 8.1.1), 
additional examinations/tests will be performed by the investigator in order to 
determine ALL possible contributing factors for each SAE.
Possible contributing factors include:

Medical history.

Other medication.

Protocol required procedure.

Other procedure not required by the protocol.

Erroneous administration

Other cause (specify).
 
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Assessment of outcomes
8.2.3.3.
The investigator will assess the outcome of all unsolicited AEs (including SAEs) 
recorded during the study as:

Recovered/resolved.

Recovering/resolving.

Not recovered/not resolved.

Recovered with sequelae/resolved with sequelae.

Fatal (SAEs only).
8.3.
Reporting of serious adverse events
8.3.1.
Prompt reporting of serious adverse events to GSK Biologicals
SAEs that occur in the time period defined in Section 8.2 will be reported promptly 
to GSK within the timeframes described in Table 10, once the investigator 
determines that the event meets the protocol definition of a SAE.
Table 10
Timeframes for submitting serious adverse event to GSK Biologicals
Type of Event
Initial Reports
Follow-up of Relevant Information on a 
Previous Report
Timeframe
Documents
Timeframe
Documents
SAEs
24 hours* ‡
electronic Expedited 
Adverse Events Report
24 hours*
electronic Expedited Adverse 
Events Report
* Timeframe allowed after receipt or awareness of the information.
‡ The investigator will be required to confirm review of the SAE causality by ticking the ‘reviewed’ box in the 
electronic Expedited Adverse Events Report within 72 hours of submission of the SAE.
8.3.2.
Contact information for reporting serious adverse events
Study Contact for Reporting SAEs
Refer to the local study contact information document.
Email address:
Back-up Study Contact for Reporting SAEs
24/24 hour and 7/7 day availability:
GSK Biologicals Clinical Safety & Pharmacovigilance
Fax: 
 or 
Email address: 
 
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PPD
PPD
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PPD

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8.3.3.
Completion and transmission of SAE reports to GSK Biologicals
Once an investigator becomes aware that a SAE has occurred in a study subject, the 
investigator (or designate) must complete the information in the electronic 
Expedited Adverse Events Report WITHIN 24 HOURS. The report will always be 
completed as thoroughly as possible with all available details of the event. Even if 
the investigator does not have all information regarding a SAE, the report should 
still be completed within 24 hours. Once additional relevant information is received, 
the report should be updated WITHIN 24 HOURS.
The investigator will always provide an assessment of causality at the time of the 
initial report. The investigator will be required to confirm the review of the SAE 
causality by ticking the ‘reviewed’ box in the electronic Expedited Adverse Events 
Report within 72 hours of submission of the SAE.
Back-up system in case the electronic reporting system does not work
8.3.3.1.
If the electronic reporting system does not work, the investigator (or designate) must 
complete, then date and sign a paper Expedited Adverse Events Report and fax it to 
the Study Contact for Reporting SAEs (refer to the Sponsor Information) or to GSK 
Biologicals Clinical Safety and Pharmacovigilance department within 24 hours. 
This back-up system should only be used if the electronic reporting system is not 
working and NOT if the system is slow. As soon as the electronic reporting system is 
working again, the investigator (or designate) must complete the electronic 
Expedited Adverse Events Report within 24 hours. The final valid information for 
regulatory reporting will be the information reported through the electronic SAE 
reporting system.
8.3.4.
Updating of SAE information after removal of write access to the 
subject’s eCRF
When additional SAE information is received after removal of the write access to 
the subject’s eCRF, new or updated information should be recorded on the 
appropriate paper report, with all changes signed and dated by the investigator. The 
updated report should be faxed to the Study Contact for Reporting SAEs (refer to 
the Sponsor Information) or to GSK Biologicals Clinical Safety and 
Pharmacovigilance department within the designated reporting time frames 
specified in Table 10.
8.3.5.
Regulatory reporting requirements for serious adverse events
The investigator will promptly report all SAEs to GSK in accordance with the 
procedures detailed in Section 8.3.1.
 
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8.4.
Follow-up of serious adverse events
8.4.1.
Follow-up during the study
After the initial SAE report, the investigator is required to proactively follow each 
subject and provide additional relevant information on the subject’s condition to 
GSK Biologicals (within 24 hours for SAEs; refer to Table 10).
All SAEs documented at a previous visit/contact and designated as not 
recovered/not resolved or recovering/resolving will be reviewed at subsequent 
visits/contacts until the last visit of the subject.
8.4.2.
Follow-up after the subject is discharged from the study
The investigator will follow subjects:

with SAEs or subjects withdrawn from the study as a result of an AE, until the 
event has resolved, subsided, stabilised, disappeared, or until the event is 
otherwise explained, or the subject is lost to follow-up. 
If the investigator receives additional relevant information on a previously reported 
SAE, he/she will provide this information to GSK Biologicals using a paper/ 
electronic Expedited Adverse Events Report and/or pregnancy report as applicable.
GSK Biologicals may request that the investigator performs or arranges the conduct 
of additional clinical examinations/tests and/or evaluations to elucidate as fully as 
possible the nature and/or causality of the AE or SAE. The investigator is obliged to 
assist. If a subject dies during participation in the study or during a recognised 
follow-up period, GSK Biologicals will be provided with any available post-mortem 
findings, including histopathology.
8.5.
Treatment of adverse events
Treatment of any AE is at the sole discretion of the investigator and according to 
current good medical practice. Any medication administered for the treatment of a 
SAE should be recorded in Expedited Adverse Event Report of the subject’s eCRF.
8.6.
Subject card
Study subjects must be provided with the address and telephone number of the 
main contact for information about the clinical study.
The investigator (or designate) must therefore provide a “subject card” to each 
subject. In an emergency situation this card serves to inform the responsible 
attending physician that the subject is in a clinical study and that relevant 
information may be obtained by contacting the investigator.
 
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Subjects must be instructed to keep subject cards in their possession at all times
during the study duration.
9.
SUBJECT COMPLETION AND WITHDRAWAL
Not applicable.
10.
STATISTICAL METHODS
10.1.
Primary endpoint

Seropositivity status and anti-V1V2 (IgG, IgG1, IgG2, IgG3, and IgG4) binding 
antibody concentration as measured by BAMA at Y14, and at historical time points 
of study PRO HIV-002 (D0, D182, and D672).
Testing of samples for PRO HIV-002 time points will depend on availability of samples.
10.2.
Secondary endpoints

Magnitude, responder status, cytokine co-expression profile of HIV-1 specific CD4+ 
T cell and CD8+ T cell responses as assessed by ICS assay at Y14, and at historical 
time points of study PRO HIV-002 (D0, D98, and D672).

Seropositivity status and anti-gp120 (IgG, IgG1, IgG2, IgG3, and IgG4) binding 
antibody concentration as measured by BAMA at Y14, and at historical time points 
of study PRO HIV-002 (D0, D182, and D672).
Testing of samples for PRO HIV-002 time points will depend on availability of samples.
Additional investigations, such as additional CMI parameters, memory B cell phenotypes 
and antibody function may be performed based on study results and availability of 
samples.
10.3.
Determination of sample size
The sample size in this study is driven by the sample size of the primary PRO HIV-002
study, in which 60 subjects were vaccinated with the gp120-NefTat/AS01B vaccine 
candidate, of which 54 were included in the Per-protocol (PP) cohort for 
immunogenicity. Based on preliminary assessment by the study site, it is expected that 
approximately 30 to 40 subjects will be eligible to enter this long-term follow-up (LTFU) 
study.
 
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10.4.
Cohorts for Analyses
10.4.1.
Total vaccinated cohort
The Total cohort will include all subjects who signed the informed consent and provided 
a blood sample.
10.4.2.
Per-protocol cohort for analysis of immunogenicity
The PP cohort for immunogenicity will include all subjects from the Total cohort:

who meet all eligibility criteria,

who present a negative HIV-RNA test at Y14.
Subjects eliminated from the PP cohort for immunogenicity will have their results 
reported as line listings.
10.5.
Derived and transformed data
Demography

Age at vaccination will be computed as the difference between the date of dose 1 
administration in primary PRO HIV-002 study and the 15 JUNE of the year of birth
(only the year of birth will be collected). Age at Y14 visit in the study will be 
computed as the difference between the date of Y14 visit and the 15 JUNE of the 
year of birth. The ages will be expressed in years. 

For a given subject and a given demographic variable, missing measurements will 
not be replaced.
Safety

For analysis of SAEs and for the analysis of concomitant medications, all subjects 
will be considered. Subjects who did not report an event or a concomitant GSK 
medication will be considered as subjects without the event or without the 
concomitant medication respectively.
Immunogenicity

Any missing or non-evaluable immunogenicity measurement will not be replaced. 
The descriptive analysis performed for each assay at each time point will exclude 
subjects with a missing or non-evaluable measurement. 

The geometric mean titers (GMTs)/geometric mean concentrations (GMCs) will be 
computed by taking the anti-logarithm of the arithmetic mean of the log10 
transformed titers/concentrations. 

A seronegative subject will be defined as a subject whose antibody 
titer/concentration is below the cut-off value of the assay. A seropositive subject is a 
subject whose antibody titer/concentration is greater than or equal to the cut-off 
value of the assay.
 
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
Vaccine response rates for HIV-1- specific CD4+ T-cells expressing at least two 
markers among CD40L, IL-2, TNF-α and IFN-γ with responders will be defined as 
subjects with:

a 2-fold increase as compared to the cut-off (limit of quantification [LOQ] of the 
assay), for subjects with pre-vaccination frequency below the cut-off;

at least 2-fold increase as compared to pre-vaccination frequency, for subjects 
with pre-vaccination frequency above the cut-off.
Other definitions of the CMI vaccine response might be explored during the analysis. 
They will be described in the SAP.
10.6.
Analysis of demographics
For the Total cohort and for the PP cohort for immunogenicity, demographic data (age at 
vaccination in years, age at Y14 visit of this current LTFU study in years, gender and 
race) will be summarized using descriptive statistics:

Frequency tables for gender and race.

Mean, median, standard error for age.
10.7.
Analysis of immunogenicity
The primary analysis will be performed on the PP cohort for immunogenicity and, if the 
percentage of vaccinated subjects with serological results excluded from the PP cohort 
for analysis of immunogenicity is at least 10%, a second analysis will be performed on 
the Total cohort. 
For the humoral immune response 
For anti-V1V2 and anti-gp120 (IgG, IgG1, IgG2, IgG3, and IgG4) antibody 
concentrations (by BAMA), at each time point [PRE(D0), PIV(D182), PIV(D672), 
PIV(Y14)] where such information is available:

Seropositivity rates with 95%CIs will be tabulated.

Geometric mean concentrations (GMCs) with 95% CIs will be tabulated and 
presented graphically.

Geometric mean of ratios (GMR) of antibody concentrations at each post-vaccination 
time point [PIV(D182), PIV(D672), PIV(Y14)] over pre-vaccination [PRE(D0)] will 
be tabulated with 95% CI.

Antibody concentrations post-vaccination will also be presented using reverse 
cumulative curves.
 
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For the cell-mediated immune response
The following parameters will be summarised using descriptive statistics (N, GM, min, 
Q1, median, Q3, max) at each time point [PRE(D0), PIII(D98), PIV(D672), PIV(Y14)] 
where CMI results are available:

Frequency of HIV-1 specific CD4+/CD8+ T-cells expressing at least two markers
among CD40L, IL-2, TNF-α and IFN-γ as measured by ICS using PBMCs.

Frequency of HIV-1 specific CD4+/CD8+ T-cells expressing at least one marker
among CD40L, IL-2, TNF-α and IFN-γ as measured by ICS using PBMCs. 

Geometric mean ratios (GMR) of frequency of HIV-1 specific CD4+/CD8+ T-cells 
expressing at least two markers among CD40L, IL-2, TNF-α and IFN-γ at each 
post-vaccination time point [PIII(D98), PIV(D672), PIV(Y14)] over pre-vaccination 
(D0) will be tabulated with 95% CI.

Vaccine response rates for HIV-1-specific CD4+ T-cells expressing at least two 
markers among CD40L, IL-2, TNF-α and IFN-γ.
These analyses will be further detailed in the SAP document.
10.8.
Analysis of safety
For the Total cohort, a detailed description of SAEs considered by the investigator to be 
related to study participation or concurrent GSK medication/product will be provided.
10.9.
Interpretation of analyses
The analyses will be descriptive with the aim to characterise the gp120-NefTat/AS01B 
group in the endpoint(s) related to the objective(s). These descriptive analyses should not 
be interpreted. 
10.10.
Conduct of analyses
Any deviation(s) or change(s) from the original statistical plan outlined in this 
protocol will be described and justified in the final study report.
10.10.1.
Sequence of analyses
All pre-planned analyses will be performed when all data of the primary and secondary 
objectives for all enrolled subjects are available and cleaned.
An integrated clinical study report containing all data will be written and made 
available to the investigators.
If additional exploratory analyses are performed, they may be reported in subsequent 
annex reports, depending on data availability. 
 
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10.10.2.
Statistical considerations for interim analyses
No interim analysis is planned for this descriptive LTFU study.
11.
ADMINISTRATIVE MATTERS
To comply with ICH GCP administrative obligations relating to data collection, 
monitoring, archiving data, audits, confidentiality, public disclosure requirements 
and publications must be fulfilled. 
11.1.
Electronic Case Report Form instructions
A validated GSK defined electronic data collection tool will be used as the method 
for data collection. 
In all cases, subject initials will not be collected nor transmitted to GSK. Subject 
data necessary for analysis and reporting will be entered/transmitted into a 
validated database or data system. Clinical data management will be performed in 
accordance with applicable GSK standards and data cleaning procedures.
While completed eCRFs are reviewed by a GSK Biologicals’ Site Monitor at the 
study site, omissions or inconsistencies detected by subsequent eCRF review may 
necessitate clarification or correction by the investigator or appropriately qualified 
designee. In all cases, the investigator remains accountable for the study data. 
The investigator will be provided with a CD-ROM of the final version of the data 
generated at the investigational site once the database is archived and the study 
report is complete and approved by all parties.
11.2.
Study Monitoring by GSK Biologicals
GSK will monitor the study to verify that, amongst other items, the: 

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol, any 
other study agreements, GCP and all applicable regulatory requirements. 
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents.
The investigator must ensure provision of reasonable time, space and qualified 
personnel for monitoring visits.
Direct access to all study-site related and source data is mandatory for the purpose 
of monitoring review. The monitor will perform a eCRF review and a Source 
Document Verification (SDV). By SDV we understand verifying eCRF entries by 
 
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comparing them with the source data that will be made available by the investigator 
for this purpose. 
The Source Documentation Agreement Form describes the source data for the 
different data in the eCRF. This document should be completed and signed by the 
site monitor and investigator and should be filed in the investigator’s study file. Any 
data item for which the eCRF will serve as the source must be identified, agreed and 
documented in the source documentation agreement form. 
Upon completion or premature discontinuation of the study, the monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations, GCP, and GSK procedures.
11.3.
Record retention
Following closure of the study, the investigator must maintain all site study records 
(except for those required by local regulations to be maintained elsewhere) in a safe 
and secure location. The records must be easily accessible, when needed (e.g. audit 
or inspection), and must be available for review in conjunction with assessment of 
the facility, supporting systems, and staff. Where permitted by applicable 
laws/regulations or institutional policy, some or all of these records can be 
maintained in a validated format other than hard copy (e.g. microfiche, scanned, 
electronic); however, caution needs to be exercised before such action is taken. The 
investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, 
including re-generating a hard copy, if required. Furthermore, the investigator must 
ensure that an acceptable back-up of the reproductions exists and that there is an 
acceptable quality control procedure in place for making these reproductions.
GSK will inform the investigator/institution of the time period for retaining these 
records to comply with all applicable regulatory requirements. The minimum 
retention time will meet the strictest standard applicable to a particular site, as 
dictated by ICH GCP, any institutional requirements, applicable laws or 
regulations, or GSK standards/procedures, otherwise, the minimum retention 
period will default to 20 years after completion of the study report.
The investigator/institution must notify GSK of any changes in the archival 
arrangements, including, but not limited to archival at an off-site facility, transfer of 
ownership of the records in the event the investigator leaves the site. 
11.4.
Quality assurance
To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance audit. Regulatory agencies may also conduct a 
regulatory inspection of this study. Such audits/inspections can occur at any time 
during or after completion of the study. If an audit or inspection occurs, the 
investigator and institution agree to allow the auditor/inspector direct access to all 
 
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relevant documents and to allocate his/her time and the time of his/her staff to the 
auditor/inspector to discuss findings and any relevant issues.
11.5.
Posting of information on publicly available clinical trial 
registers and publication policy
Interventional studies that do not evaluate vaccines/products are progressed for 
publication in the scientific literature when the results provide important scientific 
or medical knowledge or are relevant for patient care, and will be considered for 
disclosure on the GSK website and in publicly accessible regulatory registry(ies).
11.6.
Provision of study results to investigators
Where required by applicable regulatory requirements, an investigator signatory 
will be identified for the approval of the study report. The investigator will be 
provided reasonable access to statistical tables, figures, and relevant reports and will 
have the opportunity to review the complete study results at a GSK site or other 
mutually-agreeable location.
GSK Biologicals will also provide the investigator with the full summary of the 
study results. The investigator is encouraged to share the summary results with the 
study subjects, as appropriate.
11.7.
Data Sharing
Under the framework of the SHARE initiative, results of GSK studies may be 
combined with non- GSK studies, to investigate further about the study product(s) 
and other product(s), and /or the disease/condition under investigation and related 
diseases and conditions.
12.
COUNTRY SPECIFIC REQUIREMENTS
Not applicable.
 
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13.
REFERENCES
GlaxoSmithKline Biologicals Clinical Study Report 732461/002 (PRO HIV-002). A 
Phase I-II, double-blind, randomised study to compare the safety and immunogenicity of 
GSK Biologicals’ candidate HIV vaccine gp120/NefTat (20 µg/20 µg) adjuvanted to 
AS02A, AS02V or AS01B administered intramuscularly to healthy HIV seronegative 
volunteers.
Haynes BF, Gilbert PB, McElrath MJ, et al. Immune-correlates analysis of an HIV-1 
vaccine efficacy trial. NEJM. 2012; 366(14): 1275-86.
Leroux-Roels I, Koutsoukos M, Clement F, et al. Strong and persistent CD4+ T-cell 
response in healthy adults immunized with a candidate HIV-1 vaccine containing gp120, 
Nef and Tat antigens formulated in three Adjuvant Systems. Vaccine. 2010; 28(?): 7016-
24.
Rerks-Ngarm S, Pitisuttithum P, Nitayaphan S, et al. Vaccination with ALVAC and 
AIDSVAX to prevent HIV-1 infection in Thailand. NEJM. 2009; 361(23): 2209-20.
Robb ML, Rerks-Ngarm S, Nitayaphan S, Pitisuttithum P, Kaewkungwal J, Kunasol P, et 
al. Risk behaviour and time as covariates for efficacy of the HIV vaccine regimen 
ALVAC-HIV (vCP1521) and AIDSVAX B/E: a post-hoc analysis of the Thai phase 3 
efficacy trial RV 144. Lancet Infect Dis. 2012;12(7):531-7. doi: 10.1016/S1473-
3099(12)70088-9.
Yates NL, Liao HX, Fong Y, deCamp A, Vandergrift NA, Williams WT, et al. Vaccine-
induced Env V1-V2 IgG3 correlates with lower HIV-1 infection risk and declines soon 
after vaccination. Sci Transl Med. 2014;6(228):228ra39. doi: 
10.1126/scitranslmed.3007730.
Yates NL, Lucas JT, Nolen TL, Vandergrift NA, Soderberg KA, Seaton KE, et al. 
Multiple HIV-1-specific IgG3 responses decline during acute HIV-1: implications for 
detection of incident HIV infection. AIDS. 2011 Nov 13;25(17):2089-97. doi: 
10.1097/QAD.0b013e32834b348e.
 
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APPENDIX A
LABORATORY ASSAYS
Humoral immunity
Evaluation of antibody quality by BAMA avidity index or BLI
To evaluate avidity index using the BAMA protocol, samples will be incubated with 
beads in either the assay diluent above (PBS) or a sodium citrate solution at pH 4.0 (CIT) 
and then washed. Binding magnitude response will be evaluated for each, and the avidity 
index will be calculated as (MFI (CIT)/MFI (PBS))*100.
To evaluate antibody quality of total serum by BLI, HIV antigens will be immobilized 
onto Streptavidin (SA) or amine reactive (AR2G) using procedures and coupling reagents 
supplied by the manufacturer. The binding time courses will be monitored by dipping the 
antigen immobilized sensors directly into 1:50 diluted (in PBS buffer, pH 7.4) serum 
samples to monitor association for 300 seconds and then into PBS buffer to record 
dissociation for 300 seconds. Appropriate control peptide/protein immobilized sensors 
will be used as parallel reference sensors to record non-specific binding and will be 
subtracted to obtain antigen-specific binding curves. The specific binding curves will be 
analysed to obtain antigen-specific binding responses and dissociation rates of total 
antibodies. ForteBio Octet Red 384 instruments will be used for measurements. 
HIV antigen-specific total IgG avidity assays will be carried out by capturing total IgG 
antibodies from serum onto anti-human IgG Fc immobilized sensors. These sensors will 
be dipped into HIV antigen constructs and PBS buffer to read binding and dissociation, 
respectively. Appropriate control sensors will be used in parallel as reference to collect 
non-specific binding and will be subtracted to get total IgG binding of HIV antigens. The 
subtracted binding curves will be processed to obtain antigen-specific binding response 
and dissociation rates.
The BLI determined antigen-specific binding response, which is a reflective of the 
magnitude of antigen-specific antibodies in a sample, will be compared to gauge the 
difference in magnitude of antibody responses in participants’ serum samples. The 
antigen-specific dissociation rate is indicative of the quality of the antibodies present in a 
sample. The antigen-specific binding response and dissociation rate data together can be 
used to rank the serum samples of study participants from different groups with a score 
that includes both the magnitude of the antibody response and the dissociation rate of the 
antibody-antigen complex as measured by BLI. The binding response, dissociation rate 
and avidity rank for each antigen will be used in statistical analysis as described in 
Analytical Plan section. 
Cell-mediated immunity (CMI)
The role of CMI in the protective response to HIV is thought to be important for 
protection. The CMI response will be assessed on thawed PBMCs at CEVAC.
Detection of gp120, Nef and Tat, intracellular cytokine staining (ICS) technique
Intracellular cytokine staining (ICS) is a method providing information on the frequency 
of CD4+ and CD8+ T cells responding to the antigen and secreting molecules involved in 
immunity such as IFN- γ, IL-2, TNF-a and CD40-L. The assay will be performed on 
 
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thawed PBMCs. Briefly, PBMCs of the subjects are stimulated for 2 hours by the vaccine 
antigen and/or derived peptides. Then, intracellular block (Brefeldin A) is added to 
inhibit cytokine secretion for a subsequent overnight stimulation. The cells are then 
harvested and stained for cell surface markers (e.g. CD4 and CD8), to identify specific 
sub-populations of T cells. Subsequently, the cells are fixed and then treated with a 
detergent such as saponin to permeabilise the cell membrane. The cells are then incubated 
with fluorescently labelled, cytokine-specific or activation marker-specific antibody that 
forms complexes with the cytokine in the cytoplasm. The cytokine producing cells can 
then be detected by cytofluorometry. Results are expressed as a frequency of antigen-
specific CD4+ or CD8+ T cells identified as expressing cytokine/marker upon the in vitro 
stimulation.
 
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APPENDIX B
CLINICAL LABORATORIES
Table 11
GSK Biologicals’ laboratories
Laboratory
Address
GSK Biological’s Clinical Laboratory 
Sciences, Rixensart 
Biospecimen Reception - B7/44
Rue de l'Institut, 89 - B-1330 Rixensart – Belgium 
GSK Biological’s Clinical Laboratory 
Sciences, Wavre-Nord Noir Epine
Avenue Fleming, 20 - B-1300 Wavre - Belgium
Table 12
Outsourced laboratories
Laboratory
Address
Duke Human Vaccine Institute
Tomaras Laboratory
Duke Human Vaccine Institute
MSRB2 Bldg, Room 4116 / 2 Genome Ct.
Durham, NC 27710 USA
Email: 
 Ph.D. 
Lab: 
CEVAC - University of Gent
De Pintelaan, 185 Gent
Belgium
 
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PPD
PPD
PPD
PPD

PPD