Clinical trial NCT04711356
Conditions: Ebola Virus Disease
Keywords: Ad26.ZEBOV booster vaccination children
Sponsor: London School of Hygiene and Tropical Medicine
Start date: 2021-07-08; Completion date: 2022-03-31
Enrollment: 50 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
This is an open-label study evaluating the safety and immunogenicity of a booster dose of Ad26.ZEBOV administered to children who were previously vaccinated with Ad26.ZEBOV followed by MVA-BN-Filo 56 days later.

Detailed description:
RATIONALE:

Over 600 children have received the adenovirus serotype 26 expressing the Ebola virus Mayinga glycoprotein (Ad26.ZEBOV), modified Vaccinia Ankara Bavarian Nordic vector expressing multiple filovirus proteins (MVA- BN-Filo) Ebola vaccine regimen in the EBL2002 and EBL3001 clinical trials. The vaccine regimen was well tolerated and highly immunogenic in children; however, the durability of vaccine-induced immune responses is not known. In adults previously vaccinated with the Ad26.ZEBOV and MVA-BN-Filo regimen, a booster vaccination with Ad26.ZEBOV was safe and induced a strong anamnestic response within seven days of the booster vaccination. It is important to establish if a booster dose of Ad26.ZEBOV is safe and immunogenic also in children, as this can guide the clinical use of the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen in this age group. For example, it could support the strategy of boosting immunised children at the start of an Ebola outbreak.

STUDY OBJECTIVES AND HYPOTHESIS:

This study aims to evaluate the safety and immunogenicity of an Ad26.ZEBOV booster dose in healthy children who were previously (\>2 years) vaccinated with the Ad26.ZEBOV (dose 1) followed by MVA-BN-Filo (dose 2) 56 days later, by monitoring adverse events (AEs) following the booster vaccination and by assessing binding antibody responses using the Filovirus Animal Non-Clinical Group (FANG) Enzyme-Linked Immunosorbent Assay (ELISA).

Primary Objectives:

* To assess the safety and tolerability of a booster dose of Ad26.ZEBOV at a dose of 5x10\^10 viral particles (vp) in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen with a 56-day interval. Safety and tolerability will be assessed by measuring:

  1. Incidence of solicited local (at the administration site) and systemic adverse events as assessed on the day of booster vaccination and by diary card for 7 days after booster vaccination.
  2. Incidence of unsolicited adverse events from the day of the booster vaccination to 28 days post booster vaccination.
* To assess vaccine-induced humoral immune responses to the Ebola virus glycoprotein (EBOV GP), as measured by FANG ELISA (EU/ml), at 7 and 21 days following a booster dose of Ad26.ZEBOV at a dose of 5x10\^10 vp in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen with a 56-day interval.

Exploratory Objectives

• To assess neutralising antibody responses directed against the Ad26 vector before booster vaccination as measured by a virus neutralization assay (VNA).

Hypothesis: as this study is designed to provide descriptive information regarding safety and immunogenicity without formal treatment comparisons, no formal statistical hypothesis testing is planned.

OVERVIEW OF THE STUDY:

This is an open-label study evaluating the immune response to a booster dose of Ad26.ZEBOV administered to children who were previously vaccinated with Ad26.ZEBOV followed by MVA-BN-Filo 56 days later. Only subjects who received the Ad26.ZEBOV and MVA-BN-Filo regimen during their participation in the VAC52150EBL3001 (EBOVAC-Salone) vaccine trial (ClinicalTrials.gov Identifier: NCT02509494), are eligible for enrolment in this study. Participants will be recruited in two age groups: children aged 4-11 years at the time of dose 1 vaccination and children aged 1-3 years at the time of dose 1 vaccination in the EBOVAC-Salone trial. Approximately 25 subjects will be enrolled in each of these two age groups. Parents/guardians will be asked to consent for the participation of their children in the study. Children aged 7 years and older at the time of enrolment in this study will be asked to give positive assent for their participation. Participants will be followed up to 28 days after their booster vaccination. The study will be conducted in Kambia, Sierra Leone.

Study Population: potential participants must be healthy children (based on physical examination, medical history, a haematological assessment and clinical judgment) who received the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen in the EBOVAC-Salone trial and were aged ≥1 to ≤11 years at the time of dose 1 vaccination. They must also be enrolled in the long-term follow-up study to the EBOVAC-Salone trial, VAC52150EBL3005 (EBOVAC-Salone Extension study, ClinicalTrials.gov Identifier: NCT03820739) but not in the immunogenicity subset.

Dosage and administration: a single dose of Ad26.ZEBOV at a dose of 5x10\^10 vp administered intramuscularly.

Safety evaluations: solicited local (at the administration site) and systemic AEs will be assessed on the day of vaccination and using a diary for a period of 7 days following the booster vaccination. Unsolicited AEs will be tracked for 28 days following booster vaccination, while serious adverse events will be tracked for the duration of the study.

Immunogenicity evaluations: blood will be drawn for assessments of immune responses at 7 and 21 days post booster vaccination.

STATISTICAL METHODS:

The primary analysis will be done when all subjects have completed their 28-day post-booster visit or discontinued earlier. This analysis will include all available data up to this point.

Sample Size Determination: the sample size is a convenience sample and is not based on formal hypothesis testing considerations.

Safety analysis: no formal statistical testing of safety data is planned. Safety data will be analysed descriptively by age group, 1-3 years and 4-11 years (age the participant was when they received dose 1 vaccination in the EBOVAC-Salone trial).

Immunogenicity analysis: no formal hypothesis on immunogenicity will be tested. Descriptive statistics (i.e. geometric mean and 95% confidence interval, as appropriate) will be calculated for continuous immunologic parameters at all available time points. Graphical representations of immunologic parameters will be made as applicable.

Frequency tabulations will be calculated for discrete (qualitative) immunologic parameters (i.e. responder rate), as applicable. Responders rate defined as \>2.5-time increase over baseline value (or lower limit of quantitation \[LLOQ\]) pre-dose 1 vaccination in the EBOVAC-Salone trial, will be calculated depending on availability of sample results from the EBOVAC-Salone trial.

Arms: arm 1: All study participants will receive an Ad26.ZEBOV vaccine at a dose of 5x10\^10 vp given via IM injection
Interventions: intervention 1: Ad26.ZEBOV booster vaccination, given at a dose of 5x10^10 vp, via IM injection
Intervention descriptions: intervention 1: Ad26.ZEBOV is a monovalent, replication-incompetent adenovirus serotype 26-based vector that expresses the full-length Ebola virus Mayinga glycoprotein

Protocol documents:
Prot_000.pdf: 
 
Page 1 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
 
 
 
 
 
 
 
 
 
 
 
VAC52150EBL2011 
 
 
 
An open label study to evaluate the safety and immunogenicity of an Ad26.ZEBOV booster dose in 
children previously vaccinated with the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen 
 
 
Protocol VAC52150EBL2011; Phase 2 
 
 
Innovative Medicines Initiative 
 
London School of Hygiene and Tropical Medicine and  
 and Janssen Vaccines and Prevention B.V. 
 
VAC52150 (Ad26.ZEBOV/MVA-BN-Filo [MVA-mBN226B]) 
 
 
 
 
Version 2.0, 15 June 2021 
 
 
NUMBER 
DATE 
FINAL VERSION 
1.0 
30/10/2020 
AMENDMENT (if any) 
2.0 
15/06/2021 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


 
Page 2 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
 
 
 
 
Protocol authorised by: 
Name: 
Professor Deborah Watson-Jones 
Role: 
Chief Investigator 
Signature: 
 
Date: 
 
Name: 
Dr Bailah Leigh 
Role: 
Principal Investigator 
Signature: 
 
Date: 
 
Name: 
Mrs Patricia Henley 
Role: 
Sponsor Representative 
Signature: 
 
Date: 
 
 
 
 
Sponsor 
London School of Hygiene & Tropical Medicine is the main research sponsor for this study. For further 
information regarding the sponsorship conditions, please contact: 
 
The Research Governance and Integrity Office, 
London School of Hygiene & Tropical Medicine 
Keppel Street 
London WC1E 7HT 
Tel: +44 207 927 2626 
Email: rgio@lshtm.ac.uk 
 
Funder 
This project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking under grant 
agreement EBOVAC1 (grant nr. 115854). This Joint Undertaking receives support from the European Union’s 
Horizon 2020 research and innovation programme and the European Federation of Pharmaceutical Industries 
and Associations. 
This study will adhere to the principles outlined in the International Conference on Harmonisation Good Clinical Practice 
guidelines, protocol and all applicable local regulations.  
 
 


 
Page 3 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
Table of Contents 
 
ABBREVIATIONS ................................................................................................................................................................. 5 
PROTOCOL AMENDMENTS ................................................................................................................................................ 6 
SYNOPSIS ............................................................................................................................................................................ 8 
TIME AND EVENTS SCHEDULE .......................................................................................................................................... 11 
1 
INTRODUCTION ................................................................................................................................................... 13 
1.1 
BACKGROUND ................................................................................................................................................ 13 
1.2 
BENEFITS/RISKS OF PARTICIPATION ............................................................................................................... 14 
1.3 
OVERALL RATIONALE FOR THE STUDY ............................................................................................................ 16 
2 
OBJECTIVES, ENDPOINTS, AND HYPOTHESIS ...................................................................................................... 17 
2.1 
OBJECTIVES AND ENDPOINTS ......................................................................................................................... 17 
2.2 
HYPOTHESIS .................................................................................................................................................... 17 
3 
STUDY DESIGN .................................................................................................................................................... 17 
3.1 
OVERVIEW OF STUDY DESIGN ........................................................................................................................ 17 
4 
PARTICIPANT POPULATION ................................................................................................................................. 18 
4.1 
INCLUSION CRITERIA ...................................................................................................................................... 18 
4.2 
EXCLUSION CRITERIA ...................................................................................................................................... 19 
4.3 
PROHIBITIONS AND RESTRICTIONS ................................................................................................................ 20 
5 
INTERVENTION ALLOCATION AND BLINDING ..................................................................................................... 20 
5.1 
BLINDING ........................................................................................................................................................ 20 
6 
DOSAGE AND ADMINISTRATION ........................................................................................................................ 20 
6.1 
CRITERIA FOR POSTPONEMENT OF VACCINATION ........................................................................................ 21 
7 
CONCOMITANT THERAPY ................................................................................................................................... 21 
8 
STUDY EVALUATIONS .......................................................................................................................................... 21 
8.1 
STUDY PROCEDURES ...................................................................................................................................... 22 
8.2 
PROCEDURES IN CASE OF A STUDY PAUSE ..................................................................................................... 23 
8.3 
SAFETY EVALUATIONS .................................................................................................................................... 23 
8.4 
IMMUNOGENICITY ASSESSMENTS ................................................................................................................. 26 
8.5 
SAMPLE COLLECTION AND HANDLING ........................................................................................................... 27 
9 
PARTICIPANT COMPLETION/WITHDRAWAL FROM THE STUDY .......................................................................... 27 
9.1 
COMPLETION .................................................................................................................................................. 27 
9.2 
WITHDRAWAL FROM THE STUDY ................................................................................................................... 27 
10 
STATISTICAL METHODS .................................................................................................................................. 28 
10.1 
ANALYSIS SETS ................................................................................................................................................ 28 
10.2 
SAMPLE SIZE DETERMINATION ...................................................................................................................... 28 
10.3 
PARTICIPANT INFORMATION .......................................................................................................................... 28 
10.4 
SAFETY ANALYSES ........................................................................................................................................... 28 
10.5 
IMMUNOGENICITY ANALYSIS ......................................................................................................................... 29 
10.6 
INTERIM ANALYSIS .......................................................................................................................................... 29 
10.7 
INDEPENDENT DATA MONITORING COMMITTEE .......................................................................................... 29 
11 
ADVERSE EVENT REPORTING .......................................................................................................................... 29 
11.1 
DEFINITIONS ................................................................................................................................................... 29 
11.2 
SPECIAL REPORTING SITUATIONS ................................................................................................................... 32 
11.3 
PROCEDURES .................................................................................................................................................. 32 
11.4 
CONTACTING SPONSOR REGARDING SAFETY ................................................................................................. 34 
12 
PRODUCT QUALITY COMPLAINT HANDLING .................................................................................................. 34 
12.1 
PROCEDURES .................................................................................................................................................. 34 
12.2 
CONTACTING SPONSOR REGARDING PRODUCT QUALITY .............................................................................. 35 
13 
STUDY INTERVENTION INFORMATION ........................................................................................................... 35 
13.1 
PACKAGING .................................................................................................................................................... 35 
13.2 
LABELLING ...................................................................................................................................................... 35 
13.3 
PREPARATION, HANDLING, STORAGE ............................................................................................................ 35 
13.4 
INTERVENTION ACCOUNTABILITY .................................................................................................................. 35 
14 
STUDY -SPECIFIC MATERIALS .......................................................................................................................... 36 
15 
ETHICAL ASPECTS ............................................................................................................................................ 36 
15.1 
STUDY-SPECIFIC DESIGN CONSIDERATIONS ................................................................................................... 36 
15.2 
REGULATORY ETHICS COMPLIANCE ............................................................................................................... 37 


 
Page 4 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
16 
ADMINISTRATIVE REQUIREMENTS ................................................................................................................. 40 
16.1 
PROTOCOL AMENDMENTS ............................................................................................................................. 40 
16.2 
REGULATORY DOCUMENTATION ................................................................................................................... 40 
16.3 
SUBJECT IDENTIFICATION AND ENROLMENT LOGS ........................................................................................ 41 
16.4 
SOURCE DOCUMENTATION ............................................................................................................................ 41 
16.5 
CASE REPORT FORM COMPLETION ................................................................................................................ 42 
16.6 
DATA QUALITY ASSURANCE/QUALITY CONTROL ........................................................................................... 42 
16.7 
RECORDS RETENTION ..................................................................................................................................... 42 
16.8 
MONITORING ................................................................................................................................................. 43 
16.9 
STUDY COMPLETION/TERMINATION ............................................................................................................. 43 
16.10 
ON-SITE AUDITS ......................................................................................................................................... 44 
16.11 
USE OF INFORMATION AND PUBLICATION ................................................................................................ 44 
APPENDIX 1 ...................................................................................................................................................................... 45 
APPENDIX 2: ..................................................................................................................................................................... 49 
REFERENCES ..................................................................................................................................................................... 50 
 
 


 
Page 5 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
ABBREVIATIONS 
 
Ad26.ZEBOV 
adenovirus serotype 26 expressing the Ebola virus Mayinga glycoprotein 
AE 
 
adverse event 
CRF 
 
case report form 
DMID  
Division of Microbiology and Infectious Diseases 
EBOV  
Ebola virus 
eDC 
 
electronic data capture  
eCRF 
 
electronic case report form 
ELISA 
 
enzyme-linked immunosorbent assay  
EPI 
 
Expanded Program on Immunisation 
EVD 
 
Ebola virus disease 
FANG  
Filovirus Animal Non-Clinical Group  
GCP 
 
Good Clinical Practice 
GP 
 
glycoprotein 
IB 
 
Investigator’s Brochure 
ICF 
 
informed consent form 
ICH 
 
International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use 
ICMJE  
International Committee of Medical Journal Editors 
IDMC  
Independent Data Monitoring Committee  
IEC 
 
Independent Ethics Committee 
IM 
 
intramuscular 
LLOQ 
 
lower limit of quantitation 
LSHTM  
London School of Hygiene & Tropical Medicine 
MedDRA 
Medical Dictionary for Regulatory Activities 
mL 
 
millilitres 
MVA-BN-Filo 
Modified Vaccinia Ankara Bavarian Nordic vector expressing multiple filovirus proteins 
PCR 
 
polymerase chain reaction 
PI 
 
Principal Investigator 
PQC 
 
Product Quality Complaint 
RBC 
 
red blood cell 
SAE 
 
serious adverse event 
SUSAR  
suspected unexpected serious adverse reaction 
TOU 
 
Test of Understanding 
VISP 
 
vaccine induced seropositivity 
vp 
 
viral particle(s) 
WBC 
 
white blood cell 
WHO 
 
World Health Organisation 
 
 
 
 
 


 
Page 6 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
PROTOCOL AMENDMENTS  
 
 
 
 
Amendments below are listed beginning with the most recent amendment.  
Amendment 1 (This document) 
The overall reasons for the amendment:  
- 
To include the assessment of neutralising antibody responses directed against the Ad26 vector 
before booster vaccination as an exploratory outcome of the study.  
- 
To add safety information on the Ad26-based vaccines, following the latest update of the 
Investigator’s Brochure and Addendum.  
- 
To add Adverse Events of Special Interest 
The changes made to the clinical protocol VAC52150EBL2011 are listed below, including the rationale of 
each change and a list of all applicable sections.  
Rationale: the assessment of neutralising antibody responses directed against the Ad26 vector will be 
useful to understand if previous immunity against the Ad26 vector influences the immune response to the 
Ad26.ZEBOV booster. The assessment of neutralising antibody responses directed against the Ad26 vector 
before booster vaccination has been included as an exploratory outcome of the study. 
SYNOPSIS 
Time and Events Schedule 
2.1 OBJECTIVES AND ENDPOINTS 
8.4 IMMUNOGENICITY ASSESSMENTS 
------------------------------------------------------ 
Rationale: new information on the safety of the Ad26-based vaccines, in particular, the risk of thrombosis 
with thrombocytopenia syndrome (TTS), has been included following the latest update of the Investigator’s 
Brochure Addendum.  
Clinical Safety Experience with Ad26-based Vaccines in section 1.2.3 Potential Risks 
------------------------------------------------------ 
Rationale: history of thrombotic thrombocytopaenia syndrome (TTS) or heparin-induced 
thrombocytopaenia and thrombosis (HITT) have been mentioned as conditions that would increase the risk 
of an adverse outcome from participation in the study in exclusion criterion 10.  
4.2 EXCLUSION CRITERIA 
------------------------------------------------------ 
Rationale: Thrombotic events and symptomatic thrombocytopenia have been added to the protocol as 
Adverse Events of Special Interest. 
Protocol Versions 
DATE 
Original Protocol, Version 1.0 
30/10/2020 
Amendment 1, Protocol Version 2.0 
15/06/2021 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
Adverse Events of Special Interest in section 11.1.1 Adverse Event Definitions and Classifications 
APPENDIX 2 
------------------------------------------------------ 
Rationale: inconsistent information about retesting of values, rescreening and rescheduling of the 
vaccination (Day 1) have been corrected.  
8.1.3 Vaccination Period 
------------------------------------------------------ 
Rationale: Minor editorial changes have been made.  
Throughout the document  
------------------------------------------------------ 
Rationale: References have been updated.  
REFERENCES 
 
 


 
Page 8 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
SYNOPSIS 
TITLE  
An open label study to evaluate the safety and immunogenicity of an Ad26.ZEBOV booster dose in children 
previously vaccinated with the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen.  
RATIONALE 
Over 600 children have received the adenovirus serotype 26 expressing the Ebola virus Mayinga glycoprotein 
(Ad26.ZEBOV), modified Vaccinia Ankara Bavarian Nordic vector expressing multiple filovirus proteins (MVA-
BN-Filo) Ebola vaccine regimen in the EBL2002 and EBL3001 clinical trials. The vaccine regimen was well-
tolerated and highly immunogenic in children; however, the durability of vaccine-induced immune responses 
is not known. In adults previously vaccinated with the Ad26.ZEBOV and MVA-BN-Filo regimen, a booster 
vaccination with Ad26.ZEBOV was safe and induced a strong anamnestic response within seven days of the 
booster vaccination. It is important to establish if a booster dose of Ad26.ZEBOV is safe and immunogenic 
also in children, as this can guide the clinical use of the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen in this 
age group. For example, it could support the strategy of boosting immunised children at the start of an Ebola 
outbreak. 
STUDY OBJECTIVES AND HYPOTHESIS 
This study aims to evaluate the safety and immunogenicity of an Ad26.ZEBOV booster dose in healthy 
children who were previously (>2 years) vaccinated with the Ad26.ZEBOV (dose 1) followed by MVA-BN-Filo 
(dose 2) 56 days later, by monitoring adverse events (AEs) following the booster vaccination and by assessing 
binding antibody responses using the Filovirus Animal Non-Clinical Group (FANG) Enzyme-Linked 
Immunosorbent Assay (ELISA). 
Primary Objectives 
• 
To assess the safety and tolerability of a booster dose of Ad26.ZEBOV at a dose of 5x1010 viral 
particles (vp) in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen 
with a 56-day interval. 
• 
To assess vaccine-induced humoral immune responses to the Ebola virus glycoprotein (EBOV GP), as 
measured by FANG ELISA, at 7 and 21 days following a booster dose of Ad26.ZEBOV at a dose of 
5x1010 vp in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen with 
a 56-day interval. 
Exploratory Objectives 
• 
To assess neutralising antibody responses directed against the Ad26 vector before booster  
vaccination as measured by a virus neutralization assay (VNA). 
 
Hypothesis 
As this study is designed to provide descriptive information regarding safety and immunogenicity without 
formal treatment comparisons, no formal statistical hypothesis testing is planned. 
OVERVIEW OF STUDY DESIGN 
This is an open-label study evaluating the immune response to a booster dose of Ad26.ZEBOV administered 
to children who were previously vaccinated with Ad26.ZEBOV followed by MVA-BN-Filo 56 days later. Only 
subjects who received the Ad26.ZEBOV and MVA-BN-Filo regimen during their participation in the 


 
Page 9 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
VAC52150EBL3001 (EBOVAC-Salone) vaccine trial are eligible for enrolment in this study. Participants will 
be recruited in two age groups: children aged 4-11 years at the time of dose 1 vaccination and children 
aged 1-3 years at the time of dose 1 vaccination in the EBOVAC-Salone trial. Approximately 25 subjects will 
be enrolled in each of these two age groups.  
Parents/guardians will be asked to consent for the participation of their children in the study. Children aged 
7 years and older at the time of enrolment in this study will be asked to give positive assent for their 
participation. Participants will be followed up to 28 days after their booster vaccination. 
The Principal Investigator, together with the sponsor’s medical safety officer, will be responsible for the 
safety monitoring of the study.  
The study will be conducted in Kambia, Sierra Leone.  
SUBJECT POPULATION 
Potential participants must be healthy children (based on physical examination, medical history, a 
haematological assessment and clinical judgment) who received the Ad26.ZEBOV and MVA-BN-Filo vaccine 
regimen in the EBOVAC-Salone trial and were aged ≥1 to ≤11 years at the time of dose 1 vaccination. They 
must also be enrolled in the long-term follow-up study to the EBOVAC-Salone trial, VAC52150EBL3005 
(EBOVAC-Salone Extension study) but not in the immunogenicity subset. 
DOSAGE AND ADMINISTRATION 
A single dose of Ad26.ZEBOV at a dose of 5x1010 vp administered intramuscularly.  
SAFETY EVALUATIONS 
Solicited local (at the administration site) and systemic AEs will be assessed on the day of vaccination and 
using a diary for a period of seven days following the booster vaccination. Unsolicited AEs will be tracked for 
28 days following booster vaccination, while serious adverse events will be tracked for the duration of the 
study. 
IMMUNOGENICITY EVALUATIONS 
Blood will be drawn for assessments of immune responses at the time points indicated in the TIME AND 
EVENTS SCHEDULE. The site staff will perform sample collection and processing according to current versions 
of approved standard operating procedures.  
Future scientific research may be conducted to further investigate Ebola vaccine- and disease-related 
questions and to study other infections of public health importance in Sierra Leone and neighbouring 
countries. This may include the development of new, or the improvement of, existing techniques to 
characterize EBOV-directed immune responses or diagnostic tests. No additional samples will be taken for 
these analyses, however, residual samples from the study tests may be retained for these purposes and 
analysed after the end of the study. 
STATISTICAL METHODS 
The primary analysis will be done when all subjects have completed their 28-day post-booster visit or 
discontinued earlier. This analysis will include all available data up to this point.  
 
Sample Size Determination 
The sample size is a convenience sample and is not based on formal hypothesis testing considerations. 
SAFETY ANALYSES 
No formal statistical testing of safety data is planned. Safety data will be analysed descriptively by age 
group, 1-3 years and 4-11 years (age the participant was when they received dose 1 vaccination in the 
EBOVAC-Salone trial). 


 
Page 10 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
IMMUNOGENICITY ANALYSES 
No formal hypothesis on immunogenicity will be tested. Descriptive statistics (i.e. geometric mean and 95% 
confidence interval, as appropriate) will be calculated for continuous immunologic parameters at all 
available time points (i.e. Day 1, 8 and 22). Graphical representations of immunologic parameters will be 
made as applicable.  
 
Frequency tabulations will be calculated for discrete (qualitative) immunologic parameters (i.e. responder 
rate), as applicable. Responders rate defined as >2.5-time increase over baseline value (or lower limit of 
quantitation [LLOQ]) pre-dose 1 vaccination in the EBOVAC-Salone trial, will be calculated depending on 
availability of sample results from the EBOVAC-Salone trial. 
 
 
 
 
 
 
 


 
Page 11 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
TIME AND EVENTS SCHEDULE 
Study Procedures 
Screening 
(≤28 days)a 
Study Period 
Day 1 
Booster 
Days 2-7 
Day 8 
(+ 3 days)  
Day 22 
(± 3 days)  
Day 29b 
(± 3 days) 
Test of Understanding (TOU)c 
X 
 
 
 
 
 
Informed consent and assent (if applicable) d 
X 
 
 
 
 
 
Medical history and demographics  
X 
 
 
 
 
 
Inclusion/exclusion criteriae 
X 
X 
 
 
 
 
Urine pregnancy testf 
X 
Xg 
 
 
 
 
Physical examinationh 
X 
Xf 
 
X 
X 
Xb 
Vital signs i 
X 
X 
 
X 
X 
Xb 
Vaccine Administrationd 
 
X 
 
 
 
 
30 minutes post-vaccination observation j 
 
X 
 
 
 
 
Distribution of participant diary 
 
X 
 
 
 
 
Completion of the diary at homek 
 
X 
X 
 
 
 
Completion and review of the diary by study 
site personnel 
 
 
 
X 
 
 
Solicited adverse events recording j 
 
X 
X 
X 
 
 
Unsolicited adverse events recording 
 
From booster vaccination (Day 1) onwards until 28 days post booster  
Serious Adverse Events l 
Continuous 
Concomitant medications m 
X 
X 
X 
X 
X 
X 
Blood draw 
 
 
 
 
 
 
Haematology (i.e. full blood count) 
X 
 
 
X 
X 
 
Immunogenicity (serum) n  
 
X 
 
X 
X 
 
Approximate blood draw volumes 
 
 
 
 
 
 
Haematology: 2.0 mL per blood draw 
2.0 
 
 
2.0 
2.0 
 
Immunogenicity:  
Participants aged<6 years, 2.5 mL per blood 
draw 
Participcant aged 6 years and older, 5.0 mL 
per blood draw 
 
2.5 
5.0 
 
2.5 
5.0 
2.5 
5.0 
 
Total (Haematology + Immunogenicity) 
Participants aged <6 years 
Participant aged 6 years and older 
 
2.0 
2.0 
 
2.5 
5.0 
 
 
4.5 
7.0 
 
4.5 
7.0 
 
NOTE: In case of early withdrawal due to an adverse event (AE), the investigator or clinical designee will collect all information 
relevant to the AE and safety of the participant, and will follow the participant until resolution of the AE or until reaching a clinically 
stable endpoint. Parents/guardians who withdraw consent, or participants who wish to withdraw assent, will be offered an optional 
visit for safety follow-up (before the formal withdrawal of consent). Participants and/or parents/guardians have the right to refuse 
such a visit for themselves/their child. 
 
 
 
a Screening may be split into multiple days or visits. Retesting of values (eg, safety laboratory) that lead to exclusion is 
allowed once using an unscheduled visit during the screening period, provided there is an alternative explanation for the out 
of range value. The safety laboratory assessments at screening are to be performed within 28 days prior of the vaccination 
(including Day 1 before vaccination) and may be repeated if they fall outside this time window. If retesting is required, all 
screening procedures (except TOU) should be repeated 
b The Day 29 visit can be conducted via phone call if the participant is unable, or does not wish, to attend the clinic for this 
visit. If Day 29 visit is conducted over the phone, no physical exam and no vital signs will be collected or recorded for this 
visit.  
c The TOU should be administered to the parent or guardian who will provide consent, and will be administered after reading 
but before signing the informed consent form. 
d Informed consent must be obtained before any study-related activities are performed. 
e The investigators should ensure that all study enrollment criteria have been met at the end of the enrolment period and 
before the booster vaccination on Day 1. If a participant’s clinical status changes (including available laboratory results or the 
receipt of additional medical records) after screening but before Day 1 such that the participant no longer meets all eligibility 
criteria, then the participant will be excluded from participating in the study. 


 
Page 12 of 50 
VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
 
 
f For all adolescent girls who are ≥12 years of age at the time of screening.  
g Prior to study vaccine administration. 
h A full physical examination including also body weight and height will be conducted at screening. At following vists, a brief 
physical examination will be symptom-directed. Physical examination findings (i.e. abnormalities) prior to vaccination are to 
be recorded as medical history, after vaccination as adverse event(s).  
i Vital signs include blood pressure, pulse/heart rate (at rest), respiratory rate, and body temperature. 
j After the vaccination, participants will remain under observation at the study site for at least 30 minutes for presence of any 
acute reactions, or longer if deemed necessary by the investigator. Solicited local (at the injection site) and systemic adverse 
events and unsolicited adverse events emerging during the observation period will be recorded in the CRF. 
k Diaries will be completed at home by either a project field worker who will visit the participant during daily visits or by the 
parent/guardian, to document symptoms of solicited local (at the injection site) and systemic adverse events in the evening 
after the vaccination and then daily for the next 6 days at approximately the same time each day. Day 8 of the diary will be 
completed at the study clinic by a study doctor or nurse. 
l Serious adverse events and/or special reporting situations that are related to study procedures will be reported from the 
time a signed and dated ICF is obtained onwards until the end of the study. All other serious adverse events and/or special 
reporting situations will be reported from the day of vaccination onwards until the end of the study. 
mConcomitant therapies must be recorded from screening onwards until 28 days post-vaccination.  
nSerum aliquots for FANG ELISA at all timepoints (D1, D8, D22). Serum aliquot for Ad26 VNA testing at D1. 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
 
1 
INTRODUCTION  
Ebola viruses belong to the Filoviridae family and cause Ebola virus disease (EVD), which can induce severe 
haemorrhagic fever in humans and nonhuman primates. Case fatality rates in EVD range from 25% to 90% 
(average: 50%) according to the World Health Organisation (WHO). [1] 
Janssen Vaccines & Prevention B.V., in collaboration with Bavarian Nordic GmbH Denmark and in conjunction 
with an Innovative Medicines Initiative consortium led by the London School of Hygiene and Tropical 
Medicine (LSHTM), including the Institut National de la Santé et de la Recherche Médicale (INSERM), the 
University of Oxford and the Sierra Leone College of Medicine and Allied Health Sciences as partners, is 
investigating the potential of a prophylactic Ebola vaccine regimen (VAC52150) comprised of the following 
two candidate vaccines: 
Ad26.ZEBOV is a non-replicating monovalent vaccine expressing the full-length Mayinga glycoprotein (GP) of 
the Ebola virus (formerly known as Zaire ebolavirus), and is produced in the human cell line PER.C6®. 
MVA-mBN226B, further referred to as MVA-BN-Filo, is a non-replicating multivalent vaccine expressing the 
Sudan virus GP, the EBOV GP, the Marburg virus Musoke GP, and the Tai Forest virus (formerly known as Côte 
d’Ivoire ebolavirus) nucleoprotein, and is produced in chicken embryo fibroblast cells. The EBOV GP expressed 
by MVA-BN-Filo has 100% homology to the one expressed by Ad26.ZEBOV. 
The GP of the Ebola virus (EBOV) responsible for the 2013-2016 epidemic in West Africa had 97% homology 
to the EBOV GP used in this vaccine regimen. 
For the most up-to-date nonclinical and clinical information regarding Ad26.ZEBOV and MVA-BN-Filo, please 
refer to the latest versions of the Investigator’s Brochures (IBs) and Addenda (if applicable) [2, 3]. A brief 
summary of the nonclinical and clinical information available at the time of the protocol writing is provided 
below. 
1.1 
BACKGROUND 
The safety, reactogenicity, and immunogenicity of the Ad26.ZEBOV and MVA-BN-Filo vaccines have been 
evaluated in phase 1, 2 and 3 clinical trials. More than 100,000 participants, including children (1–17 years 
old) have received the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen in a number of completed and ongoing 
clinical studies and in a large scale vaccination campaign in Rwanda. Data from these studies have shown 
that the two-dose heterologous Ad26.ZEBOV, MVA-BN-Filo vaccine regimen is generally well tolerated and 
able to induce humoral immune responses persisting for at least two years in adults and for at least one year 
in children. 
 
Safety and immunogenicity of the Ad26.ZEBOV and MVA-BN-Filo vaccines in paediatric participants 
In the VAC52150EBL2002 (Burkina Faso, Cote D’Ivoire, Uganda, Kenya) and VAC52150EBL3001 (Sierra Leone) 
studies, three age cohorts (12–17 years, 4–11 years and 1–3 years) were vaccinated with Ad26.ZEBOV or 
control followed by MVA-BN-Filo or control, 28 or 56 days later. The control used for Dose 1 and 2 in 
VAC52150EBL2002 was a placebo (i.e. saline injection). In VAC52150EBL3001, Dose 1 consisted of a 
meningococcal conjugate vaccine (MenACWY) and Dose 2 of placebo.  
 
In these studies, the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen was well tolerated with no safety signals 
identified. Most solicited (local and systemic) AEs were mild to moderate. Overall, the frequency of grade 3 
solicited AEs was low (<2%) in all children assigned to Ad26.ZEBOV, MVA-BN-Filo regimen. The most 
frequently reported solicited local AE was injection site pain; frequency was similar between the age cohorts, 
but higher in participants assigned to active vaccines than to placebo. The most frequently reported solicited 
systemic AEs was headache in the 12–17 years and 4–11 years age cohorts, and decreased appetite in the 1-
3 years age cohort. The frequency of unsolicited AEs was higher in the 1-3 years age cohort compared to 
older children. The frequency of Grade 3 unsolicited AEs was similar across age cohorts. No serious adverse 
events (SAEs) were considered related to Ad26.ZEBOV or MVA-BN-Filo vaccines. The Ad26.ZEBOV, MVA-BN-


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
Filo vaccine regimen induced robust binding antibody responses, which persisted for at least up to 12 months. 
Younger children (1-3 years) had higher antibody responses compared to older children (4-11 years and 12-
17 years age cohorts). [4, 5] 
 
Safety and immunogenicity of an Ad26.ZEBOV booster dose in adult participants 
An Ad26.ZEBOV booster vaccination was given to 29 adult participants, who were vaccinated approximately 
2 years before with Ad26.ZEBOV followed by MVA-BN-Filo 56 days later in the VAC52150EBL3001 (EBOVAC-
Salone) trial in Sierra Leone. The booster vaccination was safe and induced a strong anamnestic response in 
96% of participants at seven days post-booster vaccination and in all 29 participants at 21 days post-booster 
vaccination. [6] 
 
1.2 
BENEFITS/RISKS OF PARTICIPATION 
1.2.1 
Potential Benefits 
The 2-dose Ebola vaccine regimen (Ad26.ZEBOV followed by MVA-BN-Filo 56 days later) has received 
marketing authorisations for prophylactic use in adults and children ≥1 years old in the European Union.[7] 
The marketing authorisation also includes the possibility for an Ad26.ZEBOV booster dose to be given to 
subjects who received the 2-dose regimen more than 4 months earlier and are at imminent risk of infection 
with Ebola virus.[8] This vaccine regimen was previously shown to provide protection in vaccinated non-
human primates against an EBOV challenge, which is fully lethal in unvaccinated control animals. Although 
clinical efficacy data are not available for this vaccine regimen, the marketing authorisation was granted on 
the basis of the potential clinical benefit induced by vaccination by correlating the magnitude of vaccine-
elicited immune parameters in non-human primates with those observed in vaccinated humans in phase 1, 
2 and 3 clinical studies.[9]  
If the Ad26.ZEBOV booster dose is shown to be safe and immunogenic in children as in adults, the participants 
enrolled in this study may benefit from the potential protective benefit of the booster vaccination in the case 
of a future exposure to EBOV.  
Participants will also benefit from clinical testing and physical examination; others may benefit from the 
knowledge that they may aid in the development of an Ebola vaccine.  
1.2.2 
Known Risks 
Ad26.ZEBOV  
The safety, reactogenicity, and immunogenicity of the Ad26.ZEBOV vaccine has been evaluated in a number 
of completed and ongoing clinical studies in adults and children. The vaccine was well tolerated, with no 
safety concerns identified. The vaccine mainly elicited some solicited local and systemic reactions, as 
expected with injectable vaccines, and there were no serious safety concerns in study participants. For 
details, see the safety data presented in Section 1.1. For the most up-to-date nonclinical and clinical 
information regarding Ad26.ZEBOV, refer to the latest versions of the IB and Addenda (if applicable) [2, 3]. 
1.2.3 
Potential Risks 
The following potential risks will be monitored during the study and are specified below:  
Risks Related to Vaccination 
In general, intramuscular (IM) injection may cause local itching, warmth, pain, tenderness, erythema, 
swelling, arm discomfort or bruising of the skin at vaccine injection sites. 
Participants may exhibit general signs and symptoms associated with the administration of a vaccine, 
including fever, chills, rash, nausea/vomiting, general itching, headache, myalagia, arthralgia, and fatigue. 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
Young children may also experience loss of appetite, diarrhoea, decreased activity/lethargy, and 
irritability/crying. These side effects will be monitored, but are generally short-term and do not require 
treatment. 
Participants may have an allergic reaction to the vaccine. An allergic reaction may cause a rash, hives or even 
anaphylaxis. Severe reactions are rare. Medications must be available in the clinic to treat serious allergic 
reactions. 
Risks from Blood Draws 
As with all clinical studies requiring blood sampling, there are risks associated with venipuncture and multiple 
blood sample collection. Blood drawing may cause pain, tenderness, bruising, bleeding, dizziness, vaso-vagal 
response, syncope, and, rarely, infection at the site where the blood is taken. The total blood volume to be 
collected is considered to be an acceptable amount of blood over this time period from the population in this 
study (see Section 15.1). 
Concomitant Vaccination 
Concomitant vaccination might have an influence on both the safety profile and immunogenicity of 
Ad26.ZEBOV. Likewise, the study intervention might have an influence on both the safety profile and 
immunogenicity of any concomitant vaccination. Therefore, a participant should not receive a live-
attenuated vaccine from 30 days before the vaccination until 30 days after the vaccination unless a vaccine 
preventable disease such as measles emerges which would warrant administration of live-attenuated 
vaccines. Immunizations with inactivated vaccines should be administered at least 15 days before or after 
administration of any study intervention in order to avoid any potential interference with the efficacy of the 
routine immunizations or the interpretation of immune responses to study intervention, as well as to avoid 
potential confusion with regard to attribution of adverse reactions. However, if a vaccine is indicated in a 
post-exposure setting (e.g., rabies or tetanus), it must take priority over the study intervention. Otherwise, a 
participant will not postpone, forego or delay the receipt of any recommended vaccine according to local 
schedules (e.g., Expanded Program on Immunization [EPI] schedule according to the WHO regional office for 
West Africa).  
Clinical Safety Experience with Ad26-based Vaccines 
Thrombosis in combination with thrombocytopenia (thrombosis with thrombocytopenia syndrome [TTS]), 
in some cases accompanied by internal bleeding, has been observed very rarely following vaccination with 
the Janssen COVID-19 (Ad26.COV2.S) vaccine. Reports include severe cases of venous thrombosis at unusual 
sites such as cerebral venous sinus thrombosis (CVST), splanchnic vein thrombosis and arterial thrombosis, 
in combination with thrombocytopenia. The associated symptoms began approximately 1 to 2 weeks after 
vaccination, mostly in women under 60 years of age. Thrombosis in combination with thrombocytopenia 
can be fatal. The exact pathophysiology of TTS is unclear. This event has not been observed to date with 
any other Janssen Ad26-based vaccines. Participants should be instructed to seek immediate medical 
attention if they develop symptoms such as shortness of breath, chest pain, leg swelling, persistent 
abdominal pain, severe or persistent headaches, blurred vision, skin bruising or petechiae beyond the site 
of vaccination. 
Vaccine Induced Seropositivity 
The potential of a participant becoming polymerase chain reaction (PCR)-positive after vaccination was 
assessed in study VAC52150EBL1002. The risk for false positives is low and expected to decrease rapidly over 
time after administration of Ad26.ZEBOV. 
Uninfected participants in Ebola vaccine studies may develop Ebola-specific antibodies as a result of an 
immune response to the candidate Ebola vaccine, referred to as vaccine induced seropositivity (VISP). These 
antibodies may be detected in Ebola serologic tests, causing the test to appear positive even in the absence 
of actual Ebola infection. VISP may become evident during the study, or after the study has been completed.  
Unknown Risks 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
There may be other risks that are not known. If any significant new risks are identified, the Principal 
Investigator and participants (parents/guardians) will be informed. 
1.2.3 
Overall Benefit/Risk Assessment 
Based on the available data and proposed safety measures, the overall benefit/risk assessment for this 
clinical study is considered acceptable for the following reasons: 
• 
To date, safety data from the studies in the clinical development program revealed no significant 
safety issues (see Section 1.1). Further experience from Ad26.ZEBOV Filo will be gained from 
currently ongoing clinical studies. 
• 
For all participants, there are pre-specified pausing rules that would result in pausing of further 
vaccination if predefined conditions occur, preventing exposure of new participants to study 
intervention until an independent medical reviewer evaluates all safety data (see Sections 3.1, 8.3, 
10.7). 
• 
Only participants who meet all inclusion criteria and none of the exclusion criteria (specified in 
Section 4) will be allowed to participate in this study. The selection criteria include adequate 
provisions to minimize the risk and protect the well-being of participants in the study. 
• 
Several safety measures are included in this protocol to minimize the potential risk to participants, 
including the following: 
- Participants will remain at the site for at least 30 minutes after each vaccination to monitor the 
development of any acute reactions, or longer if deemed necessary by the investigator (e.g. in 
case of grade 3 AEs). Refer to Section 6 for more information on emergency care.  
- Safety evaluations (physical examinations and vital sign measurements) will be performed at 
scheduled visits during the study, as indicated in the TIME AND EVENTS SCHEDULE. 
- The investigator or clinical designee will document unsolicited AEs from the vaccination (Day 1) 
onwards until 28 days post-vaccination (Day 29). The investigator or clinical designee will 
document serious AEs and/or special reporting situations that are related to study procedures 
from the time a signed and dated informed consent form (ICF) is obtained onwards until the end 
of the study. 
- Any clinically significant abnormalities (including those persisting at the end of the study/early 
withdrawal) will be followed by the investigator until resolution or until a clinically stable 
endpoint is reached. 
- If acute illness (excluding minor illnesses such as diarrhoea or mild upper respiratory tract 
infection) or axillary temperature ≥38°C is present at the scheduled time for vaccination, the 
participant may be rescheduled for vaccination at a later time point within the window allowed 
for screening, or be withdrawn from vaccination at the discretion of the investigator and after 
consultation with the sponsor (see Section 6.1). 
1.3 
OVERALL RATIONALE FOR THE STUDY 
Over 600 paediatric participants have received the Ad26.ZEBOV, MVA-BN-Filo Ebola vaccine regimen in the 
EBL2002 and EBL3001 clinical trials. [4, 5] The vaccine regimen was well-tolerated and highly immunogenic 
in children, however, the durability of the immune response is not known. In adults previously vaccinated 
with the Ad26.ZEBOV and MVA-BN-Filo regimen, an Ad26.ZEBOV booster vaccination was safe and induced 
a strong anamnestic response. [6] It is important to establish the safety and immunogenicity of a booster 
dose of Ad26.ZEBOV in children, as this can guide the clinical use of the vaccine regimen in this group. For 
example, it could support the strategy of boosting immunised children at the start of an Ebola outbreak. 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
2 
OBJECTIVES, ENDPOINTS, AND HYPOTHESIS 
2.1 
OBJECTIVES AND ENDPOINTS 
Primary Objectives 
• 
To assess the safety and tolerability of a booster dose of Ad26.ZEBOV at a dose of 5x1010 viral 
particles (vp) in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen 
with a 56-day interval. 
• 
To assess vaccine-induced humoral immune responses to the Ebola virus glycoprotein (EBOV GP), as 
measured by FANG ELISA, at 7 and 21 days following a booster dose of Ad26.ZEBOV at a dose of 
5x1010 vp in children previously vaccinated with the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen with 
a 56-day interval. 
Exploratory Objectives 
• 
To assess neutralising antibody responses directed against the Ad26 vector before booster  
vaccination as measured by a virus neutralization assay (VNA). 
 
2.2 
HYPOTHESIS 
As this study is designed to provide descriptive information regarding the safety and immunogenicity of a 
booster dose of Ad26.ZEBOV in children, no formal statistical hypothesis testing is planned. 
 
3 
STUDY DESIGN 
3.1 
OVERVIEW OF STUDY DESIGN 
This is an open label phase 2 study evaluating the immune response to a booster dose of Ad26.ZEBOV 
administered to children who were previously vaccinated with Ad26.ZEBOV followed by MVA-BN-Filo 56 days 
later. Only subjects who received the Ad26.ZEBOV and MVA-BN-Filo regimen during their participation in the 
EBOVAC-Salone trial are eligible for enrolment in this study. Participants will be recruited in two age groups: 
children aged 4-11 years at the time of dose 1 vaccination and children aged 1-3 years at the time of dose 1 
vaccination. Up to 25 subjects will be enrolled in each age group.  
The study will consist of a screening period of up to 28 days, a booster vaccination (Day 1) and a post-booster 
vaccination follow-up period until 28 days post-vaccination (Day 29). 
Parents/guardians will be asked to consent for the participation of their children in the study. Children aged 
7 years and older at the time of enrolment in this study will be asked to give positive assent for their 
participation.  
After informed consent and assent (if applicable) have been obtained, investigators should ensure that all 
study eligibility criteria have been met prior to the study vaccination on Day 1 (see list of inclusion and 
exclusion criteria in Section 4). Eligibility will be based on medical history, physical examinations (including 
body height and weight), vital sign measurements, a haematological (i.e. full blood count) assessment, and 
urine pregnancy test in all adolescent girls who are ≥12 years of age at the time of screening.  
After vaccination, participants will remain under observation at the study site for at least 30 minutes for 
surveillance for any acute reactions, or longer if deemed necessary by the investigator. Following the 
vaccination, any unsolicited, solicited local or systemic AEs, and vital signs will be documented by study-site 
personnel at the end of this observation period. 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
The participant’s parent/guardian will be given a thermometer, ruler and participant diary with instructions 
for the proper recording of events occurring after the booster vaccination. Diaries will be completed at home 
by either a project field worker who will visit the participant or by the parent/guardian to document solicited 
local (at the injection site) and systemic AEs and body temperature, beginning on the evening of the 
vaccination, and then daily for the next 6 days. Temperatures should be taken at approximately the same 
time each day, preferably in the evening and additionally whenever the child feels warm. Study-site 
personnel will collect, review, and complete the participant diary information at the 7-day post-vaccination 
visit (Day 8). 
Unsolicited AEs will be recorded from the booster vaccination (Day 1) onwards until 28 days post-booster 
vaccination (Day 29). SAEs and/or special reporting situations that are related to study procedures will be 
reported from the time a signed and dated ICF is obtained onwards until the end of the study (Day 29).  
Blood samples will be collected for immunogenicity assessments at Day 1 (i.e. the baseline sample before 
vaccination), 7 days post-vaccination (Day 8), and 21 days post-dose-vaccination (Day 22). At Day 8 and Day 
22, a sample for haematology will also be collected.  
Participants will exit the study after 28 days post-vaccination (Day 29). The study is considered completed at 
the site at final database lock, which will occur after the last participant has completed the last study visit or 
left the study. 
 
4 
PARTICIPANT POPULATION  
The study will be open to healthy children (based on physical examination, medical history, a haematological 
assessment and clinical judgement) who received the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen in the 
EBOVAC-Salone trial. The children must be enrolled in the long-term follow-up study to the EBOVAC-Salone 
trial, the VAC52150EBL3005 (EBOVAC-Salone Extension) study but not in the immunogenicity subset.  
The inclusion and exclusion criteria for enrolling participants in this study are described in the following two 
subsections. If there is a question about the inclusion and exclusion criteria, the investigator must consult 
with the appropriate sponsor representative and resolve any issues before enrolling a participant in the 
study. Waivers are not allowed. 
4.1 
INCLUSION CRITERIA 
Each potential participant must satisfy all of the following criteria to be enrolled in this current study: 
1. Child must be enrolled in the VAC52150EBL3005 (EBOVAC-Salone Extension) study but not in the 
immunogenicity subset of EBOVAC-Salone Extension study.  
2. Child must be a former participant in the VAC52150EBL3001 (EBOVAC-Salone) trial, and have received 
Ad26.ZEBOV (dose 1) vaccination followed by the MVA-BN-Filo (dose 2) vaccination within the EBOVAC-
Salone trial window for dose 2 vaccination. 
3. Child must have been aged 1 to 11 years old at the time of dose 1 vaccination in the EBOVAC-Salone 
trial. 
4. The parent/guardian must consent for their child to participate in the VAC52150EBL2011 study by 
signing (or thumb printing, if illiterate) an ICF, indicating that he or she understands the purpose of, and 
procedures required for, the study, understands the potential risks and benefits of the study, and is 
willing to allow their child to participate in the study. If the parent/guardian cannot read or write, the 
procedures must be explained, and informed consent must be witnessed by a literate third party not 
involved in the conduct of the study. Children aged 7 years and older will be asked to give positive assent 
for their participation in the study and the assent procedure must be witnessed by an adult, literate 
parent/guardian/third party not involved in the conduct of the study, and documented. 
5. The parent/guardian is willing/able to ensure that their child adheres to the prohibitions and restrictions 
specified in this protocol (see Section 4.3) 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
6. Child must be healthy in the investigator’s clinical judgement (and the parent/guardian’s judgement) on 
the basis of medical history, physical examination, vital signs, and a haematological assessment (i.e. full 
blood count) performed at screening. Subjects must meet the following haematology parameters within 
28 days before Day 1:  
• 
Haemoglobin ≥8.0 g/dL for children aged 1 to <5 years, ≥9g/dL for children aged 5 or older 
• 
Platelet count ≥100 x 109/L 
• 
White blood cell count ≥5.0 x 109/L 
Note: The haematological assessment at screening is to be performed within 28 days prior to vaccination 
on Day 1 and may be repeated if it falls outside this time window. 
Note: If haematological screening assessment is out of range and deemed clinically significant, repeating 
screening test to assess eligibility is permitted once during the screening period, using an unscheduled 
visit. 
7. Adolescent girls who have started their menstrual periods and/or are ≥12 years of age at the time of 
screening, must have a negative urine β-hCG pregnancy test at screening and immediately prior to the 
booster vaccination on Day 1.  
8. The parent/guardian is available and willing to have their child participate for the duration of the study 
visits. 
9. The parent/guardian must have the means to be contacted. 
10. The parent/guardian must pass the Test of Understanding (TOU).  
Note: If the parent/guardian fails the TOU on the first attempt, he/she must be retrained on the purpose 
of the study and must take the test again (2 repeats are allowed). If he/she fails on the third attempt, 
he/she should not continue with enrolling or consenting procedures. 
 
4.2 
EXCLUSION CRITERIA 
Any potential participant who meets any of the following criteria will be excluded from participating in the 
study: 
1. Participants in the EBOVAC-Salone trial who were allocated to the control arm receiving the WHO-
prequalified Meningococcal Group A, C, W135 and Y conjugate vaccine. 
2. Participants in the EBOVAC-Salone trial who were age 12 years and older at the time of dose 1 
vaccination. 
3. Known allergy or history of anaphylaxis or other serious adverse reactions to vaccines or vaccine 
products (including any of the constituents of the study vaccine, e.g., polysorbate 80, 
ethylenediaminetetraacetic acid or L-histidine for Ad26.ZEBOV vaccine), including known allergy to 
chicken or egg proteins and aminoglycosides (gentamicin). 
4. Presence of acute illness (this does not include minor illnesses such as mild diarrhoea or mild upper 
respiratory tract infection) or axillary temperature ≥38ºC on Day 1. Participants with such symptoms 
will be excluded from enrolment at that time but may be rescheduled for enrolment at a later date 
within the screening window. 
5. Clinically significant history of skin disorder (e.g., psoriasis, contact dermatitis), allergy, symptomatic 
immunodeficiency, cardiovascular disease, respiratory disease, endocrine disorder, liver disease, renal 
disease, gastrointestinal disease, neurological illness as judged by the investigator or other delegated 
individual. 
 
6. Adolescent girls who are known to be pregnant or breastfeeding at screening.  


 
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7. Received a blood transfusion or other blood products within 8 weeks of vaccination day. 
8. Children who have been vaccinated with live-attenuated vaccines within 30 days before the study 
vaccination, and with inactivated vaccine within 15 days before the study vaccination. 
9. Children who, in the opinion of the investigator, are unlikely to adhere to the requirements of the study 
or are unlikely to complete the vaccination and observation 
10. Any other finding which in the opinion of the investigator or other delegated individual would increase 
the risk of an adverse outcome from participation in the study, e.g. history of thrombotic 
thrombocytopaenia syndrome (TTS) or heparin-induced thrombocytopaenia and thrombosis (HITT). 
NOTE: Investigators should ensure that all study eligibility criteria have been met prior to the study 
vaccination on Day 1. If a subject’s clinical status changes (including receipt of additional medical records 
or available laboratory results) after enrolment but before the vaccination (Day 1) so that he or she no 
longer meets all eligibility criteria, then the subject should be excluded from participation in the study. 
4.3 
PROHIBITIONS AND RESTRICTIONS 
The parent/guardian must be willing and able to adhere to the following prohibitions and restrictions during 
the course of the study for their child to be eligible for participation: 
 
5 
INTERVENTION ALLOCATION AND BLINDING 
5.1 
BLINDING 
As this is an open-label study, blinding procedures are not applicable.  
 
6 
DOSAGE AND ADMINISTRATION 
All participants will receive the Ad26.ZEBOV vaccine, at a concentration of 5x1010vp, as a 0.5 millilitres (mL) 
IM injection into the deltoid muscle. 
Study intervention will be prepared by a pharmacist or qualified staff member with primary responsibility for 
study intervention preparation and dispensing of the vaccine.  
Ad26.ZEBOV will be administered as a 0.5 mL IM injections in the deltoid muscle, by a study intervention 
administrator. The injection site should be free from any injury, local skin conditions, or other issue that might 
interfere with the evaluation of local reactions. No local or topical anaesthetic will be used prior to the 
injection. 
Participants will remain at the site for at least 30 minutes after the vaccination for the detection  of any acute 
reactions, or longer if deemed necessary by the investigator (e.g. in case of a grade 3 AEs). As with any 
vaccine, allergic reactions following vaccination with the study intervention are possible. Therefore, 
appropriate drugs and medical equipment to treat acute anaphylactic reactions must be immediately 
available, and a medically qualified member of study-site personnel trained to recognize and treat 
1. 
In case of a new Ebola outbreak: participants and their parent/guardian must not travel to an 
area with an Ebola outbreak while the participant is enrolled in the study from the start of 
screening onwards until the last study visit. If applicable, any travelling to an area with an 
Ebola outbreak should be documented in the case report form (CRF). The date of travel and 
the destination should be clearly identified. 
2. 
Ensure that their child does not receive any experimental medication (including experimental 
vaccines other than the study intervention) as described in Section 7. 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
anaphylaxis must be present in the clinic during the entire vaccination procedure and post-vaccination 
monitoring period. 
The investigator must provide emergency care as needed for any participant who experiences a life-
threatening event. The site will have facilities, equipment and the ability to manage an anaphylactic reaction. 
If additional therapy is required, the investigator will arrange for transport to the closest appropriate facility 
for continuing care. 
The Site Investigational Product Procedure Manual specifies the procedures for administration of the study 
intervention. 
Ad26.ZEBOV will be manufactured by Janssen Vaccines & Prevention B.V. and provided under the 
responsibility of the sponsor, LSHTM. Please refer to the IB for a list of excipients. 
6.1 
CRITERIA FOR POSTPONEMENT OF VACCINATION 
A participant will not be given any vaccination if he/she experiences any of the following events at the 
scheduled time for vaccination:  
- 
An acute illness at the time of vaccination (this does not include minor illnesses such as diarrhoea or 
mild upper respiratory tract infection). 
- 
An axillary temperature ≥38°C at the time of vaccination. 
Participants experiencing any of these events may be rescheduled for vaccination at a later time point within 
the window allowed for screening, or be withdrawn from vaccination at the discretion of the investigator and 
after consultation with the sponsor. 
 
7 
CONCOMITANT THERAPY 
Concomitant therapies that the participant is taking at the time of screening must be recorded in the CRF at 
screening. 
Should immunisation with an inactivated vaccine be required, it should be administered at least 15 days 
before or after administration of the study intervention in order to avoid any potential interference in efficacy 
of the routine immunizations or the interpretation of immune responses to study intervention, as well as to 
avoid potential confusion with regard to attribution of adverse reactions. However, if a vaccine is indicated 
in a post-exposure setting (e.g., rabies or tetanus), it must take priority over the study intervention. A 
participant will not postpone, forego, or delay the receipt of any recommended vaccine according to local 
schedules (e.g. EPI schedule according to the WHO regional office for West Africa). 
Analgesic/antipyretic medications and non-steroidal anti-inflammatory drugs may be used post-vaccination 
in case of medical need (e.g., fever or pain). Use of these medications as routine prophylaxis prior to study 
intervention administration is not recommended. The use of these medications must be documented.  
Concomitant therapies must be recorded from screening onwards until 28 days post-vaccination.  
Use of any experimental medication (including experimental vaccines other than the study intervention) 
during the study is not allowed. 
The sponsor must be notified in advance (or as soon as possible thereafter) of any instances in which 
prohibited therapies are administered. 
 
8 
STUDY EVALUATIONS 
Prior to any study-related activities being performed, participants, or participant’s parent/guardian, must 
have signed a study ICF (see Section 15.2.3 Consent) 


 
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8.1 
STUDY PROCEDURES 
8.1.1 
Overview 
The TIME AND EVENTS SCHEDULE summarises the frequency and timing of all study procedures, which are 
provided in the following sections. Additional unscheduled study visits may be required if, in the 
investigator’s opinion, further clinical or laboratory evaluation is needed. 
Visit Windows 
Visit windows are provided in the TIME AND EVENTS SCHEDULE. If a participant did not receive the study 
intervention on the planned day of vaccination, the timings of the next visits post-vaccination will be 
determined relative to the actual day of vaccination. The participant should be encouraged to come within 
these required windows. 
Blood Sampling Volume 
The approximate blood volumes collected in this study are indicated in the TIME AND EVENTS SCHEDULE. 
These study-related blood volumes obtained at each study visit (including any losses during phlebotomy) will 
not exceed 3% of the total blood volume. The total volume of blood is estimated at 80 to 90 mL/kg body 
weight; 3% is 2.4 mL blood per kg body weight. [10] The allowable blood volume calculations are based on 
the 3rd percentile for growth charts for 1 to 5-year-old [11] and 6- to 10-year-old children. [12] 
Repeat or unscheduled samples may be taken for safety reasons or for technical issues with the samples. 
8.1.2 
Screening Period 
After signing and dating the ICF (see Section 16.2.3) and up to 28 days before Day 1 (day of vaccination), 
screening assessments will be performed as indicated in the TIME AND EVENTS SCHEDULE. Screening may 
be split into multiple days or visits.  
Only participants complying with the criteria specified in Section 4 will be included in the study. The 
investigator will provide detailed information on the study to the participant and his/her parent/guardian 
will obtain written informed consent prior to study participation of the child. 
After reading but before signing the ICF, the TOU will be administered to the parent/guardian. If the 
parent/guardian fails the TOU, they may repeat the test twice (and have to pass the third time for their child 
to be eligible) (for details, see Section 15.1). 
The overall eligibility of the child to participate in the study will be assessed once all the screening results are 
available. Retesting of values (i.e. haematology) that lead to exclusion is allowed once using an unscheduled 
visit. If rescreening is required, all screening procedures (except TOU) should be repeated. Rescreening is 
allowed one time at the discretion of the study doctor. Study participants who qualify for inclusion will be 
contacted and scheduled for vaccination within 28 days. 
A serum sample will be taken before vaccination at D1, to serve as a pre-vaccination baseline sample for 
immunogenicity assessments (see Section 8.4). 
8.1.3 
Vaccination Period 
If eligible, the participant will be invited to come for the vaccination visit (Day 1). The investigator should 
ensure that all eligibility criteria have been met during the screening period. If a participant’s clinical status 
changes (including available laboratory results or receipt of additional medical records) after screening but 
before the vaccination (Day 1) such that he/she no longer meets all eligibility criteria, then the participant 
should not be enrolled. Retesting of values and rescreening might be considered (see Section 8.1.2 for further 
information). Rescheduling of the vaccination (Day 1) visit, if the participant has an acute illness, is allowed 
within the screening window. 
Before vaccination, a brief physical examination and measurement of vital signs will be performed.  
Participants will be vaccinated as described in Section 6. After vaccination, participants will remain under 
observation at the study site for at least 30 minutes for surveillance for any acute reactions, or longer if 
deemed necessary by the investigator. Following the vaccination, any unsolicited, solicited local or systemic 
AEs, and vital signs will be documented by study-site personnel at the end of this observation period.  


 
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Upon discharge from the site, the parent/guardian will be provided with a thermometer (to measure body 
temperature), a ruler (to measure local injection site reactions), and a participant diary to record body 
temperature and solicited local (at the injection site) and systemic symptoms and will be trained on how to 
collect this information. Symptoms of solicited local and systemic AEs will be collected in the diary in the 
evening after the vaccination and then daily for the next 7 days at approximately the same time each day. 
Diaries will be completed at home by either a project field worker who will visit the participant during daily 
visits or by the parent/guardian and checked by a project field worker. The investigator or clinical designee 
will review information from the participant’s diary. 
Participants will come to the site 7 days after the vaccination (Day 8) as indicated in the TIME AND EVENTS 
SCHEDULE. The participant’s diary will be reviewed by study-site personnel. The investigator will examine the 
injection site for occurrences of erythema, swelling, or tenderness at these visits in order to complete the 
relevant parts of the CRF.  
Unsolicited AEs will be reported from the first day of vaccination until 28 days post-vaccination (Day 29). 
SAEs and/or special reporting situations that are related to study procedures will be reported from the time 
a signed and dated ICF is obtained onwards until the end of the study (Day 29). All other SAEs and/or special 
reporting situations will be reported from the day of vaccination onwards until the end of the study (Day 29). 
Participants will come to the site 7 and 21 days after the vaccination (Day 8 and Day 22) for safety and 
immunogenicity assessments. Please refer to 8.4 for details on the immunogenicity evaluations. 
The parent/guardian will be instructed to contact the investigator anytime during the course of the study if 
their child experiences any AE or intercurrent illness that they perceive as relevant and/or can be possibly 
related to study intervention in their opinion.  
When an enrolled participant completes or withdraws from the study, the investigator will complete an end-
of-study form for the individual participant and provide a specific date for the end-of-study observation(s). 
When a participant withdraws before completing the study, the reason for withdrawal (if available) will be 
documented in the CRF and in the source documents. 
Participants who appear to be lost to follow-up will be contacted, as per local practice, and the data recorded 
in the CRF. Participants lost to follow-up will have all data included for analysis. 
8.2 
PROCEDURES IN CASE OF A STUDY PAUSE 
A study pause can affect participants that are awaiting the booster vaccination. After approval is granted to 
restart the study, participants who are awaiting the booster vaccination and whose screening period is longer 
than the protocol-defined 28 days as a result of a study pause, will be allowed to rescreen once (following 
the screening procedures described in Section 8.1.2, excluding TOU). Participants that are rescreened due to 
a pause must have new safety assessments (including full blood count, physical examination, and vital signs) 
within 28 days of the booster vaccination. The TOU does not need to be repeated. After screening, these 
participants will follow the same study procedures as those participants who were unaffected by a study 
pause (described in Section 8.1.3). 
8.3 
SAFETY EVALUATIONS 
8.3.1 
Safety Assessments 
The Principal Investigator, together with the Chief Investigator and the sponsor’s medical safety officer or 
delegate, will be responsible for the safety monitoring of the study, and will halt vaccination of further 
participants in case any of the pre-specified pausing rules described in Section 8.3.2 have been met. Further 
safety measures with regards to vaccination are described in Section 6.1. 
An independent medical reviewer will be appointed by the sponsor before the start of the study to perform 
review of the safety data during the study. Details regarding this role are provided in Section 10.7. 
Symptoms of solicited local and systemic AEs will be collected in the diary in the evening after the vaccination 
and then daily for the next 7 days. Unsolicited AEs will be collected from vaccination until 28 days post-
vaccination (Day 29). Serious adverse events and/or special reporting situations that are related to study 


 
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procedures will be reported from the time a signed and dated ICF is obtained onwards until the end of the 
study (Day 29). All other SAEs and/or special reporting situations will be reported from the day of vaccination 
onwards until 28 days post-vaccination. 
Any clinically relevant changes must be recorded on the AE section of the CRF.  
Any clinically significant abnormalities persisting at the end of the study/early withdrawal will be followed by 
the investigator until resolution or until a clinically stable condition is reached.  
All AEs will be coded for severity according to the criteria presented in Section 11.1.3. 
The study will include the following evaluations of safety and reactogenicity according to the time points 
provided in the Time and Events Schedule: 
Adverse Events 
Adverse events will be reported as specified in Section 11.3.1. 
Solicited Adverse Events 
After vaccination, participants will remain under observation at the study site for at least 30 minutes for 
surveillance for  any acute reactions, or longer if deemed necessary by the investigator. Symptoms of solicited 
local and systemic AEs will be collected in the diary in the evening after the vaccination and then daily for the 
next 7 days. Diaries will be completed at home by either a project field worker who will visit the participant 
during daily visits or by the parent/guardian and checked by a project field worker for 6 days. Day 8 of the 
diary will be completed by study site staff at the clinic. Diary information will be transcribed by the study 
personnel in the diary CRF pages. Once a solicited symptom from a diary is considered to be of severity Grade 
1 or above, it will be referred to as a solicited AE. 
Solicited Injection Site (Local) Adverse Events 
Parents/guardians (or the project field worker) will be asked to note in the diary occurrences of tenderness, 
erythema and swelling at the study intervention injection site daily for 6 days post-vaccination (day of 
vaccination and the subsequent 6 days). The extent (largest diameter) of any erythema and swelling should 
be measured (using the ruler supplied) and recorded daily. Day 8 of the diary will be completed by study staff 
at the site clinic. 
• 
Injection Site Tenderness 
Injection site tenderness is a painful sensation localized at the injection site upon palpation or 
movement of the limb. Due to the subjective nature of the reaction, the severity assessment of 
tenderness is self-reported (if a participant is unable to provide self-report, other reporters include 
parent/care giver or health care provider).  
• 
Injection Site Erythema 
Injection site erythema is a redness of the skin caused by dilatation and congestion of the capillaries 
localized at the injection site. It can best be described by looking and measuring. 
• 
Injection Site Swelling 
Injection site swelling is a visible enlargement of the site of injection. It may be either soft (typically) 
or firm (less typical).  
Note: Any other injection site events not meeting the above case definitions should be reported 
separately as unsolicited AEs. 
Solicited Systemic Adverse Events 
Parents/guardians will be instructed on how to record daily temperature using a thermometer provided for 
home use. The axillary temperature of the participant should be recorded in the diary in the evening of the 
day of vaccination, and then daily for the next 6 days at approximately the same time each day. The 
participant’s temperature on Day 8 will be taken by study staff at the site clinic. If more than one 
measurement is made on any given day, the highest temperature of that day will be used in the CRF. 


 
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In this protocol, fever is defined as an endogenous elevation of body temperature ≥38° C, as recorded in at 
least one measurement.  
Parents/guardians will also be instructed on how to note daily in the diary symptoms for 6 days post-
vaccination (day of vaccination and the subsequent 6 days) any of the following events:  
• 
body temperature 
• 
fatigue/malaise 
• 
chills 
• 
headache 
• 
nausea/vomiting 
• 
muscle pain 
• 
joint pain 
• 
loss of appetite, in young children 
Symptoms 7 days post vaccination (Day 8) will be assessed and recorded by study staff at the site clinic. 
Physical Examination 
A full physical examination will be performed at the screening visit. At subsequent study visits a brief, 
symptom-directed examination will be performed based on any clinically relevant issues, clinically relevant 
symptoms and medical history. The symptom-directed physical examination may be repeated if deemed 
necessary by the investigator. Physical examinations will be performed by the investigator or by a designated 
medically-trained clinician. Physical examination findings (i.e. abnormalities) prior to vaccination (Day 1) are 
to be recorded as medical history, but after vaccination as an AE. 
Vital Signs 
Axillary temperature, blood pressure, pulse/heart rate (beats per minute), and respiratory rate (breaths per 
minute) will be assessed.  
Blood pressure and pulse/heart rate measurements will be assessed with a completely automated device. 
Manual techniques will be used only if an automated device is not available. Pulse/heart rate measurements 
should be preceded by at least 5 minutes of rest in a quiet setting without distractions.  
Clinical Laboratory Tests 
A haematological assessment (i.e., full blood count) will be performed by the local laboratory at the time 
points indicated in the TIME AND EVENTS SCHEDULE. The investigator must review the laboratory report, 
document this review, and record any clinically relevant changes on the AE page and/or the screening page 
of the CRF. Laboratory reports must be filed with the source documents. 
A full blood count will include:  
-haemoglobin 
-haematocrit 
-red blood cell (RBC) count 
-white blood cell (WBC) count with differential 
-platelet count 
Note: a WBC evaluation may include any abnormal cells, which will then be reported by the laboratory. An RBC 
evaluation may include abnormalities in the RBC count, or RBC morphology, which will then be reported by the
laboratory. 
8.3.2 
Study Pausing Rules 
The investigators and the sponsor’s medical safety officer or delegate will review the safety of enrolled 
participants on an ongoing basis and will halt vaccination of further participants in case any of the pre-
specified pausing rules described in this section are met. The sponsor’s medical safety officer or delegate will 
be involved in all discussions and decisions.  


 
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If any of the following events occur in any participant who received the study intervention, the site 
investigator will halt the vaccination of further participants in the study, and the sponsor’s medical safety 
officer or delegate will be notified immediately: 
1. 
Death of a participant considered related to the study intervention, or if the causal relationship to 
the study intervention cannot be excluded; OR 
 
Note: All cases of death will be sent to the independent medical reviewer for information. Upon their 
review, the independent medical reviewer may then also advise whether a study pause is required. 
2. 
One or more participants experience a SAE (solicited or unsolicited) that is determined to be related 
to study intervention; OR 
3. 
One or more participants experience anaphylaxis or generalized urticaria within 24 hours of 
vaccination, clearly not attributable to causes other than vaccination with the study intervention. 
To enable prompt response to a situation that could trigger pausing rules, the investigator should notify the 
sponsor’s medical safety officer or delegate immediately and no later than 24 h after becoming aware of any 
related AE of grade 3 or above AND update the CRF with relevant information on the same day the AE 
information is collected. A thorough analysis of all grade 3 cases will be carried out by the sponsor’s medical 
safety officer or designee, irrespective of whether the criteria for pausing the study are met. Based on the 
pausing criteria, the sponsor’s medical safety officer or delegate then decides whether a study pause is 
warranted. All investigator(s) will be notified immediately in case of a study pause. The sponsor’s medical 
safety officer or delegate is responsible for the immediate notification of independent medical reviewer and 
coordination of a meeting with the independent medical reviewer in case of a study pause.  
The sponsor’s medical safety officer or delegate or the investigator (upon consultation with the sponsor’s 
medical safety officer or delegate) may contact the independent medical reviewer in any case in which, in 
their professional opinion, the safety of the participants or the reliability of the data could be affected. 
Vaccinations for the study may be suspended for safety concerns other than those described above, or before 
pausing rules are met, if, in the judgment of the independent medical reviewer, participant safety may be 
threatened. 
Resumption of vaccinations will start only upon receipt of written recommendations by the independent 
medical reviewer. The clinical site(s) will be allowed to resume activities upon receipt of written notification 
from the sponsor. The communications from the independent medical reviewer will be forwarded by the 
investigator to the Independent Ethics Committee (IEC) and by the sponsor to the relevant health authorities, 
according to local standards and regulations.  
8.4 
IMMUNOGENICITY ASSESSMENTS 
Venous blood samples for the determination of immune responses will be collected at the time points 
indicated in the TIME AND EVENTS SCHEDULE. Serum samples will be analysed to determine binding 
antibodies against EBOV GP using the FANG ELISA and neutralising antibodies against the Ad26 vector using 
the Ad26 VNA. 
Sample collection and processing will be performed by the study-site personnel according to current versions 
of approved standard operating procedures. The Laboratory Manual contains further details regarding the 
collection, handling, labeling, and shipment of blood samples to the relevant laboratories. 
8.4.1 
Future scientific research 
Future scientific research may be conducted to further investigate Ebola vaccine- and disease-related 
questions and to study other infections of public health importance in Sierra Leone and neighbouring 
countries. This may include the development of new, or the improvement of existing, techniques to 
characterise EBOV-directed immune responses or other diagnostic tests. No additional samples will be taken 
for these analyses, however, residual samples from the study tests may be retained for these purposes and 
analysed after the end of the study.  


 
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8.5 
SAMPLE COLLECTION AND HANDLING 
The actual dates and times of sample collection must be recorded in the CRF or laboratory requisition form.  
Refer to the TIME AND EVENTS SCHEDULE for the timing and frequency of all sample collections. 
Instructions for the collection, handling, storage, and shipment of samples are found in the laboratory manual 
that will be provided. Collection, handling, storage, and shipment of samples must be under the specified, 
and where applicable, controlled temperature conditions as indicated in the laboratory manual. 
 
9 
PARTICIPANT COMPLETION/WITHDRAWAL FROM THE STUDY 
9.1 
COMPLETION 
A participant will be considered to have completed the study if he or she has completed all assessments at 
the 28-day post-vaccination visit (Day 29). 
Participants who prematurely discontinue the study intervention for any reason before completion of the 
28-day post-vaccination visit will not be considered to have completed the study 
9.2 
WITHDRAWAL FROM THE STUDY 
A parent/guardian has the right to withdraw their child from the study at any time for any reason. The 
investigator should make an attempt to contact the parent/guardian of a child who did not return for 
scheduled visits. Although a participant and/or parent/guardian are not obliged to give reason(s) for 
withdrawing prematurely, the investigator should make a reasonable effort to ascertain the reason(s) while 
fully respecting the participant’s rights.  
A participant will be withdrawn from the study for any of the following reasons: 
• 
Lost to follow-up 
• 
Withdrawal of consent 
• 
Death 
• 
Repeated failure to comply with protocol requirements 
• 
Decision by the sponsor or the investigator to stop or cancel the study 
• 
Decision by local regulatory authorities or IEC to stop or cancel the study 
If a participant is lost to follow-up, every reasonable effort must be made by the study-site personnel to 
contact 
the 
participant 
and/or 
the 
parent/guardian 
and 
determine 
the 
reason 
for 
discontinuation/withdrawal. The measures taken to follow up must be documented. 
When a participant withdraws, or a parent/guardian withdraws a participant, before completing the study, 
the reason for withdrawal is to be documented in the CRF and in the source document.  
A participant who wishes to withdraw consent, or a participant whose parent/guardian wishes to withdraw 
consent, will be offered an optional visit for safety follow-up (before the formal withdrawal of consent). The 
participant or participant’s parent/guardian has the right to refuse this optional visit. 
Withdrawal of Consent for the Use of Samples in Future Research 
A participant, or a participant’s parent/guardian, may withdraw consent for the use of samples for research 
(refer to Section 15.2.5). In such a case, every possible effort will be made to destroy samples after they are 
no longer needed for the study. Details of the sample retention for research are presented in the ICF. 


 
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10 
STATISTICAL METHODS 
Statistical analysis will be done by the sponsor or under the authority of the sponsor. A general description 
of the statistical methods to be used to analyze the safety and immunogenicity data is outlined below. 
Specific details will be provided in the Statistical Analysis Plan. 
The final analysis will be performed at study completion, defined as the date of the final database lock, which 
will occur after all participants have completed the last study-related visit or left the study. 
10.1 
ANALYSIS SETS 
Full Analysis Set: The full analysis set will include all participants with study intervention administration 
documented. This will be used for safety analysis.  
Per-protocol Immunogenicity Population: The per-protocol immunogenicity population will include all 
vaccinated participants for whom immunogenicity data are available, excluding participants with major 
protocol deviations expecting to impact the immunogenicity outcomes. 
10.2 
SAMPLE SIZE DETERMINATION 
Approximately 50 participants are expected to be enrolled into this trial (approximately 25 for each age 
group). The sample size is a convenience sample and is not based on formal hypothesis testing 
considerations. 
10.3 
PARTICIPANT INFORMATION 
For all participants, demographic characteristics (e.g. age and sex), and other baseline characteristics (e.g. 
vital signs) will be tabulated and summarized with descriptive statistics. 
10.4 
SAFETY ANALYSES 
No formal statistical testing of safety data is planned. Safety data will be analysed descriptively (including 
95% CIs, if applicable). 
Adverse Events (Including Reactogenicity) 
The verbatim terms used in the CRF by investigators to identify AEs will be coded using the Medical Dictionary 
for Regulatory Activities (MedDRA). All reported AEs and events-related diary information (solicited local at 
injection site and systemic, and unsolicited) with onset within 28 days after the vaccination will be included 
in the analysis. For each AE, the number and percentage of participants who experience at least 1 occurrence 
of the given event will be summarized. 
Summaries, listings, datasets, or participant narratives may be provided, as appropriate, for those 
participants who die, who discontinue study intervention due to an AE, or who experience a severe or a 
serious adverse event.  
Physical Examination 
Because only abbreviated, symptom-directed examinations are performed per the discretion of the 
investigator, physical examination findings (i.e., abnormalities) after vaccination are to be recorded as AEs, 
and will be analysed and presented as indicated above. When reported prior to vaccination, they will be 
recorded as medical history. 
Vital Signs 
Descriptive statistics of temperature, blood pressure, pulse/heart rate, and respiratory rate values will not 
be summarized at each scheduled time point. A listing of participants with clinically significant abnormal 
values will be provided. 
Clinical Laboratory Tests 
Laboratory abnormalities will be determined according to the United States National Institutes of Health, 
National Institute of Allegy and Infectious Disease, Division of Microbiology and Infectious Diseases (DMID) 


 
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paediatric toxicity table (November 2007) (see Appendix 1) and in accordance with the normal ranges of the 
clinical laboratory. The most severe laboratory abnormalities following vaccination will be listed. 
10.5 
IMMUNOGENICITY ANALYSIS 
No formal hypothesis on immunogenicity will be tested. Descriptive statistics (e.g. geometric mean and 95% 
CI) will be calculated for continuous immunologic parameters at all available time points. Graphical 
representations of immunological parameters will be made as applicable. Frequency tabulations will be 
calculated for discrete (qualitative) immunologic parameters (i.e. responder rate),  as applicable. Responders 
rate defined as >2.5-time increase over baseline value (or LLOQ) pre-dose 1 vaccination in the EBOVAC-Salone 
trial, will be calculated depending on availability of sample results from the EBOVAC-Salone trial. 
10.6 
INTERIM ANALYSIS  
No interim analysis will be performed for this study. 
10.7 
INDEPENDENT DATA MONITORING COMMITTEE 
An independent data monitoring committee (IDMC) will not be appointed for this study. The safety of the 
Ad26.ZEBOV vaccine has already been shown in children in previous studies. A booster dose of Ad26.ZEBOV 
was also shown to be safe in adults. There are no planned interim analyses. Thus the IDMC’s role will be 
designated to an independent medical reviewer. The independent medical reviewer will be identified by 
the sponsor to review the accumulating safety data on an ongoing basis to ensure the continuing safety of 
the participants enrolled in this study.  
The independent medical reviewer will be consulted periodically to review newly generated data. Ad hoc 
meetings may be requested via the sponsor if any of the pre-specified pausing rules for this study are met 
(see Section 8.2.) or in any situation that could affect the safety of the participants.   
After the review, the independent medical reviewer will make recommendations regarding the continuation 
of the study. The independent medical reviewer responsibilities, authorities, frequency and timing of the 
evaluations and procedures will be documented in a role and responsibility description.  
The independent medical reviewer will be external and independent of the sponsor. He or she will be a 
medical expert in the relevant field. 
11 
ADVERSE EVENT REPORTING 
Timely, accurate, and complete reporting and analysis of safety information from clinical studies are crucial 
for the protection of participants, investigators, and the sponsor, and are mandated by regulatory agencies 
worldwide. The sponsor has established standard operating procedures in conformity with regulatory 
requirements worldwide to ensure appropriate reporting of safety information; all clinical studies conducted 
by the sponsor or its affiliates are conducted in accordance with those procedures. 
Method of Detecting AEs and Serious Adverse Events 
Care will be taken not to introduce bias when detecting AEs or SAEs. Open-ended and nonleading verbal 
questioning of the participant’s parent(s)/guardian is the preferred method to inquire about AE occurrence.  
Solicited Adverse Events 
Solicited AEs are predefined local (at the injection site) and systemic events for which the participant’s 
parent/guardian is specifically questioned, and which are noted in the participant’s diary. 
Unsolicited Adverse Events 
Unsolicited AEs are all AEs for which the participant’s parent/guardian is not specifically questioned in the 
participant diary. 
11.1 
DEFINITIONS 
11.1.1 Adverse Event Definitions and Classifications 


 
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Adverse Event  
An AE is any untoward medical occurrence in a clinical study subject administered a medicinal (investigational 
or non-investigational) product. An AE does not necessarily have a causal relationship with the treatment. 
An AE can therefore be any unfavourable and unintended sign (including an abnormal finding), symptom, or 
disease temporally associated with the use of a medicinal (investigational or non-investigational) product, 
whether or not related to that medicinal (investigational or non-investigational) product. (Definition per 
International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use 
[ICH].) 
This includes any occurrence that is new in onset or aggravated in severity or frequency from the baseline 
condition, or abnormal results of diagnostic procedures, including laboratory test abnormalities. 
Note: For the time period of AE collection, see Section 11.3. 
Serious Adverse Event  
A SAE, based on ICH and EU Guidelines on Pharmacovigilance for Medicinal Products for Human Use, is any 
untoward medical occurrence that at any dose: 
• 
Results in death 
• 
Is life-threatening (the participant was at risk of death at the time of the event. It does not refer to an 
event that hypothetically might have caused death if it were more severe.) 
• 
Requires inpatient hospitalisation or prolongation of existing hospitalisation 
• 
Results in persistent or significant disability/incapacity 
• 
Is a congenital anomaly/birth defect 
• 
Is a suspected transmission of any infectious agent via a medicinal product 
• 
Is medically important* 
*Medical and scientific judgment should be exercised in deciding whether expedited reporting is also 
appropriate in other situations (other than those listed above) , such as important medical events that may 
not be immediately life-threatening or result in death or hospitalization but may jeopardize the participant 
or may require intervention to prevent one of the other outcomes listed in the definition above. These 
should usually be considered serious. 
If a serious and unexpected AE occurs for which there is evidence suggesting a causal relationship between 
the study intervention and the event (e.g., death from anaphylaxis), the event must be reported as a 
suspected unexpected serious adverse reaction (SUSAR) (even after the study is over, if the sponsor, the 
independent medical reviewer, or the investigator becomes aware of it). 
Unlisted (Unexpected) Adverse Event/Reference Safety Information 
An AE is considered unlisted if the nature or severity is not consistent with the applicable product reference 
safety information. For Ad26.ZEBOV the expectedness of an AE will be determined by whether or not it is 
listed in the IB.  
Adverse Event Associated With the Use of the Intervention 
An AE is considered associated with the use of the intervention if the attribution is related by the 
definitions listed in Section 11.1.2. 
An AE is considered not associated with the use of the intervention if the attribution is unrelated by the 
definitions listed in Section 11.1.2. 
Adverse Events of Special Interest 
AESIs (including potential AESIs) are significant AEs that are judged to be of special interest because of 
clinical importance, known or suspected effects of similar vaccines, or based on nonclinical signals. AESIs 
and potential AESIs will be carefully monitored during the study by the sponsor. 


 
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AESIs and potential AESIs must be reported to the sponsor within 24 hours of awareness, irrespective of 
seriousness (i.e., serious and nonserious AEs) or causality following the procedure described above for SAEs. 
Specific requirements for the AESI are described below. 
 
Thrombosis with Thrombocytopenia Syndrome (TTS) 
TTS has been observed very rarely following vaccination with Janssen COVID vaccine and is considered 
to be an AESI in this study. TTS is a syndrome characterized by a combination of both a thrombotic 
event and thrombocytopenia. [13, 14]  
Because this syndrome is rare and not completely understood, all cases of thrombosis and/or 
symptomatic thrombocytopenia will be considered a potential case of TTS and should be reported to 
the sponsor within 24 hours of awareness. Each potential AESI will be reviewed to identify a TTS case. 
A potential TTS case is defined as: 
• 
Thrombotic events: suspected deep vessel venous or arterial thrombotic events as detailed in 
Appendix 2 
and/or 
• 
Symptomatic thrombocytopenia, defined as platelet count below LLN for the testing lab 
Symptoms, signs, or conditions suggestive of a thrombotic event or symptomatic thrombocytopenia 
should be recorded and reported as a potential AESI even if the final or definitive diagnosis has not yet 
been determined, and alternative diagnoses have not yet been eliminated or shown to be less likely. 
Follow-up information and final diagnoses, if applicable, should be submitted to the sponsor as soon 
as they become available. 
In the event of symptomatic thrombocytopenia, study site personnel should report the absolute value 
for the platelet count and the reference range for the laboratory test used. 
For either a thrombotic event or symptomatic thrombocytopenia, it is recommended to test for anti-
Platelet Factor 4 antibodies (anti-PF4) at the local laboratory or substitute local laboratory; repeat 
testing may be requested for confirmation upon sponsor discretion. 
AESIs, including potential AESIs, will require enhanced data collection and evaluation. Every effort 
should be made to report as much information as possible about the event to the sponsor in a 
reasonable timeframe. 
Treatment and Follow-up Recommendation 
The medical management of thrombotic events with thrombocytopenia is different from the 
management of isolated thromboembolic diseases. Study site personnel and/or treating physicians 
should follow available guidelines for treatment of thrombotic thrombocytopenia (e.g., American 
Society of Hematology 2021; British Society of Haematology 2021; CDC 2021). The use of heparin may 
be harmful and alternative treatments may be needed. Consultation with a hematologist is strongly 
recommended. 
 
11.1.2 Attribution Definitions 
Every effort should be made by the investigator to explain any AE and to assess its potential causal 
relationship, i.e., to administration of the study intervention or to alternative causes (e.g., natural history of 
an underlying diseases, concomitant therapies). This applies to all AEs, whether serious or non-serious. 
Causality of AEs should be assessed by the investigator based on the following: 
Related 


 
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There is suspicion that there is a relationship between study intervention and AE (without determining the 
extent of that probability); there is a reasonable possibility that the study intervention contributed to the AE.  
All AEs assessed as possibly, probably or definitely related to the study intervention will be considered related 
to the study intervention. 
Unrelated 
There is no suspicion that there is a relationship between the study intervention and the AE; there are other 
more likely causes and administration of the study intervention is not suspected to have contributed to the 
AE.  
All AE assessed as unrelated or doubtfully related to the study intervention will be considered unrelated to 
the study intervention. 
By definition, all solicited AEs at the injection site (local) will be considered related to the study intervention 
administration. 
11.1.3 Severity Criteria 
All AEs, except for solicited AEs, will be coded for severity using a modified version of the November 2007 
DMID paediatric toxicity table (see Appendix 1). 
For AEs not identified in the table, the following guidelines will apply: 
Mild 
Grade 1 
Symptoms causing no or minimal interference with usual social 
and functional activities. 
Moderate 
Grade 2 
Symptoms causing greater than minimal interference with usual 
social and functional activities. 
Severe 
Grade 3 
Symptoms causing inability to perform usual social and 
functional activities. 
Potentially Life-
threatening 
Grade 4 
Any grade 3 symptom that requires hospitalization/in-patient 
medical intervention.  
Note: Only clinically significant abnormalities in laboratory data occurring from signing of the ICF onwards 
will be reported as adverse events and graded using the table above. 
The investigator should use clinical judgment in assessing the severity of events not directly experienced by 
the participant (e.g. laboratory abnormalities). 
11.2 
SPECIAL REPORTING SITUATIONS 
Safety events of interest on a study intervention that may require expedited reporting or safety evaluation 
include, but are not limited to: 
• 
Overdose of a study intervention 
• 
Suspected abuse/misuse of a study intervention 
• 
Medication error involving a product (with or without participant exposure to the study intervention, 
e.g., name confusion) 
Special reporting situations should be recorded in the CRF. Any special reporting situation that meets the 
criteria of a SAE should be recorded on the serious adverse event page of the CRF. 
11.3 
PROCEDURES 
Depending on the nature of the event, the reporting procedures below will be followed. Any questions 
concerning AE reporting will be directed to the sponsor. Further details on AE reporting can be found in the 
AE reporting flowchart. 


 
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11.3.1 All Adverse Events 
Symptoms of solicited local and systemic AEs will be collected in the diary in the evening after the 
vaccination and then daily for the next 7 days. Unsolicited AEs will be reported from vaccination until 28 
days post-vaccination. 
Serious adverse events and/or special reporting situations that are related to study procedures will be 
reported from the time a signed and dated ICF is obtained onwards until the end of the study. All other 
SAEs and/or special reporting situations will be reported from the day of vaccination onwards until 28 days 
post-vaccination. The sponsor will evaluate any safety information that is spontaneously reported by an 
investigator beyond the time frame specified in the protocol. 
The investigator will monitor and analyse the study data including all AE and clinical laboratory data as they 
become available and will make determinations regarding the severity of the adverse experiences and their 
relation to study intervention. All AEs will be deemed related to study intervention or not related to study 
intervention, according to Section 11.1.  
The investigator or clinical designee must review both post-injection reactogenicity and other AE CRFs to 
insure the prompt and complete identification of all events that require expedited reporting as SAEs, invoke 
pausing rules, or are other serious and unexpected events. 
All AEs, regardless of seriousness, severity, or presumed relationship to study intervention, must be 
recorded using medical terminology in the source document and the CRF. Whenever possible, diagnoses 
should be given when signs and symptoms are due to a common aetiology (e.g., cough, runny nose, 
sneezing, sore throat, and head congestion should be reported as "upper respiratory infection"). 
Investigators must record in the CRF their opinion concerning the relationship of the AE to study therapy. 
All measures required for AE management must be recorded in the source document and reported 
according to sponsor instructions. 
The LSHTM assumes responsibility for appropriate reporting of AEs to the regulatory authorities. The 
LSHTM will also report to the vaccine manufacturer and investigator (and the head of the investigational 
institute, as required) all SUSARs. The investigator (or sponsor, as required) must report SUSARs to the 
appropriate IEC that approved the protocol unless otherwise required and documented by the IEC.  
Janssen Vaccines & Prevention B.V., as the vaccine manufacturer, will report any SUSAR to all the 
investigators of studies using the experimental vaccine. 
The parent/guardian will be provided with a "wallet (study) card" and instructed to carry this card with 
their child for the duration of the study indicating the following:  
• 
Study number 
• 
Statement, in the local language(s), that the child is participating in a clinical study.  
• 
Investigator's name and 24-hour contact telephone number 
• 
Local sponsor's name and 24-hour contact telephone number (for medical staff only) 
• 
Participant number 
11.3.2 Serious Adverse Events 
All SAEs occurring during the study must be reported to the appropriate person nominated by the sponsor 
by study-site personnel within 24 hours of their knowledge of the event and to IEC and local regulatory 
authority, as required. 
Information regarding SAEs will be transmitted to the sponsor and to IEC and local regulatory authority, as 
required, using the Serious Adverse Event Form, which must be completed and signed by a physician from 
the study site, and transmitted to the sponsor, IEC and local regulatory authority, within 24 hours. The initial 
and follow-up reports of a SAE should scanned and sent by email. 


 
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All SAEs that have not resolved by the end of the study, or that have not resolved upon discontinuation of 
participation in the study, must be followed until any of the following occurs: 
• 
The event resolves 
• 
The event stabilizes 
• 
The event returns to baseline, if a baseline value/status is available 
• 
The event can be attributed to agents other than the study intervention or to factors unrelated to 
study conduct 
• 
It becomes unlikely that any additional information can be obtained (participant or health care 
practitioner refusal to provide additional information, lost to follow-up after demonstration of due 
diligence with follow-up efforts) 
Suspected transmission of an infectious agent by a medicinal product will be reported as a SAE. Any event 
requiring hospitalisation (or prolongation of hospitalisation) that occurs during the course of a subject's 
participation in a study must be reported as a SAE, except hospitalisations for the following: 
• 
Hospitalisations not intended to treat an acute illness or AE (e.g., social reasons such as pending 
placement in a long-term care facility) 
• 
Surgery or procedure planned before entry into the study (this must be documented in the CRF).  
Note: Hospitalisations that were planned before the signing of the ICF, and where the underlying 
condition, for which the hospitalisation was planned, has not worsened, will not be considered SAEs. 
Any AE that results in a prolongation of the originally planned hospitalisation is to be reported as a 
new SAE. 
During the entire study, the cause of death of a participant in a study, whether or not the event is expected 
or associated with the study intervention, is considered a SAE. 
11.4 
CONTACTING SPONSOR REGARDING SAFETY 
The names (and corresponding telephone numbers) of the individuals who should be contacted regarding 
safety issues or questions regarding the study are listed in the Contact Information page(s), which will be 
provided as a separate document. 
12 
PRODUCT QUALITY COMPLAINT HANDLING 
A product quality complaint (PQC) is defined as any suspicion of a product defect related to manufacturing, 
labeling, or packaging, i.e., any dissatisfaction relative to the identity, quality, durability, or reliability of a 
product, including its labeling or package integrity. A PQC may have an impact on the safety and efficacy of 
the product. Timely, accurate, and complete reporting and analysis of PQC information from studies are 
crucial for the protection of participants, investigators, and the sponsor, and are mandated by regulatory 
agencies worldwide. Janssen Vaccines & Prevention B.V. and the sponsor have established procedures in 
conformity with regulatory requirements worldwide to ensure appropriate reporting of PQC information; all 
studies conducted by the sponsor or its affiliates will be conducted in accordance with those procedures. 
12.1 
PROCEDURES 
All initial PQCs must be reported to the sponsor by the study-site personnel within 24 hours after being made 
aware of the event. 
If the defect is combined with a SAE, the study-site personnel must report the PQC to the sponsor according 
to the SAE reporting timelines (refer to Section 11.3). A sample of the suspected product should be 
maintained for further investigation if requested by the sponsor and Janssen Vaccines & Prevention B.V. 


 
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12.2 
CONTACTING SPONSOR REGARDING PRODUCT QUALITY 
The names (and corresponding telephone numbers) of the individuals who should be contacted regarding 
product quality issues are listed in the Contact Information page(s), which will be provided as a separate 
document. 
13 
STUDY INTERVENTION INFORMATION 
Ad26.ZEBOV 
Ad26.ZEBOV is a monovalent, replication-incompetent adenovirus serotype 26-based vector that expresses 
the full-length EBOV Mayinga GP and is produced in the human cell line PER.C6®. 
The Ad26.ZEBOV vaccine will be supplied at a concentration of 1x1011 vp/mL in 2-mL single-use glass vials as 
a frozen liquid to be thawed before use. Each vial contains an extractable volume of 0.5 mL. Refer to the IB 
for a list of excipients. 
The Ad26.ZEBOV vaccine is manufactured by IDT Biologika GmbH for Janssen Vaccines & Prevention B.V., The 
Netherlands. 
13.1 
PACKAGING 
All study intervention will be manufactured and packaged in accordance with Good Manufacturing Practice. 
All study intervention will be packaged and labeled under the responsibility of the sponsor. No study 
intervention can be repacked or re-labeled without prior approval from the sponsor. 
Further details for study intervention packaging and labeling can be found in the Site Investigational Product 
Procedures Manual. 
13.2 
LABELLING 
Study intervention labels will contain information to meet the applicable regulatory requirements. 
13.3 
PREPARATION, HANDLING, STORAGE 
All study intervention must be stored at controlled temperatures. Guidance on storage temperature is 
provided in the Site Investigational Product Procedures Manual. 
Vials must be stored in a secured location with no access for unauthorized personnel. All equipment for 
storage of the study intervention (including refrigerators, freezers) must be equipped with a continuous 
temperature monitor and alarm, and with back-up power systems. In the event that the study intervention 
is exposed to temperatures outside the specified temperature ranges, all relevant data will be sent to the 
sponsor to determine if the affected study intervention can be used or will be replaced. The affected study 
intervention must be quarantined and not used until further instruction from the sponsor is received. 
A pharmacist/qualified staff member will prepare all doses for vaccine administration and provide it for 
dispensing. 
Full details on the preparation, the holding time, and storage conditions from the time of preparation to 
delivery of Ad26.ZEBOV are provided in the Site Investigational Product Procedures Manual. 
13.4 
INTERVENTION ACCOUNTABILITY 
The investigator is responsible for ensuring that all study intervention received at the site is inventoried and 
accounted for throughout the study. The study intervention administered to the participant must be 
documented on the intervention accountability form. All study intervention will be stored and disposed of 
according to the sponsor's instructions. Study-site personnel must not combine contents of the study 
intervention containers. 
Study intervention must be handled in strict accordance with the protocol and the container label, and must 
be stored at the study site in a limited-access area or in a locked cabinet under appropriate environmental 


 
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conditions. Unused study intervention must be available for verification by the sponsor's study site monitor 
or delegate during on-site monitoring visits. The return to the sponsor of unused study intervention will be 
documented on the Investigational Product Destruction Form. When the study site is an authorized 
destruction unit and study intervention supplies are destroyed on-site, this must also be documented on the 
Investigational Product Destruction Form. 
Potentially hazardous materials such as used ampules, needles, syringes, and vials containing hazardous 
liquids, should be disposed of immediately in a safe manner and therefore will not be retained for 
intervention accountability purposes.  
Study intervention should be dispensed under the supervision of the investigator or a qualified member of 
the study-site personnel, or by a hospital/clinic pharmacist. Study intervention will be supplied only to study 
participants. The investigator agrees neither to dispense the study intervention from, nor store it at, any site 
other than the study site agreed upon with the sponsor. 
14 
STUDY -SPECIFIC MATERIALS 
The investigator will be provided with the following supplies: 
• 
IB and Addendum (if applicable) for Ad26.ZEBOV 
• 
Site Investigational Product Procedures Manual 
• 
Laboratory manual 
• 
Electronic Data Capture (eDC) Manual/electronic CRF Completion Guidelines 
• 
Sample ICF 
• 
Participant diaries 
• 
TOU 
• 
Rulers, thermometers 
• 
Participant wallet card  
15 
ETHICAL ASPECTS 
15.1 
STUDY-SPECIFIC DESIGN CONSIDERATIONS 
Potential participants, and/or their parent/guardian, will be fully informed of the risks and requirements of 
the study. During the study, participants and/or their parent/guardian will be given any new information that 
may affect their decision to continue participation. They will be told that their consent to participate in the 
study is voluntary and may be withdrawn at any time with no reason given and without penalty or loss of 
benefits to which they would otherwise be entitled. Only parents/guardians of participants who are fully able 
to understand the risks, benefits, and potential AEs of the study, and provide their consent voluntarily will 
be allowed to enrol their child. 
The primary ethical concern is the safety of the enrolled children. 
When referring to the signing of the ICF, the term guardian refers to the traditionally or legally appointed 
guardian of the child with authority to authorise participation in research. For each participant, his or her 
parent or guardian, must give permission and written consent (according to local requirements) after the 
nature of the study has been fully explained and before any study-related activities are performed. Only 
parents/guardians who are fully able to understand the risks and benefits, and who provide their consent 
voluntarily, can enrol their child into the study. Assent must be obtained from children capable of 
understanding the nature of the study, typically potential participants 7 years of age and older. For the 
purposes of this study, all references to participants who have provided consent (and assent as applicable) 
refer to the participant and his or her parent/guardian who has provided consent according to this process. 
Children who assent to a study and later withdraw that assent will not be maintained in the study against 
their will, even if their parent/guardian still want them to participate. 


 
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Test of Understanding 
The TOU is a short assessment of the parent/guardian of the potential participant’s understanding of key 
aspects of the study. The test will help the study staff to determine how well the parent/guardian 
understands the study and their requirements for participation of their child. 
The parent/guardian must pass the TOU, indicating that he or she understands the purpose of, and 
procedures required for the study, after reading the informed consent and after the investigator or designee 
has provided detailed information on the study and has answered the questions of the parent/guardian. The 
parent/guardian must subsequently sign the ICF, indicating that he or she is willing to allow their child to 
participate in the study. 
If a parent/guardian fails to achieve the passing score on an attempt, further information and counselling will 
be provided to the parent/guardian by a study team member. The parent/guardian is allowed to retake the 
test twice to achieve the passing score (≥90%) required for participation of their child in the study. If the 
parent/guardian fails to achieve the passing score on the third attempt they will not be able to re-take the 
test again, and their child will not be allowed to participate in the study. 
Any parent/guardian of a potential participant not capable of understanding the key aspects of the study, 
and their requirements for participation, should not be allowed to enrol their child. 
15.2 
REGULATORY ETHICS COMPLIANCE 
All references to the IEC refer to the LSHTM Ethics Committee and the Sierra Leone Ethics and Scientific 
Review Committee.  
15.2.1 Investigator Responsibilities 
The investigator will be responsible for ensuring that the study is performed in accordance with the protocol, 
current ICH guidelines on Good Clinical Practice (GCP), and applicable regulatory and country-specific 
requirements. 
GCP is an international ethical and scientific quality standard for designing, conducting, recording, and 
reporting studies that involve the participation of human subjects. Compliance with this standard provides 
public assurance that the rights, safety, and well-being of study subjects are protected, consistent with the 
principles that originated in the Declaration of Helsinki, and that the study data are credible. 
15.2.2 Independent Ethics Committee 
Before the start of the study, the investigator (or sponsor where required) will provide the IEC with current 
and complete copies of the following documents (as required by local regulations): 
• 
Final protocol and, if applicable, amendments 
• 
Sponsor-approved ICF (and any other written materials to be provided to the study subjects, e.g. 
diary) 
• 
IB (or equivalent information) and amendments 
• 
Sponsor-approved subject recruiting materials 
• 
Information on compensation for study-related injuries or payment to study participants for 
participation in the study, if applicable 
• 
Investigator's curriculum vitae or equivalent information (unless not required, as documented by 
the IEC) 
• 
Information regarding funding, name of the sponsor, institutional affiliations, other potential 
conflicts of interest, and incentives for study participants 
• 
Any other documents that the IEC requests to fulfil its obligation 
This study will be undertaken only after the IEC has given full approval of the final protocol, amendments (if 
any, excluding the ones that are purely administrative, with no consequences for participants, data or study 


 
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conduct, unless required locally), the ICF, applicable recruiting materials, and subject compensation 
programmes, and after the sponsor has received a copy of this approval. This approval letter must be dated 
and must clearly identify the IEC and the documents being approved. 
During the study the investigator (or sponsor where required) will send the following documents and updates 
to the IEC for their review and approval, where appropriate: 
• 
Protocol amendments (excluding the ones that are purely administrative, with no consequences 
for participants, data, or study conduct) 
• 
Revision(s) to ICF and any other written materials to be provided to participants 
• 
New or revised participant recruiting materials approved by the sponsor (when applicable) 
• 
Revisions to compensation for study-related injuries or payment to participants for participation in 
the study, if applicable 
• 
New edition(s) of the IB and amendments/addenda 
• 
Summaries of the status of the study at intervals stipulated in guidelines of the IEC (at least 
annually) 
• 
Reports of AEs that are serious, unlisted/unexpected, and associated with the study vaccine 
• 
New information that may adversely affect the safety of the participants or the conduct of the study 
• 
Deviations from or changes to the protocol to eliminate immediate hazards to the participants 
• 
Report of deaths of participants under the investigator's care 
• 
Notification if a new investigator is responsible for the study at the centre 
• 
Development Safety Update Report and Line Listings, where applicable 
• 
Any other requirements of the IEC 
For all protocol amendments (excluding the ones that are purely administrative, with no consequences for 
participants, data, or study conduct), the amendment and applicable ICF revisions will be submitted promptly 
to the IEC for review and approval before implementation of the change(s). 
At least once a year, the IEC will be asked to review and reapprove this study, where required. 
At the end of the study, the investigator (or sponsor where required) will notify the IEC about the study 
completion. 
15.2.3 Informed Consent  
In this community-based study, informed consent will take place at several levels ranging from approval from 
the government authorities, followed by community level consent, and finally, individual consent. 
Approval from Respective Health Authorities 
Approval for this project will be obtained from the respective health authorities in the country where the 
study is conducted. 
Additionally, during the planning process of the study, approval will be sought from other authorities such as 
district or local councillors, political leaders, and traditional leaders. 
Consent at the Community Level 
Documented community-level consent by a community leader must be available. 
Consent at the Individual Level 
The parent/guardian of a participant (in this section referred to as the legally acceptable representative) must 
give written consent according to local requirements after the nature of the study has been fully explained. 
The ICF must be signed before performance of any study-related activity. The ICF(s) that is/are used must be 


 
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approved by both the sponsor and by the reviewing IEC and be in a language that the legally acceptable 
representative and can read and understand. The informed consent should be in accordance with principles 
that originated in the Declaration of Helsinki, current ICH and GCP guidelines, applicable regulatory 
requirements, and sponsor policy. 
Before enrolment in the study, the investigator or an authorized member of the study-site personnel must 
explain to the subject and/or the subject’s legally acceptable representative the aims, methods, reasonably 
anticipated benefits, and potential hazards of the study, and any discomfort participation in the study may 
entail. The subject and/or the legally acceptable representative will be informed that their participation/the 
participation of their child is voluntary and that they may withdraw consent to participate at any time. They 
will be told that the investigator will maintain a participant identification register if needed and that their 
records may be accessed by health authorities and authorized sponsor personnel without violating the 
confidentiality of the participant, to the extent permitted by the applicable law(s) or regulations. By signing 
the ICF the legally acceptable representative is authorizing such access, which includes permission to obtain 
information about their child’s health status. It also denotes that the legally acceptable representative agrees 
to allow his or her child’s study physician to recontact the them for the purpose of obtaining consent for 
additional safety evaluations.  
The legally acceptable representative will be given sufficient time to read the ICF and the opportunity to ask 
questions. After this explanation and before their child’s entry into the study, consent should be 
appropriately recorded by means of the legally acceptable representative's personally dated signature. After 
having obtained the consent, a copy of the ICF must be given to the legally acceptable representative. 
If the legally acceptable representative is unable to read or write, an impartial witness should be present for 
the entire informed consent process (which includes reading and explaining all written information) and the 
impartial witness should personally date and sign the ICF after the consent of the legally acceptable 
representative is obtained. 
Children will be enrolled only after obtaining consent of a parent/guardian. Assent will be obtained from 
children capable of understanding the nature of the study, typically subjects 7 years of age and older. Written 
assent will be obtained from subjects who are able to write. Children who assent to a study and later 
withdraw that assent will not continue in the study against their will, even if their parent/guardian still want 
them to participate. 
Children who turn 7 years of age will be asked to provide assent, if it is deemed they are able to comprehend 
the information provided, for continuation in the study at the first available opportunity. 
15.2.4 Privacy of Personal Data 
The collection and processing of personal data from participants enrolled in this study will be limited to those 
data that are necessary to fulfill the objectives of the study. 
These data must be collected and processed with adequate precautions to ensure confidentiality and 
compliance with applicable data privacy protection laws and regulations. Appropriate technical and 
organizational measures to protect the personal data against unauthorized disclosures or access, accidental 
or unlawful destruction, or accidental loss or alteration must be put in place. Sponsor personnel whose 
responsibilities require access to personal data agree to keep the identity of participants confidential. 
The informed consent obtained from the legally acceptable representative includes explicit consent for the 
processing of personal data of the participant and for the investigator/institution to allow direct access to his 
or her original medical records (source data/documents) for study-related monitoring, audit, IEC review, and 
regulatory inspection. This consent also addresses the transfer of the data to other entities and to other 
countries. 
The legally acceptable representative has the right to request through the investigator access to the personal 
data of their child and the right to request rectification of any data that are not correct or complete. 
Reasonable steps will be taken to respond to such a request, taking into consideration the nature of the 
request, the conditions of the study, and the applicable laws and regulations. 


 
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Exploratory research is not conducted under standards appropriate for the return of data to subjects. In 
addition, the sponsor cannot make decisions as to the significance of any findings resulting from exploratory 
research. Therefore, exploratory research data will not be returned to subjects or investigators, unless 
required by law or local regulations. Privacy and confidentiality of data generated in the future on stored 
samples will be protected by the same standards applicable to all other clinical data. 
15.2.5 Long-Term Retention of Samples for Additional Future Research 
Samples collected in this study may be stored for up to 15 years (or according to local regulations) for 
additional research. Future scientific research may be conducted to further investigate Ebola vaccine- and 
disease-related questions and to study other infections of public health importance in Sierra Leone and 
neighbouring countries. This may include the development of new or the improvement of existing techniques 
to characterize EBOV-directed immune responses or diagnostic tests. No additional samples will be taken for 
these analyses, however, residual samples from other tests may be retained for these purposes and analysed 
after the end of the study. 
Parents/guardians will be asked to consent voluntarily for their child’s blood samples to be stored for other 
research studies that may be done after this study is completed. Participants for whom such consent is not 
given, can participate in the study without having their blood samples stored for future testing (see also 
Section 9.2). In such case, their blood samples will be destroyed after all the immunogenicity tests have been 
concluded (as agreed by the sponsor). 
All samples, for which consent has been obtained and for which additional material is available after study-
specified testing is complete, will be stored for future testing.  
Stored samples will be coded throughout the sample storage and analysis process and will not be labeled 
with personal identifiers. Parents/guardians may withdraw consent for their child’s samples to be stored for 
research at any time during the study. 
16 
ADMINISTRATIVE REQUIREMENTS 
16.1 
PROTOCOL AMENDMENTS 
Neither the investigator nor the sponsor will modify this protocol without a formal amendment by the 
sponsor. All protocol amendments will be issued by the sponsor, and signed and dated by the relevant 
investigator. Protocol amendments will not be implemented without prior IEC approval, or when the relevant 
competent authority has raised any grounds for non-acceptance, except when necessary to eliminate 
immediate hazards to the participants, in which case the amendment will be promptly submitted to the IEC 
and relevant competent authority. Documentation of amendment approval by the investigator and IEC will 
be provided to the sponsor. When the change(s) involves only logistic or administrative aspects of the study, 
the IEC (where required) will be notified. 
During the course of the study, in situations where a departure from the protocol is unavoidable, the 
investigator or other physician in attendance will contact the appropriate sponsor representative (listed in 
the Contact Information page(s), which will be provided as a separate document). Except in emergency 
situations, this contact will be made before implementing any departure from the protocol. In all cases, 
contact with the sponsor will be made as soon as possible to discuss the situation and agree on an appropriate 
course of action. The data recorded in the CRF and source documents will reflect any departure from the 
protocol, and the source documents will describe this departure and the circumstances requiring it. 
16.2 
REGULATORY DOCUMENTATION 
16.2.1 Regulatory Approval and Notification 
This protocol and any amendment(s) must be submitted to the appropriate regulatory authority in the 
country where the study is being conducted. A study may not be initiated until all local regulatory 
requirements are met. All references to the local Regulatory Authority refer to the Pharmacy Board of Sierra 
Leone.  


 
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16.2.2 Required Pre-study Documentation  
The following documents will be provided to the sponsor before shipment of study intervention to the study 
site: 
• 
Protocol and amendment(s), if any, signed and dated by the Principal Investigator 
• 
A copy of the dated and signed (or sealed, where appropriate per local regulations), written IEC 
approval of the protocol, amendments, ICF, any recruiting materials, and if applicable, participant 
compensation programs. This approval must clearly identify the specific protocol by title and number 
and must be signed (or sealed, where appropriate per local regulations) by the chairman or 
authorised designee. 
• 
Name and address of the IEC, including a current list of the IEC members and their function, with a 
statement about how it is organised and operates according to GCP and the applicable laws and 
regulations. If accompanied by a letter of explanation, or equivalent, from the IEC, a general 
statement may be substituted for this list. If an investigator or a member of the study-site personnel 
is a member of the IEC, documentation must be obtained to state that this person did not participate 
in the deliberations or in the vote/opinion of the study. 
• 
Regulatory authority approval or notification 
• 
Signed and dated statement of investigator (e.g. Form FDA 1572), If applicable 
• 
Documentation of investigator qualifications (e.g. curriculum vitae) 
• 
Completed investigator financial disclosure form from the Principal Investigator, where required 
• 
Signed and dated clinical trial agreement, which includes financial agreement 
• 
Any other documentation required by local regulations 
 
The following documents must be provided to the sponsor before enrolment of the first participant: 
• 
Completed investigator financial disclosure forms from all sub investigators 
• 
Documentation of sub-investigator qualifications (e.g., curriculum vitae) 
• 
Name and address of any local laboratory conducting tests for the study, and a dated copy of current 
laboratory normal ranges for these tests, if applicable 
16.3 
SUBJECT IDENTIFICATION AND ENROLMENT LOGS 
The investigator agrees to complete a subject identification and enrolment log to permit easy identification 
of each subject during and after the study. This document will be reviewed by the sponsor study-site 
representative for completeness. 
The participant identification and enrolment log will be treated as confidential and will be filed by the 
investigator in the study file. To ensure participant confidentiality, no copy will be made. All reports and 
communications relating to the study will identify subjects by participant identification and age at initial 
informed consent.  
The investigator must also complete a participant screening log, which reports on all participants who were 
seen to determine eligibility for inclusion in the study. 
16.4 
SOURCE DOCUMENTATION 
At a minimum, source documents consistent in the type and level of detail with that commonly recorded at 
the study site as a basis for standard medical care must be available for the following: subject identification, 
eligibility, and study identification; study discussion and date of signed informed consent; dates of visits; 
results of safety and efficacy parameters as required by the protocol; record of all AEs and follow-up of AEs; 
concomitant medication; intervention receipt/dispensing/return records; study intervention administration 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
information; and date of study completion and reason for early discontinuation of study intervention or 
withdrawal from the study, if applicable.  
The author of an entry in the source documents should be identifiable. 
Specific details required as source data for the study and source data collection methods will be reviewed 
with the investigator before the study and will be described in the monitoring guidelines (or other equivalent 
document). 
The TOU and participant’s diary used to collect information regarding solicited symptoms after vaccination 
will be considered source data. 
Inclusion and exclusion criteria not requiring documented medical history must be verified at a minimum by 
participant interview or other protocol required assessment (e.g., physical examination, laboratory 
assessment) and documented in the source documents. 
16.5 
CASE REPORT FORM COMPLETION 
Case report forms are prepared and provided by the sponsor for each subject in electronic format. All CRF 
entries, corrections, and alterations must be made by the investigator or authorized study-site personnel. 
The investigator must verify that all data entries in the CRF are accurate and correct. 
The study data will be transcribed by study-site personnel from the source documents onto an electronic 
case report form (eCRF). Study-specific data will be transmitted in a secure manner to the sponsor within the 
timeframe agreed upon between the sponsor and the study site.  
Worksheets may be used for the capture of some data to facilitate completion of the eCRF. Any such 
worksheets will become part of the participant's source documentation. Data must be entered into eCRFs in 
English. Study site personnel must complete the eCRF promptly after a participant visit, and the forms should 
be available for review at the next scheduled monitoring visit.  
If necessary, queries will be generated in the eDC tool. If corrections to a CRF are needed after the initial 
entry into the eCRF, this can be done in either of the following ways: 
• 
Investigator and study site personnel can make corrections in the eDC tool at their own initiative or as a 
response to an auto query (generated by the eDC tool). 
• 
Sponsor or sponsor delegate can generate a query for resolution by the investigator and study site 
personnel. 
16.6 
DATA QUALITY ASSURANCE/QUALITY CONTROL 
Steps to be taken to ensure the accuracy and reliability of data include the selection of qualified investigators 
and appropriate study sites, review of protocol procedures with the investigator and study-site personnel 
before the study, and periodic monitoring visits by the sponsor and/or remote monitoring by the sponsor. 
Written instructions will be provided for collection, handling, storage, and shipment of samples. 
Guidelines for CRF completion will be provided and reviewed with study-site personnel before the start of 
the study. The sponsor will review CRFs for accuracy and completeness during onsite monitoring visits and 
after transmission to the sponsor; any discrepancies will be resolved with the investigator or designee, as 
appropriate. The data will be entered into the study database and verified for accuracy and consistency with 
the data sources. 
Representatives of the sponsor may visit the participating site at any time during or after completion of the 
study to conduct an audit of the study in compliance with regulatory guidelines and/or LSHTM policy. Similar 
procedures may also be conducted by a regulatory authority. Further details of on-site audit policies are 
presented in Section 16.10. 
16.7 
RECORDS RETENTION 
In compliance with ICH/GCP guidelines, the investigator/institution will maintain all CRFs and all source 
documents that support the data collected from each participant, as well as all study documents as specified 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
in ICH/GCP Section 8, Essential Documents for the Conduct of a Clinical Trial, and all study documents as 
specified by the applicable regulatory requirement(s). The investigator/institution will take measures to 
prevent accidental or premature destruction of these documents. 
Data and all appropriate documentation will be stored for a minimum of 10 years after the completion of 
the study, as per LSHTM SOP (LSHTM-SOP-049 Site Close out).  
If it becomes necessary for the sponsor or the appropriate regulatory authority to review any documentation 
relating to this study, the investigator/institution must permit access to such reports. 
16.8 
MONITORING 
The sponsor, or their delegate, will perform study site visits to monitor this study. 
The sponsor, or their delegate will perform on-site monitoring visits as frequently as necessary. The monitor 
will record dates of the visits in a study site visit log that will be kept at the study site. The first post-initiation 
visit will be made as soon as possible after enrolment has begun. At these visits, the monitor will compare 
the data entered into the CRF with the vaccination unit and/or clinic records (source documents) (e.g., 
hospital / clinic / physician’s office medical records). The nature and location of all source documents will be 
identified to ensure that all sources of original data required to complete the CRF are known to the sponsor 
and study-site personnel and are accessible for verification by the sponsor study-site contact. If electronic 
records are maintained at the study site, the method of verification must be discussed with the study-site 
personnel. 
Direct access to source documents (medical records) must be allowed for the purpose of verifying that the 
recorded data are consistent with the original source data. Findings from this review will be discussed with 
the study-site personnel. The sponsor expects that, during monitoring visits, the relevant study-site 
personnel will be available, the source documents will be accessible, and a suitable environment will be 
provided for review of study-related documents. The monitor will meet with the investigator on a regular 
basis during the study to provide feedback on the study conduct. 
In addition to on-site monitoring visits, remote contacts can occur. It is expected that during these remote 
contacts, study-site personnel will be available to provide an update on the progress of the study at the site.  
16.9 
STUDY COMPLETION/TERMINATION 
16.12.1 Study Completion/End of Study 
The study is considered completed at final database lock, which will occur after the last participant in the 
study has completed their last study-related visit or left the study. The final data from the study site will be 
sent to the sponsor (or designee) after completion of the final participant visit at that study site, in the time 
frame specified in the Clinical Trial Agreement. 
16.12.2 Study Termination 
The sponsor reserves the right to close the study for data collection or terminate the study at any time for 
any reason at the sole discretion of the sponsor. The study will be closed upon study completion. The study 
is considered closed when all the required documents and study supplies have been collected and a study-
site closure visit has been performed. 
The investigator may initiate study-site closure at any time, provided there is reasonable cause and sufficient 
notice is given in advance of the intended termination.  
Reasons for the early closure of the study site by the sponsor or investigator may include but are not limited 
to: 
• 
Failure of the investigator to comply with the protocol, the requirement of the IEC or local health 
authorities, the sponsor’s procedures, or GCP guidelines 
• 
Inadequate recruitment of participants by the investigator 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
• 
Discontinuation of further study intervention development 
16.10 ON-SITE AUDITS 
Representatives of the sponsor's clinical quality assurance department may visit the study site at any time 
during or after completion of the study to conduct an audit of the study in compliance with regulatory 
guidelines and company policy. These audits will require access to all study records, including source 
documents, for inspection. Subject privacy must, however, be respected. The investigator and study-site 
personnel are responsible for being present and available for consultation during routinely scheduled study-
site audit visits conducted by the sponsor or its designees.  
Similar auditing procedures may also be conducted by agents of any regulatory body, either as part of a 
national GCP compliance program or to review the results of this study in support of a regulatory submission. 
The investigator should immediately notify the sponsor if he or she has been contacted by a regulatory 
agency concerning an upcoming inspection. 
16.11 USE OF INFORMATION AND PUBLICATION 
All information, including but not limited to information regarding the Ad26.ZEBOV vaccine or the sponsor's 
operations (e.g., manufacturing processes, basic scientific data, prior clinical data, formulation information) 
supplied by the sponsor to the investigator and not previously published, and any data, including exploratory 
research data, generated as a result of this study, are considered confidential. The investigator agrees to 
maintain this information in confidence and use this information only to accomplish this study, and will not 
use it for other purposes without the sponsor's prior written consent. 
The investigator understands that the information developed in the study will be used by the sponsor or the 
vaccine manufacturer, Janssen Vaccines & Prevention B.V., in connection with the continued development 
of the Ad26.ZEBOV and MVA-BN-Filo vaccine regimen, and thus may be disclosed as required to other clinical 
investigators or regulatory agencies. To permit the information derived from the clinical studies to be used, 
the investigator is obligated to provide the sponsor with all data obtained in the study. 
Consistent with Good Publication Practices and International Committee of Medical Journal Editors 
guidelines, the sponsor shall have the right to publish such primary data and information without approval 
from the investigator. Authorship of publications resulting from this study will be based on the guidelines on 
authorship, such as those described in the International Committee of Medical Journal Editors (ICMJE), 
Recommendations for the Conduct, Reporting, Editing and Publication of Scholarly Work in Medical Journals, 
which state that the named authors must have made a significant contribution to the conception or design 
of the work; or the acquisition, analysis, or interpretation of the data for the work; and drafted the work or 
revised it critically for important intellectual content; and given final approval of the version to be published; 
and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy 
or integrity of any part of the work are appropriately investigated and resolved. 
Registration of Clinical Studies and Disclosure of Results 
The sponsor will register and disclose the existence of, and the results of, clinical studies as required by law. 
 
 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
APPENDIX 1 
 
Toxicity Grading Scale for Healthy Paediatric Participants (older than 3 months) Enrolled in Preventive Vaccine 
Clinical Trials 
Adapted from: DMID° Pediatric Toxicity Tables (November 2007, draft). For adverse events not included in the 
tables below, refer to the severity criteria guidelines in section 11.1.3 Severity Criteria. Local lab references take 
preference over the DMD table and the different grades.   
 
The abbreviations used in the following tables are: LLN: lower limit of normal; IV: intravenous; ULN: upper limit of 
normal. 
 
LOCAL REACTIONS 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Tenderness 
Mild discomfort to 
touch; minimal to no 
limitation of use of 
limb 
 
Notable discomfort 
to touch; Greater 
than minimal 
limitation of use of 
limb 
 
Significant 
discomfort at rest; 
Severe limitation of 
use of limb 
 
Hospitalization or ER 
visit for treatment 
Erythema  
<10 mm 
10-25 mm 
26-50 mm 
>50 mm or any 
grade 3 with 
hospitalization or ER 
visit for treatment 
Swelling 
<10 mm 
10-25 mm 
26-50 mm 
>50 mm or any 
grade 3 with 
hospitalization or ER 
visit for treatment 
HEMATOLOGY 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Hemoglobin for children 
greater than 3 months and 
less than 2 years of age 
9.0-9.9 gm/dL 
7.0-8.9 gm/dL 
<7.0 gm/dL 
Cardiac Failure 
secondary to 
Anemia 
Hemoglobin for children 
greater than 2 years of age 
10.0-10.9 gm/dL 
7.0-9.9 gm/dL 
<7.0 gm/dL 
Cardiac Failure 
secondary to 
Anemia 
Absolute Neutrophil Count 
750-1200/mm3 
400-749/mm3 
250-399/mm3 
<250/mm3 
Platelets 
75,000-99,999/mm3 
50,000-74,999/mm3 
25,000-49,999/mm3 
<25,000/mm3 
Prothrombin Time (PT) 
1.1-1.2 x ULN 
1.3-1.5 x ULN 
1.6-3.0 x ULN 
>3.0 x ULN 
Partial Thromboplastin 
Time (PTT) 
1.1-1.6 x ULN 
1.7-2.3 x ULN 
2.4-3.0 x ULN 
>3.0 x ULN 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
GASTROINTESTINAL 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Bilirubin (Fractionated bilirubin test must be performed when total bilirubin is elevated) 
Bilirubin for children 
greater than 3 months of 
age (when accompanied by 
any increase in other liver 
function test) 
1.1- <1.25 x ULN 
1.25- <1.5 x ULN 
1.5-1.75 x ULN 
>1.75 x ULN 
Bilirubin for children 
greater than 3 months of 
age (when other liver 
functions are in the normal 
range) 
1.1- <1.5 x ULN 
1.5- <2.0 x ULN 
2.0-3.0 x ULN 
>3.0 x ULN 
AST (SGOT) 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
ALT (SGPT) 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
GGT 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
Pancreatic Amylase 
1.1- 1.4 x ULN 
1.5- 1.9 x ULN 
2.0-3.0 x ULN 
>3.0 x ULN 
Uric Acid 
7.5-9.9 mg/dL 
10.0-12.4 mg/dL 
12.5-15.0 mg/dL 
>15.0 mg/dL 
Loss of Appetite 
Feeding minimally 
reduced 
Feeding reduced by 
more than 50% of 
normal for the child 
Refusing all feeds 
 
No solid or liquid 
taken orally for in 
the last 24 hours; 
requires intravenous 
fluids 
Diarrhea 
Change in 
consistency of stools 
OR increase of 1-3 
stools over baseline 
per 24-hour period 
liquid/watery stools 
OR increase of 4 to 6 
stools over baseline 
per 24-hour period 
Increase of ≥7 stools 
over baseline per 
24-hour period  
Requires IV fluid 
resuscitation and 
electrolytes 
repletion OR 
hypotensive shock 
 
Constipation 
Slight change in 
consistency and/or 
frequency of stools 
Hard, dry stools with 
a change in 
frequency 
Intestinal 
obstruction 
accompanied with 
abdominal pain 
Hospitalization; 
Severe abdominal 
distention and 
vomiting accompani
ed with severe 
abdominal pain 
Vomiting 
1 episode/ day (24h) 
2-3 episodes per day 
(24h) 
4-6 episodes per day 
(24h) 
Greater than 
6 episodes per day 
(24h) OR intractable 
vomiting 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
ELECTROLYTES 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Creatinine 
3 Months – 2 Years of age 
0.6-0.8 x ULN 
0.9-1.1 x ULN 
1.2-1.5 x ULN 
> 1.5 x ULN 
2 Years – 3 Years of age 
0.7-1.0 x ULN 
1.1-1.6 x ULN 
1.7-2.0 x ULN 
>2.0 x ULN 
Hypernatremia 
---- 
145-149 mEq/L 
150-155 mEq/L 
>155 mEq/L or 
abnormal sodium 
AND mental status 
changes 
Hyponatremia 
---- 
130-135 mEq/L 
129-124 mEq/L 
<124 mEq/L or 
abnormal sodium 
AND mental status 
changes 
Hyperkalemia 
5.0-5.9 mEq/L 
6.0-6.4 mEq/L 
6.5-7.0 mEq/L 
>7.0 mEq/L or 
abnormal potassium 
AND cardiac 
arrhythmia 
Hypokalemia 
3.0-3.5 mEq/L 
2.5-2.9 mEq/L 
2.0-2.4 mEq/L 
<2.0 mEq/L or 
abnormal potassium 
AND cardiac 
arrhythmia 
Hypercalcemia 
10.5-11.2 mg/dL 
11.3-11.9 mg/dL 
12.0-12.9 mg/dL 
>13.0 mg/dL 
Hypocalcemia 
7.8-8.4 mg/dL 
7.0-7.7 mg/dL 
6.0-6.9 mg/dL 
>6.0 mg/dL 
Hypomagnesemia 
1.2-1.4 mEq/L 
0.9-1.1 mEq/L 
0.6-0.8 mEq/L 
<0.6 mEq/L or 
abnormal 
magnesium AND 
cardiac arrhythmia 
hypoglycemia 
55-65 mg/dL 
40-54 mg/dL 
30-39 mg/dL 
<30 mg/dL or 
adnormal glucose 
AND mental status 
changes 
Hyperglycemia 
116-159 mg/dL 
160-249 mg/dL 
250-400 mg/dL 
>400 mg/dL or 
ketoacidosis 
Proteinuria 
Tr-1+ or <150 
mg/day 
2+ or 150-499 
mg/day 
3+ or 500-1000 
mg/day 
4+ or Nephrotic 
syndrome >1000 
mg/day 
Hematuria 
Microscopic <25 
cells/hpf 
Microscopic >25 
cells/hpf 
Gross hematuria 
Hospitalization; Life-
threatening 
consequences 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
Neurologic 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Irritability 
Easily consolable; 
minimal or no 
interference with 
activity. Episodes of 
continuous crying 
<60 min. 
Difficult to console. 
Episodes of 
continuous crying 
>60 min <120 min 
Inconsolable, 
prevents daily 
activity. 
Episodes of 
continuous crying 
>120 min 
Hospitalization or ER 
visit for treatment 
Decreased Activity  
Minimal decrease in 
alertness, minimal 
or no interference 
with activity 
Some interference 
with activity, slightly 
subdued 
unable to achieve 
normal level of 
alertness, lethargic 
ER visit or 
hospitalization for 
treatment or life-
threatening 
consequences 
Neuropathy/ Lower Motor 
Neuropathy 
---- 
Mild transient 
Paresthesia only 
Persistent or 
progressive 
paresthesia, burning 
sensation in feet, or 
mild dysesthesia; no 
weakness; mild to 
moderate deep 
tendon reflex 
changes; no sensory 
loss 
Onset of significant 
weakness, decrease 
or loss of DTRs, 
sensory loss in 
"stocking glove" 
distribution, 
radicular sensory 
loss, multiple cranial 
nerve involvement; 
bladder or bowel 
dysfunction, 
fasciculations, 
respiratory 
embarrassment 
from chest wall 
weakness. 
Myopathy or 
Neuromuscular Junction 
Impairment 
Normal or mild (<2 x 
ULN) CPK elevation 
Mild proximal 
weakness and/or 
atrophy not 
affecting gross 
motor function. Mild 
myalgias, +/- mild 
CPK elevation (<2 x 
ULN) 
Proximal muscle 
weakness and/or 
atrophy affecting 
motor function +/- 
CPK elevation; or 
severe myalgias with 
CPK >2 x ULN; 
Onset of 
myasthenia-like 
symptoms (fatigable 
weakness with 
external, variable 
ophthalmoplegia 
and/or ptosis), or 
neuromuscular 
junction blockade 
(acute paralysis) 
symptoms 
OTHER 
 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Fever/pyrexia 
38.0 - 38.4 °C or  
100.4 - 101.1 °F 
38.5 - 38.9 °C or  
101.2 - 102.0 °F 
39.0 - 40.0 °C or  
102.1 - 104.0 °F 
Greater than 40 °C 
or 104.0 °F 
Acute allergic reaction 
Pruritus without 
Rash 
Pruritic Rash 
Mild Urticaria 
Severe Urticaria 
Anaphylaxis, 
Angioedema 
Stomatitis 
Mild discomfort 
Painful, difficulty 
swallowing, but able 
to eat and drink 
Painful: unable to 
swallow solids 
Painful: unable to 
swallow liquids; 
requires IV fluids 
Illness or clinical adverse 
event (as defined according 
to applicable regulations) 
No interference with 
activity 
Some interference 
with activity not 
requiring medical 
intervention 
Prevents daily 
activity and requires 
medical intervention 
Hospitalization 
 
 
 
 
 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
APPENDIX 2:  
 
THROMBOTIC EVENTS TO BE REPORTED AS POTENTIAL ADVERSE EVENTS OF SPECIAL INTEREST 
 
The list of thrombotic events to be reported to the sponsor as potential AESIs is provided below. Further 
guidance may become available on thrombotic events of interest. 
 
• MedDRA PTs for large vessel thrombosis and embolism 
Aortic embolus, aortic thrombosis, aseptic cavernous sinus thrombosis, brain stem embolism, brain stem 
thrombosis, carotid arterial embolus, carotid artery thrombosis, cavernous sinus thrombosis, cerebral artery 
thrombosis, cerebral venous  sinus thrombosis, cerebral venous thrombosis, superior sagittal sinus thrombosis, 
transverse sinus thrombosis, mesenteric artery embolism, mesenteric artery thrombosis, mesenteric vein 
thrombosis, splenic artery thrombosis, splenic embolism, splenic thrombosis, thrombosis mesenteric vessel, 
visceral venous thrombosis, hepatic artery embolism, hepatic artery thrombosis, hepatic vein embolism, hepatic 
vein thrombosis, portal vein embolism, portal vein thrombosis, portosplenomesenteric venous thrombosis, 
splenic vein thrombosis, spontaneous heparin-induced thrombocytopenia syndrome, femoral artery embolism, 
iliac artery embolism, jugular vein embolism, jugular vein thrombosis, subclavian artery embolism, subclavian 
vein thrombosis, obstetrical  pulmonary embolism, pulmonary artery thrombosis, pulmonary thrombosis, 
pulmonary venous thrombosis, renal artery thrombosis, renal embolism, renal vein embolism, renal vein 
thrombosis, brachiocephalic vein thrombosis, vena cava embolism, vena  cava thrombosis, truncus coeliacus 
thrombosis 
 
• MedDRA PTs for more common thrombotic events 
Axillary vein thrombosis, deep vein thrombosis, pulmonary embolism, MedDRA PTs for acute myocardial 
infarction*, MedDRA PTs for stroke* 
 
Source: Shimabukuro T. CDC COVID-19 Vaccine Task Force. Thrombosis with thrombocytopenia syndrome (TTS) 
following Janssen COVID-19 vaccine. Advisory Committee on Immunization Practices (ACIP). April 23, 2021. 
https://www.cdc.gov/vaccines/acip/meetings/slides-2021-04-23.html. 
 
*Vaccine Adverse Event Reporting System (VAERS)Standard Operating Procedures for COVID-19 (as of 29 January 
2021) https://www.cdc.gov/vaccinesafety/pdf/VAERS-v2-SOP.pdf 
 
 


 
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VAC52150EBL2011 Protocol Version 2.0                                                  Protocol Version Date: 15/06/2021 
 
REFERENCES 
 
1. 
WHO. Ebola virus disease - key facts. 2021; Available from: http://www.who.int/news-room/fact-
sheets/detail/ebola-virus-disease (Accessed 27 May 2021). 
2. 
Investigator’s Brochure: VAC52150 (Ad26.ZEBOV, MVA-BN-Filo [MVA-mBN226B]) Edition 1. Janssen 
Vaccines & Prevention B.V. (18 December 2020). 
3. 
Addendum 2 to Investigator's Brochure VAC52150 (Ad26.ZEBOV, MVA-BN-Filo [MVA-mBN226B]) 
Edition 1. Janssen Vaccines & Prevention (06 May 2021). 
4. 
Afolabi M.O., Mutua G., Barry H. et al. Safety and Immunogenicity of a 2-Dose Heterologous 
Vaccine Regimen Against Ebola in African Children and Adolescents. American Society of Tropical 
Medicine and Hygiene 68th Annual Meeting, National Harbor, Maryland USA November 20-24, 
2019. 
5. 
Afolabi M.O., Ishola D., Manno D. at al. Safety and immunogenicity of a two-dose heterologous 
vaccine regimen against Ebola in children in Sierra Leone: a randomised, double-blind, placebo-
controlled trial. Lancet Infect Dis 2021; (accepted). 
6. 
Ishola D., Manno D., Afolabi M.O., et al. Safety and long-term immunogenicity of a two-dose 
Ad26.ZEBOV, MVA-BN-Filo Ebola vaccine regimen in adults: a randomised trial initiated during the 
Ebola outbreak in Sierra Leone. Lancet Infect Dis 2021; (accepted). 
7. 
European Commission. Vaccine against Ebola: Commission grants new market authorisations. 
Accessible at: https://ec.europa.eu/commission/presscorner/detail/en/ip_20_1248 (Accessed 27 
May 2021). 
8. 
European Medicines Agency. Zabdeno. Accessible at: 
https://www.ema.europa.eu/en/medicines/human/EPAR/zabdeno (Accessed 27 May 2021). 
9. 
Bockstal V., Roozendaal R., Van Effelterre T. et al. Immunobridging approach to assess clinical 
benefit as the basis for licensure of the monovalent Ebola vaccine. Accessible at: 
https://www.imi.europa.eu/sites/default/files/events/2018/ScientificSymposium/37-
%20Viki%20Bockstal.pdf (Accessed 27 May 2021). 
10. 
Ethical Considerations for Clinical Trials on Medicinal Products Conducted With the Pediatric 
Population: Recommendations of the Ad Hoc Group for the Development of Implementing 
Guidelines for Directive 2001/20/EC Relating to Good Clinical Practice in the Conduct of Clinical 
Trials on Medicinal Products for Human Use. 2008. 
11. 
WHO. Weight-for-age: Birth to 5 years in girls. Available from: 
http://www.who.int/childgrowth/standards/cht_wfa_girls_p_0_5.pdf?ua=1 (Accessed 27 May 
2021). 
12. 
WHO. Weight-for-age (5-10 years) percentiles in girls. Available from: 
http://www.who.int/growthref/cht_wfa_girls_perc_5_10years.pdf?ua=1 (Accessed 27 May 2021). 
13. 
American Society of Hematology. COVID-19 resources. Thrombosis with Thrombocytopenia 
Syndrome (also termed Vaccine-induced Thrombotic Thrombocytopenia). Updated 6 May 2021. 
https://www.hematology.org/covid-19/vaccine-induced-immune-thrombotic-thrombocytopenia  
(Accessed 27 May 2021). 
14. 
Brighton Collaboration. Interim Case Definition of Thrombosis with Thrombocytopenia Syndrome 
(TTS). 21 April 2021. 
https://brightoncollaboration.us/thrombosis-with-thrombocytopenia-
syndrome-interim-case-definition/ (Accessed 29 April 2021) 
 
 

SAP_001.pdf: 
Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
 
Statistical analysis plan 
 
An open label study to evaluate safety and immunogenicity of Ad26.ZEBOV 
booster dose in children previously vaccinated with the Ad.26ZEBOV and MVA-
BN-Filo vaccine regimen 
 
Protocol VAC52150EBL2011; Phase 2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Status: version 1.1 
Date: 6 January 2022 
Prepared by: LSHTM 
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
TABLE OF CONTENTS 
 
AMENDMENT HISTORY .......................................................................................................................... 4 
ABBREVIATIONS ..................................................................................................................................... 5 
1. 
INTRODUCTION .............................................................................................................................. 6 
1.1. 
Trial Objectives ..................................................................................................................... 6 
1.2. 
Trial Design ............................................................................................................................ 6 
1.3. 
Statistical Hypotheses for Trial Objectives ............................................................................ 6 
1.4. 
Sample Size Justification ....................................................................................................... 6 
1.5. 
Randomization and Blinding ................................................................................................. 7 
2. 
GENERAL ANALYSIS DEFINITIONS ................................................................................................... 7 
2.1. 
Analysis Window and Analysis Periods ................................................................................. 7 
2.2. 
Pooling of Data ...................................................................................................................... 8 
2.3. 
Analysis Sets .......................................................................................................................... 9 
2.3.1. 
Full Analysis Set ................................................................................................................ 9 
2.3.2. 
Per Protocol Immunogenicity Population ......................................................................... 9 
2.4. 
Definition of Subgroups ........................................................................................................ 9 
3. 
CHANGES TO THE PLANNED ANALYSIS ........................................................................................... 9 
4. 
PARTICIPANT INFORMATION ....................................................................................................... 10 
4.1. 
Demographics and Baseline Characteristics ....................................................................... 10 
4.1.1. 
Description of the procedure to determine Weight-for-Age percentile ........................ 10 
5. 
IMMUNOGENICITY ....................................................................................................................... 12 
5.1. 
Immunogenicity against EBOV GP ...................................................................................... 12 
5.1.1. 
Humoral Immune response ............................................................................................ 12 
5.1.1.1. 
Parameters ..................................................................................................................... 12 
5.1.1.2. 
Handling of Missing/ Invalid Data ................................................................................... 12 
5.2. 
Immunogenicity Against Ad26 Vector ................................................................................ 13 
5.2.1. 
Humoral Immune Response ........................................................................................... 13 
5.2.1.1. 
Handling Missing/ Invalid Data ....................................................................................... 13 
5.3. 
Analysis ............................................................................................................................... 13 
6. 
SAFETY .......................................................................................................................................... 14 
6.1. 
Adverse events .................................................................................................................... 14 
6.1.1. 
Definitions ...................................................................................................................... 14 
6.1.1.1. 
Solicited Local (Injection Site) Reactions ........................................................................ 15 
6.1.1.2. 
Solicited Systemic Adverse Events .................................................................................. 15 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
6.1.2. 
Imputation ...................................................................................................................... 16 
6.1.3. 
Analysis ........................................................................................................................... 16 
6.2. 
Clinical Laboratory Tests ..................................................................................................... 16 
6.2.1. 
Analysis Methodology .................................................................................................... 17 
6.3. 
Vital Signs and Physical Examination Findings .................................................................... 17 
REFERENCES ......................................................................................................................................... 18 
ATTACHMENTS ..................................................................................................................................... 18 
1. 
PERIODIC ALLOCATION OF ADVERSE EVENTS .............................................................................. 18 
2. 
Toxicity Grading Scale for Healthy Paediatric Participants (older than 3 months) Enrolled in 
Preventive Vaccine Clinical Trials ......................................................................................................... 20 
3. 
TRANSFORMING ON-SITE ASSESSMENTS AND DIARIES ASSESSMENTS OF SOLICITED AES INTO 
ANALYSIS FORMAT ............................................................................................................................... 24 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
 
AMENDMENT HISTORY 
 
Amendment 1  
Version 1.1 Dated 6 January 2022: 
- 
The interpretation of ELISA results for values above the upper limit of quantification 
(ULOQ) has been added under section 5.1.1.2 Handling of Missing/ Invalid Data 
- 
The protocol version and date were updated in the reference section.   
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
ABBREVIATIONS 
 
AE 
 
Adverse events 
aMLV   
Amphotropic murine leukemia virus 
BMI  
 
Body mass index 
CDC  
 
Center for Disease Control and Prevention 
CI 
 
Confidence interval 
CRF 
 
Case report form 
CSR 
 
Clinical study report 
IDMC  
Independent Data Monitoring Committee 
DPS 
 
Data programming specifications 
EBOV GP 
Ebola virus (formerly known as Zaire ebolavirus) Mayinga glycoprotein 
eCRF  
Electronic case report form 
ELISA  
Enzyme-linked immunosorbent assay 
EU/mL  
Elisa units/mL 
FAS 
 
Full analysis set 
FDA 
 
Food and Drug Administration 
IRE  
 
Immediate Reportable Event 
LLOQ  
Lower limit of quantification 
MeDRA 
Medical Dictionary of Regulatory Activities 
NSAIDs 
Non-steroidal anti-inflammatory drugs 
PP  
 
Per protocol 
PRNT  
Plaque reduction neutralization test 
SAE 
 
Serious adverse event 
SAP 
 
Statistical analysis plan 
SD 
 
Standard deviation 
ULOQ  
Upper limit of quantification 
VNA 
 
Virus neutralization assay 
 
WHO   
World Health Organization 
 
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
1. INTRODUCTION 
This is the statistical analysis plan (SAP) for the phase 2 VAC52150EBL2011 study. This SAP 
will be applicable to the study results based on version 2.0 of the clinical protocol of 15 June 
2021. 
1.1. Trial Objectives 
The primary objectives of the study are: 
• To assess the safety and tolerability of a booster dose of Ad26.ZEBOV at a dose of 
5×1010 viral particles (vp) in children previously vaccinated with Ad26.ZEBOV, MVA-
BN-Filo vaccine regimen with a 56-day interval. 
• To assess the vaccine-induced humoral immune response to the Ebola virus 
glycoprotein (EBOV GP), as measured by FANG ELISA, at 7 and 21 days following a 
booster dose of Ad26.ZEBOV at a dose of  5×1010  vp in children previously vaccinated 
with Ad26.ZEBOV, MVA-BN-Filo vaccine regimen with a 56 day interval. 
1.2. Trial Design 
This is a phase 2 open label study to evaluate the immune response to a booster dose of 
Ad26.ZEBOV administered to children who were previously vaccinated with Ad26.ZEBOV 
followed by MVA-BN-Filo 56 days later. Only children who received Ad26.ZEBOV and MVA-
BN-Filo regimen during their participation in VAC52150EBL3001 (EBOVAC-Salone) vaccine 
trial are eligible for enrolment in this study. Participants will be recruited in two age groups: 
children aged 4-11 years at the time of dose 1 vaccination and children aged 1-3 years at the 
time of dose 1 vaccination in the EBOVAC-Salone trial. Approximately 25 participants will be 
enrolled in each of these two age groups. 
Participants will be followed up to 28 days after their booster vaccination.  
 
1.3. Statistical Hypotheses for Trial Objectives 
As this study is designed to provide descriptive information regarding safety and 
immunogenicity without formal treatment comparisons, no formal statistical hypothesis 
testing is planned. 
  
1.4. Sample Size Justification 
Approximately 50 participants are expected to be enrolled in this trial (approximately 25 for 
each age group). The sample size is a convenience sample and is not based on formal 
hypothesis testing considerations. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
1.5. Randomization and Blinding 
Randomisation 
There will be no randomisation in this study with all participants receiving the booster dose 
of Ad26.ZEBOV vaccine.  
Blinding 
This is an open label study in which all the participants will receive the booster dose of 
Ad26.ZEBOV vaccine. 
 
2. GENERAL ANALYSIS DEFINITIONS 
Data will be shown by the study age groups (1-3 and 4-11 years). A booster baseline (or 
reference) value will be described as the value of the last available assessment performed 
prior to booster vaccination on Day 1. 
Unless otherwise specified, the results of the statistical analysis will be presented by period, 
except for participant information sections. For immunogenicity, vital signs, and physical 
examination, data will be presented per time point as appropriate. Assessments over time 
will be reported for each period or visit (see Section 2.1). 
• Categorical variables will be summarised using the following statistics: number of 
observations, percentages and Exact Clopper-Pearson 95% confidence interval (CI), as 
applicable. 
• Continuous variables will be summarised using the following statistics as appropriate: 
number of observations, geometric mean, arithmetic mean (mean), 95% confidence 
interval (CI) for the mean, standard deviation (SD), standard error (SE), median, 
quartiles (Q1 and Q3), minimum and maximum. In addition, reverse cumulative 
distribution plots will be used for exploratory analysis of antibody data. 
Unless otherwise specified, listings will only be generated for final analysis.  
2.1. Analysis Window and Analysis Periods 
For descriptive statistics over time, assessments (regardless of the investigated parameter) 
will be allocated to an analysis visit based on the actual date, visit slotting will be done as 
specified below. 
The number of days in the phase (relday) will be defined as: 
relday = visit day – reference day+1 for visits on or after the reference day, 
relday = visit day – reference day+1 for visits before the reference day, 
where the reference day equals the date of vaccination.  


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
All dates will be allocated to the following time points, based on relday (Table 1). A per 
protocol time interval is defined for immunogenicity analysis only. The windows are defined 
as per protocol. 
Table 1: Visit windows 
Analysis time point 
Target day 
Time interval 
Per Protocol Set 
(days) 
Time interval 
Full Analysis Set 
(days) 
Day 1 (Booster 
Baseline)* 
1 
≤ 1 
≤ 1 
Day 8 (7 days post 
booster dose) 
8 
[8-11] 
[8-14] 
Day 22 (21 days post 
booster dose) 
22 
[19-25] 
[15-29] 
Day 29 (28 days post 
booster dose) 
29 
NA 
NA 
NA: Not applicable, no immunogenicity assessment. 
*For safety assessments, the date and time of the baseline assessment should fall before the date and 
time of booster vaccination 
All assignments will be made in chronological order. Once an assessment is assigned to an 
analysis window it will no longer be used for a later window. 
For adverse events, vital signs abnormalities and concomitant therapies, analysis periods will 
be constructed as follows: 
Table 2: Period definitions 
Analysis period 
Interval 
From 
To  
Screening 
00:00 of the date of signing 
the informed consent 
One minute prior to booster dose 
on Day 1 
Post booster 
vaccination follow 
up (FU) 
One minute after the end of 
post booster vaccination 
period 
23:59 on the date of last 
contact (ie, study 
completion or early 
discontinuation) 
 
 
 
2.2. Pooling of Data 
Results will be pooled over sites. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
2.3. Analysis Sets 
2.3.1. Full Analysis Set 
The full analysis set will include all participants with the study intervention administration 
documented. All safety analysis will be performed on the full analysis set. In case a significant 
number of subjects are excluded from per protocol set (≥ 10%), then the immunogenicity 
analysis conducted on the full analysis set will be presented as sensitivity analysis, to 
investigate the impact of major protocol deviations on immune response. 
2.3.2. Per Protocol Immunogenicity Population 
The per-protocol immunogenicity population will include all vaccinated participants for whom 
immunogenicity data are available, excluding participants with major protocol deviations 
expected to impact the immunogenicity outcomes. The primary immunogenicity analysis will 
be performed on the per protocol immunogenicity population. 
2.4. Definition of Subgroups 
Data will be shown by age groups (1-3 years and 4-11 years).  
These subgroups will be used for showing the data for both safety analysis (full analysis set) 
and immunogenicity analysis (per protocol immunogenicity population).  
3. CHANGES TO THE PLANNED ANALYSIS 
No changes have been made to the planned analysis contained in the protocol.  
INTERIM ANALYSIS AND DATA MONITORING COMMITTEE REVIEW 
No interim analysis will be performed for this study. 
An IDMC will not be appointed for this study. The safety of Ad26.ZEBOV has already been 
shown in children in previous studies. A booster dose of Ad26.ZEBOV was also shown to be 
safe in adults. Thus the role of the IDMC will be designated to an independent medical 
reviewer. The independent medical reviewer will be identified by the sponsor to review the 
accumulating safety data on an ongoing basis to ensure the continuing safety of the 
participants enrolled in this study.  
The independent medical reviewer will be consulted periodically to review newly generated 
data. Ad hoc meetings may be requested via the sponsor if any of the pre-specified pausing 
rules for this study are met or in any situation that could affect the safety of the participants.  
For more details, refer to section 10.7 of the protocol. (1) 
  


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
4. PARTICIPANT INFORMATION  
Participant information will be analysed based on the full analysis set. 
4.1. Demographics and Baseline Characteristics 
Demographic 
characteristics 
and 
screening/baseline 
characteristics 
(e.g. 
physical 
examination, medical history) will be tabulated and summarized with descriptive statistics. 
The following demographic and baseline characteristics will be summarized: 
• Sex (Male/ Female) 
• Age (years) 
• Race 
• Ethnicity 
• Height 
• Weight 
• Weight-for-age for children 2-5 years and 5-10 years according to WHO growing charts 
• Height-for-age for children 2-5 years and 5-19 years according to WHO growing charts 
• Weight-for-height for children 2-5 years according to WHO growing charts 
• BMI-for-age for children 5-19 years according to WHO growing charts 
• Time between vaccination with Ad26.ZEBOV/ MVA-BA-Filo regimen and Ad26.ZEBOV 
booster vaccination (Median time, Q1, Q3, and range) 
The charts are available from the link below: 
https://www.who.int/tools/child-growth-standards/standards/weight-for-age 
https://www.who.int/tools/growth-reference-data-for-5to19-years/indicators/weight-for-
age-5to10-years  
https://www.who.int/tools/child-growth-standards/standards/length-height-for-age 
https://www.who.int/tools/growth-reference-data-for-5to19-years/indicators/height-for-
age  
https://www.who.int/tools/child-growth-standards/standards/weight-for-length-height 
https://www.who.int/tools/growth-reference-data-for-5to19-years/indicators/bmi-for-age 
4.1.1. Description of the procedure to determine Weight-for-Age percentile 
Reference material in Excel format can be found at this link for boys and girls: 
https://www.who.int/docs/default-source/child-growth/growth-reference-5-19-
years/weight-for-age-(5-10-years)/wfa-girls-perc-who2007-exp.xlsx?sfvrsn=5c5825c4_4 
https://www.who.int/docs/default-source/child-growth/growth-reference-5-19-
years/weight-for-age-(5-10-years)/wfa-boys-perc-who2007-exp.xlsx?sfvrsn=97ab852c_4  
.  
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Consult https://www.who.int/growthref/computation.pdf for more information on the data 
contained in the Excel file and instructions on how to calculate the exact percentile. 
 
First determine the age point to use. Age at the date of the corresponding weight assessments 
is to be used. Age is to be used at the half month point for the entire month; for example, 1.5 
months represents 1.0-1.99 months or 1.0 month up to but not including 2.0 months of age. 
Using the appropriate chart (boys vs. girls), determine the values of L, M, and S to use.  
4.2. Disposition Information 
The number and percentage of participants screened, vaccinated and included in each 
analysis set will be tabulated. 
Number and percentage of participants in the full analysis set that complete the study and 
those who are going to be discontinued together with reasons for discontinuation will be 
tabulated.   
4.3. Protocol Deviations 
All major protocol deviations will be tabulated by deviation category. Major protocol 
deviations that may have an influence on immune response will be flagged in a listing. 
4.4. Medical History 
Medical history will be listed. 
4.5. Concomitant Medications 
The analysis of concomitant therapies will be done using generic drug names. 
Based on their start and stop date, concomitant therapies will be reported in each analysis 
period during which they were applied. For missing or partial start and/or stop dates (time 
and/or day and/or month and/or year) the following allocation rules will be applied: 
• In case of partial start or stop dates, the concomitant therapy records will be allocated 
to periods using the available partial information, without imputations. If, for 
example, only month and year are available this will be compared to the month and 
the year of the periods, and concomitant therapy will be allocated to the period(s) 
where these date parts match. 
• In case of a completely missing start date, the concomitant therapy will be considered 
as having started before the trial. 
• In case of a completely missing end date, the concomitant therapy will be considered 
as ongoing at the end of the trial. 
Concomitant therapies will be tabulated per period (including follow up periods). A listing of 
all concomitant therapies will be provided. All therapies taken within 30 days prior to signing 
the informed consent form will also be listed. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
There will be special attention to analgesics/ antipyretics such as acetaminophen, non-
steroidal anti-inflammatory drugs (NSAIDS) and aspirin, administered until day 8 post-
vaccination (including day of vaccination). 
Remarks: 
In addition to the date information, time information is considered to allocate concomitant 
therapies to periods, if available. 
 
5. IMMUNOGENICITY 
 
The primary analysis of immunogenicity will be done on the per protocol immunogenicity 
population (as defined in section 2.3). 
 
5.1. Immunogenicity against EBOV GP 
 
5.1.1. Humoral Immune response 
5.1.1.1. 
Parameters 
Humoral immune response as measured by the following assays will be analysed. 
• Binding antibody responses using Filovirus Animal Nonclinical Group (FANG) 
enzyme-linked immunosorbent assay (ELISA): Quantification of antibodies binding to 
EBOV GP using ELISA units/mL (EU/mL). 
 
In addition, the following will be defined for ELISA (EU/mL): 
• Sample interpretation: a sample will be considered positive, if the value is above the 
assay-specific lower limit of quantification (LLOQ). 
• Responder: 
o If sample interpretation is negative at baseline before dose 1 vaccination in 
EBL3001 but positive post-baseline and the 
post-baseline value is greater than 2.5 x LLOQ; OR 
o If sample interpretation is positive at both baseline before dose 1 vaccination in 
EBL3001 and post-baseline and there is a greater than 2.5-fold increase from 
baseline (2.5-fold increase on the original scale) 
 
 
5.1.1.2. 
Handling of Missing/ Invalid Data 
ELISA: 
For ELISA binding antibody responses, values below the LLOQ will be imputed with LLOQ/2 
EU/mL. For the calculation of fold increases, values of binding antibody responses below the 
LLOQ will be imputed with LLOQ EU/mL. The LLOQ is provided in the database. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Values above the upper limit of quantification (ULOQ) will be imputed as ULOQ EU/mL. This 
will be used for the calculation of fold increases. The ULOQ is provided in the database. 
 
5.2. Immunogenicity Against Ad26 Vector 
 
5.2.1. Humoral Immune Response 
  
For immunogenicity against the Ad26 vector, a vector-specific reference value will be 
defined as the value closest but prior to the administration of the vector. The analysis of the 
immunogenicity against the vector will be performed only if the assay is available. 
Sample interpretation: a sample will be considered positive, if the value is above the assay 
LLOQ. 
The immune response values will be log10-transformed before any further handling. Results 
will be presented on the log10-scale, geometric mean titers and geometric mean increases 
with corresponding 95% CIS will be provided in the tables.  
 
5.2.1.1. 
Handling Missing/ Invalid Data 
For all outcomes (Ad26) values below the corresponding LLOQ will be imputed with half of 
the LLOQ (LLOQ/2). 
5.3. Analysis 
Summary statistics of observed values (on the log10 scale) will be presented for all 
immunogenicity parameters at each time point (Day 1, 8 and 22). Geometric mean 
concentrations with corresponding 95% CI and fold increase from pre-booster values will be 
calculated for ELISA antibody responses (EU/mL) on Day 8 and Day 22.  
A linear mixed model with the unstructured variance-covariance and unstructured mean (i.e. 
including time as a categorical variable) will be used to estimate the geometric mean 
concentrations and the associated 95% CI at these time points. 
Humoral immune response will be graphically presented using regimen profile plots using also 
data from the parent trial EBL3001.  
Reverse cumulative distribution plots will be used to display all the data for each 
immunogenicity parameter (on the log10 scale) against the percent of participants having at 
least that level of the parameter. The plots will be used to visually assess a value’s rank among 
all other values and to simplify comparison of immunogenicity parameter distribution by age 
group. 
Percentage of responders will be presented at different time points. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
 
6. SAFETY 
 
No formal statistical testing of safety data is planned. Safety data analysis will be performed 
on the full analysis set.  
Safety data will be analysed descriptively (including 95% CI, if applicable). Baseline for all 
safety parameters will be defined as the last value before the booster dose. 
The safety and tolerability data will include the following: 
Solicited local and systemic adverse events (reactogenicity) until 7 days post vaccination. 
Unsolicited adverse events, from provision of informed consent onwards until 28 days after 
booster vaccination. 
Serious adverse events (SAEs) until the end of the study (Day 29). 
 
6.1. Adverse events 
 
The analysis of adverse events will be done using the solicited adverse event terms pre-
specified in the diary card.  
All reported adverse events (solicited local, solicited systemic and unsolicited) after the 
booster vaccination will be attributed to analysis period as defined in Attachment 1. For each 
adverse event, the number and percent of subjects who experience at least one occurrence 
of the given event will be summarized. Summaries, listings, datasets and /or participant 
narratives may be provided as appropriate, for those participants who die, discontinue study 
vaccinations due to an adverse event, or experience a severe or serious adverse event. 
 
6.1.1. Definitions 
Solicited AEs are precisely defined events (local and systemic) that participants are specifically 
asked about and which are noted by participants or study staff in the diary. All the other AEs 
are considered unsolicited. Please refer Clinical Protocol (1), Section 11 for further details. 
Causality 
Solicited AEs will be considered as related to the study vaccine. Unsolicited AEs will be 
considered to be related to the use of the study vaccine if attribution is possibly, probably or 
very likely. An AE will be considered not related to the use of the study vaccine if the 
attribution is not related or doubtful. Please refer to Clinical Protocol (1), Section 11.1.2 for 
further details. 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Severity 
The severity of AEs is classified as mild, moderate, severe or potentially life threatening. 
Please refer to CTP (1), section 11.1.3 for further details. Solicited events that are graded less 
than mild, are not considered as AE. 
For induration/swelling both the diameter and grading as reported by the investigator are 
reported in the eCRF. Diameter will be used to derive toxicity grading. The highest grading 
should be used when both the diameter grading and the investigator-reported grading are 
available. 
6.1.1.1. 
Solicited Local (Injection Site) Reactions 
The analysis of solicited AEs will include: 
• Erythema (redness of skin) 
• Swelling/ Induration 
• Pain/Tenderness 
• Itching at the injection site 
6.1.1.2. 
Solicited Systemic Adverse Events 
The analysis of systemic solicited adverse events will include: 
• Fatigue/tiredness 
• Headache 
• Nausea/vomiting 
• Myalgia (muscle pain) 
• Arthralgia (joint pain) 
• Fever (defined as body temperature of 380 or higher) 
Adverse Events of Special Interest  
AESIs (including potential AESIs) are significant AEs that are judged to be of special interest 
because of clinical importance, known or suspected effects of similar vaccines, or based on 
nonclinical signals.  
- 
Thrombosis with Thrombocytopenia Syndrome (TTS)  
- 
Thrombotic events: suspected deep vessel venous or arterial thrombotic events as 
detailed in Appendix 2  
and/or  
- 
Symptomatic thrombocytopenia, defined as platelet count below LLN for the testing 
lab  
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
6.1.2. Imputation 
Missing data will not be imputed. If relationship of AEs to the study vaccine could not be 
derived (i.e. missing or unknown) it will be considered as unknown, for analysis purposes. 
Solicited local AEs will be considered as related with the use of study vaccine. Missing 
relationships will not be imputed. 
Period allocation of AEs is defined in Attachment 1. 
6.1.3. Analysis 
AEs (solicited and unsolicited) will be tabulated by presenting the number and percentage of 
participants having at least one of the observed AEs. 
The unsolicited AEs will be summarised by System Organ Classification (SOC) and Preferred 
Term (PT). The solicited AEs will be summarized by class (local, systemic) and PT. For solicited 
as well as unsolicited AEs, tables focussing on severity and relatedness to vaccine will be 
created. 
SAEs, AEs with fatal outcomes, AEs leading to permanent discontinuation of vaccine, Grade 3 
AEs and AESIs will be listed. Tables, summarizing all those parameters will also be created.  
For the solicited local and systemic AEs, the duration and time to onset of the events will also 
be summarized. If a participant experience more than one occurrence of a solicited event, the 
maximum duration of the events will be used. The time to first onset is defined as: 
[date of first onset – reference date+1] 
The reference date is the start date of each vaccination period. Duration and time to onset of 
AEs will be expressed in days. 
Summaries, listings, datasets, and/or participant narratives may be provided as appropriate, 
for those participants who die, or experience a SAE. 
The analysis of AEs will be based on MeDRA coded terms as provided in the clinical database. 
A listing of all SAEs during the study period will be provided. A listing of all AESIs (if flagged in 
the database) will be presented as well. 
 
6.2. Clinical Laboratory Tests 
 
In case a laboratory result is censored (no numeric value is available, but only a verbatim term) 
then a numeric value will be imputed by a value exceeding the cut-off value with one unit. 
(<x: subtract 1 unit from x, >x: add 1 unit to x)  
Toxicity grades will be determined according to the scales in Attachment 2 and 3. DMID 
Paediatric toxicity scale will be used. In case no toxicity grades are defined for a test, the 
abnormalities (below/ above normal range) will be used. In determining toxicity grades, the 
following rules will be applied: 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Highest grades/worst abnormalities are determined over the entire period. 
The abnormalities “abnormally low” and “abnormally high” are considered equally important 
and both abnormalities are shown in tables. (This mean that the sum of percentages can 
exceed 100%) 
If a laboratory value falls within the grading as specified in the grading table but also within 
the local laboratory normal limits, the value is considered as normal or Grade 0. 
Laboratory results falling between the grading scales will be allocated to the adjacent worst 
case grade (because the scale for some parameters in the grading table is not continuous as 
there may be zones where toxicity grades definitions do not exist) 
Definition emerging abnormalities following vaccination: An abnormality (toxicity grade or 
abnormality based on normal ranges) will be considered as emerging in a particular period if 
it is worse than baseline value. A shift from “abnormally low” at baseline to “abnormally high” 
post baseline or vice versa is also emerging. 
   
6.2.1. Analysis Methodology 
Laboratory data will be analysed based on the full analysis set. The data will be summarized 
by the type of laboratory test. 
Unless specified otherwise, percentages are calculated versus the number of participants in 
the analysis set with non-missing data for the parameter, period (if applicable) and 
vaccination group under evaluation. 
Tabulation of the highest emerging graded abnormalities per period and overall will be 
provided. Special attention will be given to participants who develop Grade 3 toxicities. For 
tests that have no grading, tabulations of the worst emerging abnormalities (below/above) 
will be performed.  
6.3. Vital Signs and Physical Examination Findings 
Vital sign abnormalities will be determined in accordance with the DMID Vital Signs Toxicity 
Grading (attachment 2.3.) for adults and adolescents. The worst abnormalities (following 
vaccination) of vital signs will be tabulated (i.e., showing number and percentage) and listed. 
 
For children, vital signs will be summarized and listed without grading. This summary will be 
presented per time point and showing the mean, standard deviation, median, Q1, Q3, 
minimum and maximum. Post-baseline assessments will be allocated to an analysis visit based 
on the eCRF visit; no visit slotting will be done. Scheduled visits will be used whenever 
available. If only unscheduled visits are available for a time point, the one closest to the 
scheduled visit will be used (in case of equidistance, the latest assessment will be selected). 
The baseline is the last available assessment prior to first vaccination. 
 
It is important to note that a full physical examination is only conducted at screening. At other 
visits, a brief, symptom-directed examination (including body length/height and weight in 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
children aged 12 months up to and including 24 months at the time of the boost vaccination) 
will be performed based on any clinically relevant issues, clinically relevant symptoms and 
medical history. Therefore, only a listing of participants with worst physical examination 
findings (i.e., abnormalities) following vaccination will be provided. 
 
REFERENCES 
1. Clinical Protocol VAC52150EBL2011: Version 2.0. An open label study to evaluate safety and 
immunogenicity of an Ad26.ZEBOV booster dose in children previously vaccinated with the 
Ad26.ZEBOV and MVA-BN-Filo vaccine regimen (15 June 2021). 
 
ATTACHMENTS 
 
1. PERIODIC ALLOCATION OF ADVERSE EVENTS 
Solicited AEs are always allocated to the Post boost vaccination period. Unsolicited AEs are 
allocated to the different periods according to the following rules: 
Step 1: Allocation of events to the periods: 
Adverse events present in the SDTM database are allocated to periods based on their start 
date/time. If the start date/time of an event falls between (or on) the start and stop date/time 
of a period, the AE is attributed to that period (treatment-emergent principle). 
Incomplete dates (i.e. time and/or day and/or month and/or year missing): 
- In case of partial start or stop dates, the events are allocated to the periods using the 
available partial information on start and end date; no imputation will be done. If, for 
instance, for the AE start date only month and year are available, these data are compared to 
the month and year information of the periods. This rule may lead to multiplication of the 
event because of its assignment to multiple periods (see below example). 
- In case of a completely missing end date (i.e., only for the calculation of duration): 
o In case the AE is not flagged as ongoing, the end date is considered as unknown, 
therefore the date will remain missing 
o In case the AE is flagged as ongoing the date is imputed by the cut-off date of the 
analysis for participants still ongoing in the study, and by the end date of the last 
period for participants who discontinued or completed the trial. 
Examples: 
Screening period: start date: 14JUN2021 - stop date: 28JUN2021 
Post-boost vaccination period: start date: 28JUN2021 - stop date: 19JUL2021 
1) Adverse event: start date: JUN2021- stop date: 15JUL2021 
As the start date only has information about month and year, only this information will be 
used from the periods (i.e. assuming any day of Jun is possible) and therefore the AE will be 
assigned to the Screening Period as well as to the Post-boost vaccination Period. 
2) Adverse event: start date: JUL2021- stop date 14JUL2021 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
As the AE starts after the Screening Period and after the start of the Post-boost vaccination 
period, it is only assigned to the Post-boost vaccination period. 
Remarks: 
o In addition to the date information, time information is considered to allocate AEs to 
periods, if available. 
o The imputation of missing end dates of ongoing AEs will only be used to derive the 
duration of the event (i.e., to give an indication of the minimum duration). The 
imputed end dates will not be shown in the data listings. 
 
Step 2: Combination of events: 
Overlapping/consecutive events are defined as events in the same participant with the same 
preferred term which have at least 1 day overlap or for which the start date of an event is 1 
day after the end date of the preceding event. Overlapping/consecutive events may be 
combined into one AE or not, according to the following rules: 
1) If overlapping/consecutive events start in one of the following periods - Screening or 
Post boost dose FU (i.e. non-active periods) - followed by an AE in - Post-boost dose 
(active period) - they are allocated to their respective periods and are considered as 
separate events. 
2) In case overlapping/consecutive events start within a single period, they are 
considered as one and the same AE. The individual events which contribute to this AE 
are retained as individual records in the ADaM database but are assigned the same 
onset, period, and total duration. 
3) In case overlapping/consecutive events start in both an active period followed by a 
nonactive period, they are allocated to the active period only and are considered as 
one and the same AE. The individual events which contribute to this AE are retained 
as individual records in the ADaM database but are assigned the same onset, 
treatment period, and total duration. 
4) In case an active period is followed by an active period, and the 
overlapping/consecutive events start in both periods, they are allocated to their 
respective period and are considered as separate AEs. The same rule applies for 2 non-
active periods. 
Remarks: 
1) Events can only be combined into one and the same AE if their start and stop dates 
are known. 
2) In case the completely missing end date is imputed (for period allocation), this date is 
also considered as a complete date. 
3) Time is not considered when determining overlap of events. 
4) Solicited AEs will never be attributed to the Screening Period 
 
Examples: 
Screening period: start date: 14JUN2021 - stop date: 28JUN2021 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Post-boost dose period: start date: 28JUN2021 - stop date: 26JUL2021 
Post-boost dose FU period: start date: 27JUL2021 - stop date: 15AUG2021 
Example for the above scenario 1 
AE1: start date: 20JUN2021 - stop date: 10JUL2021 
AE2: start date: 08JUL2021 - stop date: 18JUL2021 
AE1 will be attributed to the screening period and AE2 to the Post-boost dose period. 
Example for the above scenario 3 
AE1: start date: 18JUL2021- stop date: 28JUL2021 
AE2: start date: 28JUL2021- stop date: 08AUG2021 
As AE1 starts in the active period (Post-boost dose) and overlaps with AE2 which starts in a 
non-active period (Post-boost dose FU), this AE is considered as a single AE in the AE analysis 
starting on 18JUL2021 and ending on 08AUG2021 and is attributed to the post-boost dose 
period. 
 
2. Toxicity Grading Scale for Healthy Paediatric Participants (older 
than 3 months) Enrolled in Preventive Vaccine Clinical Trials 
Adapted from: DMID° Pediatric Toxicity Tables (November 2007, draft). For adverse events 
not included in the tables below, refer to the severity criteria guidelines. Reference source 
not found. Local lab references take preference over the DMD table and the different grades. 
The abbreviations used in the following tables are: LLN: lower limit of normal; IV: intravenous; 
ULN: upper limit of normal. 
 
LOCAL REACTIONS 
  
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Tenderness 
Mild discomfort to 
touch; minimal to 
no limitation of use 
of limb 
Notable discomfort 
to touch; Greater 
than minimal 
limitation of use of 
limb 
Significant 
discomfort at rest; 
Severe limitation of 
use of limb 
Hospitalization or ER 
visit for treatment 
Erythema 
<10 mm 
10-25 mm 
26-50 mm 
>50 mm or any 
grade 3 with 
hospitalization or ER 
visit for treatment 
Swelling 
<10 mm 
10-25 mm 
26-50 mm 
>50 mm or any 
grade 3 with 
hospitalization or ER 
visit for treatment 
HEMATOLOGY 
  
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Hemoglobin for 
children greater than 
3 months and less 
than 2 years of age 
9.0-9.9 gm/dL 
7.0-8.9 gm/dL 
<7.0 gm/dL 
Cardiac Failure 
secondary to 
Anemia 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Hemoglobin for 
children 
greater than 2 years 
of age 
10.0-10.9 gm/dL 
7.0-9.9 gm/dL 
<7.0 gm/dL 
Cardiac Failure 
secondary to 
Anemia 
Absolute Neutrophil 
Count 
750-1200/mm3 
400-749/mm3 
250-399/mm3 
<250/mm3 
Platelets 
75,000-
99,999/mm3 
50,000-74,999/mm3 
25,000-49,999/mm3 
<25,000/mm3 
Prothrombin Time 
(PT) 
1.1-1.2 x ULN 
1.3-1.5 x ULN 
1.6-3.0 x ULN 
>3.0 x ULN 
Partial 
Thromboplastin 
Time (PTT) 
1.1-1.6 x ULN 
1.7-2.3 x ULN 
2.4-3.0 x ULN 
>3.0 x ULN 
 
 
GASTROINTESTINAL 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Bilirubin (Fractionated bilirubin test must be performed when total bilirubin is elevated) 
Bilirubin for children 
greater than 3 months of 
age (when accompanied 
by any increase in other 
liver function test) 
1.1- <1.25 x ULN 
1.25- <1.5 x ULN 
1.5-1.75 x ULN 
>1.75 x ULN 
Bilirubin for children 
greater than 3 months of 
age (when other liver 
functions are in the 
normal range) 
1.1- <1.5 x ULN 
1.5- <2.0 x ULN 
2.0-3.0 x ULN 
>3.0 x ULN 
AST (SGOT) 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
ALT (SGPT) 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
GGT 
1.1- <2.0 x ULN 
2.0- <3.0 x ULN 
3.0-8.0 x ULN 
>8.0 x ULN 
Pancreatic Amylase 
1.1- 1.4 x ULN 
1.5- 1.9 x ULN 
2.0-3.0 x ULN 
>3.0 x ULN 
Uric Acid 
7.5-9.9 mg/dL 
10.0-12.4 mg/dL 
12.5-15.0 mg/dL 
>15.0 mg/dL 
Loss of Appetite 
Feeding minimally 
reduced 
Feeding reduced by 
more than 50% of 
normal for the 
child 
Refusing all feeds 
No solid or liquid 
taken orally for in 
the last 24 hours; 
requires intravenous 
fluids 
Diarrhea 
Change in 
consistency of stools 
OR increase of 1-3 
stools over baseline 
per 24-hour period 
liquid/watery 
stools 
OR increase of 4 to 
6 
stools over 
baseline 
per 24-hour period 
Increase of ≥7 
stools 
over baseline per 
24-hour period 
Requires IV fluid 
resuscitation and 
electrolytes 
repletion OR 
hypotensive shock 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
Constipation 
Slight change in 
consistency and/or 
frequency of stools 
Hard, dry stools 
with 
a change in 
frequency 
Intestinal 
obstruction 
accompanied with 
abdominal pain 
Hospitalization; 
Severe abdominal 
distention and 
vomiting accompani 
ed with severe 
abdominal pain 
Vomiting 
1 episode/ day (24h) 
2-3 episodes per 
day 
(24h) 
4-6 episodes per 
day 
(24h) 
Greater than 
6 episodes per day 
(24h) OR intractable 
vomiting 
 
ELECTROLYTES 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Creatinine 
3 Months – 2 Years of age 
0.6-0.8 x ULN 
0.9-1.1 x ULN 
1.2-1.5 x ULN 
> 1.5 x ULN 
2 Years – 3 Years of age 
0.7-1.0 x ULN 
1.1-1.6 x ULN 
1.7-2.0 x ULN 
>2.0 x ULN 
Hypernatremia 
---- 
145-149 mEq/L 
150-155 mEq/L 
>155 mEq/L or abnormal 
sodium AND mental 
status changes 
Hyponatremia 
---- 
130-135 mEq/L 
129-124 mEq/L 
<124 mEq/L or abnormal 
sodium AND mental 
status changes 
Hyperkalemia 
5.0-5.9 mEq/L 
6.0-6.4 mEq/L 
6.5-7.0 mEq/L 
>7.0 mEq/L or abnormal 
potassium AND cardiac 
arrhythmia 
Hypokalemia 
3.0-3.5 mEq/L 
2.5-2.9 mEq/L 
2.0-2.4 mEq/L 
<2.0 mEq/L or abnormal 
potassium AND cardiac 
arrhythmia 
Hypercalcemia 
10.5-11.2 mg/dL 
11.3-11.9 mg/dL 
12.0-12.9 mg/dL 
>13.0 mg/dL 
Hypocalcemia 
7.8-8.4 mg/dL 
7.0-7.7 mg/dL 
6.0-6.9 mg/dL 
>6.0 mg/dL 
Hypomagnesemia 
1.2-1.4 mEq/L 
0.9-1.1 mEq/L 
0.6-0.8 mEq/L 
<0.6 mEq/L or abnormal 
magnesium AND cardiac 
arrhythmia 
hypoglycemia 
55-65 mg/dL 
40-54 mg/dL 
30-39 mg/dL 
<30 mg/dL or adnormal 
glucose AND mental 
status changes 
Hyperglycemia 
116-159 mg/dL 
160-249 mg/dL  
250-400 mg/dL 
>400 mg/dL or 
ketoacidosis 
Proteinuria 
Tr-1+ or <150 
mg/day 
2+ or 150-499 
mg/day 
3+ or 500-1000 
mg/day 
4+ or Nephrotic 
syndrome >1000 mg/day 
Hematuria 
Microscopic <25 
cells/hpf 
Microscopic >25 
cells/hpf 
Gross hematuria 
Hospitalization; 
Lifethreatening 
consequences 
 
 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
NEUROLOGIC 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Irritability 
Easily consolable; 
minimal or no 
interference with 
activity. Episodes of 
continuous crying 
<60 min. 
Difficult to console. 
Episodes of 
continuous crying 
>60 min <120 min 
Inconsolable, 
prevents daily 
activity. 
Episodes of 
continuous crying 
>120 min 
Hospitalization or ER 
visit for treatment 
Decreased Activity 
Minimal decrease in 
alertness, minimal 
or no interference 
with activity 
Some interference 
with activity, slightly 
subdued 
unable to achieve 
normal level of 
alertness, lethargic 
ER visit or 
hospitalization for 
treatment or life-
threatening 
consequences 
Neuropathy/ Lower 
Motor 
Neuropathy 
---- 
Mild transient 
Paresthesia only 
Persistent or 
progressive 
paresthesia, burning 
sensation in feet, or 
mild dysesthesia; no 
weakness; mild to 
moderate deep 
tendon reflex 
changes; no sensory 
loss 
Onset of significant 
weakness, decrease or 
loss of DTRs, sensory loss 
in "stocking glove" 
distribution, radicular 
sensory loss, multiple 
cranial nerve 
involvement; bladder or 
bowel dysfunction, 
fasciculations, 
respiratory 
embarrassment from 
chest wall weakness. 
Myopathy or 
Neuromuscular 
Junction 
Impairment 
Normal or mild (<2 x 
ULN) CPK elevation 
Mild proximal 
weakness and/or 
atrophy not 
affecting gross 
motor function. Mild 
myalgias, +/- mild 
CPK elevation (<2 x 
ULN) 
Proximal muscle 
weakness and/or 
atrophy affecting 
motor function +/- 
CPK elevation; or 
severe myalgias with 
CPK >2 x ULN; 
Onset of 
myasthenia-like 
symptoms (fatigable 
weakness with 
external, variable 
ophthalmoplegia 
and/or ptosis), or 
neuromuscular 
junction blockade 
(acute paralysis) 
symptoms 
OTHER 
  
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Fever/pyrexia 
38.0 - 38.4 °C or 
100.4 - 101.1 °F 
38.5 - 38.9 °C or 
101.2 - 102.0 °F 
39.0 - 40.0 °C or 
102.1 - 104.0 °F 
Greater than 40 °C 
or 104.0 °F 
Acute allergic 
reaction 
Pruritus without 
Rash 
Pruritic Rash 
Mild Urticaria 
Severe Urticaria 
Anaphylaxis, 
Angioedema 
Stomatitis 
Mild discomfort 
Painful, difficulty 
swallowing, but able 
to eat and drink 
Painful: unable to 
swallow solids 
Painful: unable to 
swallow liquids; 
requires IV fluids 
Illness or clinical 
adverse 
event (as defined 
according 
to applicable 
regulations) 
No interference with 
activity 
Some interference 
with activity not 
requiring medical 
intervention 
Prevents daily 
activity and requires 
medical intervention 
Hospitalization 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
3. TRANSFORMING ON-SITE ASSESSMENTS AND DIARIES 
ASSESSMENTS OF SOLICITED AES INTO ANALYSIS FORMAT 
 
When creating the analysis dataset for solicited AEs, solicited AEs (recorded by day) need to 
be converted into the format of unsolicited AEs (recorded by event).  
 
All diary data will be considered, as well as any post-dose on-site assessment (scheduled as 
well as unscheduled) within 8 days after vaccination. For solicited local AEs for which a 
diameter is measured, the maximum of diameter derived grade and investigator severity will 
be used.  
 
The start date of the AE will be considered as the date of first occurrence of the solicited AE 
(both local and systemic). If on subsequent day(s), the same grade is reported, the last 
reported date is used as the end date of the AE. A new record is created in case the grade of 
the event changes. If there is a time gap of at least one day between two (or more) 
occurrences of the particular solicited AE, then the second (and/or next) occurrence will be 
considered as a new AE. In case no data is reported for a day, this is analyzed as no event 
reported. If the on-site assessment differs in grade or relatedness (if collected) with the 
corresponding diary data, only the highest grade and highest level of relatedness per AE will 
be kept in the analysis database and used in the tables and listings. If relatedness is not 
collected for an on-site temperature assessment, then relatedness collected on the diary will 
be used. 
 
The example below shows how the solicited AE should be converted into a format of 
unsolicited AEs: 
 
Data from the Participant Diary 
Participant: 0001 
Solicited systemic AE: Headache 
 
On site 
assessment 
DIARY DATA 
Solicited 
AE 
Day 1 
01Aug21 
Day 1 
01 
Aug21 
Day 2 
02 
Aug21 
Day 3 
03 
Aug21 
Day 4 
04 
Aug21 
Day 5 
05 
Aug21 
Day 6 
06 
Aug21 
Day 7 
07 
Aug21 
Day 8 
08 
Aug21 
Grade 
2 
1 
1 
0 
3 
3 
1 
0 
0 
Relatedness Doubtful 
Probable 
 
The data should be converted and stored in the AE dataset as follows: 
Participant 
No. 
AE 
Start 
Date 
(Char) 
Stop 
Date 
(Char) 
Severity 
Relatedness AEID 
Duration 
0001 
Headache 
01Aug21 02Aug21 
2 Probable 
1 
6 
0001 
Headache 
04Aug21 05JAug21 
3 Probable 
1 
6 
0001 
Headache 
06Aug21 06Aug21 
1 Probable 
1 
6 


Statistical Analysis Plan VAC52150EBL2011  
 
version 1.1 date 6 January 2022 
 
If a solicited AE ends after day 8: 
 
- The stop date of the event is the ‘Date of Last day of symptom’ as recorded in the eCRF and 
the ‘Maximum severity’ after Day 8 as recorded in the CRF. A separate record starting on day 
9 is created for this, in case this severity deviates from the previous record. 
 
Of note, to complete the start and end-date based on diary data, the date will be calculated 
based on the day the AE is reported relative to vaccination and not on the reported date. For 
example, if the vaccination is on 1st August 2021, and the AE starts on DAY 3, the start date 
will be set to the 3rd of August 2021 independent of the reported actual date. For the on-site 
assessments, the actual date as reported in the database will be used. 
 
For the calculation of duration, the first and last day is used, even in case interruptions 
occurred in between by missing reporting days or 0 grade. In the above example, the 4 records 
contribute to the same AE, therefore AEID is set to the same value and the duration of the AE 
is set to 6 for all records.