Immunogenicity and Safety Study of Two Different Formulations of GlaxoSmithKline (GSK) Biologicals' Oral Live Attenuated Human Rotavirus (HRV) Vaccine, Rotarix in Healthy Infants
Phase: PHASE3
Conditions: Rotavirus
Sponsor: GlaxoSmithKline

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Clinical Study Protocol
Sponsor:
GlaxoSmithKline Biologicals SA
Rue de l’Institut 89
1330 Rixensart, Belgium
Primary Study vaccine and 
number
Liquid formulation of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus 
(HRV) vaccine, Rotarix (444563)
Other study vaccine
Lyophilized formulation of GSK Biologicals’ oral 
live attenuated HRV vaccine (444563)
eTrack study number and 
Abbreviated Title
116566 (ROTA-083)
EudraCT number
2012-001875-35
Date of protocol
Final Version 1: 11 April 2012
Date of protocol 
amendments
Amendment 1 Final: 24 July 2013
Amendment 2 Final: 19 September 2016
Amendment 3 Final: 31 October 2017
Amendment 4: Final: 30 October 2019
Title
Immunogenicity and safety study of two different 
formulations of GlaxoSmithKline (GSK) Biologicals’ 
oral live attenuated human rotavirus (HRV) vaccine, 
Rotarix in healthy infants.
Detailed Title
A phase III, randomized, open study to assess the 
immunogenicity, reactogenicity and safety of two 
different formulations of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus 
(HRV) vaccine, Rotarix, when given as a two-dose
primary vaccination, in healthy infants with no 
previous history of rotavirus illness or vaccination.
Co-ordinating author
Contributing authors
(Amended: 30 October 2019)

, Clinical & Epidemiology Project 
Lead (CEPL)

, 
, Clinical
Research and Development Leads (CRDL)

,
, Statisticians

, 
, Lead 
Statisticians

, 
, 
 
, 
, Study Delivery 
Leads
 
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PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD

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eTrack study number and 
Abbreviated Title
116566 (ROTA-083)
EudraCT number
2012-001875-35
Detailed Title
A phase III, randomized, open study to assess the 
immunogenicity, reactogenicity and safety of two 
different formulations of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus 
(HRV) vaccine, Rotarix, when given as a two-dose 
primary vaccination, in healthy infants with no 
previous history of rotavirus illness or vaccination.
Contributing authors
(Amended: 30 October 2019)
(Continued…)

, 
, Project 
Delivery Leads

,
,
 
,
, Clinical Laboratory 
Sciences (CLS), Clinical Readout Team Leaders

,
, Laboratory Study 
Managers

,
,
, Clinical 
Safety representatives

, 
, 
Oversight Data Managers

, 
, 
,
Study Data Managers

, Global Regulatory Affairs 
representative

, 
, Global Patents 
representatives

, 
, 
 
, 
,
, 
Clinical Trial Supply Managers

, 
, Local Medical
Leads

, 
, Local 
Delivery Leads
GSK Biologicals’ Protocol DS v 15.0
© 2012-2019 GSK group of companies or its licensor.
 
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PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD

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Protocol Amendment 4 Sponsor Signatory Approval
eTrack study number and 
Abbreviated Title
116566 (ROTA-083)
EudraCT number
2012-001875-35
Date of protocol amendment
Amendment 4 Final: 30 October 2019
Detailed Title
A phase III, randomized, open study to assess the 
immunogenicity, reactogenicity and safety of two 
different formulations of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus 
(HRV) vaccine, Rotarix, when given as a two-dose 
primary vaccination, in healthy infants with no 
previous history of rotavirus illness or vaccination.
Sponsor signatory
Paul Gillard, MD
Clinical & Epidemiology Project Lead,
Live Viral Vaccines, RDC Belgium
GlaxoSmithKline Biologicals, SA.
Signature
Date
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Protocol Amendment 4 Rationale
Amendment number:
Amendment 4
Rationale/background for changes: 
As per Protocol Amendment 3 (31 October 2017), subjects seropositive before 
vaccination are to be eliminated from the Per-Protocol Set (PPS) for immunogenicity.
Laboratory testing of the pre-vaccination blood samples from the first 141 subjects 
enrolled in the study indicated a seropositivity rate of 47.5 % in these subjects. This 
suggests an overall high seropositivity rate at baseline in the total study population.
Therefore, an elimination rate exceeding the 35% rate of non-evaluable subjects
predefined using historical data from Rota-044 study [Narang, 2009] is expected.
Because (1) the pre-vaccination seropositivity rate is expected to be high and (2) the 
effect of the vaccine can be measured in seropositive subjects, as previously 
demonstrated (see Table 1 below), the following changes have been made to the 
protocol:

The PPS was modified to include subjects seropositive at baseline.

The statistical method to derive the 95% confidence interval (CI) for the geometric
mean concentration (GMC) group ratio was revised to be an ANCOVA.

The seroconversion threshold was redefined to account for seropositive subjects at
pre-vaccination:

for subjects with a pre-vaccination anti-rotavirus IgA antibody concentration
<20 U/mL, seroconversion is achieved when the post-vaccination
concentration is ≥20 U/mL.

for subjects with a pre-vaccination anti-rotavirus IgA antibody concentration
≥20 U/mL, seroconversion is achieved when the post-vaccination
concentration is ≥2 times the pre-vaccination concentration.
Table 1: Distribution of fold increase in anti-rotavirus IgA antibody 
concentrations from pre-vaccination to one month post-dose 2 (Total vaccinated 
cohort for seropositive subjects at pre-vaccination, Rota-044) [Narang, 2009]
HRV 
N = 43
Placebo 
N = 40
Ratio
n
%
n
%
≥2
26
60.5
3
7.5
≥3
21
48.8
2
5.0
≥4
17
39.5
2
5.0
N = number of subjects seropositive at baseline for anti-rotavirus IgA antibodies and with one month post-dose 2 
data available; n/% = number/percentage of subject in a given category.
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Protocol Amendment 4 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 vaccines 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 vaccines, 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
116566 (ROTA-083)
EudraCT number
2012-001875-35
Date of protocol amendment
Amendment 4 Final: 30 October 2019
Detailed Title
A phase III, randomized, open study to assess the 
immunogenicity, reactogenicity and safety of two 
different formulations of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus 
(HRV) vaccine, Rotarix, when given as a two-dose 
primary vaccination, in healthy infants with no 
previous history of rotavirus illness or vaccination.
Investigator name
Signature
Date
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For internal use only
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Sponsor Information
1.
Sponsor
GlaxoSmithKline Biologicals SA
Rue de l’Institute 89, 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 III, randomized, open study to assess the 
immunogenicity, reactogenicity and safety of two different 
formulations of GlaxoSmithKline (GSK) Biologicals’ oral 
live attenuated human rotavirus (HRV) vaccine, Rotarix,
when given as a two-dose primary vaccination, in healthy 
infants with no previous history of rotavirus illness or 
vaccination.
Indication
Active immunization of infants against gastroenteritis (GE) 
due to rotavirus (RV).
Rationale for the 
study and study 
design
The lyophilized formulation of the HRV vaccine was 
licensed in India in February 2008. Following the request of 
the Regulatory Authority in India, this study will be 
conducted to generate additional clinical data for the liquid 
formulation of the HRV vaccine in India, as recommended 
by New Drug Advisory Committee on Vaccines (NDAC-
Vaccines) of Drug Controller General of India (DCGI).
This study will be conducted to evaluate the 
immunogenicity, reactogenicity and safety of GSK 
Biologicals’ HRV liquid vaccine compared to GSK 
Biologicals’ HRV lyophilized vaccine. The study is planned 
to be conducted in healthy infants aged 6-10 weeks at the 
time of the first dose. Two oral doses of the study vaccine 
will be administered to the subjects, according to a 0, 1 
month schedule. The blood samples for immunogenicity 
assessment will be collected from all subjects before the first 
dose and at one month post dose 2.
Objectives
(Amended: 30
October 2019)
Primary
To evaluate non-inferiority of GSK Biologicals' HRV liquid 
vaccine compared to GSK Biologicals' HRV lyophilized 
vaccine in terms of geometric mean concentrations (GMCs) 
for anti-RV antibodies, one month post dose 2 of HRV liquid 
vaccine and HRV lyophilized vaccine.
Criterion: Non-inferiority will be stated if the lower limit of 
the two-sided 95% confidence interval (CI) for the ratio of 
anti RV IgA antibody GMCs between HRV liquid vaccine 
over the HRV lyophilized vaccine, one month after dose 2 is 
greater than or equal to 0.5.
 
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Secondary

To assess the immunogenicity of the HRV liquid 
vaccine and HRV lyophilized vaccine, in terms of 
seroconversion* rates, one month post dose 2 of HRV 
vaccine.
*Definition:

for subjects with a pre-vaccination anti-RV
IgA antibody concentration <20 U/mL, 
seroconversion is achieved when the post-
vaccination concentration is ≥20 U/mL.

for subjects with a pre-vaccination anti-RV
IgA antibody concentration ≥20 U/mL, 
seroconversion is achieved when the post-
vaccination concentration is ≥2 times the pre-
vaccination concentration.

To assess the reactogenicity of the HRV liquid vaccine 
and the HRV lyophilized vaccine in terms of solicited 
adverse events (AEs), during the 8-day (Day 1–Day 8) 
follow-up period after each vaccination.

To assess the safety of the HRV liquid vaccine and the 
HRV lyophilized vaccine in terms of unsolicited AEs, 
during the 31-day (Day 1–Day 31) follow-up period 
after each vaccination and serious adverse events 
(SAEs), during the entire study period.
Study design

Experimental design: Phase III, open-label, randomized
(1:1), multi-centric, single-country study with two
parallel groups.

Duration of the study: The intended duration of the 
study, per subject, is approximately two months.

Epoch 001: Primary starting at Visit 1 (Day 1) and 
ending at Visit 3 (Month 2).

Primary Completion Date (PCD): Last subject attending 
Visit 3.

End of Study (EoS): Last testing results released for 
samples collected at Visit 3. Please refer to GLOSSARY 
OF TERMS for the definition of EoS.

Study groups: 
The study groups and epoch foreseen in the study are 
provided in Synopsis Table 1.
 
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The study groups and treatment foreseen in the study are 
provided in Synopsis Table 2.
Synopsis Table 1
Study groups and epoch foreseen in the study
Study groups
Number of subjects
Age at Dose 1 (Min/Max)
Epoch 001
HRV Liq
225
6 weeks- 10 weeks
●
HRV Lyo
225
6 weeks- 10 weeks
●
Synopsis Table 2
Study groups and treatment foreseen in the study
Treatment name
Vaccine/Product name
Study Groups
HRV Liq
HRV Lyo
HRV Liquid
GSK Biologicals’ HRV liquid vaccine
x
HRV Lyophilized
GSK Biologicals’ HRV lyophilized vaccine
x 
GSK Biologicals’ calcium carbonate buffer
x 

Control: active control (HRV lyophilized vaccine)

Vaccination schedule:

Two oral doses of the HRV vaccine are to be given 
according to a 0, 1 month schedule.

All subjects are allowed to receive routine childhood 
vaccinations according to the local immunization 
practice. The administration of all routine childhood 
vaccinations given since birth will be recorded in the 
electronic Case Report Form (eCRF).

Treatment allocation: Randomized (1:1). Treatment 
number allocation using GSK Biologicals’ Randomization 
System on Internet (SBIR).

Blinding: open
The blinding of study epoch is provided in Synopsis Table 3.
Synopsis Table 3
Blinding of study epoch
Study Epoch
Blinding
Epoch 001
open

Sampling schedule: Details of the samples to be 
collected are as follows:

Blood samples will be collected from all subjects at 
Visit 1 and Visit 3 to measure serum anti-RV 
Immunoglobulin A (IgA) antibody concentrations 
using Enzyme Linked Immunosorbent Assay 
(ELISA).

Type of study: self-contained.
 
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
Data collection: eCRF
Number of subjects
The target will be to enrol 450 subjects who will be 
randomly assigned to one of the two study groups in a 1:1 
ratio, to obtain at least 292 evaluable subjects
(approximately 146 subjects in each group).
Endpoints
(Amended: 30
October 2019)
Primary

Anti-RV IgA antibody concentrations

Serum anti-RV IgA antibody concentrations, 
expressed as GMCs, one month post dose 2 of HRV 
vaccine.
Secondary

Anti-RV IgA antibody concentrations

Anti-RV IgA antibody seroconversion* rate, one 
month post dose 2 of HRV vaccine.
*Definition:

for subjects with a pre-vaccination anti-RV IgA 
antibody concentration <20 U/mL, seroconversion 
is achieved when the post-vaccination 
concentration is ≥20 U/mL.

for subjects with a pre-vaccination anti-RV IgA 
antibody concentration ≥20 U/mL, seroconversion 
is achieved when the post-vaccination 
concentration is ≥2 times the pre-vaccination 
concentration.

Solicited general symptoms

Occurrence of each type of solicited general 
symptom within the 8-day (Day1-Day 8) solicited 
follow-up period, after each dose of HRV 
lyophilized vaccine and HRV liquid vaccine.

Unsolicited adverse events

Occurrence of unsolicited AEs within 31 days 
(Day 1- Day 31) after any dose of HRV lyophilized 
vaccine and HRV liquid vaccine according to 
Medical Dictionary for Regulatory Activities 
(MedDRA) classification.
 
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
Serious adverse events

Occurrence of SAEs from dose 1 of HRV 
lyophilized vaccine and HRV liquid vaccine up to 
study end.
 
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TABLE OF CONTENTS
PAGE
SPONSOR INFORMATION ............................................................................................7
SYNOPSIS......................................................................................................................8
LIST OF ABBREVIATIONS...........................................................................................19
GLOSSARY OF TERMS ...............................................................................................21
TRADEMARK................................................................................................................25
1.
INTRODUCTION....................................................................................................26
1.1.
Background (Amended: 30 October 2019)..................................................26
1.2.
Rationale for the study and study design ....................................................26
1.3.
Benefit: Risk Assessment ...........................................................................27
1.3.1.
Risk Assessment .........................................................................27
1.3.2.
Benefit Assessment .....................................................................28
1.3.3.
Overall Benefit: Risk Conclusion..................................................28
2.
OBJECTIVES.........................................................................................................28
2.1.
Primary objective ........................................................................................28
2.2.
Secondary objectives (Amended: 30 October 2019) ...................................28
3.
STUDY DESIGN OVERVIEW ................................................................................29
4.
STUDY COHORT...................................................................................................31
4.1.
Number of subjects/centers ........................................................................31
4.2.
Inclusion criteria for enrolment....................................................................31
4.3.
Exclusion criteria for enrolment...................................................................31
5.
CONDUCT OF THE STUDY ..................................................................................33
5.1.
Regulatory and ethical considerations, including the informed
consent process..........................................................................................33
5.2.
Subject identification and randomization.....................................................34
5.2.1.
Subject identification....................................................................34
5.2.2.
Randomization of treatment.........................................................34
5.2.2.1.
Randomization of supplies..........................................34
5.2.2.2.
Treatment allocation to the subject.............................34
5.2.2.2.1.
Study group and treatment 
number allocation ...................................34
5.2.2.2.2.
Treatment number allocation for 
subsequent doses ..................................35
5.3.
Method of blinding ......................................................................................35
5.4.
General study aspects ................................................................................35
5.5.
Outline of study procedures ........................................................................35
5.6.
Detailed description of study procedures ....................................................37
5.6.1.
Informed consent.........................................................................37
5.6.2.
Check inclusion and exclusion criteria .........................................37
5.6.3.
Collect demographic data ............................................................37
5.6.4.
Medical and vaccination history ...................................................37
 
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5.6.5.
Physical examination ...................................................................37
5.6.6.
Check contraindications, warnings and precautions to 
vaccination...................................................................................38
5.6.7.
Assess pre-vaccination body temperature ...................................38
5.6.8.
Study group and treatment number allocation..............................38
5.6.9.
Sampling......................................................................................38
5.6.9.1.
Blood sampling for immune response 
assessment ................................................................38
5.6.10.
Study Vaccine administration.......................................................38
5.6.11.
Record regurgitation ....................................................................39
5.6.12.
Check and record concomitant medication/vaccination and 
intercurrent medical conditions ....................................................39
5.6.13.
Recording of AEs and SAEs ........................................................39
5.6.14.
Study conclusion..........................................................................40
5.7.
Biological sample handling and analysis.....................................................40
5.7.1.
Use of specified study materials ..................................................41
5.7.2.
Biological sample.........................................................................41
5.7.3.
Laboratory assays .......................................................................41
5.7.4.
Biological samples evaluation......................................................42
5.7.4.1.
Immunological read-outs ............................................42
5.7.5.
Immunological correlates of protection.........................................42
6.
STUDY VACCINES AND ADMINISTRATION ........................................................42
6.1.
Description of study vaccines......................................................................42
6.2.
Storage and handling of study vaccine .......................................................43
6.3.
Dosage and administration of study vaccine...............................................44
6.4.
Replacement of unusable vaccine doses....................................................45
6.5.
Contraindications to subsequent vaccination ..............................................45
6.6.
Warnings and precautions ..........................................................................46
6.7.
Concomitant medications/products and concomitant vaccination................46
6.7.1.
Recording of concomitant medications/products and 
concomitant vaccinations.............................................................46
6.7.2.
Concomitant medications/products/vaccines that may lead 
to the elimination of a subject from per-protocol analyses............47
6.8.
Intercurrent medical conditions that may lead to elimination of a 
subject from per-protocol analyses .............................................................48
7.
HEALTH ECONOMICS ..........................................................................................48
8.
SAFETY.................................................................................................................48
8.1.
Safety definitions ........................................................................................48
8.1.1.
Definition of an adverse event......................................................48
8.1.2.
Definition of a serious adverse event ...........................................49
8.1.3.
Solicited adverse events..............................................................50
8.1.3.1.
Solicited general adverse events................................50
8.1.4.
Clinical laboratory parameters and other abnormal 
assessments qualifying as adverse events or serious 
adverse events ............................................................................51
8.2.
Detecting and recording adverse events and serious adverse events.........51
8.2.1.
Time period for detecting and recording adverse events 
and serious adverse events .........................................................51
8.2.2.
Post-Study adverse events and serious adverse events..............52
 
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8.2.3.
Evaluation of adverse events and serious adverse events...........52
8.2.3.1.
Active questioning to detect adverse events 
and serious adverse events........................................52
8.2.3.2.
Assessment of adverse events...................................53
8.2.3.2.1.
Assessment of intensity..........................53
8.2.3.2.2.
Assessment of causality .........................55
8.2.3.3.
Assessment of outcomes............................................56
8.2.3.4.
Medically attended visits.............................................56
8.3.
Reporting of serious adverse events...........................................................57
8.3.1.
Prompt reporting of serious adverse events to GSK 
Biologicals ...................................................................................57
8.3.2.
Contact information for reporting serious adverse events ............57
8.3.3.
Completion and transmission of SAE reports to GSK 
Biologicals ...................................................................................57
8.3.3.1.
Back-up system in case the electronic reporting 
system does not work.................................................58
8.3.4.
Updating of SAE information after removal of write access 
to the subject’s eCRF...................................................................58
8.3.5.
Regulatory reporting requirements for serious adverse 
events..........................................................................................58
8.4.
Follow-up of adverse events and serious adverse events ...........................58
8.4.1.
Follow-up during the study...........................................................58
8.4.2.
Follow-up after the subject is discharged from the study..............59
8.5.
Treatment of adverse events ......................................................................59
8.6.
Subject card................................................................................................59
9.
SUBJECT COMPLETION AND WITHDRAWAL.....................................................60
9.1.
Subject completion .....................................................................................60
9.2.
Subject withdrawal......................................................................................60
9.2.1.
Subject withdrawal from the study................................................60
9.2.2.
Subject withdrawal from study vaccine ........................................61
10. STATISTICAL METHODS......................................................................................61
10.1.
Primary Endpoint ........................................................................................61
10.2.
Secondary Endpoints (Amended: 30 October 2019) ...................................61
10.3.
Determination of sample size (Amended: 30 October 2019) .......................62
10.4.
Cohorts for Analyses...................................................................................63
10.4.1.
Exposed Set ................................................................................63
10.4.2.
Per-Protocol Set for analysis of immunogenicity 
(Amended: 30 October 2019).......................................................63
10.5.
Derived and transformed data (Amended: 30 October 2019)......................63
10.6.
Analysis of demographics ...........................................................................64
10.7.
Analysis of immunogenicity.........................................................................65
10.7.1.
Within group assessment (Amended: 30 October 2019)..............65
10.7.2.
Between groups assessment (Amended: 30 October 
2019) ...........................................................................................65
10.8.
Analysis of safety........................................................................................66
10.8.1.
Within groups assessment...........................................................66
10.9.
Interpretation of analyses............................................................................66
10.10. Statistical Methods (Amended: 30 October 2019).......................................67
10.11. Conduct of analysis ....................................................................................67
10.11.1. Sequence of analyses..................................................................67
 
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10.11.2. Statistical considerations for interim analyses..............................67
11. ADMINISTRATIVE MATTERS ...............................................................................67
11.1.
electronic Case Report Form instructions ...................................................67
11.2.
Study Monitoring by GSK Biologicals..........................................................68
11.3.
Record retention .........................................................................................68
11.4.
Quality assurance.......................................................................................69
11.5.
Posting of information on publicly available clinical trial registers and 
publication policy ........................................................................................69
11.6.
Provision of study results to investigators ...................................................69
11.7.
Data Sharing...............................................................................................70
12. COUNTRY SPECIFIC REQUIREMENTS...............................................................70
13. REFERENCES (AMENDED: 30 OCTOBER 2019).................................................71
 
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LIST OF TABLES
PAGE
Table 1
Study groups and epoch foreseen in the study.......................................30
Table 2
Study groups and treatment foreseen in the study .................................30
Table 3
Blinding of the study epoch ....................................................................30
Table 4
List of study procedures .........................................................................36
Table 5
Intervals between study visits.................................................................37
Table 6
Biological samples .................................................................................41
Table 7
Humoral Immunity (Antibody determination)...........................................41
Table 8
Immunological read-outs........................................................................42
Table 9
Study vaccine (Amended: 30 October 2019)..........................................43
Table 10
Dosage and administration.....................................................................45
Table 11
Solicited general adverse events............................................................50
Table 12
Reporting periods for collecting safety information .................................52
Table 13
Intensity scales for solicited symptoms in infants....................................54
Table 14
Timeframes for submitting serious adverse event and other 
events reports to GSK Biologicals ..........................................................57
Table 15
Power according to different scenarios for the % of non-
evaluable subjects, with respect to a total sample size of 450 
subjects* (Amended: 30 October 2019)..................................................62
Table 16
GSK Biologicals’ laboratories .................................................................73
Table 17
Outsourced laboratories.........................................................................73
 
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LIST OF APPENDICES
PAGE
APPENDIX A LABORATORY ASSAYS .......................................................................72
APPENDIX B CLINICAL LABORATORIES ..................................................................73
APPENDIX C AMENDMENTS AND ADMINISTRATIVE CHANGES TO THE 
PROTOCOL...........................................................................................74
 
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LIST OF ABBREVIATIONS
AE:
Adverse Event
ATP:
According-To-Protocol
CCID50:
Median Cell Culture Infective Dose (quantity of virus 
causing infection in 50% of exposed cells)
CDISC:
Clinical Data Interchange Standards Consortium
CI:
Confidence Interval
CLS:
Clinical Laboratory Sciences
cm: 
centimeter
DCGI: 
Drug Controller General of India
DMEM:
Dulbecco’s Modified Eagle Medium
eCRF:
electronic Case Report Form
ELISA:
Enzyme Linked Immunosorbent Assay
EoS:
End of Study
ES:
Exposed Set
eTDF:
electronic Temperature Excursion Decision Form
GCP:
Good Clinical Practice
GE:
Gastroenteritis
GMC:
Geometric Mean Concentration
GSK:
GlaxoSmithKline
HRV:
Human Rotavirus
IB:
Investigator Brochure
ICF: 
Informed Consent Form
ICH:
International Conference on Harmonization
IEC: 
Independent Ethics Committee
IgA:
Immunoglobulin A
 
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IMP:
Investigational Medicinal Product
IRB: 
Institutional Review Board
IS:
Intussusception
Kg:
Kilograms
LAR:
Legally Acceptable Representative
LSLV:
Last Subject Last Visit
m:
meter
MATEX:
MATerial EXcellence
MedDRA:
Medical Dictionary for Regulatory Activities
mg:
Milligrams
mL:
Milliliter
NDAC:
New Drug Advisory Committee
PCD:
Primary Completion Date
PPS:
Per-Protocol Set
RCC:
Reverse Cumulative Curve
RT-PCR:
Reverse Transcription Polymerase Chain reaction
RV:
Rotavirus
SAE:
Serious Adverse Event
SAS:
Statistical Analysis System
SBIR:
Randomization System on Internet
SCID:
Severe Combined Immunodeficiency
SmPC: 
Summary of Product Characteristics
SPM:
Study Procedures Manual
U:
Unit
WHO:
World Health Organization
 
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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.
Blinding:
A procedure in which one or more parties to the trial are 
kept unaware of the treatment assignment in order to 
reduce the risk of biased study outcomes. The level of 
blinding is maintained throughout the conduct of the 
trial, and only when the data are cleaned to an 
acceptable level of quality will appropriate personnel be 
unblinded or when required in case of a Serious Adverse 
Event (SAE). In an open-label study, no blind is used. 
Both the investigator and the subject know the identity 
of the treatment assigned.
Child in care:
A child who has been placed under the control or 
protection of an agency, organization, institution or 
entity by the courts, the government or a government 
body, acting in accordance with powers conferred on 
them by law or regulation. The definition of a child in 
care can include a child cared for by foster parents or 
living in a care home or institution, provided that the 
arrangement falls within the definition above. The 
definition of a child in care does not include a child who 
is adopted or has an appointed legal guardian.
Diarrhea:
Passage of three or more looser than normal stools 
within a day.
Eligible:
Qualified for enrolment into the study based upon strict 
adherence to inclusion/exclusion criteria.
 
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End of Study (EoS):
(Synonym of End of Trial)
For studies without collection of human biologicals 
samples or imaging data EoS is the Last Subject Last 
Visit (LSLV).
For studies with collection of Human Biologicals 
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 is 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.
Typical examples of epochs are screening, primary 
vaccinations, boosters, yearly immunogenicity 
follow-ups, and surveillance periods for efficacy or 
safety.
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 Sections 6.7.2
and 10.4 for details on criteria for evaluability).
Gastroenteritis:
Diarrhea with or without vomiting.
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.
Investigational vaccine:
(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.
 
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Legally acceptable 
representative (LAR):
(The terms legal 
representative or legally 
authorized representative 
are used in some settings.)
An individual or juridical or other body authorized under 
applicable law to consent, on behalf of a prospective 
subject, to the subject's participation in the clinical trial.
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 Harmonization (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.
Randomization:
Process of random attribution of treatment to subjects in 
order to reduce bias of selection.
Self-contained study:
Study with objectives not linked to the data of another 
study.
Site monitor:
An individual assigned by the sponsor who is 
responsible for assuring proper conduct of clinical 
studies at one or more investigational sites.
Seroconversion:
(Amended: 30 October 
2019)
For subjects with a pre-vaccination anti-RV IgA 
antibody concentration <20 U/mL, seroconversion is 
achieved when the post-vaccination concentration is 
≥20 U/mL.
For subjects with a pre-vaccination anti-RV IgA 
antibody concentration ≥20 U/mL, seroconversion is 
achieved when the post-vaccination concentration is ≥2 
times the pre-vaccination concentration.
Seronegative:
A seronegative subject for anti-rotavirus IgA antibody is
defined as a subject who has an antibody concentration 
below the clinically meaningful threshold of 20 U/ml.
 
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Seropositive:
A seropositive subject is defined as a subject who has an 
antibody concentration greater than or equal to the
clinically meaningful threshold of 20 U/ml.
Solicited adverse event:
AEs to be recorded as endpoints in the clinical study. 
The presence/occurrence/intensity of these events is 
actively solicited from the subject or an observer during 
a specified post-vaccination follow-up period.
Study vaccine:
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.
Treatment number:
A number identifying a treatment to a subject, according 
to the treatment allocation.
Unsolicited adverse event:
Any AE reported in addition to those solicited during the 
clinical study. Also any ‘solicited’ symptom with onset 
outside the specified period of follow-up for solicited 
symptoms will be reported as an unsolicited AE.
Vomiting:
One or more episodes of forceful emptying of partially 
digested stomach contents ≥1 hour after feeding within a 
day.
 
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TRADEMARK
The following trademark is used in the present protocol.
Note: In the body of the protocol (including the synopsis), the name of the vaccine will be 
written without the superscript symbol ™ or ® and in italics.
Trademarks of the GSK group of 
companies
Generic description
Rotarix
Human rotavirus vaccine
 
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1.
INTRODUCTION
1.1.
Background (Amended: 30 October 2019)
Rotavirus (RV) infection is the leading cause of acute Gastroenteritis (GE) and severe 
diarrhea in infants and young children <5 years of age [Atherly, 2009]. It has been 
estimated that in 2013, approximately 215,000 deaths were caused due to RV. India, 
Nigeria, Pakistan, and Democratic Republic of Congo accounted for approximately half 
(49%) of all the estimated RV deaths in 2013 [Tate, 2016].
Hospital-based surveillance studies in Asia have shown that 20% to 50% of the 
hospitalizations for diarrhea among children <5 years of age, are associated with RV 
infection [Kang, 2006]. In 2013, an estimated 47,100 RV deaths occurred in India, which 
was 22% of all deaths that occurred globally due to RV in the same year [Tate, 2016].
In 2013, WHO recommended inclusion of RV vaccination in the national immunization 
programs of all countries, particularly in countries where RV GE associated fatality rates 
are high among children aged <5 years (e.g., south and south-eastern Asia and sub-
Saharan Africa) [WHO position paper, 2013].
Rotarix is currently registered in over 130 countries and about 552 446 066 doses of the 
vaccine (lyophilized and liquid formulations) are estimated to have been distributed 
worldwide since its launch until July 2019.
GSK Biologicals’ lyophilized Human Rotavirus (HRV) vaccine has been extensively 
tested in clinical studies in infants and the vaccine efficacy has been demonstrated in 
several clinical studies conducted in Europe, North America, Latin America and the 
Caribbean, Asia and Africa [Vesikari, 2007; Linhares, 2008; Salinas, 2005; Phua, 2009;
Madhi, 2010]
Please refer to the current Investigator Brochure (IB) for information regarding the 
summary of potential risks and benefits of the Rotarix vaccine.
1.2.
Rationale for the study and study design
The lyophilized formulation of the HRV vaccine was licensed in India in February 2008. 
This study will be conducted to generate additional clinical data for the liquid formulation 
of the HRV vaccine in India, as recommended by New Drug Advisory Committee on 
Vaccines (NDAC-Vaccines) of Drug Controller General of India (DCGI).
This study will be conducted to evaluate the immunogenicity, reactogenicity and safety of 
the GSK Biologicals’ HRV liquid vaccine compared to the GSK Biologicals’ HRV 
lyophilized vaccine. The study is planned to be conducted in healthy infants aged 
6-10 weeks at the time of the first dose. Two oral doses of the study vaccine will be 
administered to the subjects, according to a 0, 1 month schedule. The blood samples for 
immunogenicity assessment will be collected from all subjects before the first dose and at 
one month after the second dose of the vaccine.
 
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1.3.
Benefit: Risk Assessment
Please refer to the current IB and prescribing information (lyophilized vaccine) for 
information regarding the summary of potential risks and benefits of the Rotarix vaccine.
The following section outlines the risk assessment and mitigation strategy for this study 
protocol:
1.3.1.
Risk Assessment
Important 
Potential/Identified Risk
Data/Rationale for Risk
Mitigation Strategy
Investigational study vaccine (Rotarix)
Intussusception (IS)
Spontaneous data

Subjects will be followed up
to the end of the study after 
the receipt of the vaccine to 
check for any safety signal.

Parents/ Legally acceptable 
representatives (LARs) of
subjects should report any 
untoward symptoms 
experienced after receiving 
the vaccine immediately to 
the investigator.

All Serious Adverse Events 
(SAEs) should be reported 
by the investigator 
immediately to GSK.

Subjects with SCID will be 
excluded from participating 
in this study (Refer to
Section 4.3 and Section 6
for more details).
Hematochezia
Spontaneous data
Gastroenteritis with vaccine 
viral shedding in infants with 
severe combined 
immunodeficiency (SCID)
Spontaneous data
Kawasaki disease
Based on signal observed 
for RotaTeq vaccine
Study Procedures
Allergic reaction to the 
vaccine
Spontaneous data
Subjects will be observed for at 
least 30 minutes after the 
vaccine administration, with 
medical attention available in 
case of anaphylactic reactions.
 
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1.3.2.
Benefit Assessment
By receiving the HRV vaccine, the subject may become protected against the RV disease. 
In addition, the subject’s participation will benefit other children in the future, since 
information collected during this study will help in the evaluation of the safety, 
reactogenicity and immunogenicity of the HRV liquid vaccine.
In addition, the subject will undergo a physical examination at the first study visit. In case 
the study doctor discovers any medical condition, the subject will be referred to the local 
healthcare system.
1.3.3.
Overall Benefit: Risk Conclusion
Considering the measures taken to minimize the risk to subjects participating in this 
study, the potential or identified risks in association with [HRV vaccine] are justified by 
the potential benefits (prevention/treatment) that may be afforded to subjects receiving 
the vaccine for immunization against RV.
2.
OBJECTIVES
2.1.
Primary objective

To evaluate non-inferiority of GSK Biologicals' HRV liquid vaccine compared to 
GSK Biologicals' HRV lyophilized vaccine in terms of geometric mean 
concentrations (GMCs) for anti-RV antibodies, one month post dose 2 of HRV liquid
vaccine and HRV lyophilized vaccine.

Criterion: Non-inferiority will be stated if the lower limit of the two-sided 95% 
confidence interval (CI) for the ratio of anti RV IgA antibody GMCs between 
HRV liquid vaccine over the HRV lyophilized vaccine, one month after dose 2 is 
greater than or equal to 0.5.
Refer to Section 10.1 for the definition of the primary endpoint.
2.2.
Secondary objectives (Amended: 30 October 2019)

To assess the immunogenicity of the HRV liquid vaccine and HRV lyophilized
vaccine, in terms of seroconversion* rates, one month post dose 2 of HRV vaccine.
*Definition: 

for subjects with a pre-vaccination anti-RV IgA antibody concentration 
<20 U/mL, seroconversion is achieved when the post-vaccination 
concentration is ≥20 U/mL.

for subjects with a pre-vaccination anti-RV IgA antibody concentration 
≥20 U/mL, seroconversion is achieved when the post-vaccination 
concentration is ≥2 times the pre-vaccination concentration.

To assess the reactogenicity of the HRV liquid vaccine and the HRV lyophilized
vaccine in terms of solicited adverse events (AEs), during the 8-day (Day 1–Day 8) 
follow-up period after each vaccination.
 
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
To assess the safety of the HRV liquid vaccine and the HRV lyophilized vaccine in 
terms of unsolicited AEs, during the 31-day (Day 1–Day 31) follow-up period after 
each vaccination and serious adverse events (SAEs), during the entire study period.
Refer to Section 10.2 for the definition of the secondary endpoints.
3.
STUDY DESIGN OVERVIEW
N= number of subjects planned to be enrolled; HRV= Human Rotavirus; Pre-Vacc= Pre-vaccination; 
Post-Vacc=Post-Vaccination; BS=Blood sample
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 III, open-label, randomized (1:1), multi-centric, 
single-country study with two parallel groups.

Duration of the study: The intended duration of the study, per subject, is
approximately two months.

Epoch 001: Primary starting at Visit 1 (Day 1) and ending at Visit 3 (Month 2).

Primary Completion Date (PCD): Last subject attending Visit 3.
Refer to glossary of terms for the definition of PCD.

End of Study (EoS): Last testing results released for samples collected at Visit 3.
Refer to glossary of terms for the definition of EoS.

Study groups: The study groups and epoch foreseen in the study are provided in
Table 1.
Vaccination visits
Follow-up visit
HRV Liq (N=225)
HRV Lyo (N=225)
Randomization 
(1:1)
Visit 1-Day 1
Dose 1
Pre-vacc BS
Visit 2-Month 1
Dose 2
Visit 3-Month 2
Post- vacc BS
 
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Table 1
Study groups and epoch foreseen in the study
Study groups
Number of subjects
Age at Dose 1 (Min/Max)
Epoch 001
HRV Liq
225
6 weeks-10 weeks
●
HRV Lyo
225
6 weeks- 10 weeks
●

Treatment groups: Table 2 presents the study groups and the vaccine to be 
administered in the study.

HRV Liquid vaccine group (also referred to as HRV Liq)

HRV Lyophilized vaccine group (also referred to as HRV Lyo).
Table 2
Study groups and treatment foreseen in the study
Treatment name
Vaccine/Product name
Study Groups
HRV Liq
HRV Lyo
HRV Liquid
GSK Biologicals’ HRV liquid vaccine
x
HRV Lyophilized
GSK Biologicals’ HRV lyophilized vaccine
x
GSK Biologicals’ calcium carbonate buffer
x

Control: active control (Lyophilized HRV vaccine)

Vaccination schedule:

Two oral doses of the HRV vaccine to be given according to a 0, 1 month 
schedule.

All subjects are allowed to receive routine childhood vaccinations according to 
the local immunization practice. Administration of all routine childhood 
vaccinations given since birth will be recorded in the electronic Case Report 
Form (eCRF).

Treatment allocation: Randomized (1:1). Treatment number will be allocated using 
GSK Biologicals’ Randomization System on Internet (SBIR).

Blinding: open
The blinding of study epoch is provided in Table 3.
Table 3
Blinding of the study epoch
Study Epoch
Blinding
Epoch 001
open

Sampling schedule: Details of the samples to be collected are as follows:

Blood samples (approximately 2 ml) will be collected from all subjects at Visit 1 
and Visit 3 to measure serum anti-RV Immunoglobulin A (IgA) antibody 
concentrations using Enzyme Linked Immunosorbent Assay (ELISA).

Type of study: self-contained.

Data collection: eCRF.
 
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4.
STUDY COHORT
4.1.
Number of subjects/centers
The target enrolment will be 450 subjects (225 subjects in each study group) to obtain at 
least 292 evaluable subjects (146 subjects in each study group). Refer to Section 10.3 for
the detailed description of the criteria used in the estimation of the sample size.
Overview of recruitment plan:

The study will be conducted in multiple centers across India.

Enrolment will be terminated when 450 subjects have been enrolled.

The recruitment and randomization will be monitored by SBIR.
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’ parent(s)/LAR(s) who, in the opinion of the investigator, can and will 
comply, with the requirements of the protocol (e.g. completion of the diary cards, 
return for follow-up visits).

Written informed consent obtained from the parent(s)/LAR(s) of the subject prior to 
performing any study specific procedure.

A male or female between, and including, 6 and 10 weeks of age at the time of the 
first vaccination.

Healthy subjects as established by medical history and clinical examination before 
entering into the study.

Birth weight >2000 grams.
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.
 
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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:

Child in care
Please refer to the glossary of terms for the definition of child in care.

Use of any investigational or non-registered product (drug or vaccine) other than the 
study vaccines during the period starting 30 days before the first dose of study 
vaccines) (Day-29 to Day 1), or planned use during the study period.

Chronic administration (defined as more than 14 days in total) of 
immunosuppressants or other immune-modifying drugs since birth. For 
corticosteroids, this will mean prednisone 0.5 mg/kg/day, or equivalent. Inhaled and 
topical steroids are allowed.

Administration of any chronic drug therapy to be continued during the study period.

Planned administration/administration of a vaccine not foreseen by the study 
protocol in the period starting 30 days before the first dose of vaccine administration 
and ending at Visit 3; with the exception of the inactivated influenza vaccine, which 
is allowed at any time during the study, and other licensed routine childhood 
vaccinations, according to the local immunization practice.

Concurrently participating in another clinical study, at any time during the study 
period, in which the subject has been or will be exposed to an investigational or a 
non-investigational product (pharmaceutical product or device).

History of confirmed RV GE.

Previous vaccination against RV.

Any confirmed or suspected immunosuppressive or immunodeficient condition, 
(including Severe Combined Immunodeficiency [SCID] disorder) based on medical 
history and physical examination (no laboratory testing required).

Uncorrected congenital malformation (such as Meckel’s diverticulum) of the 
gastrointestinal tract that would predispose for Intussusception (IS).

History of IS.

Very prematurely born infants (born ≤28 weeks of gestation).

Hypersensitivity to latex.

Family history of congenital or hereditary immunodeficiency.

History of any reaction or hypersensitivity likely to be exacerbated by any 
component of the vaccine.

Major congenital defects or serious chronic illness.

History of any neurological disorders or seizures.
 
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
Acute disease and/or fever at the time of enrolment. This warrants deferral of 
vaccination.

Fever is defined as temperature 38.0°C/100.4°F. The preferred location for 
measuring temperature in this study will be the oral cavity, the axilla or the 
rectum.

Subjects with a minor illness (such as mild diarrhea, mild upper respiratory 
infection) without fever, may be enrolled at the discretion of the investigator.

Acute or chronic, clinically significant pulmonary, cardiovascular, hepatic or renal 
functional abnormality, as determined by physical examination or medical history.

Administration of immunoglobulins and/or any blood products since birth or planned 
administration during the study period.

Administration of long-acting immune-modifying drugs at any time during the study 
period (e.g., infliximab).

GE within 7 days preceding the study vaccine administration (warrants deferral of 
the vaccination).
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 International Conference on 
Harmonization (ICH) Guideline for Good Clinical Practice (GCP), all applicable subject 
privacy requirements and the guiding principles of the Declaration of Helsinki.
The study has been designed and will be conducted in accordance with the ICH 
Harmonized Tripartite Guideline for clinical investigation of medicinal products in the 
paediatric population (ICH E11) and all other applicable ethical guidelines.
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’s parent(s)/LAR(s) 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.
 
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Freely given and written or witnessed/thumb printed informed consent must be obtained 
from each subject’s parent(s)/LAR(s) as appropriate, 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.
5.2.
Subject identification and randomization
5.2.1.
Subject identification
Subject identification numbers will be assigned sequentially to the subjects whose 
parent(s)/LAR(s) have consented to their participation in the study, according to the range 
of subject identification numbers allocated to each study center.
5.2.2.
Randomization of treatment
5.2.2.1.
Randomization of supplies
The numbering of supplies will be performed at GSK Biologicals, using a block scheme 
randomization in MATerial EXcellence (MATEX), a program developed for use in 
Statistical Analysis System (SAS®) (Cary, NC, US) by GSK Biologicals. Entire blocks 
will be shipped to the study center(s)/warehouse(s).
To allow GSK Biologicals to take advantage of greater rates of recruitment than 
anticipated at individual centers in this multi-center study and to thus reduce the overall 
study recruitment period, additional supplies will be prepared.
5.2.2.2.
Treatment allocation to the subject
The treatment numbers will be allocated by dose.
5.2.2.2.1.
Study group and treatment number allocation
The target will be to enrol 450 subjects who will be randomly assigned to one of the two
study groups in a (1:1) ratio (225 subjects in each group).
Allocation of the subject to a study group at the investigator site will be performed using 
SBIR. The randomization algorithm will use a minimization procedure accounting for 
center and the study as minimization factors. Minimization factors will have equal weight 
in the minimization algorithm.
 
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After obtaining the signed and dated ICF from the subject’s parent/LAR and having 
checked the eligibility of the subject, the site staff in charge of the vaccine administration 
will access SBIR. Upon providing the subject identification number, SBIR will provide 
the treatment number to be used for the first dose.
The number of each administered treatment must be recorded in the eCRF on the Vaccine 
Administration screen.
When SBIR is not available, please refer to the SBIR user guide or the Study Procedures 
Manual (SPM) for specific instructions.
5.2.2.2.2.
Treatment number allocation for subsequent doses 
For the second dose, the study staff in charge of the vaccine administration will access 
SBIR, provide the subject identification number, and the system will provide a treatment 
number consistent with the allocated study group.
The number of each administered treatment must be recorded in the eCRF on the Vaccine 
Administration screen.
5.3.
Method of blinding
This is an open-label study.
The laboratory in charge of the laboratory testing will be blinded to the treatment, and 
codes will be used to link the subject and study (without any link to the treatment 
attributed to the subject) to each sample.
5.4.
General study aspects
Supplementary study conduct information not mandated to be present in this protocol is 
provided in the accompanying 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
The list of study procedures is detailed in Table 4.
 
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Table 4
List of study procedures
Age at enrolment
6-10 weeks
Epoch
Epoch 001
Type of contact
Visit 1
Visit 2
Visit 3
Time points
Day 1
Month 1
Month 2
Sampling time points
Pre-Vacc
Post-Vacc
Informed consent
●
Check inclusion/exclusion criteria
●
Collect demographic data
●
Check contraindications and warnings and precautions
●
●
Medical history
●
Physical examination
●
History of previous vaccination from birth
●
Pre-vaccination body temperature
●
●
Vaccines
Study group and treatment number allocation
O
Treatment number allocation for subsequent doses
O
Recording of administered treatment number
●
●
HRV vaccine administration
●
●
Record regurgitation
●
●
HRV vaccine replacement dose administration in case of regurgitation *
●
●
Laboratory Assays
Blood sampling for antibody determination (2 ml)
●
●
Safety assessments
Distribution of diary cards
O
O
Recording of solicited adverse events within 8 days after each dose of HRV 
vaccine (Day 1–Day 8)
●
●
Recording of non-serious adverse events within 31 days (Day 1–Day 31) post-
vaccination
●
●
●
Record any concomitant medication/vaccination
●
●
●
Record any intercurrent medical conditions
●
●
●
Recording of AEs/SAEs leading to withdrawal from study
●
●
●
Recording of SAEs
●
●
●
Recording of SAEs related to study participation, or to a concurrent GSK 
medication/vaccine
●
●
●
Return of diary cards
O
O
Diary card transcription by investigator
●
●
Study Conclusion
●
● is used to indicate a study procedure that requires documentation in the individual eCRF.
○ is used to indicate a study procedure that does not require documentation in the individual eCRF.
LAR = Legally Acceptable Representative; AE = Adverse Events; SAE = Serious Adverse Events
* If regurgitation or vomiting occurs after vaccination, a single replacement dose may be given at the same vaccination 
visit at the discretion of the Investigator.
Note: All subjects are allowed to receive routine childhood vaccinations according to the local immunization practice.
 
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Whenever possible the investigator should arrange the study visits within the interval 
provided in Table 5.
Table 5
Intervals between study visits
Interval
Optimal length of interval 1
Allowed interval
Visit 1 Visit 2
1 month
28-48 days after Dose 1
Visit 2 Visit 3
1 month
31-48 days after Dose 2
1 Whenever possible the investigator should arrange the study visits within this interval.
Note: The date of the previous visit serves as the reference date for the intervals between the study visits.
5.6.
Detailed description of study procedures
5.6.1.
Informed consent
The signed/witnessed/thumb printed informed consent of the subject’s parent(s)/LAR(s) 
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 age, sex and geographical ancestry 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 any pre-existing conditions or signs and/or 
symptoms present in a subject, prior to the first study vaccination in the eCRF.
5.6.5.
Physical examination
Perform a physical examination of the subject, including assessment of 
oral/axillary/rectal body temperature, height and weight. Collected information needs to 
be recorded in the eCRF.
Physical examination at each study visit subsequent to the first vaccination visit will be 
performed only if the subjects’ parent(s)/LAR(s) indicates during questioning that there 
might be some underlying pathology(ies) or if deemed necessary by the Investigator or 
delegate.
If the investigator determines that the subject’s health on the day of vaccination 
temporarily precludes vaccination, the visit will be rescheduled.
 
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Treatment of any abnormality observed during physical examination has to be performed 
according to local medical practice outside this study or by referral to an appropriate 
health care provider.
5.6.6.
Check contraindications, warnings and precautions to 
vaccination 
Contraindications, warnings and precautions to vaccination must be checked at the
beginning of each vaccination visit. Refer to Sections 6.5 and 6.6 for more details.
5.6.7.
Assess pre-vaccination body temperature
The oral/axillary/rectal body temperature of each subject needs to be measured prior to 
any study vaccine administration. If the subject has fever on the day of vaccination, the 
vaccination visit will be rescheduled within the allowed interval for this visit 
(see Table 5).
5.6.8.
Study group and treatment number allocation
Study group and treatment number allocation will be performed as described in Section 
5.2.2. The number of each administered treatment must be recorded in the eCRF.
5.6.9.
Sampling
Refer to the Module on Biospecimen Management in the SPM for detailed instructions 
for the collection, handling and processing of the samples.
5.6.9.1.
Blood sampling for immune response assessment
Blood samples will be taken at Visit 1 and Visit 3 as specified in 
Section 5.5, outline of study procedures.

A volume of approximately 2 mL of whole blood (to provide approximately 
0.7 mL of serum) should be drawn from all subjects for anti-RV antibody 
determination at Visit 1 and Visit 3. After centrifugation, serum samples should 
be kept at –20°C/–4°Fahrenheit or below until shipment. Refer to the SPM for 
more details on sample storage conditions.
5.6.10.
Study Vaccine administration

After completing all prerequisite procedures prior to vaccination, two oral doses of 
the study vaccine will be administered at an approximate one-month interval to 
subjects, according to the immunization schedule for HRV vaccine administration 
(refer to Section 6.3 for detailed description of the vaccines administration 
procedure). If the investigator or delegate determines that the subject’s health on the
day of administration temporarily precludes vaccines administration, the visit will be 
rescheduled within the allowed interval for this visit (refer to Table 5).
 
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
The subjects will be observed closely for at least 30 minutes following the 
administration of the vaccine, with appropriate medical treatment readily available in 
case of anaphylaxis.
5.6.11.
Record regurgitation
If regurgitation or vomiting occurs after vaccination, a single replacement dose may be 
given at the same vaccination visit at the discretion of the Investigator. This information 
should be recorded in the eCRF. The subject may continue to participate in the study.
5.6.12.
Check and record concomitant medication/vaccination and 
intercurrent medical conditions
Concomitant medication/vaccination must be checked and recorded in the eCRF as 
described in Section 6.7.
Intercurrent medical conditions must be checked and recorded in the eCRF as described 
in Section 6.8.
5.6.13.
Recording of AEs and SAEs

Refer to Section 8.2 for procedures for the investigator to record AEs and SAEs. 
Refer to Section 8.3 for guidelines and how to report SAE reports to GSK 
Biologicals.

The subjects’ parent(s)/LAR(s) will be instructed to contact the investigator 
immediately should the subjects manifest any signs or symptoms they perceive as 
serious.

At each vaccination visit, diary cards will be provided to the subject’s 
parent(s)/LAR(s). The subject’s parent(s)/LAR(s) will be instructed to measure and 
record the oral, axillary or rectal body temperature, and any solicited general AEs 
(i.e., on the day of vaccination and during the next seven days) or any unsolicited 
AEs (i.e., on the day of vaccination and during the next 30 days after vaccination). 
The subject’s parent(s)/LAR(s) will be instructed to return the completed diary card 
to the investigator at the next study visit.

Collect and verify completed diary cards during discussion with the subject’s 
parent(s)/LAR(s).

Any unreturned diary cards will be sought from the subject’s parent(s)/LAR(s) 
through telephone call(s) or any other convenient procedure.

The investigator will transcribe the collected information into the eCRF in English.
 
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5.6.14.
Study conclusion
The investigator will:

review data collected to ensure accuracy and completeness, and

complete the study conclusion screen in 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 India and will only be performed once an IEC 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’s 
parent(s)/LAR(s).
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).
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.
 
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5.7.1.
Use of specified study materials
When materials are provided by GSK Biologicals, it is MANDATORY that all clinical 
samples (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
analyzed). 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 sample
The biological samples to be collected from subjects are described in Table 6.
Table 6
Biological samples
Sample type
Quantity
Unit
Time points
Blood
Approximately 2
ml
Visit 1 and Visit 3
ml-milliliter
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 and/or sample management.
All serological assays for the determination of antibodies against RV will be performed 
by ELISA at GSK Biologicals’ laboratory or in a laboratory designated by GSK 
Biologicals using standardized and validated procedures (refer to Table 7).
Table 7
Humoral Immunity (Antibody determination)
System
Component
Method
Kit/Manufacturer
Unit
Cut-off
Laboratory
SER
Rotavirus Ab.IgA
ELI
NA
U/ml
13*
GSK Biologicals**
SER = Serum
IgA = Immunoglobulin A
ELI = ELISA (Enzyme Linked Immunosorbent Assay)
U = Units; ml = milliliters
*This corresponds to the technical cut-off of the revalidated laboratory assay.
**GSK Biologicals laboratory refers to the Clinical Laboratory Sciences (CLS) in Rixensart, Belgium; Wavre, Belgium
and Marburg, Germany
Additional exploratory testing on the vaccine and/or on 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.
 
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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
5.7.4.1.
Immunological read-outs
The immunological read-outs are presented in Table 8.
Table 8
Immunological read-outs
Blood sampling time point
Type of contact and 
time point
Sampling time point
No. subjects
Component
Components 
priority rank
Visit 1 (Day 1)
Pre-Vacc
All
Rota Ab.IgA
None
Visit 3 (Month 2) 
Post-Vacc
All
Rota Ab.IgA
None
Pre-Vacc: Pre-vaccination; Post-Vacc: Post Vaccination; Ab: Antibody IgA: Immunoglobulin A
5.7.5.
Immunological correlates of protection
No immunological correlate of protection has been demonstrated so far for the antigen 
used as part of the HRV vaccine. However, a study by Cheuvart et al., in 2014 indicated 
that post-vaccination anti-RV IgA seropositivity (antibody concentration 20 U/mL) 
may serve as a useful correlate of vaccine efficacy in clinical trials of Rotarix [Cheuvart, 
2014].
The investigator is encouraged to share the immunological assay results for non-
responders with the study subjects’ parent(s)/LAR(s).
6.
STUDY VACCINES AND ADMINISTRATION
6.1.
Description of study vaccines
The study vaccines to be used have been developed and manufactured by GSK 
Biologicals.
The Quality Control Standards and Requirements for each study vaccines are described in 
separate Quality Assurance documents (e.g., release protocols, certificate of analysis) and 
the required approvals have been obtained.
The vaccines are labelled and packed according to applicable regulatory requirements.
Commercial vaccines are assumed to comply with the specifications given in the 
manufacturer’s Summary of Product Characteristics (SmPC).
The study vaccines to be utilized in the study are detailed in Table 9.
 
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Table 9
Study vaccine (Amended: 30 October 2019)
Treatment 
name
Vaccine 
name
Formulation
Presentation
Volume to 
be 
administered
orally
Number 
of doses
HRV Liquid
HRV Liquid 
vaccine
Active substance: 
HRV RIX4414 live 
attenuated 
>=10⁶⋅⁰CCID₅₀
Excipients:
Sucrose=1.073g; Di-
sodium 
Adipate=132.74mg; 
DMEM=2.26mg; water 
for injection=1.5ml
Liquid vaccine in a pre-filled 
oral applicator
1.5 ml
2
HRV 
Lyophilized
HRV 
Lyophilized 
vaccine
HRV RIX4414 live 
attenuated 
>=10⁶⋅⁰CCID₅₀
Lyophilized vaccine in a 
monodose glass vial. Diluent 
(calcium carbonate buffer) 
supplied separately
1 ml
2
HRV 
Diluent
CaCO₃=60mg
Diluent for lyophilized 
vaccine (calcium carbonate 
liquid antacid) supplied 
separately in a prefilled oral 
applicator
HRV=Human Rotavirus; CCID50 = Median Cell Culture Infective Dose (quantity of virus causing infection in 50% of 
exposed cells); DMEM = Dulbecco's Modified Eagle Medium; ml = milliliter;
6.2.
Storage and handling of study vaccine
The study vaccines must be stored at the label storage temperature conditions in a safe 
and locked place. Access to the storage space should be limited to authorized study 
personnel. The storage conditions will be assessed during pre-study activities under the 
responsibility of the sponsor study contact. The storage temperature should be 
continuously monitored with calibrated (if not validated) temperature monitoring 
device(s) and recorded. Refer to the Module on Clinical Trial Supplies in the SPM for 
more details on storage of the study vaccines.
Temperature excursions must be reported in degree Celsius.
Any temperature excursion outside the range of 0.0 to +8.0Celsius (for +2 to +8°C/+36 
to +46°Fahrenheit label storage condition) impacting investigational medicinal products 
(IMPs) must be reported in the appropriate (electronic) temperature excursion decision 
form ([e]TDF). The impacted IMPs must not be used and must be stored in quarantine at 
label temperature conditions until usage approval has been obtained from the sponsor.
In case of temperature excursion below +2.0°Celsius down to 0.0°Celsius impacting 
IMP(s) there is no need to report in (e)TDF, but adequate actions must be taken to restore 
the +2 to +8°Celsius/+36 to +46°Fahrenheit label storage temperature conditions. The 
impacted IMP(s) may still be administered, but the site should avoid re-occurrence of 
such temperature excursion. Refer to the Module on Clinical Trial Supplies in the SPM 
for more details on actions to take.
 
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Refer to the Module on Clinical Trial Supplies in the SPM for details and instructions on 
the temperature excursion reporting and usage decision process, packaging and 
accountability of the study vaccines.
6.3.
Dosage and administration of study vaccine
Liquid formulation of HRV vaccine
The pre-filled oral applicator should be shaken well before use. The vaccine 
(approximately 1.5 mL) should then be administered orally as a single dose. 
Lyophilized formulation of HRV vaccine
To prepare GSK Biologicals' HRV lyophilized vaccine for administration, the entire 
content of the supplied diluent (calcium carbonate buffer) should be transferred from the 
oral applicator into the vial of the lyophilized product via the intermediate device. The 
vial should be shaken well to re-suspend the vaccine. The entire volume of the 
re-suspended product (approximately 1 mL) should be withdrawn into the same oral 
applicator and the re-suspended product should then be administered promptly as a single 
oral dose.
Administration of the vaccines
In order to allow the swallowing of the entire volume of the single oral dose, the 
administration should occur in a quiet environment. The child should be seated in a 
reclining position. Administer orally (i.e., into the child’s mouth towards the inner cheek) 
the entire content of oral applicator. Sufficient time should be allowed for the baby to 
swallow the vaccine solution, to avoid regurgitation or vomiting. Should however the 
subject regurgitate or vomit after study vaccine administration, a single replacement dose 
may be given at the same vaccination visit at the discretion of the Investigator. This 
information should be recorded in the eCRF. The subject may continue to participate in 
the study.
The vaccinees will be observed closely for at least 30 minutes, with appropriate medical 
treatment readily available in case of a rare anaphylactic reaction following the 
administration of vaccines.
The vaccination regimen is summarized in Table 10.
 
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Table 10
Dosage and administration
Type of contact 
and time point
Study group
Treatment 
name
Volume to be 
administered 
orally
Route
Site
Location
Directionality
Laterality
Visit 1 (Day 1),
Visit 2 (Month 1)
HRV Liq
HRV Liquid
1.5 ml
O
Not 
applicable
Not applicable
Not 
applicable
Visit 1 (Day 1),
Visit 2 (Month 1)
HRV Lyo
HRV 
Lyophilized
1 ml
O
Not 
applicable
Not applicable
Not 
applicable
O= Oral; ml-millilitre; HRV- Human Rota Virus
6.4.
Replacement of unusable vaccine doses
In addition to the vaccine doses provided for the planned number of subjects (including
over-randomization when applicable), at least 30% additional vaccine doses will be 
supplied to replace those that are unusable.
6.5.
Contraindications to subsequent vaccination
The following events constitute absolute contraindications to further administration of 
HRV vaccine. If any of these events occur during the study, the subject must not receive 
additional doses of vaccines but may continue other study procedures at the discretion of 
the investigator (see Section 8.4).

Anaphylaxis following the administration of vaccines.

Hypersensitivity reaction following the administration of the HRV vaccine.

Any uncorrected congenital malformation of the gastrointestinal tract (such as 
Meckel’s diverticulum) that would predispose for IS.

Any history of IS.

SCID.
The following events constitute contraindications to administration of the study vaccines 
at that point in time; if any of these events occur at the time scheduled for vaccination, 
the subject may be vaccinated at a later date, within the time window specified in the 
protocol (see Section 5.5), or the subject may be withdrawn at the discretion of the 
investigator (see Section 8.4). 

Acute disease and/or fever at the time of vaccination. 

Fever is defined as temperature 38.0°C/100.4°F. The preferred location for 
measuring temperature in this study will be the oral cavity, the axilla and the 
rectum.

Subjects with a minor illness (such as mild diarrhoea, mild upper respiratory 
infection) without fever may be vaccinated at the discretion of the investigator).

GE within seven days preceding the study vaccine administration.
 
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6.6.
Warnings and precautions
The Rotarix vaccine should under no circumstances be injected.
There is no data on the safety and efficacy of Rotarix in infants with gastrointestinal 
illnesses. Administration of Rotarix may be considered with caution in such infants when, 
in the opinion of the physician, withholding the vaccine entails a greater risk.
Post-marketing safety data indicate a transient increased risk of IS after vaccination, 
mostly within 7 days following the administration of the first dose of Rotarix and, to a 
lesser extent, the second dose. The overall incidence of IS remains rare. Whether Rotarix
affects the overall risk of IS, has not been established.
Therefore, parents/LARs should be advised to promptly report any symptoms indicative 
of IS (severe abdominal pain, persistent vomiting, bloody stools, abdominal bloating 
and/or high fever).
Excretion of the vaccine virus in the stools is known to occur after vaccination and lasts 
for 10 days on average with peak excretion around the seventh day. In clinical trials, 
cases of transmission of excreted vaccine virus to seronegative contacts of vaccinees have 
been observed without causing any clinical symptoms. Rotarix should be administered 
with caution to individuals with immunodeficient close contacts, such as individuals with 
malignancies, or who are otherwise immunocompromised or receiving 
immunosuppressive therapy. Contacts of recent vaccinees should be advised to observe 
careful hygiene (including washing their hands) when changing children’s diapers.
The tip caps of the prefilled oral applicators of diluent may contain natural rubber latex 
which may cause allergic reactions in individuals who are sensitive to latex.
Refer to the approved product label/package insert or IB for more details.
6.7.
Concomitant medications/products and concomitant 
vaccination
At each study visit, the investigator or delegate should question the subject’s 
parent(s)/LAR(s) about any medications/products taken and vaccinations received by the 
subject.
6.7.1.
Recording of concomitant medications/products and 
concomitant vaccinations
The following concomitant medication(s)/product(s)/vaccine(s) must be recorded in 
the eCRF.

All concomitant medications/products, except vitamins and dietary supplements, 
administered during the period starting 30 days before and following each dose of 
study vaccine.
 
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
Any concomitant vaccination administered in the period from first study vaccination 
(Visit 1) and ending at study Visit 3.

Prophylactic medication (i.e., medication administered in the absence of ANY 
symptom and in anticipation of a reaction to the vaccination).
E.g., an anti-pyretic is considered to be prophylactic when it is given in the absence 
of fever and any other symptom, to prevent fever from occurring.

Any concomitant medications/products/vaccines listed in Section 6.7.2.

Any concomitant medications/products/vaccines relevant to a SAE to be reported as 
per protocol or administered during the study period for the treatment of a SAE. 
Concomitant medications relevant to SAEs need to be recorded on the expedited 
Adverse Event report.

Any antipyretic administered in the period starting 6 hours before vaccination and 
ending 12 hours after vaccination.
6.7.2.
Concomitant medications/products/vaccines that may lead to the 
elimination of a subject from per-protocol analyses
The use of the following concomitant medications/products/vaccines will not require 
withdrawal of the subject from the study but may determine a subject’s evaluability in the 
per-protocol analysis. See Section 10.4 for cohorts to be analyzed.

Any investigational or non-registered product (drug or vaccine) other than the study 
vaccines, used during the study period.

Immunosuppressants or other immune-modifying drugs administered chronically 
(i.e., more than 14 days) during the study period between blood sampling at Visit 1 
to blood sampling at Visit 3. For corticosteroids, this will mean prednisone 
0.5 mg/kg/day, or equivalent. Inhaled and topical steroids are allowed.

Immunoglobulins and/or any blood products administered during the study period.

Long-acting immune-modifying drugs administered at any time during the study 
period (e.g., infliximab).

A vaccine not foreseen by the study protocol administered during the period starting 
from 30 days before the first dose of HRV vaccine* administration and ending at 
Visit 3** blood sampling, with the exception of the inactivated influenza vaccine, and 
other licensed routine childhood vaccinations which are allowed at any time during 
the study.
*Administration of any HRV vaccine other than the study vaccine will lead to 
elimination (Refer to Section 4.3).
**In case an emergency mass vaccination for an unforeseen public health threat (e.g.: 
a pandemic) is organized by the public health authorities, outside the routine 
immunization program, the time period described above can be reduced if necessary 
for that vaccine provided it is licensed and used according to its Prescribing 
 
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Information and according to the local governmental recommendations and provided 
a written approval of the Sponsor is obtained.
A detailed, comprehensive list of reasons for elimination from PPS analyses will be 
established at the time of data cleaning.
6.8.
Intercurrent medical conditions that may lead to elimination 
of a subject from per-protocol analyses
At each study visit subsequent to the first vaccination visit, it must be verified if the 
subject has experienced or is experiencing any intercurrent medical condition. If it is the 
case, the condition(s) must be recorded in the eCRF.
Subjects may be eliminated from the PPS for analysis of immunogenicity if, during the 
study, they incur a condition that has the capability of altering their immune response or 
are confirmed to have an alteration of their initial immune status.
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 AE or SAE as provided in 
this protocol.
Each subject’s parent(s)/LAR(s) will be instructed to contact the investigator immediately 
should the subject manifest any signs or symptoms they perceive as serious.
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:

Significant or unexpected worsening or exacerbation of the condition/indication 
under study.
 
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
Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study vaccines administration even 
though they may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
vaccines or a concurrent medication (overdose per se should not be reported as an 
AE/SAE).

Signs, symptoms temporally associated with study vaccines administration.

Significant failure of expected pharmacological or biological action.

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).
AEs to be recorded as endpoints (solicited AEs) are described in Section 8.1.3. All other 
AEs will be recorded as UNSOLICITED AEs.
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.
c.
Requires hospitalization or prolongation of existing hospitalization,
Note: In general, hospitalization 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 hospitalization are also considered AEs. If a complication prolongs 
 
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hospitalization or fulfils any other serious criteria, the event will also be considered 
serious. When in doubt as to whether ‘hospitalization’ occurred or was necessary, the 
AE should be considered serious.
Hospitalization 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, diarrhea, 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.
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 hospitalization 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 hospitalization.
8.1.3.
Solicited adverse events
8.1.3.1.
Solicited general adverse events
The following general AEs will be solicited (Table 11):
Table 11
Solicited general adverse events
Fever
Irritability/Fussiness
Diarrhea
Vomiting
Loss of appetite
Cough/runny nose
Note: Parent(s)/LAR(s) will be instructed to measure and record the oral, axillary or 
rectal body temperature in the evening. Should additional temperature measurements be 
performed at other times of day, parent(s)/LAR(s) will be instructed to record the highest 
temperature in the diary card.
An overview of the protocol-required reporting periods for solicited general AEs is given 
in Table 12.
 
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8.1.4.
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., vital signs etc) that are 
judged by the investigator to be clinically significant will be recorded as AE or SAE if 
they meet the definition of an AE or SAE (refer to Sections 8.1.1 and 8.1.2). 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 
AEs or 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.
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 adverse events and serious 
adverse events
8.2.1.
Time period for detecting and recording adverse events and
serious adverse events
All AEs starting within 31 days following administration of each dose of study vaccines 
(Day 1 to Day 31) must be recorded into the appropriate section of the eCRF, irrespective 
of intensity or whether or not they are considered vaccination-related.
The time period for collecting and recording SAEs will begin at the receipt of study 
vaccine and will end 30 days following administration of the last dose of study vaccine 
for each subject. See Section 8.3 for instructions on reporting of SAEs.
All AEs/SAEs leading to withdrawal from the study will be collected and recorded from 
the time of the first receipt of study vaccines.
In addition to the above-mentioned reporting requirements and in order to fulfil 
international reporting obligations, SAEs that are related to study participation (i.e., 
protocol-mandated procedures, invasive tests, a change from existing therapy) or are 
related to a concurrent GSK medication/vaccine will be collected and recorded from the 
time the subject consents to participate in the study until she/he is discharged from the 
study.
An overview of the protocol-required reporting periods for AEs and SAEs is given in 
Table 12.
 
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Table 12
Reporting periods for collecting safety information
Event
Pre-
V1*
V1
7 days 
post 
vacc
30 
days
post 
vacc
V2
7 days 
post 
vacc
30 days
post 
vacc
V3
D1
D 8
D 31
M1
M2
Study 
Conclusion
Solicited general 
AEs
Unsolicited AEs
AEs/SAEs leading 
to withdrawal from 
the study
SAEs
SAEs related to 
study participation 
or concurrent GSK 
medication/vaccine
* Consent obtained. Pre-V: pre-vaccination; Post-Vacc: post-vaccination; V: Visit; D: Day, M: Month; 
AEs = Adverse Events; SAEs = Serious Adverse Events
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 defined in Table 12. 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 vaccines, the investigator will promptly notify the 
Study Contact for Reporting SAEs.
8.2.3.
Evaluation of adverse events and serious adverse events
8.2.3.1.
Active questioning to detect adverse events and serious adverse 
events
As a consistent method of collecting AEs, the subject’s parent(s)/LAR(s) should be asked 
a non-leading question such as:
‘Has your child acted differently or felt different in any way since receiving the vaccines 
or since the last visit?’
 
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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.
8.2.3.2.
Assessment of adverse events
8.2.3.2.1.
Assessment of intensity
The intensity of the following solicited AEs will be assessed as described in Table 13:
 
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Table 13
Intensity scales for solicited symptoms in infants
Infants
Adverse Event
Intensity grade
Parameter
Fever*
Record temperature in °C/°F
0
temperature <38.0°C/100.4° F
1
temperature 38.0°C/100.4° F – 38.5°C/101.3 F 
2
temperature >38.5°C/101.3 F – 39.5°C/103.1 F
3
temperature >39.5°C/103.1 F
Irritability/Fussiness
0
Behavior as usual
1
Mild: Crying more than usual/no effect on normal activity
2
Moderate: Crying more than usual/interferes with normal activity
3
Severe: Crying that cannot be comforted/prevents normal activity
Diarrhea$
Record the number of looser than normal stools/day
0
Normal (0-2 looser than normal stools/day)
1
3 looser than normal stools/day
2
4-5 looser than normal stools/day
3
6 looser than normal stools/day
Vomiting§
Record the number of vomiting episodes/day
0
Normal (no emesis)
1
1 episode of vomiting/day
2
2 episodes of vomiting/day
3
3 episodes of vomiting/day
Loss of appetite
0
Appetite as usual
1
Mild: Eating less than usual/no effect on normal activity
2
Moderate: Eating less than usual/interferes with normal activity
3
Severe: Not eating at all
Cough/runny nose
0
Normal
1
Mild: Cough/runny nose which is easily tolerated
2
Moderate: Cough/runny nose which interferes with daily activity
3
Severe: Cough/runny nose which prevents daily activity
* Fever is defined as temperature 38.0°C/100.4°F. The preferred location for measuring temperature in this study 
will be the oral cavity, the axilla and the rectum.
$ Diarrhea is defined as passage of three or more looser than normal stools within a day. 
§ Vomiting is defined as one or more episodes of forceful emptying of partially digested stomach contents ≥1 hour 
after feeding within a day.
The intensity grade for diarrhea, vomiting and fever as shown in Table 13 will be scored at GSK Biologicals.
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.
The intensity should be assigned to one of the following categories:
 
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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 
(in a young child, such an AE would, for example, prevent 
attendance at a day-care center and would cause the 
parent(s)/LAR(s) to seek medical advice.)
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 8.1.2.
8.2.3.2.2.
Assessment of causality
The investigator is obligated to assess the relationship between study vaccines and the 
occurrence of each AE/SAE using clinical judgement. In case of concomitant 
administration of multiple vaccines/products, if possible, the investigator should specify 
if the AE could be causally related to a specific vaccine/product administered (i.e., 
investigational, control/placebo or co-administered vaccine). When causal relationship to 
a specific vaccine(s)cannot be determined the investigator should indicate the AE to be 
related to all products.
Alternative plausible causes, such as natural history of the underlying diseases, 
concomitant therapy, other risk factors, and the temporal relationship of the event to the 
study vaccines will be considered and investigated. The investigator will also consult the 
IB and/or SmPC and/or Prescribing Information for marketed products to determine 
his/her assessment.
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.
 
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Causality of all AEs should be assessed by the investigator using the following question:
Is there a reasonable possibility that the AE may have been caused by the study vaccine?
YES
:
There is a reasonable possibility that the study vaccines contributed to 
the AE. 
NO
:
There is no reasonable possibility that the AE is causally related to the 
administration of the study vaccines. There are other, more likely causes 
and administration of the study vaccines is not suspected to have 
contributed to the AE.
If an event meets the criteria to be determined as ‘serious’ (see Section 8.1.2), 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.

Lack of efficacy of the vaccines, if applicable.

Erroneous administration.

Other cause (specify).
8.2.3.3.
Assessment of outcomes
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).
8.2.3.4.
Medically attended visits
For each solicited and unsolicited symptom the subject experiences, the subject’s 
parent(s)/LAR(s) will be asked if the subject received medical attention defined as 
hospitalization, or an otherwise unscheduled visit to or from medical personnel for any 
reason, including emergency room visits. This information will be recorded in the eCRF.
 
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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 14, once the investigator determines that 
the event meets the protocol definition of a SAE.
Table 14
Timeframes for submitting serious adverse event and other events 
reports 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.
Back-up Study Contact for Reporting SAEs
24/24 hour and 7/7 day availability:
GSK Biologicals Clinical Safety & Pharmacovigilance
Outside US & Canada sites:
Fax: 
 or 
Email address: 
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 
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PPD
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8.3.3.1.
Back-up system in case the electronic reporting system does not work
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 14.
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. GSK Biologicals has a legal responsibility to promptly notify, 
as appropriate, both the local regulatory authority and other regulatory agencies about the 
safety of a product under clinical investigation. Prompt notification of SAEs by the 
investigator to the Study Contact for Reporting SAEs is essential so that legal obligations 
and ethical responsibilities towards the safety of other subjects are met.
Investigator safety reports are prepared according to the current GSK policy and are 
forwarded to investigators as necessary. An investigator safety report is prepared for a 
SAE(s) that is both attributable to the study vaccines and unexpected. The purpose of the 
report is to fulfil specific regulatory and GCP requirements, regarding the product under 
investigation.
8.4.
Follow-up of adverse events and serious adverse events
8.4.1.
Follow-up during the study
After the initial AE/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 14).
 
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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.
All AEs documented at a previous visit/contact and designated as not recovered/not 
resolved or recovering/resolving will be reviewed at subsequent visits/contacts until 30 
days after the last vaccination.
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 an 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 the subject’s eCRF (refer to Section 6.7).
8.6.
Subject card
Study subjects’ parent(s)/LAR(s) 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’s 
parent(s)/LAR(s). 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.
Subjects’ parent(s)/LAR(s) must be instructed to keep subject cards in their possession at 
all times during the study duration.
 
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9.
SUBJECT COMPLETION AND WITHDRAWAL
9.1.
Subject completion
A subject who returns to the concluding visit foreseen in the protocol is considered to 
have completed the study.
9.2.
Subject withdrawal
Withdrawals will not be replaced.
9.2.1.
Subject withdrawal from the study
From an analysis perspective, a ‘withdrawal’ from the study refers to any subject who 
was did not come back for the concluding visit foreseen in the protocol.
All data collected until the date of withdrawal/last contact of the subject will be used for 
the analysis.
A subject is considered a ‘withdrawal’ from the study when no study procedure has 
occurred, no follow-up has been performed and no further information has been collected 
for this subject from the date of withdrawal/last contact.
Investigators will make an attempt to contact those subjects who do not return for 
scheduled visits or follow-up.
Information relative to the withdrawal will be documented in the eCRF. The investigator 
will document whether the decision to withdraw a subject from the study was made by 
the subject’s parent(s)/LAR(s), or by the investigator, as well as which of the following 
possible reasons was responsible for withdrawal:

SAE

Unsolicited non-serious AE

Solicited AE

Protocol violation (specify)

Consent withdrawal, not due to an AE*

Moved from the study area

Lost to follow-up

Other (specify)
*In case a subject is withdrawn from the study because the subject’s parent(s)/LAR(s) has 
withdrawn consent, the investigator will document the reason for withdrawal of consent, 
if specified by the subject’s parent(s)/LAR(s), in the eCRF.
 
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Subjects who are withdrawn from the study because of SAEs/AEs must be clearly 
distinguished from subjects who are withdrawn for other reasons. Investigators will 
follow subjects who are withdrawn from the study as result of a SAE/AE until resolution 
of the event (see Section 8.4.2).
9.2.2.
Subject withdrawal from study vaccine
A ‘withdrawal’ from the study vaccine refers to any subject who does not receive the 
complete treatment, i.e., when no further planned dose is administered from the date of 
withdrawal. A subject withdrawn from the study vaccine may not necessarily be
withdrawn from the study as further study procedures or follow-up may be performed 
(safety or immunogenicity) if planned in the study protocol.
Information relative to premature discontinuation of the study vaccines will be 
documented on the Vaccine Administration screen of the eCRF. The investigator will 
document whether the decision to discontinue further vaccination/treatment was made by 
the subject’s parent(s)/LAR(s), or by the investigator, as well as which of the following 
possible reasons was responsible for withdrawal:

SAE

Unsolicited non-serious AE

Solicited AE

Not willing to be vaccinated

Other (specify)
10.
STATISTICAL METHODS
10.1.
Primary Endpoint

Anti-RV IgA antibody concentrations

Serum anti-RV IgA antibody concentrations, expressed as GMCs, one month 
post dose 2 of HRV vaccine.
10.2.
Secondary Endpoints (Amended: 30 October 2019)

Anti-RV IgA antibody concentrations

Anti-RV IgA antibody seroconversion* rate, one month post dose 2 of HRV 
vaccine.
*Definition: 

for subjects with a pre-vaccination anti-RV IgA antibody concentration <20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥20 U/mL.
 
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
for subjects with a pre-vaccination anti-RV IgA antibody concentration ≥20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥2 times the pre-vaccination concentration.

Solicited general symptoms

Occurrence of each type of solicited general symptom within the 8-day 
(Day1-Day 8) solicited follow-up period, after each dose of HRV vaccine.

Unsolicited adverse events

Occurrence of unsolicited AEs within 31 days (Day 1-Day 31) after any dose of
HRV vaccine according to Medical Dictionary for Regulatory Activities 
(MedDRA) classification.

Serious adverse events

Occurrence of SAEs from dose 1 of HRV vaccine up to study end.
10.3.
Determination of sample size (Amended: 30 October 2019)
Considering the total sample of 450 subjects (225 subjects per arm), Table 15 provides 
the power to demonstrate non-inferiority in terms of GMCs according to the percentage 
of non-evaluable subjects.
Based on historical data from ROTA-044 study [Narang, 2009], 13.8% of subjects were 
non-evaluable for reasons other than ‘initially positive or unknown for serum anti-RV
IgA antibodies at Dose 1’. The power to reach the primary objective in a scenario of 
13% non-evaluable subjects is 96% (Table 15). Considering a scenario of 35% non-
evaluable subjects, the power to reach the primary objective is 90%. As shown in Table
15, the power decreases when the % of non-evaluable subjects increases.
Table 15
Power according to different scenarios for the % of non-evaluable 
subjects, with respect to a total sample size of 450 subjects*
(Amended: 30 October 2019)
Standard deviation
Alpha
% of non-evaluable subjects
Power
[Log10 (concentration)]
0.790**
0.025
13%
96%
15%
96%
25%
94%
30%
92%
35%
90%
40%
88%
45%
85%
50%
81%
55%
77%
*PASS one-sided non-inferiority for 2 independent means with common variance for log10(0.5) as non-
inferiority margin, for the alternative hypothesis that the means are equal and for a type I error=2.5 %
**Reference used for power computation
 
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10.4.
Cohorts for Analyses
10.4.1.
Exposed Set
The ES will include all subjects with at least one study vaccine administration 
documented.

A safety analysis based on the ES will include all vaccinated subjects.

An immunogenicity analysis based on the ES will include all vaccinated subjects for 
whom immunogenicity data is available.
10.4.2.
Per-Protocol Set for analysis of immunogenicity
(Amended: 30 October 2019)
The PPS for immunogenicity will include all eligible subjects from the ES:

who have received both doses of study vaccine according to their random 
assignment.

for whom the HRV vaccine, liquid or lyophilized formulation, is administered 
according to protocol. Note that the subjects who regurgitate after vaccination and 
receive a replacement dose are to be retained in the PPS,

who comply with the vaccination schedule for HRV vaccine (liquid or lyophilized 
formulation), as per Table 5,

who have not received a vaccine prohibited by the protocol up to Visit 3 blood 
sample,

who have not received medication prohibited by the protocol up to Visit 3 blood 
sample,

whose underlying medical condition(s) was (were) not prohibited by the protocol up 
to Visit 3 blood sample,

with no protocol violation of demographics (unknown age at study entry or outside 
protocol defined age-interval),

who comply with blood sampling schedule, as per Table 5,

for whom immunogenicity data are available at pre- and post-vaccination sampling 
time points.

for whom the post-vaccination immunogenicity data are within the 21-48 days 
interval after the second dose.

who have no other concomitant infection up to Visit 3 blood sample, which may 
influence the immune response.
10.5.
Derived and transformed data (Amended: 30 October 2019)
Demography
 
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For a given subject and a given demographic variable, missing measurements will not be 
replaced.
Safety/Reactogenicity
Subjects who missed reporting symptoms (solicited/unsolicited or concomitant
medications) will be treated as subjects without symptoms (solicited/unsolicited or
concomitant medications, respectively).
Immunogenicity (Amended: 30 October 2019)

A seronegative subject is a subject whose anti-RV IgA antibody concentration is 
below the clinically meaningful threshold of <20 U/ml*.

A seropositive subject is a subject whose anti-RV IgA antibody concentration is
greater than or equal to the clinically meaningful threshold of 20 U/ml.

Seroconversion is defined as:

for subjects with a pre-vaccination anti-RV IgA antibody concentration <20 
U/mL, seroconversion is achieved when the post-vaccination concentration is 
≥20 U/mL.

for subjects with a pre-vaccination anti-RV IgA antibody concentration ≥20 
U/mL, seroconversion is achieved when the post-vaccination concentration is 
≥2 times the pre-vaccination concentration.

The GMCs calculations are performed by taking the anti-log of the mean of the log
concentrations transformations. Antibody concentrations below the technical cut-off
(<13 U/ml*) of the assay will be given an arbitrary value of half the cut-off for the
purpose of GMC calculation.

For the immunogenicity measurement, missing or non-evaluable measurements will
not be replaced.
*Note: 20 U/ml corresponds to the clinically meaningful threshold to define 
seroconversion rate, while 13 U/ml corresponds to technical cut-off of revalidated 
laboratory assay.
10.6.
Analysis of demographics
The distribution of subjects enrolled among the study centers will be tabulated as a whole 
and for each group.
The number of subjects who withdraw from the study will be tabulated by group 
according to the reason for drop-out.
The number of subjects enrolled into the study as well as the number of subjects excluded 
from PPS analyses will be tabulated as a whole and for each group.
The deviations from specifications for age and intervals between study visits will be 
tabulated by group.
 
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The median, mean, range and standard deviation of age (in weeks) at each HRV vaccine 
dose will be computed by group. The median, mean and standard deviation of height in 
centimeter (cm) and weight in kilograms (kg) at Visit 1 will be computed by group. The 
geographical ancestry and sex composition will be presented.
Summary of co-administered vaccinations (i.e., vaccinations given on the day of each 
HRV vaccine dose) and intercurrent vaccinations (i.e., vaccinations other than the HRV 
lyophilized and HRV liquid vaccine administered from birth up to Visit 3, excluding 
vaccination given on the day of HRV vaccine doses) will be summarized by group for the 
ES.
10.7.
Analysis of immunogenicity
The primary analysis will be based on the PPS for analysis of immunogenicity. If, in any 
vaccine group, the percentage of vaccinated subjects with serological results excluded 
from the PPS for analysis of immunogenicity is 5% or more, a second analysis based on 
the ES will be performed to complement the PPS analysis.
10.7.1.
Within group assessment (Amended: 30 October 2019)
The following calculations will be performed for each group

For each group, at each time point that anti-rotavirus IgA is measured,

GMCs and their 95% CIs will be computed.

Seropositivity/seroconversion rates and their exact 95% CI will be computed,

The distribution of anti-RV IgA antibody concentrations at Visit 1 and Visit 3 
will be displayed using Reverse Cumulative Curves (RCCs).
10.7.2.
Between groups assessment (Amended: 30 October 2019)

The 95% CI for the ratio of anti-RV IgA antibody GMCs at Visit 3 between the HRV 
liquid vaccine over the HRV lyophilized vaccine will be computed using an 
ANCOVA model on the logarithm-transformed concentrations. This model will 
include the vaccine group and the logarithm of the baseline concentration as 
covariables. The GMC ratios and their 95% CI will be derived by exponential 
transformation of the corresponding group contrast in the model (primary 
objective).

The asymptotic standardized 95% CI for the difference in seroconversion rate at 
Visit 3 between the HRV liquid vaccine and HRV lyophilized vaccine will be 
computed (secondary objective).
Refer to the Section 2.1 for the success criteria of the primary objective.
 
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10.8.
Analysis of safety
The ES will be used for the analysis of safety.
10.8.1.
Within groups assessment
The following calculations will be performed for each group:
The percentage of doses and of subjects reporting at least one symptom (solicited or
unsolicited) during the 8-day (Day 1-Day 8) solicited follow-up period will be computed, 
along with exact 95% CI. The same calculations will be done for symptoms (solicited or 
unsolicited) rated as grade 3 in intensity, for symptoms (solicited or unsolicited) assessed 
as causally related to vaccination and for symptoms resulting in medically attended visit.
The percentage of doses and of subjects reporting each individual solicited general
symptom will be computed, over the 8-day (Day 1-Day 8) solicited follow-up period, 
post vaccination, along with exact 95% CI. The same calculations will be done for each 
individual solicited general symptom rated as grade 3 in intensity, for each individual 
solicited general symptom assessed as causally related to vaccination and for symptoms 
resulting in medically attended visit.
The verbatim reports of unsolicited AEs will be reviewed by a physician and the signs
and symptoms will be coded according to MedDRA. Every verbatim term will be
matched with the appropriate Preferred Term. The percentage of subjects with unsolicited
AEs occurring within 31-day (Day 1-Day 31) follow-up period after any dose with its 
exact 95% CI will be tabulated by preferred term. Similar tabulation will be done for
unsolicited AEs rated as grade 3, for unsolicited AEs with causal relationship to 
vaccination, for unsolicited AE resulting in medically attended visit and for AEs or SAEs 
leading to drop out.
The percentages of subjects who started taking at least one concomitant medication, by 
type, from Day 1 to Day 8 after vaccinations will be tabulated with exact 95% CI. The 
percentages of subjects who started taking at least one concomitant medication, by type, 
during the study period will also be tabulated with exact 95% CI.
SAEs reported during the study period will be described in detail.
10.9.
Interpretation of analyses 
Except for analyses addressing the primary objectives referred as confirmatory analyses, 
all the analyses will be descriptive/exploratory in nature. These descriptive analyses 
should be interpreted with caution
 
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10.10.
Statistical Methods (Amended: 30 October 2019)

The exact CIs for a proportion within a group will be calculated using SAS 
[Clopper, 1934].

The CI for GMCs will be obtained within each group separately. The CI for the 
mean of log-transformed concentration will be first obtained assuming that log-
transformed values were normally distributed with unknown variance. The CI for the 
GMCs will then be obtained by exponential-transformation of the CI for the mean of 
log-transformed concentration.

The standardized asymptomatic CI for the group difference in proportion will be 
calculated using SAS. The method used within GSK Biologicals is Method 6 
[Newcombe, 1998].
10.11.
Conduct of analysis
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.
10.11.1.
Sequence of analyses
A final analysis of all data collected up to Visit 3 will be conducted. This will include 
final analysis of immunogenicity, reactogenicity and safety. A clinical report will be 
written at this stage.
10.11.2.
Statistical considerations for interim analyses
No interim analysis is planned for this 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 
 
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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 an eCRF review and Source Document 
Verification (SDV). By SDV we understand verifying eCRF entries by 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 
 
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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 25 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 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
GSK assures that the key design elements of this protocol will be posted on the GSK 
website and in publicly accessible database(s) such as clinicaltrials.gov, in compliance 
with the current regulations.
GSK also assures that results of this study will be posted on the GSK website and in 
publicly accessible regulatory registry(ies) within the required time-frame, in compliance 
with the current regulations. The minimal requirement is to have primary endpoint 
summary results disclosed at latest 12 months post PCD and to have secondary endpoint 
disclosed at latest 12 months after the LSLV as described in the protocol.
GSK also aims to publish the results of these studies in searchable, peer reviewed 
scientific literature and follows the guidance from the International Committee of 
Medical Journal Editors.
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 
 
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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 (AMENDED: 30 OCTOBER 2019)
Atherly D, Dreibelbis R, Parashar U D, et al. Rotavirus Vaccination: Cost-Effectiveness 
and Impact on Child Mortality in Developing Countries The Journal of Infectious 
Diseases 2009; 200:S28–38.
Clopper C. J, Pearson E.S. The Use Of Confidence Or Fiducial Limits Illustrated In The 
Case Of The Binomial. Biometrika. 1934: 26, 404-413.
Cheuvart B, Neuzil KM, Steele AD, et al. Association of serum anti-rotavirus 
immunoglobulin A antibody seropositivity and protection against severe rotavirus 
gastroenteritis: Analysis of clinical trials of human rotavirus vaccine. Hum Vaccin 
Immunother. 2014; 10: 505-511.
Kang G. Rotavirus vaccines. Indian J Med Microbiol. 2006; 24(4): 252-257.
Linhares AC, Velázquez FR, Pérez-Schael I, et al. Efficacy and safety of an oral live 
attenuated human rotavirus vaccine against rotavirus gastroenteritis during the first 2 
years of life in Latin American infants: a randomized, double-blind, placebo-controlled 
phase III study. Lancet. 2008;371(9619):1181-9.
Madhi SA, Cunliffe NA, Duncan Steele D, et al. Effect of Human Rotavirus Vaccine on 
Severe Diarrhea in African Infants. N Engl J Med. 2010 Jan 28;362(4):289-98.
Narang A, Bose A, Pandit AN, Dutta P, Kang G, Bhattacharya SK, Datta SK, 
Suryakiran PV, Delem A, Han HH, Bock HL. Immunogenicity, reactogenicity and 
safety of human rotavirus vaccine (RIX4414) in Indian infants. Hum Vaccin. 2009 
Jun;5(6):414-9.
Newcombe R G. Two-sided confidence intervals for the single proportion: Comparison 
of seven methods. Statistics in Medicine. 1998; 17(8):857–72.
Phua K.B, Lim F.S, Lau Y.L et al. Safety and efficacy of human rotavirus vaccine during 
the first 2 years of life in Asian infants: Randomized, double-blind, controlled study. 
Vaccine. 2009: 5936-5941.
Salinas B, Perez-Schael I, Linhares AC. Evaluation of safety, immunogenicity and 
efficacy of an attenuated rotavirus vaccine RIX4414: a randomized, placebo-controlled 
trial in Latin American infants. Pediatr Infect Dis J. 2005:807-16.
Tate JE, Burton AH, Boschi-Pinto C. Global, Regional, and National Estimates of 
Rotavirus Mortality in Children <5 Years of Age, 2000-2013. Clin Infect Dis. 2016: S96-
S105.
Vesikari T, Karvonen A, Prymula R. et al. Efficacy of human rotavirus vaccine against 
rotavirus gastroenteritis during the first 2 years of life in European infants: randomised, 
double-blind controlled study. Lancet 2007; 370:1757–63.
WHO position paper. Weekly epidemiological record. Rotavirus vaccines WHO position 
paper-January 2013.2013; 88: 49-64.
 
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APPENDIX A
LABORATORY ASSAYS
Rotavirus Ab IgA Determination
The anti-RV antibody concentrations are determined by a validated anti-RV IgA ELISA. 
Microtiter plates (96-well) are coated with an anti-RV monoclonal antibody. The wells 
are washed and incubated with (positive wells) or without (negative wells) RV. 
Following incubation, the plates are washed and serum, standard and control dilutions are 
incubated in both types of wells (positive and negative). Bound 
anti-RV IgA in the well are detected by incubation with peroxidase conjugated 
anti-human IgA polyclonal antibodies. Color development proportional to the quantity of 
bound anti-RV IgA occurs in the presence of a chromogen, TetraMethylBenzidine
(TMB), and measured spectrophotometrically. Specific optical densities are calculated for 
each sample/control/standard dilution by measuring the difference between positive and 
negative wells, the use of negative wells allowing to assess non-specific IgA binding. The 
concentrations of the samples expressed in units per milliliter are calculated relative to 
the four-parameter logistic function generated from the standard curve.
 
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APPENDIX B
CLINICAL LABORATORIES
Table 16
GSK Biologicals’ laboratories
Laboratory
Address
GSK Biological’s Clinical 
Laboratory Sciences (CLS), 
Rixensart 
Biospecimen Reception-B7/44
Rue de l'Institut, 89
B-1330 Rixensart
Belgium
GSK Biological’s CLS, Wavre-Nord 
Noir Epine
Avenue Fleming, 20
B-1300 Wavre
Belgium
GSK Vaccines GmbH
Emil-von-Behring-Str. 76
35041 Marburg
Deutschland/ Germany
Table 17
Outsourced laboratories
Laboratory
Address
Q² Solutions Clinical Trials (US)
27027 Tourney Road, Suite 2E
Valencia, CA 91355
US
Q² Solutions Clinical Trials (UK)
1 Simpson Parkway
The Alba Campus
Rosebank
Livingston
EH54 7EG
UK
 
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APPENDIX C
AMENDMENTS AND ADMINISTRATIVE 
CHANGES TO THE PROTOCOL
GlaxoSmithKline Biologicals SA
Vaccines R &D
Protocol Amendment 1
eTrack study number 
and Abbreviated Title 
116566 (ROTA-083)
EudraCT number
2012-001875-35
Amendment number:
Amendment 1
Amendment date:
24 July 2013
Co-ordinating author:
Rationale/background for changes: NDAC- Vaccines (New Drug Advisory 
Committee on Vaccines) of Drugs Controller General of India (DCGI) recommended 
to conduct this study as a Phase IV study instead of phase III as initially planned 
because Rotarix liquid file application is considered as a line extension of Rotarix
Lyophilized formulation. The protocol was amended to adapt the change recommended 
by NDAC-Vaccines of DCGI.
Amended text has been included in bold italics and deleted text in strikethrough in 
the following sections:
Contributing authors

, Clinical Regulatory Representative 

 and 
Clinical Safety
Copyright 2012-2013 the GlaxoSmithKline group of companies. All rights reserved. 
Unauthorised copying or use of this information is prohibited.
Detailed Title 
A phase III IV, randomized, open study to assess the immunogenicity, reactogenicity 
and safety of two different formulations of GlaxoSmithKline (GSK) Biologicals’ oral 
live attenuated human rotavirus (HRV) vaccine, Rotarix given as a two dose primary 
vaccination, in healthy infants with no previous history of RV illness or vaccination.
Rationale for the study and study design
The lyophilized formulation of the HRV vaccine was licensed in India in November 
2007. Following the request of the Regulatory Authority in India, this study will be 
conducted to generate the additional clinical data for the liquid formulation of HRV 
vaccine in India as recommended by NDAC- Vaccines (New Drug Advisory 
Committee on Vaccines) of DCGI.
 
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Exclusion criteria for enrolment 

Acute disease and/or fever at the time of enrolment.

Fever is defined as temperature ≥37.5°C /99.5°F on oral, axillary or tympanic 
setting, or ≥38.0°C /100.4°F on rectal setting. The preferred route for 
recording temperature in this study will be axillary.

Subjects with a minor illness (such as mild diarrhoea, mild upper respiratory 
infection) without fever may, be enrolled at the discretion of the investigator 
(warrants deferral of the vaccination).
 
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GlaxoSmithKline Biologicals SA
Vaccines R &D
Protocol Amendment 2
eTrack study number 
and Abbreviated Title 
116566 (ROTA-083)
EudraCT number
2012-001875-35
Amendment number:
Amendment 2
Amendment date:
19 September 2016
Co-ordinating author:
Rationale/background for changes The ROTA-083 (116566) protocol and protocol 
amendment 1 were written in the protocol template edition 13.2. The protocol 
amendment 2 is a rewriting of the protocol to comply with the latest version of the 
protocol template (edition 15.0, effective 1st April, 2016).
The study design has not been impacted by this amendment.
The main changes included in the current amendment are listed below:

The addition of the definitions for End of Study (EoS) and Primary Completion Date 
(PCD),

addition of the section for Benefit and Risk assessment, 

the definition of fever has been updated and

other minor changes to align the protocol with the specific control terminology used 
in Clinical Data Interchange Standards Consortium (CDISC). This includes renaming 
of the study cohorts and numbering of day, so that Day 1 corresponds to the date of 
vaccination
 
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GlaxoSmithKline Biologicals SA
Vaccines R &D
Protocol Amendment 3
eTrack study number 
and Abbreviated Title 
116566 (ROTA-083)
EudraCT number
2012-001875-35
Amendment number:
Amendment 3
Amendment date:
31 October 2017
Co-ordinating author:
Rationale/background for changes: Based on the feedback from the regulatory 
authorities in India on the protocol amendment 2, the following changes have been 
made to the protocol:

The phase of this clinical trial is revised from Phase IV to Phase III.

The study objectives have been updated to include a confirmatory primary 
objective and the study has been powered accordingly.

The statistical considerations have been revised to define the success criteria for 
primary objective.

Collection and testing of stool samples from subjects who develop gastroenteritis
(GE) during the study period is removed in order to simplify the study procedure 
and considering no subjects were eliminated from the according-to-protocol (ATP)
cohort due to HRV GE in study ROTA-044. The ROTA-044 study was a Phase 
IIIB, randomised, multicentre, double blind, placebo controlled study conducted in 
India to assess the immunogenicity and safety of two doses of GSK Biologicals’
lyophilized formulation of HRV vaccine.

The study procedures have been updated accordingly.

The list of assays and section on biological samples evaluation have been updated.

The laboratory assay section has been updated to account for the characteristics of 
revalidated GSK assay.

The list of contributing authors has been updated and some typographic errors 
have been corrected.
Amended text has been included in bold italics and deleted text in strikethrough in 
the following sections:
Detailed Title: A phase III,IV randomized, open study to assess the immunogenicity, 
reactogenicity and safety of two different formulations of GlaxoSmithKline (GSK) 
Biologicals’ oral live attenuated human rotavirus (HRV) vaccine, Rotarix, when given 
as a two dose primary vaccination, in healthy infants with no previous history of 
rotavirus illness or vaccination.
 
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Contributing authors

, Statistician

, Lead Statistician

, Study Delivery Lead

,
Clinical Laboratory Sciences (CLS), Clinical 
Readout Team Leaders

,
, Clinical Safety representatives

, Oversight Data Manager

, Study Data Manager

, Global Patents representative

, 
Clinical Trial Supply Managers
© 2012-2017 GSK group of companies or its licensor Copyright 2012-2016 the 
GlaxoSmithKline group of companies. All rights reserved. Unauthorised copying 
or use of this information is prohibited
Synopsis and Section 1.2- Rationale for the study and study design
The lyophilized formulation of the HRV vaccine was licensed in India in November 
2007 February 2008.
The study is planned to be conducted in healthy infants aged ≥6 6-10 weeks at the time 
of the first dose. Two oral doses of the study vaccine will be administered to the 
subjects, according to a 0, 1 month schedule. The blood samples for immunogenicity 
assessment will be collected from all subjects before the first dose and at one month 
post dose 2. In addition, stool samples will be collected from the subjects if they 
develop GE, to detect the presence of wild type RV
Please refer to the current Investigator Brochure (IB) for information regarding the 
summary of potential risks and benefits of the Rotarix vaccine.
 
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Section 1.3 Benefit: Risk Assessment
Please refer to the current IB and prescribing information (lyophilized vaccine) for 
information regarding the summary of potential risks and benefits of the Rotarix
vaccine.
The following section outlines the risk assessment and mitigation strategy for this 
study protocol
Section 1.3.1- Risk assessment
Important 
Potential/Identified Risk
Data/Rationale for Risk
Mitigation Strategy
Investigational study vaccine (Rotarix)
Intussusception (IS)
Spontaneous data

Subjects will be followed up 
to the end of the study after 
the receipt of the vaccine to 
check for any safety signal.

Parents/legally acceptable 
representatives (LARs) of
subjects should report any 
untoward symptoms 
experienced after receiving 
the vaccine immediately to 
the investigator.

All Serious Adverse Events 
(SAEs) should be reported by 
the investigator immediately 
to GSK.

Subjects with SCID will be 
excluded from participating 
in this study (Refer to 
Section 4.3 and Section 6
for more details)
Hematochezia
Spontaneous data
Gastroenteritis with vaccine 
viral shedding in infants
with severe combined 
immunodeficiency (SCID)
Spontaneous data
Kawasaki disease
Based on signal observed 
for RotaTeq vaccine
 
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Synopsis and Section 2- Objectives
Primary objective:

To evaluate non-inferiority of GSK Biologicals’ HRV liquid vaccine compared 
to GSK Biologicals’ HRV lyophilized vaccine in terms of geometric mean 
concentrations (GMCs) for anti-RV antibodies, one month post dose 2 of HRV 
vaccine.

Criterion: Non-inferiority will be stated if the lower limit of the two-sided 
95% confidence interval (CI) for the ratio of anti RV IgA antibody GMCs 
between HRV liquid vaccine over the HRV lyophilized vaccine, one month 
post dose 2 is greater than or equal to 0.5

To assess the immunogenicity of GSK Biologicals’ HRV liquid vaccine and 
GSK Biologicals’ HRV lyophilized vaccine, in terms of seroconversion rates, 
one month post dose 2 of HRV vaccine
Secondary objectives

To assess the immunogenicity of the HRV liquid vaccine and HRV lyophilized 
vaccine, in terms of seroconversion rates, one month post dose 2 of HRV 
vaccine.
*Seroconversion rate is defined as the percentage of subjects with anti-RV IgA 
antibody concentration ≥20 U/mL one-month post dose 2 among subjects with 
anti-RV IgA antibody concentration < 20 U/mL at pre-vaccination.

To assess the immunogenicity of the two formulations of GSK Biologicals’ HRV 
vaccine, in terms of Geometric Mean Concentration (GMC), one month post Dose 
2 of HRV vaccine.

To assess the presence of wild type RV in GE stool samples collected from Dose 1 
of the HRV vaccine up to Visit 3.
 
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Section 3- Study Design Overview
HRV= Human Rota Virus; N = Number of subjects planned to be enrolled; Pre-Vacc= Pre-vaccination; Post-Vac= Post-
vaccination; BS = Blood Sample
Study design: (Synopsis and Section 3)

Experimental design: Phase III IV open-label, randomized (1:1), multi-centric, 
single-country study with two parallel groups.

End of Study (EoS): Last testing results released for samples collected at Visit 3.
Please refer to GLOSSARY OF TERMS for the definition of EoS.
Vaccination visits
Follow-up visit
HRV Liq (N=225)
HRV Lyo (N=225)
Randomization 
(1:1)
Visit 1-Day 1
Dose 1
Pre-vacc BS
Visit 2-Month 1
Dose 2
Visit 3-Month 2
Post- vacc BS
 
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Synopsis Table 4 and Table 1
Study groups and epoch foreseen in the study
Study groups
Number of 
subjects
Age at Dose 1 (Min/Max)
Epoch 001
HRV Liq
225 154
6 weeks-10 weeks
HRV Lyo
225 154
6 weeks-10 weeks

Sampling schedule: Any GE episodes occurring from 
Dose 1 of the HRV vaccine up to Visit 3 will be 
recorded in the diary card. Parents/Legally Acceptable 
Representative(s) (LARs) will be instructed to collect 
the stool sample(s) if the subject develops GE during 
this period, from Dose 1 of HRV vaccine up to Visit 3. 
A stool sample will be collected as soon as possible, 
after illness begins, and preferably not later than 7 days 
after the start of GE symptoms, to identify the presence 
of wild-type RV exposure during vaccination and to 
eliminate such cases from Per-Protocol Set (PPS) 
analysis. Two occurrences of diarrhea will be classified 
as separate episodes if five or more diarrhea-free days 
occur between the episodes. Refer to glossary of terms 
for the definitions of GE and diarrhea
Section 4.1- Number of subjects/centers
The target will be to enrol 450 308 eligible subjects (225 in each of the study groups) 
who will be randomly assigned to two study groups in a (1:1) ratio (approximately 154 
subjects in each group), to obtain at least 292 200 evaluable subjects (approximately
146100 subjects in each group)
Overview of recruitment plan:

Enrolment will be terminated when 308450 eligible subjects have been enrolled.
Section 4.3- Exclusion criteria

Acute disease and/or fever at the time of enrolment. This warrants deferral of 
vaccination. 
 
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Section 5.2.2.2- Study group and treatment number allocation to the subject: 
The target will be to enrol 308 450 eligible subjects who will be randomly assigned to 
two study groups in a (1:1) ratio (approximately 225 154 subjects in each group), to 
obtain 200 evaluable subjects. (approximately 100 subjects in each group).
Allocation of the subject to a study group at the investigator site will be performed 
using SBIR. The randomization algorithm will use a minimization procedure 
accounting for center and the study as minimization factors. Minimization factors 
will have equal weight in the minimization algorithm.
After obtaining the signed and dated ICF from the subject’s parent/LAR and having 
checked the eligibility of the subject, the site staff in charge of the vaccine 
administration will access SBIR. Upon providing the subject identification number, 
SBIR will provide the treatment number to be used for the first dose.
 
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Table 4- List of study procedures
Age at enrolment
6–10 weeks
2 months
3 months
Epoch
Epoch 001
Type of contact
Visit 1
Visit 2
Visit 3
Time points
Day 1
Month 1
Month 2
Sampling time points
Pre-Vacc 
Post-Vacc
Informed consent
●
Check inclusion/exclusion criteria
●
Collect demographic data
●
Check contraindications and warnings and precautions
●
●
Medical history
●
Physical examination
●
History of previous vaccination from birth
●
Pre-vaccination body temperature
●
●
Vaccines
Study group and treatment number allocation
O
Treatment number allocation for subsequent doses
O
Recording of administered treatment number
●
●
HRV vaccine administration
●
●
Record regurgitation
●
●
HRV vaccine replacement dose administration in case of regurgitation *
●
●
Laboratory Assays
Blood sampling for antibody determination (2 ml)
●
●
Collection of stool sample post vaccination if the child develops GE
Safety assessments
Distribution of diary cards
O
O
Recording of solicited adverse events within 8 days after each dose of HRV 
vaccine (Day 1–Day 8)
●
●
Recording of non-serious adverse events within 31 days (Day 1–Day 31) post-
vaccination
●
●
●
Recording of GE episodes for all subjects 
●
●
●
Record any concomitant medication/vaccination
●
●
●
Record any intercurrent medical conditions
●
●
●
Recording of AEs/SAEs leading to withdrawal from study
●
●
●
Recording of SAEs
●
●
●
Recording of SAEs related to study participation, or to a concurrent GSK 
medication/vaccine
●
●
●
Return of diary cards
O
O
Diary card transcription by investigator
●
●
Study Conclusion
●
● is used to indicate a study procedure that requires documentation in the individual eCRF.
○ is used to indicate a study procedure that does not require documentation in the individual eCRF.
LAR = Legally Acceptable Representative; GE = Gastroenteritis; AE = Adverse Events; SAE = Serious Adverse Events
* If regurgitation or vomiting occurs after vaccination, a single replacement dose may be given at the same vaccination 
visit at the discretion of the Investigator.
Note: All subjects are allowed to receive routine childhood vaccinations according to the local immunization practice.
 
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Table 5- Interval between study visits
Interval
Optimal length of interval 1
Allowed interval2
Visit 1 Visit 2
1 month
28-48 days after Dose 1
Visit 2 Visit 3
1 month
2131-48 days after Dose 2
1 Whenever possible the investigator should arrange the study visits within this interval.
2 Subjects will not be eligible for inclusion in the Per-Protocol Set (PPS) for analysis of immunogenicity if they make 
the study visit outside the 21-48 day interval.
Note: The date of the previous visit serves as the reference date for the intervals between the study visits.
Section 5.6.3: Collect demographic data 
Record demographic data such as age, sex and race geographical ancestry in the 
subject’s eCRF
Section 5.6.9.2- Other biological samples
Collection of stool samples
Parents/LARs of all subjects will be instructed to collect stool sample(s) from the 
subject if the subject develops GE from Dose 1 of the HRV vaccine up to Visit 3. A 
stool sample should be collected as soon as possible, after the illness begins, and 
preferably not later than 7 days after the start of GE symptoms. A second GE stool 
sample should be collected if the first sample is insufficient. A stool sample should be 
collected for each GE episode. Two occurrences of diarrhea will be classified as 
separate episodes five or more diarrhea-free days occur between the episodes.
Refer to the Module on Biospecimen Management in the SPM for detailed instructions 
for the collection, handling and processing of the samples
Table 6- Biological samples
Sample type
Quantity
Unit
Time points
Blood
Approximately 2
ml
Visit 1 and Visit 3
Stool
NA
NA
From Dose 1 of the HRV vaccine up to Visit 3
ml-milliliter; NA-Not Applicable
Section 5.6.10- Study vaccine administration
After completing all prerequisite procedures prior to vaccination, two oral doses of the 
study vaccine will be administered at an approximate one-month interval to subjects, 
according to the immunization schedule for HRV vaccine administration in the 
participating country (refer to Section 6.3 for detailed description of the vaccines 
administration procedure). If the investigator or delegate determines that the subject’s 
health on the day of administration temporarily precludes vaccines administration, the 
visit will be rescheduled within the allowed interval for this visit (refer to Table 5)
 
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Section 5.6.13; Recording of AEs and SAEs
At each vaccination visit, diary cards will be provided to the subject’s 
parent(s)/LAR(s). The subject’s parent(s)/LAR(s) will be instructed to measure and 
record the oral, axillary or rectal body temperature, and any solicited general AEs (i.e., 
on the day of vaccination and during the next 7 days) or any unsolicited AEs (i.e., on 
the day of vaccination and during the next 30 days occurring after vaccination) or any 
GE episodes occurring from Dose 1 of the HRV vaccine up to Visit 3. The subject’s 
parent(s)/LAR(s) will be instructed to return the completed diary card to the 
investigator at the next study visit.
Section 5.7.3- Laboratory assays
GE Stool analysis
All GE stool samples collected from Dose 1 of the HRV vaccine up to Visit 3 will be 
tested at GSK Biologicals’ laboratory or in a validated laboratory designated by GSK 
Biologicals using standardized and validated procedures, for the purpose of identifying
wild-type RV exposure during vaccination and to eliminate such cases from PPS 
immunogenicity analysis.
All GE stool samples will be analyzed by ELISA for detection of RV antigen. If a stool 
sample tests positive for RV antigen, the sample will be tested by Reverse 
Transcription Polymerase Chain reaction (RT-PCR) followed by sequencing to 
determine the G and P genotype.
Table 7- Humoral Immunity (Antibody determination)
System
Component
Method
Kit/Manufacturer
Unit
Cut-off
Laboratory
SER
Rotavirus Ab.IgA
ELI
N/A
U/ml
13* 20
GSK Biologicals**
SER = Serum
IgA = Immunoglobulin A
ELI =ELISA (Enzyme Linked Immunosorbent Assay)
U = Units; ml = milliliters
* This corresponds to the technical cut-off of the revalidated laboratory assay.
**GSK Biologicals laboratory refers to the Clinical Laboratory Sciences (CLS) in Rixensart, Belgium; Wavre, 
Belgium and Marburg, Germany
Additional exploratory testing on the vaccine and/or on 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
Table 8- Immunological read-outs
Blood sampling time point
Type of contact and 
time point
Sampling time point
No. subjects
Component
Components 
priority rank
Visit 1 (Day 1)
Pre-Vacc
All
Rotavirus Ab.IgA
None
Visit 3 (Month 2) 
Post-Vacc
All
Rotavirus Ab.IgA
None
GE analysis†
Visit 1 (Day 1) to Visit 3
(Month 2)
From Dose 1 of the HRV 
vaccine up to Visit 3
All subjects with 
GE
RV antigen
None
Pre-Vacc- Pre-vaccination; Post-Vacc- Post Vaccination; Ab- Antibody RV = Rotavirus; IgA = Immunoglobulin A
 
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† Stool analysis will be performed for subjects who experience GE episodes.
Section 5.7.5- Immunological correlates of protection
No immunological correlate of protection has been demonstrated so far for the antigen 
used as part of the HRV vaccine. However, a study by Cheuvart et al., in 2014 indicated
that post-vaccination anti-RV IgA seropositivity (antibody concentration 20 U/mL) 
may serve as a useful correlate of vaccine efficacy in clinical trials of Rotarix
[Cheuvart, 2014].
The investigator is encouraged to share the immunological assay results for non-
responders with the study subjects’ parent(s)/LAR(s).
Table 12- Reporting periods for collecting safety information
Event
Pre-
V1*
V1
7 days 
post 
vacc
30 
days
post 
vacc
V2
7 days 
post 
vacc
30 days
post 
vacc
V3
D1
D 8
D 31
M1
M2
Study 
Conclusion
Solicited general 
AEs
Unsolicited AEs
GE episodes**
AEs/SAEs leading 
to withdrawal from 
the study
SAEs
SAEs related to 
study participation 
or concurrent GSK 
medication/vaccine
* Consent obtained. Pre-V: pre-vaccination; Post-Vacc: post-vaccination; V: Visit; D: Day, M: Month; 
GE = Gastroenteritis; AEs = Adverse Events; SAEs = Serious Adverse Events
** If GE appears during the solicited period, then it will be recorded on the solicited AEs screen of eCRF as diarrhea
and if it appears outside the solicited period up to Visit 3, then it will be recorded as an unsolicited AE.
 
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Table 13- Intensity scales for solicited symptoms in infants
Infants
Adverse Event
Intensity grade
Parameter
Fever*
Record temperature in °C/°F
0
temperature <38.0°C/100.4° F
1
temperature 38.0°C/100.4° F – 38.5°C/101.3 F 
2
temperature >38.5°C/101.3 F – 39.5°C/103.1 F
3
temperature >39.5°C/103.1 F
Irritability/Fussiness
0
Behavior as usual
1
Mild: Crying more than usual/no effect on normal activity
2
Moderate: Crying more than usual/interferes with normal activity
3
Severe: Crying that cannot be comforted/prevents normal activity
Diarrhea$
Record the number of looser than normal stools/day
0
Normal (0-2 looser than normal stools/day)
1
3 looser than normal stools/day
2
4-5 looser than normal stools/day
3
6 looser than normal stools/day
Vomiting§
Record the number of vomiting episodes/day
0
Normal (no emesis)
1
1 episode of vomiting/day
2
2 episodes of vomiting/day
3
3 episodes of vomiting/day
Loss of appetite
0
Appetite as usual
1
Mild: Eating less than usual/no effect on normal activity
2
Moderate: Eating less than usual/interferes with normal activity
3
Severe: Not eating at all
Cough/runny nose
0
Normal
1
Mild: Cough/runny nose which is easily tolerated
2
Moderate: Cough/runny nose which interferes with daily activity
3
Severe: Cough/runny nose which prevents daily activity
* Fever is defined as temperature 38.0°C/100.4°F. The preferred location for measuring temperature in this study 
will be the oral cavity, the axilla and the rectum.
$ Diarrhea is defined as passage of three or more looser than normal stools within a day. 
§ Vomiting is defined as one or more episodes of forceful emptying of partially digested stomach contents ≥1 hour 
after feeding within a day.
The intensity grade for diarrhea, vomiting and fever as shown in Table 13 will be scored at GSK 
Biologicals.
 
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Section 6.5: Contraindications to subsequent vaccination

Subjects with a minor illness (such as mild diarrhoea, mild upper respiratory 
infection) without fever may be vaccinated enrolled at the discretion of the 
investigator.
Section 6.7.2: Concomitant medications/products/vaccines that may lead to the 
elimination of a subject from per-protocol analyses

A vaccine not foreseen by the study protocol administered during the period 
starting from 30 days before the first dose of HRV vaccine* administration and 
ending at Visit 3** blood sampling, with the exception of the inactivated influenza 
vaccine, and other licensed routine childhood vaccinations which are allowed at 
any time during the study.
*Administration of any HRV vaccine other than the study vaccine will lead to 
elimination (refer to Section 4.3).
**In case an emergency mass vaccination for an unforeseen public health threat 
(e.g.: a pandemic) is organized by the public health authorities, outside the routine 
immunization program, the time period described above can be reduced if 
necessary for that vaccine provided it is licensed and used according to its 
Prescribing Information and according to the local governmental recommendations 
and provided a written approval of the Sponsor is obtained.
A detailed, comprehensive list of reasons for elimination from PPS analyses will be 
established at the time of data cleaning
Synopsis, Section 10.1 and Section 10.2- Endpoints
Primary Endpoint

Anti-RV IgA antibody concentrations

Serum anti-RV IgA antibody concentrations, expressed as GMCs, one 
month post dose 2 of HRV vaccine.
Secondary

Anti-RV antibody concentrations

Anti-RV IgA antibody seroconversion rate, one month post dose 2 of HRV 
vaccine.

Presence of wild type RV in GE stool samples

Presence of wild type RV in GE stools collected from Dose 1 of the HRV
vaccine up to Visit 3
Section 10.3 Determination of sample size
A total sample of 450 subjects (225 subjects per arm) is based on a 90% power to 
detect non-inferiority on GMCs using PASS 2014, under the alternative hypothesis 
 
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of equal means and variances (Table 15). Based on historical data (ROTA- 044) it is 
assumed that up to 35% of the subjects will not be evaluable for the analysis of the 
primary endpoint, therefore the sample was increased accordingly.
Table 15: Probability that the lower limit of the 95% CI around the anti-RV IgA 
antibody GMC ratio (HRV Liq/HRV Lyo), one month after dose 2 of HRV vaccine, is 
0.5
Endpoint
Standard deviation
N evaluable
Power
Alpha
[Log10 (titre)]
(each HRV group)
Anti-RV IgA 
antibody 
concentration
0.790*
146
90%
0.025
*=Reference used for power computation
Considering a target sample size of 308 enrolled subjects (1:1 randomization, 154 
subjects in each group) and assuming that up to 35% of the subjects will not be 
available for the analysis of the primary endpoint (Reference: Rota-044), it can be 
considered that at least 200 subjects (100 in each group) will be available for the 
analysis.

Table 15 presents the exact two-sided 95% Confidence Interval (CI) with 
the sample size for different values of seroconversion rates, for subjects in 
HRV Liq group.
 
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Table 15 Exact 95 percent CI with the sample size for different values of 
seroconversion rates, for subjects in HRV Liq group
Number of evaluable 
subjects
(N)
Anti rotavirus IgA seroconversion rate with 95%CI
n
%*
Exact 95%CI
LL
UL
100
50
50
39.8
60.2
100
60
60
49.7
69.7
100
70
70
60.0
78.8
100
80
80
70.8
87.3
100
85
85
76.5
91.4
100
90
90
82.4
95.1
*Range of seroconversion rates in agreement with following reference studies: 
Rota-044: 58.3% [95% CI: 48.7; 67.4], Rota-036: 83.9% [95% CI: 83.9; 88.8], Rota-061: 88.6% [86.1; 90.8]
n = Number of subjects,
% = (n/N)*100, LL = Lower Limit, UL = Upper Limit
95% CI = 95% Confidence Interval
Section 10.4.2- Per-Protocol Set for analysis of immunogenicity

for whom the post-vaccination immunogenicity data are within the 21-48 days
interval after the second dose.

who have no RV other than vaccine strain in GE stool samples collected up to 
Visit 3 blood sample,
 
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Section 10.5- Derived and transformed data
Immunogenicity

A seronegative subject is a subject whose anti-RV IgA antibody concentration is 
below the clinically meaningful threshold of <20 U/ml*.

The GMCs calculations are performed by taking the anti-log of the mean of the log
concentrations transformations. Antibody concentrations below the cut-off
(<13* 20 U/ml) of the assay will be given an arbitrary value of half the cut-off for 
the purpose of GMC calculation.
*Note: 20 U/ml corresponds to clinically meaningful threshold to define 
seroconversion while 13 U/ml corresponds to technical cut off of revalidated 
laboratory assay.
Section 10.6- Analysis of demographics.
The number of subjects enrolled into the study as well as the number of subjects 
excluded from PPS analyses will be tabulated.
The median, mean, range and standard deviation of age (in weeks) at each HRV liquid 
vaccine dose will be computed. The median, mean and standard deviation of height in 
centimeter (cm) and weight in kilograms (kg) at Visit 1 will be computed. The Body 
Mass Index (BMI) at Visit 1 will also be computed as weight (in kg)/height² (in 
meters). The racial geographical ancestry and sex composition will be presented. 
Section 10.7.1- Within groups assessment
The following calculations will be performed for each group.

For each group, at each time point that anti-rotavirus IgA is measured,

GMCs and their 95% CIs will be computed.

Seropositivity/seroconversion rates and their exact 95% CI will be 
computed,

The distribution of anti-RV IgA antibody concentrations at Visit 3 will be 
displayed using Reverse Cumulative Curves (RCCs).

For each treatment group and for anti-rotavirus IgA antibody concentration,

Seroconversion rates at Visit 3 and their exact 95% CI will be computed 
[Clopper, 1934].

GMCs and their 95% CIs at Visit 3 will be computed.
Section 10.7.2- Between groups assessment

The 95% CI for the ratio of anti-RV IgA antibody GMCs at Visit 3 between the 
HRV liquid vaccine over the HRV lyophilized vaccine will be computed (primary 
objective).
 
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
The asymptotic standardized 95% CI for the difference in seroconversion rate at 
Visit 3 between the HRV liquid vaccine and HRV lyophilized vaccine will be 
computed (secondary objective).
Refer to the Section 2.1 for the success criteria of the primary objective.
Section 10.8.1- Within groups assessment
The following calculations will be performed for each group:
The percentage of doses and of subjects reporting at least one symptom (solicited or 
unsolicited) during the 8-day (Day 1-Day 8) solicited follow-up period will be 
computed, along with exact 95% CI [Clopper, 1934].The same calculations will be
done for symptoms (solicited or unsolicited) rated as grade 3 in intensity, and for 
symptoms (solicited or unsolicited) assessed as causally related to vaccination and for 
symptoms resulting in medically attended visit.
The percentage of doses and of subjects reporting each individual solicited general
symptom will be computed, over the 8-day (Day 1-Day 8) solicited follow-up period, 
post vaccination, along with exact 95% CI [Clopper, 1934]. The same calculations will 
be done for each individual solicited general symptom rated as grade 3 in intensity and 
for each individual solicited general symptom assessed as causally related to 
vaccination and for symptoms resulting in medically attended visit.
The verbatim reports of unsolicited AEs will be reviewed by a physician and the signs
and symptoms will be coded according to MedDRA. Every verbatim term will be
matched with the appropriate Preferred Term. The percentage of subjects with 
unsolicited AEs occurring within 31-day (Day 1-Day 31) follow-up period after any 
dose with its exact 95% CI [Clopper, 1934], will be tabulated by group, and by 
preferred term. Similar tabulation will be done for unsolicited AEs rated as grade 3, for 
unsolicited AEs with causal relationship to vaccination, for symptoms resulting in 
medically attended visit and for AEs or SAEs leading to drop out.
The percentage of subjects reporting GE episodes from Dose 1 of HRV lyophilized 
vaccine/HRV liquid vaccine up to Visit 3 will be tabulated by group.
The percentage of subjects with presence of wild-type RV in GE stool samples 
collected from Dose 1 of HRV lyophilized vaccine/HRV liquid vaccine up to Visit 3 
will be tabulated by group.
The percentages of GE episodes with no available stool results from Dose 1 of HRV 
vaccine up to Visit 3 will be tabulated.
Section 10.7 Interpretation of analyses
Except for analyses addressing the primary objectives referred as confirmatory 
analyses, all the analyses will be descriptive/exploratory in nature. All analyses will 
be descriptive with the aim to characterise the observed outcomes. These descriptive 
analyses should be interpreted with caution.
 
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Section 10.11- Conduct of analysis
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
Appendix A- Laboratory Assays
Antigen Detection in Stool Samples
The RV antigen in stool samples collected during GE episodes will be detected by 
ELISA at Central Lab (GSK or designated laboratory).
RV strain genotyping 
The RV RNA will be isolated from ELISA RV positive stool samples. In order to 
determine the RV P and G type, VP4 and VP7 genes will be then amplified by Reverse 
Transcriptase Polymerase Reactions (RT-PCR) using two separate primer sets. VP4 
and VP7 amplimers will be further analyzed by direct sequencing. The sequences will 
be interpreted using BLAST database searches in Genbank and phylogenetic analysis.
 
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Appendix B Clinical Laboratories
Table 16 GSK Biologicals’ laboratories
Laboratory
Address
GSK Biological’s Clinical 
Laboratory Sciences (CLS), 
Rixensart 
Biospecimen Reception-B7/44
Rue de l'Institut, 89
B-1330 Rixensart
Belgium
GSK Biological’s CLS, Wavre-
Nord Noir Epine
Avenue Fleming, 20
B-1300 Wavre
Belgium
GSK Vaccines GmbH
Emil-von-Behring-Str. 76
35041 Marburg
Deutschland/ Germany
Table 17 Outsourced laboratories
Laboratory
Address
Q² Solutions Clinical Trials (US)
27027 Tourney Road, Suite 2E
Valencia, CA 91355
US
Q² Solutions Clinical Trials (UK)
1 Simpson Parkway
The Alba Campus
Rosebank
Livingston
EH54 7EG
UK
DDL Diagnostic Laboratory
Visseringlaan 25
2288 ER Rijswijk
The Netherlands
 
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GlaxoSmithKline Biologicals SA
Vaccines R &D
Protocol Amendment 4
eTrack study number and 
Abbreviated Title 
116566 (ROTA-083)
EudraCT number
2012-001875-35
Amendment number:
Amendment 4
Amendment date:
30 October 2019
Co-ordinating author:
Rationale/background for changes: As per Protocol Amendment 3 (31 October 2017), 
subjects seropositive before vaccination are to be eliminated from the Per-Protocol Set 
(PPS) for immunogenicity.
Laboratory testing of the pre-vaccination blood samples from the first 141 subjects 
enrolled in the study indicated a seropositivity rate of 47.5 % in these subjects. This 
suggests an overall high seropositivity rate at baseline in the total study population.
Therefore, an elimination rate exceeding the 35% rate of non-evaluable subjects 
predefined using historical data from Rota-044 study [Narang, 2009] is expected.
Because (1) the pre-vaccination seropositivity rate is expected to be high and (2) the 
effect of the vaccine can be measured in seropositive subjects, as previously 
demonstrated (see Table 1 below), the following changes have been made to the protocol:

The PPS was modified to include subjects seropositive at baseline.

The statistical method to derive the 95% confidence interval (CI) for the geometric 
mean concentration (GMC) group ratio was revised to be an ANCOVA.

The seroconversion threshold was redefined to account for seropositive subjects at 
pre-vaccination:

for subjects with a pre-vaccination anti-rotavirus IgA antibody concentration 
<20 U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥20 U/mL.

for subjects with a pre-vaccination anti-rotavirus IgA antibody concentration 
≥20 U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥2 times the pre-vaccination concentration.
 
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Table 1: Distribution of fold increase in anti-rotavirus IgA antibody concentrations
from pre-vaccination to one month post-dose 2 (Total vaccinated cohort for 
seropositive subjects at pre-vaccination, Rota-044) [Narang, 2009]
HRV 
N = 43
Placebo 
N = 40
Ratio
n
%
n
%
≥2
26
60.5
3
7.5
≥3
21
48.8
2
5.0
≥4
17
39.5
2
5.0
N = number of subjects seropositive at baseline for anti-rotavirus IgA antibodies and with one month post-dose 2 data 
available; n/% = number/percentage of subject in a given category.
Amended text has been included in bold italics and deleted text in strikethrough in 
the following sections:
Contributing authors:
, Clinical Research and Development Lead (CRDL)
, 
, Study Delivery Leads
Project Delivery Lead
, Clinical Laboratory Sciences (CLS), Clinical Readout Team Leader
, Laboratory Study Manager
, Study Data Manager
, 
, Clinical Trial Supply Managers
, Local Medical Lead
, Local Delivery Lead
Synopsis and Section 2.2: Secondary objectives

To assess the immunogenicity of the HRV liquid vaccine and HRV lyophilized 
vaccine, in terms of seroconversion* rates, one month post dose 2 of HRV vaccine.
*Definition: 

for subjects with a pre-vaccination anti-RV IgA antibody concentration <20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥20 U/mL.

for subjects with a pre-vaccination anti-RV IgA antibody concentration ≥20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥2 times the pre-vaccination concentration.
 
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Seroconversion rate is defined as the percentage of subjects with anti-RV IgA 
antibody concentration ≥20 U/mL one-month post dose 2 among subjects with anti-
RV IgA antibody concentration < 20 U/mL at pre-vaccination.
Synopsis and Section 10.2: Secondary endpoints

Anti-RV IgA antibody concentrations

Anti-RV IgA antibody seroconversion* rate, one month post dose 2 of HRV 
vaccine.
*Definition: 

for subjects with a pre-vaccination anti-RV IgA antibody concentration <20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥20 U/mL

for subjects with a pre-vaccination anti-RV IgA antibody concentration ≥20 
U/mL, seroconversion is achieved when the post-vaccination concentration 
is ≥2 times the pre-vaccination concentration
Glossary of terms
Seroconversion:
For subjects with a pre-vaccination anti-RV IgA 
antibody concentration <20 U/mL, seroconversion is 
achieved when the post-vaccination concentration is 
≥20 U/mL.
For subjects with a pre-vaccination anti-RV IgA 
antibody concentration ≥20 U/mL, seroconversion is 
achieved when the post-vaccination concentration is ≥2 
times the pre-vaccination concentration.
Appearance of anti-rotavirus Immunoglobulin A (IgA) 
antibody concentration 20 units (U)/milliliter (ml) in 
subjects initially (i.e., prior to the first dose of Human 
Rotavirus [HRV] vaccine) seronegative for anti-RV IgA 
antibody.
Section 1.1: Background
Rotarix is currently registered in over at least 130 countries and about 552 446 066
405,524,425 doses of the vaccine (lyophilized and liquid formulations) are estimated to 
have been distributed worldwide since its launch until July 2019 2017.
 
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Table 9: Study vaccine
Treatment 
name
Vaccine 
name
Formulation
Presentation
Volume to 
be 
administered
orally
Number 
of doses
HRV Liquid
HRV Liquid 
vaccine
Active substance: 
HRV RIX4414
RIX4144 live 
attenuated 
>=10⁶⋅⁰CCID₅₀
Excipients:
Sucrose=1.073g; Di-
sodium 
Adipate=132.74mg; 
DMEM=2.26mg; water 
for injection=1.5ml
Liquid vaccine in a pre-filled 
oral applicator
1.5 ml
2
HRV 
Lyophilized
HRV 
Lyophilized 
vaccine
HRV RIX4414
RIX4144 live 
attenuated 
>=10⁶⋅⁰CCID₅₀
Lyophilized vaccine in a 
monodose glass vial. Diluent 
(calcium carbonate buffer) 
supplied separately
1 ml
2
HRV 
Diluent
CaCO₃=60mg
Diluent for lyophilized 
vaccine (calcium carbonate 
liquid antacid) supplied 
separately in a prefilled oral 
applicator
HRV=Human Rotavirus; CCID50 = Median Cell Culture Infective Dose (quantity of virus causing infection in 50% of 
exposed cells); DMEM = Dulbecco's Modified Eagle Medium; ml = milliliter;
Section 10.3: Determination of sample size
Considering the a total sample of 450 subjects (225 subjects per arm)is based on a 90% 
power to detect non-inferiority on GMCs using PASS 2014, under the alternative 
hypothesis of equal means and variances (Table 15). Based on historical data (study 
ROTA- 044) it is assumed that up to 35% of the subjects will not be evaluable for the 
analysis of the primary endpoint, therefore the sample was increased accordingly., Table 
15 provides the power to demonstrate non-inferiority in terms of GMCs according to 
the percentage of non-evaluable subjects.
Based on historical data from ROTA-044 study [Narang, 2009], 13.8% of subjects were 
non-evaluable for reasons other than ‘initially positive or unknown for serum anti-RV
IgA antibodies at Dose 1’. The power to reach the primary objective in a scenario of 
13% non-evaluable subjects is 96% (Table 15). Considering a scenario of 35% non-
evaluable subjects, the power to reach the primary objective is 90%. As shown in 
Table 15, the power decreases when the % of non-evaluable subjects increases.
 
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Table 15: Power according to different scenarios for the % of non-evaluable subjects, 
with respect to a total sample size of 450 subjects*
Standard deviation
Alpha
% of non-evaluable subjects
Power
[Log10 (concentration)]
0.790**
0.025
13%
96%
15%
96%
25%
94%
30%
92%
35%
90%
40%
88%
45%
85%
50%
81%
55%
77%
*PASS one-sided non-inferiority for 2 independent means with common variance for log10(0.5) as non-
inferiority margin, for the alternative hypothesis that the means are equal and for a type I error=2.5 %
**Reference used for power computation
Table 15: Probability that the lower limit of the 95% CI around the anti-RV IgA antibody 
GMC ratio (HRV Liq/HRV Lyo), one month after dose 2 of HRV vaccine, is 0.5. 
(Amended: 31 October 2017)
Endpoint
Standard deviation
N evaluable
Power
Alpha
[Log10 (titre)]
(each HRV group)
Anti-RV IgA 
antibody 
concentration
0.790*
146
90%
0.025
*=Reference used for power computation
Section 10.4.2: Per-Protocol Set for analysis of immunogenicity

who are seronegative for serum anti-RV IgA antibodies on the day of dose 1,
Section 10.5: Derived and transformed data
Immunogenicity

Seroconversion rate is defined as the percentage of subjects who were initially 
seronegative (i.e., with anti-RV IgA antibody concentration < 20 U/mL prior the first 
dose of HRV vaccine) and developed anti-RV IgA antibody concentration 
≥20 U/mL at Visit 3.
 
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
Seroconversion is defined as:

for subjects with a pre-vaccination anti-RV IgA antibody concentration <20 
U/mL, seroconversion is achieved when the post-vaccination concentration is 
≥20 U/mL.

for subjects with a pre-vaccination anti-RV IgA antibody concentration ≥20 
U/mL, seroconversion is achieved when the post-vaccination concentration is 
≥2 times the pre-vaccination concentration.
Section 10.7.1: Within group assessment

The distribution of anti-RV IgA antibody concentrations at Visit 1 and Visit 3 
will be displayed using Reverse Cumulative Curves (RCCs).
Section 10.7.2: Between groups assessment

The 95% CI for the ratio of anti-RV IgA antibody GMCs at Visit 3 between the HRV 
liquid vaccine over the HRV lyophilized vaccine will be computed using an 
ANCOVA model on the logarithm-transformed concentrations. This model will 
include the vaccine group and the logarithm of the baseline concentration as 
covariables. The GMC ratios and their 95% CI will be derived by exponential 
transformation of the corresponding group contrast in the model (primary 
objective).
Section 10.10: Statistical Methods

The standardized asymptomatic CI for the group difference in proportion will be 
calculated using SAS. The method used within GSK Biologicals is Method 6 
[Newcombe, 1998].
Section 13: References
Narang A, Bose A, Pandit AN, Dutta P, Kang G, Bhattacharya SK, Datta SK,
Suryakiran PV, Delem A, Han HH, Bock HL. Immunogenicity, reactogenicity and 
safety of human rotavirus vaccine (RIX4414) in Indian infants. Hum Vaccin. 2009 
Jun;5(6):414-9.
 
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