Partnership for Research on Ebola VACcinations (PREVAC)
Phase: PHASE2
Conditions: Ebola Virus Disease
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

Partnership for Research on 
Ebola VACcinnation 
(PREVAC)
Version 4.0 
10 March 2018
NCT02876328

PREVAC 
Version 4.0 
March 10, 2018 
1 
Partnership for Research on Ebola VACcination 
 (PREVAC)  
Version: 
4.0 
Date: 
March 10, 2018 
Sponsors: 
Office of Clinical Research Operations and 
Regulatory Compliance (OCRPRO) Division 
of Clinical Research (DCR) National Institute 
of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
Institut National de la Santé et de la 
Recherche Médicale (INSERM) 
London School of Hygiene & Tropical 
Medicine (LSHTM) 
Executive Committee: 
H. Clifford Lane (NIH), Yves Lévy (INSERM),
Peter Piot (LSHTM)
Coordinating Investigator: 
Yazdan Yazdanpanah MD, PhD 
Site Investigators:  
See Appendix C 
Ethics Review Boards: 
See Appendix C and Appendix F 
Protocol Number:  
C15-33 (INSERM), PREVACEBL3005 
(LSHTM) 
Study Vaccines Provided by: 
Merck & Co., Inc 
Whitehouse Station, New Jersey, United 
States 
Janssen Vaccines and Prevention BV Leiden, 
The Netherlands 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
2 
 
 
TABLE OF CONTENTS 
TABLE OF CONTENTS .................................................................................................... 2 
LIST OF ABBREVIATIONS ............................................................................................... 4 
PROTOCOL SYNOPSIS ................................................................................................... 6 
1. 
INTRODUCTION ..................................................................................................... 14 
1.1. 
Background .................................................................................................. 14 
1.1.1. 
Ebolavirus Zaire outbreak 2014 .................................................... 14 
1.1.2. 
J&J Ad26.ZEBOV/MVA-BN-Filo vaccine regimen ........................ 15 
1.1.3. 
Merck VSV-based vaccine ............................................................ 22 
1.1.4. 
Placebo ......................................................................................... 27 
1.2. 
Rationale ...................................................................................................... 27 
2. 
METHODOLOGY ..................................................................................................... 28 
2.1. 
Study design ................................................................................................ 28 
2.2. 
Primary objectives ........................................................................................ 30 
2.3. 
Primary hypotheses...................................................................................... 31 
2.4. 
Secondary objectives ................................................................................... 31 
2.5. 
Exploratory objectives to be assessed only if and when a correlate 
of protection has been established for both vaccine candidates .................. 33 
2.6. 
Endpoints ..................................................................................................... 33 
2.7. 
Sample size .................................................................................................. 34 
2.8. 
Study cohort ................................................................................................. 36 
2.8.1. 
Inclusion criteria ............................................................................ 37 
2.8.2. 
Exclusion criteria ........................................................................... 37 
2.9. 
Randomization and blinding ......................................................................... 37 
3. 
STUDY PLAN .......................................................................................................... 38 
3.1. 
Screening/Baseline data collection .............................................................. 39 
3.2. 
Vaccine administration ................................................................................. 39 
3.3. 
Follow-up data collection .............................................................................. 40 
3.4. 
Laboratory Tests .......................................................................................... 41 
3.5. 
Biometric Data .............................................................................................. 41 
4. 
ASSESSMENT OF SAFETY ................................................................................... 41 
4.1. 
SPECIFICATION OF SAFETY PARAMETERS ........................................... 42 
4.1.1. 
Definition of AE ............................................................................. 42 
4.1.2. 
Definition of an Adverse Reaction (AR) ........................................ 42 
4.1.3. 
Definition of SAE ........................................................................... 42 
4.1.4. 
Serious Adverse Reaction (SAR) .................................................. 43 
4.1.5. 
Suspected Unexpected Serious Adverse Reaction 
(SUSAR) ....................................................................................... 43 
4.1.6. 
Definition of Unanticipated Problems (UP) ................................... 43 
4.2. 
CLASSIFICATION OF AN AE ...................................................................... 44 
4.2.1. 
Relationship to study product ........................................................ 44 
4.2.2. 
Unexpectedness ........................................................................... 44 
4.3. 
TIME PERIOD AND FREQUENCY FOR EVENT ASSESSMENT 
AND FOLLOW-UP ....................................................................................... 45 
4.3.1. 
Time Period and Frequency Assessment ..................................... 45 
4.3.2. 
Time of collecting information ....................................................... 46 
4.3.3. 
Follow-up ...................................................................................... 46 
4.4. 
GENERAL REPORTING, RESPONSIBILITIES AND TIMELINES .............. 47 
4.5. 
HALTING RULES ......................................................................................... 48 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
3 
 
4.5.1. 
Study halting rules ........................................................................ 48 
4.5.2. 
Participant halting rules ................................................................. 48 
5. 
CLINICAL MANAGEMENT ...................................................................................... 48 
5.1. 
AEs and symptoms and signs of EVD .......................................................... 48 
5.2. 
HIV and syphilis counseling ......................................................................... 48 
5.3. 
Study withdrawal .......................................................................................... 48 
5.4. 
Pregnancy .................................................................................................... 48 
5.5. 
Childhood vaccinations ................................................................................ 49 
6. 
PHARMACY PROCEDURES .................................................................................. 49 
6.1. 
Study agents ................................................................................................ 49 
6.1.1. 
Ad26.ZEBOV and MVA-BN-Filo vaccines .................................... 49 
6.1.2. 
rVSV∆G-ZEBOV-GP ..................................................................... 49 
6.1.3. 
Sterile Normal Saline .................................................................... 49 
6.2. 
Study agent presentation and storage ......................................................... 50 
6.2.1. 
Study agent labels ........................................................................ 50 
6.2.2. 
Study agent storage ...................................................................... 50 
6.3. 
Preparation of study agents for injection ...................................................... 50 
6.4. 
Study agent accountability ........................................................................... 50 
7. 
EVALUATION .......................................................................................................... 51 
7.1. 
Planned data analyses ................................................................................. 51 
7.2. 
Independent DSMB ...................................................................................... 53 
7.3. 
Steering Committee...................................................................................... 54 
8. 
PROTECTION OF HUMAN SUBJECTS AND OTHER ETHICAL 
CONSIDERATIONS ................................................................................................. 55 
8.1. 
Local review of protocol and informed consent ............................................ 55 
8.2. 
Ethical conduct of the study ......................................................................... 55 
8.3. 
Informed consent of study participants ......................................................... 55 
8.4. 
Confidentiality of study participants .............................................................. 55 
8.5. 
Compensation .............................................................................................. 56 
8.6. 
Dissemination of trial results ........................................................................ 56 
8.7. 
Risk/Benefit Assessment .............................................................................. 56 
APPENDIX A TIME AND EVENTS SCHEDULE FOR BASELINE AND 
REGULARLY SCHEDULED FOLLOW-UP VISITS ................................................. 58 
APPENDIX B REFERENCE DOCUMENTS ON PREVAC STUDY WEBSITE ............... 61 
APPENDIX C SITE INVESTIGATORS AND ETHICS REVIEW BOARDS ..................... 62 
APPENDIX D PREVAC IMMUNOLOGICAL SUBSTUDY PROTOCOL ......................... 64 
APPENDIX E PREVAC SALIVA SAMPLE SUBSTUDY ................................................. 69 
APPENDIX F NIH ROLES IN THE CONDUCT OF PREVAC ......................................... 71 
REFERENCES ................................................................................................................ 72 
 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
4 
 
 
LIST OF ABBREVIATIONS 
Ad26 
Adenovirus serotype 26 
Ad26.ENVA.01 
Adenovirus serotype 26 vector expressing the human 
immunodeficiency virus type 1, Clade A envelope protein 
(vaccine) 
Ad26.ZEBOV 
Adenovirus serotype 26 vector expressing the glycoprotein of the 
Ebola virus Mayinga variant (vaccine) 
AE 
Adverse event 
AR 
Adverse reaction 
ALT 
Alanine transaminase 
AST 
Aspartate transaminase 
BDBV 
Bundibugyo ebolavirus 
BN 
Bavarian Nordic 
CBC 
Complete blood count 
CD 
Cluster of differentiation 
CFR 
Code of Federal Regulations 
ChAd3-EBO Z 
Replication deficient Chimpanzee adenovirus type 3-derived 
vector encoding the Ebola virus Zaire glycoprotein (vaccine) 
CI 
Confidence interval 
CRF 
Case report form 
DCR 
Division of Clinical Research 
DSMB 
Data and Safety Monitoring Board 
EBOV 
Ebola virus 
EC 
Ethics Committee 
eCRF 
Electronic case report form 
ELISA 
Enzyme-linked immunosorbent assay 
ELISpot 
Enzyme-linked immunospot 
EU 
European Union 
EVD 
Ebola virus disease 
FANG 
Filovirus Animal Nonclinical Group 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
GP 
Glycoprotein 
GSK 
GlaxoSmithKline plc (Brentford, UK) 
HIV 
Human Immunodeficiency Virus 
ICS 
Intracellular cytokine staining 
IFNγ 
Interferon gamma 
Ig 
Immunoglobulin 
IM 
Intramuscular 
IND  
Investigational New Drug 
INSERM 
French Institute of Health and Medical Research  
IRB 
Institutional Review Board 
J&J 
Johnson & Johnson 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
5 
 
LD50 
Median lethal dose 
LSHTM 
London School of Hygiene & Tropical Medicine 
MARV 
Marburg virus 
MCV 
Mean corpuscular volume 
MVA 
Modified Vaccinia Ankara 
MVA-BN-Filo 
Modified Vaccinia Ankara Bavarian Nordic vector expressing 
multiple filovirus proteins (vaccine) 
NHP 
Non-human primate 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
NK 
Natural killer 
NP 
Nucleoprotein 
OCRPRO 
Office of Clinical Research Operations and Regulatory 
PBMC 
Peripheral blood mononuclear cell 
PCR 
Polymerase chain reaction 
pfu 
Plaque-forming units 
PHAC 
Public Health Agency of Canada 
PID 
Participant Identification Code 
PREVAC 
Partnership for Research on Ebola VACcination 
PREVAIL 
Partnership for Research on Ebola Virus in Liberia 
RESTV 
Reston ebolavirus 
RDW 
Red cell distribution 
rHAd26/MVA 
Ad26.ZEBOV prime/MVA-BN-Filo boost (vaccine) 
RNA 
Ribonucleic acid 
RT-PCR 
Reverse transcriptase polymerase chain reaction 
rVSV 
Recombinant vesicular stomatitis virus 
rVSVΔG-ZEBOV-
GP 
Recombinant vesicular stomatitis virus-Zaire Ebola virus envelope 
glycoprotein (vaccine) 
SAE 
Serious adverse event 
SAR 
Serious adverse reaction 
SD 
Standard deviation 
SFU 
Spot forming unit 
SID 
Syringe Identification Number 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
SUDV 
Sudan ebolavirus 
TAFV 
Taï Forest ebolavirus 
TCID50 
50% tissue culture infective dose 
UK 
United Kingdom 
UP 
Unanticipated Problem 
US 
United States 
vp 
Viral particles 
VSV 
Vesicular stomatitis virus 
WHO 
World Health Organization 
ZEBOV 
Zaire ebolavirus 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
6 
 
PROTOCOL SYNOPSIS 
Full Title: 
Partnership for Research on Ebola VACcinations 
Short Title: 
PREVAC 
Clinical Phase: 
2 
Investigational New 
Drug (IND) Sponsor: NIAID/OCRPRO 
Sample Size: 
N= 2,800 with targets of 1,400 adults and 1,400 children 
to be enrolled under Version 4.0 which compares the 
Ad26.ZEBOV (adenovirus serotype 26 [Ad26] vector 
expressing the glycoprotein of the Ebola virus 
Mayinga)/MVA-BN-Filo (modified Vaccinia Ankara [MVA] 
Bavarian Nordic [BN] vector expressing multiple filovirus 
proteins) vaccine with placebo and the undiluted dose of 
the rVSVΔG-ZEBOV-GP (recombinant vesicular stomatitis 
virus [rVSV]-Zaire Ebola virus [ZEBOV] envelope 
glycoprotein [GP]) vaccine with placebo.  No participants 
were enrolled under Version 1.0; under Version 2.0, 537 
participants ≥ 12 years of age were randomized for the 
comparison of the Ad26.ZEBOV prime/MVA-BN-Filo boost 
vaccine (rHAd26/MVA) with placebo; and under Version 
3.0 an estimated 1,200 participants could be enrolled for 
the comparisons of the  rHAd26/MVA vaccine and the 
diluted dose of the rVSVΔG-ZEBOV-GP vaccine with 
placebo.  Thus, the total number randomized in PREVAC 
could be over 4,500 participants.  
Study Population: 
Adults and children, aged at least 1 year of age, will be 
invited to participate.  The trial will be conducted in 
Guinea, Liberia, Mali, and Sierra Leone  
Accrual Period: 
Estimated 6 months to enroll 2,800 participants under 
Version 4.0 
Study Design: 
Randomized, double-blind, placebo-controlled trial of 
three vaccine strategies.   
Study Duration: 
Following enrolment under Version 4.0, which is expected 
to take 6 months, participants will be seen at frequent 
follow-up visits for 12 months. After 12 months of follow-
up is complete for all participants, the study will be 
unblinded. If funding permits, all participants will be 
followed annually for 5 years.  
Background and Rationale: 
Preliminary data from an open-label, cluster randomized ring vaccination trial 
conducted in Guinea indicate that the Merck/New Link rVSVΔG-ZEBOV-GP 
vaccine prevented Ebola virus (EBOV) disease (EVD) events occurring at least 10 
days after randomization among close contacts immediately vaccinated as 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
7 
 
compared to those to be potentially vaccinated 21 days later. In that trial conducted 
in adults, most EVD events occurred shortly following vaccination; no EVD events 
occurred more than 6 days after vaccination.  The durability of the effect of the 
vaccine was not assessed. 
Two other vaccines, the GlaxoSmithKline (GSK) ChAd3-EBO Z (replication 
deficient Chimpanzee adenovirus type 3-derived vector encoding the Ebola virus 
Zaire [EBO Z] GP) vaccine and the Johnson & Johnson (J&J) prime boost vaccine, 
rHAd26/MVA, have completed phase 1 testing.  In addition, safety and 
immunogenicity after 12 months of the rVSVΔG-ZEBOV-GP and ChAd3-EBO Z 
vaccine have been evaluated in a phase 2, placebo-controlled trial of adults in 
Liberia (Partnership for Research on Ebola Virus in Liberia [PREVAIL] I).  
By the end of 2015 in West Africa, the incidence of EVD had dramatically 
decreased.  However, multiple questions regarding the safety and effectiveness of 
EVD vaccines remain unanswered, including the durability and the immediacy of 
immune responses generated by different vaccine strategies with and without a 
booster, and the safety of the different vaccines, particularly in children.   
In order to answer these questions, a randomized trial that includes adults and 
children will be carried out.  The trial will compare three vaccine strategies with 
placebo for immunogenicity and safety.  The comparisons will be carried out 
separately for adults and children and for adults and children combined.  The three 
vaccine strategies are: 1) Ad26.ZEBOV (prime) (0.5 mL) followed by an MVA-BN-
Filo boost (0.5 mL) at 56 days; 2)  rVSVΔG-ZEBOV-GP (prime) (1 mL) with a 
placebo (1 mL) boost at 56 days; and 3) rVSVΔG-ZEBOV-GP (prime) (1 mL) with 
a rVSVΔG-ZEBOV-GP boost (1 mL) at 56 days. There will be two placebo groups, 
one with a syringe fill volume of 0.5 mL to match the fill volumes of the 
Ad26.ZEBOV and MVA-BN-Filo vaccines and the other with a fill volume of 1 mL 
to match the fill volume for the rVSVΔG-ZEBOV-GP vaccine.  The two placebo 
groups will be combined for data analyses. 
Anti-GP ZEBOV antibodies will be measured and used to compare the 
immunogenicity profiles of each of the vaccine strategies with placebo. Correlates 
of protective immunity are not yet known.  Thus, blood will be stored from each 
visit for future use when a correlate of protection is identified.   
This randomized trial will provide data for the three vaccine strategies for adults 
and children on safety and durability (12 months and possibly annually thereafter), 
immediacy (14 days), and kinetics (7, 14, and 28 days after prime vaccination and 
7 and 28 to 35 days after the booster vaccination) of the immune response.  
A substudy in adults in Guinea will involve exploratory analyses of T cell and 
memory B cell responses to the three vaccine strategies (see Appendix D).  
A substudy in children will investigate viral shedding after vaccination (see 
Appendix E).  
Under Version 4.0, participants (N=2,800) will be randomly allocated to one of the 
three vaccine strategies or placebo. There will be 800 participants assigned to the 
rHAd26/MVA vaccine strategy and 800 to the rVSVΔG-ZEBOV-GP strategy with 
the placebo boost at 56 days; 400 participants will be assigned to the rVSVΔG-

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
8 
 
ZEBOV-GP arm that includes an rVSVΔG-ZEBOV-GP boost at 56 days; and 400 
participants will be assigned to each of the two placebo groups (800 total on 
placebo). 
The enrolment targets for adults and children are each 1,400 participants.  The 
study is powered to assess immune responses and safety outcomes separately in 
adults and children.   
In the event of a new Ebola epidemic, the study design, including the use of 
placebos, will be re-considered.  
Study Agent Descriptions: 
The 3 vaccine strategies to be studied are the rVSVΔG-ZEBOV-GP vaccine, with 
and without a boost at 56 days, and the rHAd26/MVA vaccine.  The rVSVΔG-
ZEBOV-GP vaccine requires a 1 mL administration from a 3 mL syringe for the 
prime and booster vaccination.  The rHAd26/MVA requires a 0.5 mL administration 
of Ad26.ZEBOV from a 3 mL syringe for the prime vaccination and a booster 
vaccination (0.5 mL from a 3 mL syringe) 56 days later of MVA-BN-Filo. 
Each study agent is briefly described below and in more detail in section 6.1. 
• The prime-boost rHAd26/MVA vaccine is comprised of an Ad26.ZEBOV 
vaccine which consists of a single recombinant, replication incompetent 
human Ad26 vector, constructed to express the Ebola virus Mayinga GP.  
The MVA-BN-Filo boost at 56 days encodes the GP of Sudan virus (SUDV; 
formerly known as Ebola Virus Sudan), EBOV (formerly known as Ebola 
Virus Zaire), and Marburg Virus (MARV) Musoke, and the nucleoprotein of 
Tai Forest virus (TAFV; formerly known as Côte d’Ivoire ebolavirus) (0.5 mL 
intramuscular [IM] administration for the Ad26.ZEBOV and MVA-BM-Filo 
vaccines). The Ad26.ZEBOV vaccine and the MVA-BN-Filo boost were 
given at the same dose in Versions 2.0, 3.0, and 4.0 of the PREVAC 
protocol.   
• The rVSVΔG-ZEBOV-GP vaccine is comprised of a single rVSV isolate (11481 
nt) modified to replace the gene encoding the VSV G envelope GP with the gene 
encoding the ZEBOV envelope GP (1 mL IM administration).  The rVSVΔG-
ZEBOV-GP vaccine was not used in Version 2.0.  The rVSVΔG-ZEBOV-
GP dose was given as a 2-fold diluted dose (approximately 5 x 107 plaque-
forming units [pfu]/mL) in Version 3.0 and is to be given as an undiluted 
dose (geometric mean of available assays 9.4 x 107 pfu/mL) in Version 4.0 
of the PREVAC protocol. The doses of rVSVΔG-ZEBOV-GP used in 
Versions 3.0 and 4.0 are referred to as the diluted and undiluted doses, 
respectively. 
For Version 4.0 of PREVAC, the primary objectives are to compare each of 
the three vaccine strategies with the pooled placebo group (3 pair-wise 
comparisons) for safety and immunogenicity.  These evaluations will be 
performed separately in adults and children.   
Separately for adults and children: 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
9 
 
• 
To compare each of the three vaccine strategies with the pooled placebo group 
(3 pair-wise comparisons) for antibody response 12 months after 
randomization (prime vaccination).  
In order to facilitate Merck regulatory filings and bridging of immune responses of 
the rVSVΔG-ZEBOV-GP vaccine from this study to other studies and between 
pediatric and adult populations, the following objective will be assessed specifically 
for those in the two rVSVΔG-ZEBOV-GP vaccine groups: 
• 
To compare the rVSVΔG-ZEBOV-GP vaccine (pooled rVSVΔG-ZEBOV-GP 
groups) with the matched placebo group for antibody response 28 days after 
randomization (prime vaccination).  
In order to facilitate Janssen regulatory filings of the rHAd26/MVA vaccine, the 
following objective will be assessed specifically for those in the rHAd26/MVA 
vaccine group: 
• 
To compare the rHAd26/MVA vaccine group with the matched placebo group 
for antibody response 3 months after randomization (approximately 28 days 
after the booster vaccination). 
Primary Endpoints 
 
• 
GP-EBOV antibody response at 12 months.  The primary analysis for adults 
and children will be performed for participants who do not have elevated 
antibody levels at baseline.  
• 
GP-EBOV antibody response at Day 28 after randomization (initial 
vaccination). For rVSVΔG-ZEBOV-GP arms only, Day 28 post first vaccination 
will be used for regulatory purposes for comparison to other studies and for 
bridging children to adults. The primary analysis for adults and children will be 
performed for participants who do not have elevated antibody levels at 
baseline. 
• 
GP-EBOV antibody response at Month 3 after randomization (approximately 
28 days after the booster vaccination).  For the rHAd26/MVA vaccine arm only, 
Month 3 will be used for regulatory purposes. The primary analysis for adults 
and children will be performed for participants who do not have elevated 
antibody levels at baseline. 
 
Primary Hypotheses 
The null hypothesis for each of the three comparisons at 12 months is that the 
vaccine strategy does not differ from the pooled placebo group (0.5 mL and 1.0 
mL). 
The following hypothesis will only apply to the rVSV∆G-ZEBOV-GP vaccine for 
the purpose of facilitating Merck regulatory filings: 
The null hypothesis is that the antibody response for the pooled rVSVΔG-
ZEBOV-GP group does not differ from the matched placebo group (0.5 mL 
and 1.0 mL groups) at Day 28 after randomization (initial vaccination). 
The following hypothesis will only apply to the rHAd26/MVA vaccine for the 
purpose of facilitating Janssen regulatory filings: 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
10 
 
The null hypothesis is that the antibody response for the rHAd26/MVA 
vaccine does not differ from the matched placebo group (0.5 mL and 1.0 
mL groups) at Month 3 after randomization. 
Secondary and Exploratory Objectives 
There are a number of secondary objectives that include comparisons of each 
vaccine strategy versus placebo for: 1) serious adverse events (SAEs; see section 
4.1.3 for definition), injection site reactions, targeted symptoms of any grade 
severity, and grade 3 or 4 adverse events (AEs), in the 28 to 35 days following 
both the prime and booster vaccinations; and 2) antibody levels measured 14 days 
after vaccination to assess the immediacy of the response and for up to 5 years (if 
funding permits) to assess the long-term durability of the antibody response.  
As a secondary objective, adults and children will be combined for comparisons of 
each vaccine strategy with placebo. 
In the event that a correlate of protection is identified, exploratory objectives that 
compare the three vaccine strategies with one another for the correlate will be 
carried out using measurements on stored blood samples. 
These secondary and exploratory objectives are stated in sections 2.4 and 2.5. 
 
Inclusion Criteria: 
• Informed consent/assent 
• Age ≥ 1 year 
• Planned residency in the area of the study site for the next 12 months 
• Willingness to comply with the protocol requirements 
Exclusion Criteria: 
• Fever > 38º Celsius 
• History of EVD (self-report) 
• Pregnancy (a negative urine pregnancy test is required for females of 
child-bearing potential, i.e., females who have experienced menarche or 
who are aged 14 years and older) 
• Positive human immunodeficiency virus (HIV) test for participants < 18 
years of age 
• Reported current breast-feeding 
• Prior vaccination against Ebola (self-report) 
• Any vaccination in the past 28 days or planned within the 28 days after 
randomization (initial vaccination) 
• In the judgement of the clinician, any clinically significant acute/chronic 
condition which would limit the ability of the participant to meet the 
requirements of the study protocol  
The demographics of those enrolled will be closely monitored to ensure that the 
targeted number of adults (N=1,400) and children (N=1,400) are enrolled. The 
steering committee may request that a gender or age group be prioritized to 
ensure broad generalizability of the findings.  
Procedures:  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
11 
 
Vaccination centers will be established in West Africa to consent, vaccinate and 
follow study volunteers.  A case report form (CRF) with a unique participant 
identification code (PID) will be completed for each participant consented and 
randomized.  The CRF may be paper or an electronic CRF (eCRF).  The 
procedures described in this protocol are in terms of a paper CRF.   
At the time of prime vaccination, a tear-off label on the syringe that includes a 
unique syringe identification code (SID) will be attached to the baseline CRF.  The 
SID is a unique number from a centrally prepared randomization schedule.  This 
will be the primary link between the blinded vaccine assignment and the PID.  
Study volunteers and clinical staff following the participants for safety and 
immunogenicity outcomes will be fully blinded.   
At 56 days, a syringe with the corresponding PID will be prepared for each 
participant on his/her scheduled 56 day follow-up visit.   
At study entry, study volunteers will be given an identification card that identifies 
them as study participants.  The identification card will include contact information 
in the event they experience vaccine side effects or signs and symptoms of Ebola. 
Participants will be instructed to avoid sharing items during the period of potential 
virus shedding. Women of childbearing potential will be told to use adequate birth 
control measures following randomization and until at least 3 months following the 
booster vaccination. Men who are sexually active with women of child-bearing 
potential will be told to use condoms following randomization and until at least 3 
months following the booster vaccination.  
Following randomization, all participants will be seen at 7, 14, 28, 56 and 63 days, 
and at 3, 6, and 12 months.  The study will be unblinded after all participants have 
completed 12 months of follow-up. If funding permits, all participants will be seen 
annually through 60 months.   
In addition, during the first week following the prime and booster vaccination, there 
will be daily contacts with participants < 18 years of age enrolled to assess injection 
site reactions and AEs graded for severity, including targeted symptoms, and 
SAEs.     
Participants will be encouraged to contact the vaccination center by telephone if 
they have any AEs.  If the participant experiences an AE requiring immediate 
attention, they will be given instructions on which hospital to go to for evaluation 
and treatment.  
Data and Safety Monitoring Board: 
An independent Data and Safety Monitoring Board (DSMB) will closely monitor 
accumulating safety data for adults and children in each age group (1-4, 5-11, 
and 12-17 years). Safety data for children will be reviewed at least monthly until 
the DSMB is satisfied that the vaccines are safe at which time less frequent 
reviews may be carried out.   
The safety reviews and strategy for sequentially enrolling increasingly younger 
children outlined below will be conducted in Version 3.0 of the PREVAC protocol 
for participants receiving Ad26.ZEBOV, diluted rVSVΔG-ZEBOV-GP, or placebo.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
12 
 
A similar strategy for safety reviews and sequentially enrolling younger children 
will be conducted for participants receiving Ad26.ZEBOV, undiluted rVSVΔG-
ZEBOV-GP, or placebo under Version 4.0.  Initially, in each version, the trial will 
only enroll adults and children ≥12 to ≤17 years of age.  After 70 children aged 
≥12 to ≤17 years have been enrolled (20 vaccinated with Ad26.ZEBOV, 30 
vaccinated with rVSVΔG-ZEBOV-GP, and 20 vaccinated with placebo) and 
followed for 28 days, the DSMB will be asked to consider whether vaccination of 
children ≥5 to ≤11 years can commence.  Similarly, after 70 children aged ≥5 to 
≤11 years have been enrolled, the DSMB will be asked to consider whether 
vaccination of children at least 1 year of age to ≤ 4 years can commence.   
Enrollment will not pause after 70 children are accrued in an age group.  Once 
the decision is made that the enrolment of children 1 to 4 years is safe, the goal 
will be to target the enrolment of the 1,400 children to achieve a similar 
distribution in the 3 age groups. 
Amendment (Version 2.0 of the PREVAC Protocol) 
Under Version 2.0 of the PREVAC protocol 537 participants ≥ 12 years of age 
were randomized to the prime-boost rHAd26/MVA vaccine strategy or to 
matching placebo.  The decision to begin the trial with the prime-boost 
rHAd26/MVA vaccine strategy and matching placebo and not include the 
rVSV∆G-ZEBOV-GP vaccine arms was reached one week before PREVAC was 
scheduled to begin with 5 randomized groups as originally planned (Version 1.0) 
because of questions regarding the occurrence of arthritis in 3 of 9 participants in 
a recent NIH study (described in Section 1.1.3) receiving vaccine from the lot of 
the rVSV∆G-ZEBOV-GP vaccine which was to be used in PREVAC.   
  
Amendment (Version 3.0 of the PREVAC Protocol) 
Under Version 3.0 of the PREVAC protocol an estimated 1,200 participants are 
to be randomized to one of five groups: 1) Ad26.ZEBOV (prime) followed by an 
MVA-BN-Filo boost at 56 days; 2)  rVSVΔG-ZEBOV-GP (prime, diluted dose) 
with a placebo boost at 56 days; 3) rVSVΔG-ZEBOV-GP (prime, diluted dose) 
with a rVSVΔG-ZEBOV-GP boost (diluted) at 56 days; 4) placebo for rVSVΔG-
ZEBOV-GP; or 5) placebo for the Ad26.ZEBOV and MVA-BN-Filo vaccines.   
 
The rVSVΔG-ZEBOV-GP vaccine was given at a 2-fold dilution in Version 3.0 
because the certificate of analysis of the vaccine lot was found to be higher than 
that used in the PREVAIL I trial in Liberia. Variation in titer/potency in live virus 
vaccines is common.  Vaccine manufacture and release for potency is based 
upon defined specifications and always encompasses a range, routinely with a 
lower and upper limit.  The lower limit, referred to as the nominal dose, is 
determined during development and is defined by the lowest dose for which there 
is demonstrated efficacy.  The lower limit for potency must still be valid at the end 
of shelf-life in order to ensure that the vaccine is still efficacious up until its 
defined expiry. 
 
Since there were limited numbers of children in previous studies of the rVSVΔG-
ZEBOV-GP vaccine, Version 3.0 of PREVAC used a measured approach with 
dilution of the rVSVΔG-ZEBOV-GP vaccine. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
13 
 
 
The Version 3.0 amendment also updated the safety information for the rVSV∆G-
ZEBOV-GP vaccine, removed language about the initial phase of PREVAC 
described in Version 2.0, and included minor edits to the Saliva Sample 
Substudy in Appendix E.  The data collection plan for Version 3.0 was the same 
as that described in Version 1.0 and Version 2.0.  
 
Amendment (Version 4.0 of the PREVAC Protocol) 
 
Version 4.0 of the protocol will not begin until at least 70 children in each of the 
three age groups (1-4, 5-11, and 12-17 years) have been enrolled in Version 3.0 
and the DSMB reviews safety data through 28 days for each age group. 
 
Under Version 4.0 of the PREVAC protocol, 2,800 participants (1,400 adults and 
1,400 children) will be randomized to one of five groups: 1) Ad26.ZEBOV (prime) 
followed by an MVA-BN-Filo boost at 56 days; 2)  rVSVΔG-ZEBOV-GP (prime, 
undiluted dose) with a placebo boost at 56 days; 3) rVSVΔG-ZEBOV-GP (prime, 
undiluted dose) with a rVSVΔG-ZEBOV-GP boost (undiluted) at 56 days; 4) 
placebo for rVSVΔG-ZEBOV-GP; or 5) placebo for the Ad26.ZEBOV and MVA-
BN-Filo vaccines (Figure 1).   
 
In addition to the dosing information for the rVSV∆G-ZEBOV-GP vaccine, 
this Version 4.0 amendment also updates the safety information for the rVSV∆G-
ZEBOV-GP vaccine, and amends the Immunological Substudy in Appendix D 
and the Saliva Substudy in Appendix E,  The Immunological Substudy is 
amended to state that up to 230 participants will be enrolled in Versions 2.0, 3.0, 
and 4.0 (in total).  The Saliva Substudy is amended to state that the target 
sample size is 140 children, with approximately an equal distribution of children 
in each of the three age groups (1-4, 5-11, and 12-17 years), for both Version 3.0 
and Version 4.0 of PREVAC. 
 
While the primary objectives of PREVAC will be accomplished with participants 
enrolled under Version 4.0, the participants randomized under Versions 2.0 and 
3.0 will contribute to the evaluation of each of the objectives that compares the 
rHAd26/MVA vaccine strategy to placebo.  In addition, participants enrolled 
under Version 3.0 of the protocol will provide information on the safety and 
immunogenicity of the diluted dose of the rVSVΔG-ZEBOV-GP vaccine 
compared to placebo for adults and children in each of the three age groups (1-4, 
5-11, and 12-17 years).   
 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
14 
 
1. 
INTRODUCTION 
1.1. 
Background 
1.1.1. 
Ebolavirus Zaire outbreak 2014 
The Zaire EVD outbreak In West Africa was first recognised on 22 March 2014 in 
Guinea. By December 1, 2014, the epidemic had spread to Liberia and Sierra 
Leone.  Through September 2015, over 28,000 confirmed cases of EVD with 
11,000 deaths had occurred in these 3 West African countries.  By the end of 2015 
in West Africa, the incidence of EVD had dramatically decreased.  Since the 
beginning of 2016, there have been flare-ups of Ebola in Sierra Leone, Guinea and 
Liberia but response teams in each country have minimized transmission.1  
The genus Ebolavirus is one of three genera in the family Filoviridae, which along 
with the genera  Marburgvirus and Cuevavirus, are known to induce viral 
haemorrhagic fever. The 5 distinct species included in the genus Ebolavirus are 
Bundibugyo (BDBV), Reston (RESTV), SUDV, TAFV, and ZEBOV. 
Ebola virus is a large, negative-strand ribonucleic acid (RNA) virus composed of 7 
genes encoding viral proteins, including a single GP. The virus is responsible for 
causing EVD in humans. In particular, BDBV, ZEBOV, and SUDV have been 
associated with outbreaks of EVD in Africa and reported case fatality rates of up 
to 90%.2 Transmission of EBOV to humans is not yet fully understood, but is likely 
due to incidental exposure to infected animals. EBOV then spreads through 
human-to-human transmission, with infection resulting from direct contact with 
blood, secretions, organs or other bodily fluids of infected people, and indirect 
contact with environments contaminated by such fluids.  
EVD has an incubation period of 2 to 21 days (mean 4 to 10 days).  The clinical 
manifestations have been summarized by Feldmann and Geisbert.3 They report 
that infection is followed by an abrupt onset of non-specific symptoms such as 
fever, chills, malaise, and myalgia.  The subsequent signs and symptoms indicate 
multisystem involvement and include systemic (prostration), gastrointestinal 
(anorexia, nausea, vomiting, abdominal pain, diarrhoea), respiratory (chest pain, 
shortness of breath, cough, nasal discharge), vascular (conjunctival injection, 
postural hypotension, edema), and neurological (headache, confusion, coma) 
manifestations.  Hemorrhagic manifestations consistent with disseminated 
intravascular coagulation may arise during the peak of the illness and include 
petechiae, ecchymoses, uncontrolled oozing from venepuncture sites, mucosal 
haemorrhages, and post-mortem evidence of visceral haemorrhagic effusions. A 
maculopapular rash associated with varying severity of erythema and 
desquamation can often be noted by day 5 to 7 of the illness; this symptom is a 
valuable differential diagnostic feature and is usually followed by desquamation in 
survivors. Abdominal pain is sometimes associated with hyperamylasemia and 
true pancreatitis. In later stages, shock, convulsions, severe metabolic 
disturbances, and, in more than half the cases, diffuse coagulopathy supervene. 
Laboratory findings include low white blood cell and platelet counts and elevated 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
15 
 
liver enzymes. In general, the symptoms last for about 7 - 14 days, after which 
recovery may occur. Death usually occurs 6 to 16 days after the onset of 
symptoms. 
The clinical manifestations of patients with EVD who were treated at an Ebola 
treatment unit in Monrovia, Liberia between August and October, 2014 have been 
described.4  Similar to the review by Feldmann and Geisbert,3 these authors 
reported that early symptoms of EVD included high fever (up to 40ºC), malaise, 
fatigue and body aches.  After 3 to 5 days, gastrointestinal symptoms began with 
epigastric pain, nausea, vomiting and diarrhea.  Other signs and symptoms 
included headache, conjunctival  infection, chest pain, abdominal pain, arthralgias, 
myalgias, and hiccups.  Clinically significant hemorrhage from the upper or lower 
gastrointestinal tract or both occurred in fewer than 5% of patients.4  Similar 
symptoms were reported for patients with laboratory confirmed EVD in Conakry, 
Guinea.  For 37 patients who presented to the hospital in March and April 2014, 
the most common symptoms were fever, fatigue, diarrhea, headache, vomiting and 
anorexia.  These patients were hospitalized a median of 5 days after symptom 
onset.5  
Among individuals who survive Ebola, the virus may persist in some body fluids 
for an extended period of time.  In 1977, a case report described an investigator 
who accidently inoculated himself with EBOV and the virus was recovered in his 
semen 61 days after onset of illness, but not on days 76, 92, and 110.6 During an 
outbreak of Ebola in the Congo in 1995, infectious virus was detected in the semen 
82 days after disease onset in one patient.6  In March 2015, it was reported that a 
woman in Liberia possibly developed Ebola through unprotected sex with an Ebola 
survivor.6  Subsequently, genomic evidence confirmed the sexual transmission 
with an Ebola survivor for whom there was evidence of persistence of infective 
EBOV in semen for 179 days or more.6  Recent reports from Liberia and Guinea 
indicate a large fraction of men have EBOV in their semen after their recovery.6,7  
An overview summarized the literature on viral persistence in survivors of Ebola.6  
1.1.2. 
J&J Ad26.ZEBOV/MVA-BN-Filo vaccine regimen  
Janssen Vaccines and Prevention BV (a Janssen pharmaceutical company of 
Johnson & Johnson), in collaboration with Bavarian Nordic GmbH, is investigating 
the potential of a prophylactic Ebola vaccine regimen comprised of the following 2 
candidate Ebola vaccines: 
Ad26.ZEBOV is a monovalent vaccine expressing the full length Mayinga GP of 
the EBOV (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 multivalent vaccine 
expressing the SUDV GP, the EBOV GP, the MARV Musoke GP, and the TAFV 
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. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
16 
 
The EBOV GP that is currently circulating in West Africa has 97% homology to the 
EBOV GP used in these vaccine regimens. 
Nonclinical Studies 
Immunogenicity and Efficacy 
Immunogenicity and efficacy of the vaccine combination Ad26.ZEBOV and MVA-
BN-Filo was evaluated in an non-human primate (NHP) model (i.e., Cynomolgus 
macaques, Macaca fascicularis). The combination was assessed in a multivalent 
filovirus setting in a small number (2 per regimen) of animals and the study 
included heterologous prime-boost regimens of Ad26, Ad35 and MVA-BN-Filo 
vectors expressing different Ebola and Marburg proteins. Full protection from EVD 
and death after wild-type EBOV Kikwit 1995 challenge was obtained with all 
heterologous regimens, including the Ad26.ZEBOV and MVA-BN-Filo vaccine 
regimen. All heterologous prime-boost regimens induced comparable immune 
responses against the EBOV Mayinga GP. Independently of the vaccine regimen, 
a strong boost effect was seen after heterologous prime-boost immunization.6 Two 
additional studies involving more animals have been performed, to strengthen the 
robustness of the nonclinical efficacy data, and also to optimize the prime-boost 
schedule so as to obtain induction of protective immunity as quickly as possible, 
to specifically respond to the EVD outbreak in West Africa. Complete survival was 
observed with 8-week heterologous regimens with Ad26.ZEBOV as prime and 
MVA-BN-Filo as boost. Partial protection was observed with both MVA-BN-Filo as 
a prime immunization and a shorter prime/boost interval.7,15  
Toxicology 
A repeated-dose toxicity study in rabbits was performed with prime-boost 
combinations of Ad26.ZEBOV and MVA-BN-Filo. The different dose regimens 
were well tolerated when administered twice by IM injection to New Zealand White 
rabbits with a 14-day interval period. All vaccine dosing regimens resulted in 
detectable EBOV GP-specific antibody titers. No significant toxicological effects 
(no adverse effects) were observed. The immune response was associated with 
transient increases in fibrinogen, C-reactive protein, globulin, decreases in 
hematocrit and hemoglobin, and microscopic findings in draining iliac lymph nodes, 
spleen and at the injection sites. The findings were noted to be recovering over a 
14-day treatment-free period and were considered to reflect a physiological 
response associated with vaccination. There were no effects noted that were 
considered to be adverse. 
In an embryo fetal and pre- and postnatal development study in female rabbits, 
there was no maternal or developmental toxicity following maternal exposure to 
the vaccine regimens during the premating and gestation period. All vaccine 
regimens elicited detectable EBOV GP-specific maternal antibody titers that were 
transferred to the fetuses. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
17 
 
Biodistribution 
Single-dose biodistribution studies in rabbits were performed using the MVA-BN 
vector or the Ad26 vector in combination with another insert (Ad26.ENVA.01: an 
experimental, prophylactic Ad26 vector expressing the HIV type 1, Clade A 
envelope protein). MVA-BN distributed to the skin, muscle, blood, spleen, lung, 
liver, and pooled lymph nodes and was rapidly cleared (within 48 hours following 
vaccination). Ad26.ENVA.01 was primarily localized in the injection site muscle, 
the regional lymph nodes and the spleen. Three months after the single IM 
injection of Ad26.ENVA.01, the vaccine was cleared from most of the examined 
tissues. As biodistribution is dependent on the vector platform (MVA-BN or Ad26) 
and not on the insert, it can be assumed that recombinant MVA-BN-Filo or 
Ad26.ZEBOV is distributed in the same way as the MVA-BN vector or 
Ad26.ENVA.01 vector, respectively. 
Clinical Studies 
Currently, the clinical program to assess the safety/tolerability and immunogenicity 
of Ad26.ZEBOV and MVA-BN-Filo consists of five Phase 1 studies: 
VAC52150EBL1001, 
VAC52150EBL1002, 
VAC52150EBL1003, 
VAC52150EBL1004 and VAC52150EBL1005 (Ad26.ZEBOV only), three Phase 2 
(VAC52150EBL2001, VAC52150EBL2002 and VAC52150EBL2003) and three 
Phase 3 
clinical 
studies 
(VAC52150EBL3001, 
VAC52150EBL3002 
and 
VAC52150EBL3003), where monovalent Ad26.ZEBOV vaccine and multivalent 
MVA-BN-Filo vaccine are combined in homologous or heterologous prime-boost 
regimens in which each vector is used to prime a filovirus-specific immune 
response followed by a boost immunization with the same or the other vector 2 to 
12 weeks later. To date, 1,176 healthy adult subjects have been enrolled in the 
ongoing studies. Two additional Phase 1 studies investigating MVA-BN-Filo have 
been performed (EBL01 and CVD-Mali Ebola Vaccine #1000).7  
Phase 1 studies 
To date, the results of the primary analysis of two Phase 1 clinical studies with 
Ad26.ZEBOV and/or MVA-BN-Filo are available (VAC52150EBL1001 and 
VAC52150EBL1002).14,15,8  
VAC52150EBL1001, a first-in-human study, is a randomized, placebo-controlled, 
observer-blind study in healthy adults evaluating the safety, tolerability and 
immunogenicity of 4 placebo-controlled regimens using MVA-BN-Filo at a dose of 
1x108 50% tissue culture infective dose (TCID50) and Ad26.ZEBOV at a dose of 
5x1010 viral particles (vp): 2 regimens with MVA-BN-Filo as prime and 
Ad26.ZEBOV as boost at a 28- or 56-day interval, and 2 regimens with 
Ad26.ZEBOV as prime and MVA-BN-Filo as boost at a 28- or 56-day interval. A 
fifth regimen, with Ad26.ZEBOV at a dose of 5x1010 vp as prime, and MVA-BN-Filo 
at a dose of 1x108 TCID50 at a 14-day interval was evaluated in an open-label, 
uncontrolled fashion in healthy adults. VAC52150EBL1001 has enrolled 
87 subjects and primary analysis data for safety and immunogenicity are available 
for 85 subjects (performed when all subjects had completed their 21-day post-

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
18 
 
boost visit or discontinued earlier). Four SAEs occurred; none of these was 
considered related to the study vaccines. Two subjects in the 14-day regimen 
group who experienced grade 3 neutropenia did not receive the boost vaccination 
as they met criteria for contraindications to boost (specified in the protocol), but 
continued scheduled assessments as planned. There were no SAEs related to the 
study vaccines.16  
In the placebo-controlled regimens, overall frequencies of solicited local and 
systemic AEs were higher after MVA-BN-Filo and Ad26.ZEBOV, relative to 
placebo. In addition, there was a trend towards higher overall frequencies of 
solicited local and solicited systemic events after Ad26.ZEBOV, relative to MVA-
BN-Filo. This trend was not observed in the open-label 14-day regimen. The most 
frequent solicited local AE was injection site pain. The most common solicited 
systemic AEs were fatigue, headache and myalgia. Grade 3 solicited local AEs 
occurred in 3 subjects (injection site pain and injection site swelling in 1 subject 
each, injection site erythema in 2 subjects). These events occurred after 
Ad26.ZEBOV vaccination. Grade 3 solicited systemic events were also reported in 
3 subjects (nausea and headache in 2 subjects each, myalgia and fatigue in 1 
subject each). All these events occurred after Ad26.ZEBOV, except fatigue which 
occurred after placebo. The systemic events after Ad26.ZEBOV were considered 
to be at least possibly related to vaccination by the investigator. In the open-label 
regimen, no subjects experienced grade 3 solicited local events. Grade 3 solicited 
systemic events occurred in 3 subjects after Ad26.ZEBOV (chills and fatigue in 3 
subjects each, headache in 2 subjects, pyrexia and nausea in 1 subject each). All 
were considered to be at least possibly related to vaccination by the investigator. 
No grade 3 solicited systemic events were reported after MVA-BN-Filo. All solicited 
(local or systemic) AEs were transient in nature and resolved without sequelae. In 
the placebo-controlled regimens, the overall frequency of unsolicited AEs was 
generally comparable between Ad26.ZEBOV, MVA-BN-Filo, and placebo 
recipients. In the open-label regimen, unsolicited AEs were more common after 
Ad26.ZEBOV than after MVA-BN-Filo.  
Overall, the reported AEs following vaccination were mild in the majority of 
subjects, transient 
in 
nature, 
and 
resolving 
without 
sequelae. 
These 
findings are consistent with the safety profiles observed for similar vaccines. The 
safety profile of the individual vaccines when used as prime was comparable to 
that when used as boost.  
There was no apparent influence of the prime-boost interval (28 days or 56 days) 
on the safety profiles of the vaccines. The 14-day prime-boost interval seemed to 
be associated with higher frequencies of solicited AEs relative to the 28-day and 
56-day prime-boost intervals; however, due to the open-label design of the 14-day 
regimen, knowledge of the treatment assignment may have biased subjects’ 
reporting.  
Both vaccination sequences tested (i.e., Ad26.ZEBOV prime followed by MVA-BN-
Filo boost and MVA-BN-Filo prime followed by Ad26.ZEBOV boost) were highly 
immunogenic and induced considerable humoral and cellular immune responses. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
19 
 
Extending the interval between the prime and boost led to increased antibody 
responses (magnitude at a 56-day prime-boost interval was about 1.8 times higher 
than at a 28-day prime-boost interval), while the effect was less pronounced or the 
reverse for T cell responses. The induced immune responses were functional, as 
demonstrated by the neutralizing activity of the antibody responses in all subjects. 
The composition of induced T cell response was favorable, with a high percentage 
of polyfunctional T cells, which generally are thought to play a role in 
immunological memory and effector functions. 
VAC52150EBL1002 is a randomized, placebo-controlled, observer-blind study in 
healthy adults evaluating the safety, tolerability and immunogenicity of 
heterologous and homologous prime-boost regimens using MVA-BN-Filo and 
Ad26.ZEBOV administered in different doses, sequences and schedules: MVA-
BN-Filo (1x108 TCID50) as prime followed by a Ad26.ZEBOV (5x1010 vp) as boost 
at 14, 28, or 56 days after prime; Ad26.ZEBOV (5x1010 vp) as prime followed by 
MVA-BN-Filo (1x108 TCID50) as boost at 28 days after prime; Ad26.ZEBOV (5x1010 
vp) as prime followed by a high dose of MVA-BN-Filo (4.4x108 TCID50) as boost at 
14 days after the prime; a high dose of Ad26.ZEBOV (1x1011 vp) as prime followed 
by a high dose of MVA-BN-Filo (4.4x108 TCID50) as boost at 28 days after the 
prime. All planned 128 subjects in VAC52150EBL1002 have received a prime 
dose, and 126 subjects have received a boost dose (2 subjects were withdrawn). 
No vaccine-related SAEs have been reported and no safety issues have been 
identified to date. 
VAC52150EBL1003 
and 
VAC52150EBL1004 
are 
randomized, 
placebo-
controlled, observer-blind studies in healthy adults evaluating the safety, 
tolerability and immunogenicity of 4 regimens using MVA-BN-Filo at a dose of 
1x108 TCID50 and Ad26.ZEBOV at a dose of 5x1010 vp: 2 regimens had MVA-BN-
Filo as prime and Ad26.ZEBOV as boost at a 28- or 56-day interval, and 
2 regimens had Ad26.ZEBOV as prime and MVA-BN-Filo as boost at a 28- or 56-
day interval. The blinded phase of studies VAC52150EBL1003 and 1004 are 
ongoing with enrolment complete in 1003. The blinded safety profile is similar to 
that reported in VAC52150EBL1001 and 1002 studies with so far, fewer reports of 
solicited local and systemic AEs. No vaccine-related SAEs have been reported in 
either study to date. 
Studies in children 
Ad26.ZEBOV and MVA-BN-Filo are being evaluated as a prime-boost vaccination 
strategy in children in a phase 2 study (VAC52150EBL2002) and in a phase 3 
study (VAC52150EBL3001). 
VAC52150EBL2002 is a randomized, observer-blind, placebo-controlled, parallel-
group, multicenter, Phase 2 study evaluating the safety, tolerability and 
immunogenicity of 3 heterologous prime-boost regimens using Ad26.ZEBOV as 
prime and MVA-BN-Filo as boost vaccination. MVA-BN-Filo boost vaccination is 
administered at 28-, 56- and 84-day intervals, in healthy and elderly participants 
and at 28- and 56-day intervals in HIV-infected participants and in children. This 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
20 
 
study is conducted in Africa and the enrolment takes place sequentially in four 
cohorts: the first cohort will consist of healthy participants (18 - 70 years); the 
second cohort (2a) will include HIV-infected participants (18 to 50 years) and 
healthy children 12 to 17 years (cohort 2b); the third cohort will include children 
aged 6 to 11 years inclusive and in the fourth cohort young children aged 1 to 5 
years will be enrolled. Within each cohort, participants will be randomized in a 5:1 
ratio to receive active vaccine versus placebo. Upon completion of the 42-day post-
boost visit those participants who received active vaccine will enter long-term 
follow-up, with visits on days 180 and 365 post-prime vaccinations to assess long-
term safety and immunogenicity. Primary outcome measures are the number of 
participants with AEs up to 42 days post-boost, the number of participants with 
SAEs in the year following vaccination and the number of participants with solicited 
local and systemic AEs in the 7 days following prime and boost vaccination. 
Secondary outcomes include GP-EBOV antibody response and T Cells function. 
The estimated enrolment is 1188 participants distributed across the 4 cohorts. The 
enrolment has begun in Burkina Faso and Uganda. 
VAC52150EBL3001 is a staged study including a double-blinded controlled phase 
to evaluate the safety and immunogenicity of Ad26.ZEBOV and MVA-BN-Filo. The 
study takes place in Sierra Leone and consists of a screening phase, an active 
phase (vaccination) and a follow-up phase. The participants will be randomized to 
Ad26.ZEBOV as prime and MVA-BN-Filo as boost vaccination at day 57 or control 
vaccination as prime and placebo as boost vaccination at day 57. The control used 
is MENVEO® World Health Organization (WHO)-prequalified Meningococcal 
Group A, C, W135 and Y conjugate vaccine. The active phase of the study will be 
conducted initially in two stages. The first stage is open-label and approximately 
40 adults aged 18 years or older will be vaccinated with Ad26.ZEBOV as prime 
and MVA-BN-Filo as boost to gain information about the safety and 
immunogenicity of the prime-boost regimen. In stage 2 a larger group of 
approximately 688 individuals will be randomized to the prime-boost or MENVEO® 
vaccines to further evaluate the safety and immunogenicity of the prime-boost 
regimen across different age groups. In this stage, children aged 1 year or older, 
adolescents and adults will be included. Safety data will be collected in stage 1 
and 2, seven days after the initial vaccination and boost vaccination. These data 
will be reviewed by an independent data monitoring committee to assess whether 
initiation of vaccination in the next stage or age group can occur. Safety 
evaluations will include assessment of AEs, which will be monitored throughout 
the study. For stages 1 and 2, the follow-up will be 360 days after prime 
vaccination.  
Safety data generated with the 2 vaccines are provided below: 
Safety Profile of Ad26.ZEBOV 
Ad26.ZEBOV is a monovalent recombinant, replication-incompetent Ad26-based 
vaccine. Only limited clinical data are available for Ad26.ZEBOV. However, 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
21 
 
adenovirus vaccine programs with other gene inserts have revealed no significant 
safety issues.14,15 
To date, a total of 9 human clinical studies with Ad26.ZEBOV are ongoing. No 
clinical studies have been completed yet. The safety/tolerability and 
immunogenicity of the Ad26.ZEBOV vaccine are being assessed in the ongoing 
Phase 
1 
(VAC52150EBL1001, 
VAC52150EBL1002, 
VAC52150EBL1003, 
VAC52150EBL1004 and VAC52150EBL1005), Phase 2 (VAC52150EBL2001) 
and Phase 3 clinical studies (VAC52150EBL3001, VAC52150EBL3002 and 
VAC52150EBL3003), where monovalent Ad26.ZEBOV vaccine and multivalent 
MVA-BN-Filo vaccine are combined in homologous or heterologous prime-boost 
regimens in which each vector is used to prime a filovirus-specific immune 
response followed by a boost immunization with the same or the other vector 2 to 
12 weeks later. Up to the cut-off date of the Investigator’ Brochure14, 1176 healthy 
adult subjects have been enrolled in these 9 ongoing studies, of which 227 
subjects, for whom the treatment assignment is known, received Ad26.ZEBOV at 
dose levels ranging from 5x1010 vp to 1x1011 vp and 32 healthy adult subjects 
received placebo. 
Safety Profile of MVA-BN 
MVA-BN is a further attenuated version of the MVA virus, which in itself is a highly 
attenuated strain of the poxvirus Chorioallantois Vaccinia Virus Ankara. MVA-BN 
induces strong cellular activity as well as a humoral (antibody) immune response 
and has demonstrated an ability to stimulate a response even in individuals with 
pre-existing immunity against Vaccinia. One of the advantages of MVA-BN is the 
virus’ inability to replicate in a vaccinated individual. The replication cycle is 
blocked at a very late stage, which ensures that new viruses are not generated 
and released. This means that the virus cannot spread in the vaccinated person 
and none of the serious side effects normally associated with replicating Vaccinia 
viruses have been seen with MVA-BN. 
MVA-BN (MVA-BN®, trade name IMVAMUNE® outside the European Union [EU], 
invented name IMVANEX® in the EU) has received marketing authorization in the 
EU for active immunization against smallpox in adults, and in Canada for adults who 
have a contraindication to the first or second generation smallpox vaccines 
including people with immune deficiencies and skin disorders. A Phase 3 clinical 
study has been performed in the United States (US) (POX-MVA-013). Results of 
completed and ongoing clinical studies of MVA-BN-based vaccines in more than 
8,100 individuals, including elderly, children and immunocompromised subjects in 
whom replicating vaccines are contraindicated, have shown that the platform 
displays high immunogenicity and a favorable safety profile.15 Across all clinical 
studies, no trends for unexpected or SAEs due to the product were detected.  
Nonclinical studies in macaques with different levels of immune suppression 
support the safety profile of the MVA-BN strain.8 Biosafety aspects of MVA-BN 
strains used for vaccinations have been reviewed.8 
Relevant Safety Information from Ongoing Trials 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
22 
 
One subject in the study VAC52150EBL2001 experienced a serious and very rare 
condition called “Miller Fisher syndrome”. This condition consists of double vision, 
pain on moving the eye, and difficulty with balance while walking. Miller Fisher 
syndrome most commonly occurs following a recent infection. The subject 
experienced these symptoms about a week after suffering from a common cold 
and fever. The event happened about a month after boost vaccination with either 
MVA-BN-Filo or placebo. This subject had to go to the hospital for treatment and 
has recovered. After an extensive investigation, the event has been considered to 
be doubtfully related to vaccine and most likely related to the previous common 
cold. 
In addition, there have been a few reports of mild transient paresthesia, especially 
in the hands and feet, or a sensation of mild muscle weakness of unknown cause 
in subjects vaccinated with Ad26.ZEBOV or placebo. These symptoms have been 
observed to last no more than 24-48 hours in the majority of cases but can last for 
several weeks to several months before going away on their own. One serious 
case of peripheral sensory neuropathy (nerve damage that may cause tingling, 
numbness, pain or loss of pain sensation) of moderate severity has occurred and 
has been ongoing for several months, interfering with some of the participant’s 
daily activities. These types of symptoms have also been reported following 
administration of other licensed vaccines and following acute viral infections of 
various types 
1.1.3. 
Merck VSV-based vaccine 
The VSV-based vaccine was originally developed by the Public Health Agency of 
Canada (PHAC) and advanced in development with the support of a number of 
partners including the US Department of Health and Human Services and the 
Department of Defense.   
The rVSVΔG-ZEBOV-GP vaccine is comprised of a single recombinant replicating 
VSV isolate (11481 nt) modified to replace the gene encoding the G envelope GP 
with the gene encoding the envelope GP from ZEBOV (Kikwit, 1995 strain).  
A brief summary of the vaccine, non-clinical, and clinical safety, immunogenicity, 
and efficacy data is presented. Detailed information may be found in the 
Investigator’s Brochure.9  
Non-clinical studies 
Immunogenicity and Efficacy 
The initial studies of the rVSVΔG-ZEBOV-GP vaccine demonstrated 100% 
protection against lethal ZEBOV challenge in 4 cynomolgus macaques following a 
single intramuscular immunization of 1 x 107 pfu.10  At the same dose, this vaccine 
was also able to protect cynomolgus macaques from homologous aerosol 
challenge.11 Protection from aerosol challenge after IM vaccination has not been 
replicated in more recent studies in which vaccinated animals experienced 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
23 
 
pulmonary inflammation with significant lung pathology; the reasons for this 
difference are being studied further.   
Delivering  2 x 107 pfu of laboratory-grade vaccine by the oral (n=4), intranasal 
(n=40), or IM (n=2) route 28 days prior to challenge protected cynomolgus 
macaques from subsequent ZEBOV challenge.12  Interestingly, the level of 
neutralizing antibodies was low by all routes of immunization, highlighting the 
uncertainty regarding immunologic correlates of protection to Ebola virus infection. 
rVSVΔG-ZEBOV-GP vaccine also protected 4 of 8 (50%) rhesus macaques when 
administered 30 minutes following challenge.13  While all macaques developed 
fever and lymphopenia, 50% of the animals did not progress to more severe 
disease. VSV viremia was detected on day 3 post-immunization in most animals. 
ZEBOV viremia in survivors was 2 to 4 logs lower than in non-survivors. Of note, 
animals treated with rVSVΔG-ZEBOV-GP did not show a decrease in natural killer 
(NK) cells following challenge but instead showed a substantial increase. 
The rVSVΔG-ZEBOV-GP vaccine has undergone challenge testing in murine and 
non-human primate models of pre- and post-exposure prophylaxis. In the murine 
model, the pre-exposure prophylaxis study include mice that  were immunized with 
20,000, 200, 2 and 0.02 pfu (n=10 per group).  The mice were challenged 28 days 
after immunization with 1000 median lethal dose (LD50) of mouse-adapted ZEBOV.  
All mice immunized with 200 or 20,000 pfu survived.  In the post-exposure 
prophylaxis study, mice were challenged with 1000 LD50 of mouse-adapted 
ZEBOV and then treated 30 minutes and 2 hours later with graded doses of 2x105, 
2x104, 2x103 and 2x102 pfu (10 per group). When treated 2 hours after the 
challenge, doses of 2x105, 2x104, and 2x103 were 80-100% effective in preventing 
death,  Treatment within 30 minutes was not effective, with the majority of mice 
dying at each dose. 
In cynomolgus macaques, one pre-exposure prophylaxis study included 4 animals 
who were inoculated IM with 1x108 pfu and 2 controls.  All animals were challenged 
IM 28 days later.  The two controls died 6 days after challenge and all 4 vaccinated 
animals survived.   
A second study included 8 cynomolgus macaques who were inoculated IM with 
3x106 pfu, 2x107 pfu, and 1x108 pfu each as well as 3 controls.  The animals were 
challenged IM 42 days later.  0/3 of the animals that received placebo, 7/8 of 
animals that received 3x106 pfu, 8/8 animals that received 2x107 pfu and 8/8 
animals that received 1x108 pfu vaccination survived. 
A third study included 4-5 cynomolgus macaques who were inoculated IM with 
3x102 pfu, 3x103 pfu, 3x104 pfu, 3x105 pfu, and 3x106 pfu each as well as 2 
controls.  The animals were challenged IM 42 days later.  0/2 of the animals that 
received placebo and all animals that received vaccination survived. 
A durability study in cynomolgus macaques is in progress to assess 
immunogenicity and protection from challenge at 42 days, three months, and one 
year following IM vaccination with 3x104 pfu (2 animals in the one year group) or 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
24 
 
3x106 pfu (the rest of the animals).  Preliminary results indicate that 0/2 of the 
animals that received placebo, 4/4 animals challenged 42 days after vaccination, 
2/6 animals challenged 3 months after vaccination, and 3/7 animals challenged 
one year after vaccination survived.  Additional studies are being planned to 
assess vaccine dose, boost, and origin of cynomolgus macaques.  
In the post-exposure prophylaxis study 12 rhesus macaques were challenged with 
3000 TCID50 of ZEBOV Kikwit.  At 30 minutes or 24 hours after challenge, groups 
of 6 macaques were vaccinated with 2x108 pfu of clinical grade rVSV∆G-ZEBOV-
GP.  Two of 6 (33%) treated 30 minutes after challenge survived but none of the 
animals treated at 24 hours survived.  Two controls died within 10 days. 19  
The kinetics of rVSVΔG-ZEBOV-GP vaccine replication in vitro suggest it 
replicates substantially faster than wild-type ZEBOV and interference with ZEBOV 
replication via competition for receptor and/or for intracellular resources is a 
potential mechanism for its effects as a strategy for post-exposure prophylaxis.14 
In contrast to control animals, which undergo a reduction in lymphocytes during 
infection, vaccinated animals either showed a stable or small increase in CD3+, 
CD4+, and CD8+ T cells. 
Toxicology 
Repeated dose toxicology studies have been performed to evaluate the toxicity 
and local tolerance of VSVΔG-ZEBOV-GP in mice and monkeys. In these studies, 
VSVΔG-ZEBOV-GP was administered via IM injection on Study Days 1 and 14 
followed by a 30-day observation period. BALB/c (Bagg albino) mice and 
cynomolgus monkeys received VSVΔG-ZEBOV-GP at up to 2.0×107 pfu/animal 
and 1.0×108 pfu/animal, respectively. There was no evidence of systemic toxicity 
based on clinical pathology and necropsy data. Developmental and reproductive 
toxicology studies are ongoing.19 The nonclinical safety of rVSV-HIV vaccines has 
also been assessed in non-Good Laboratory Practice (GLP) NHP studies, 
indicating that rVSV is highly attenuated and supports the continued use of 
rVSVΔG-ZEBOV-GP in clinical trials.15 Particularly noteworthy was the absence of 
neurovirulence in NHP following intrathalamic inoculation.16 
Clinical Studies  
Due to the public health emergency of the 2014-2016 West African outbreak, 
rVSV∆G-ZEBOV-GP vaccine has been administered to over 16,000 adults in eight 
phase 1 and four phase 2-3 clinical trials.  
Safety Profile of rVSVΔG-ZEBOV-GP 
Preliminary safety data collected to date have demonstrated rVSVΔG-ZEBOV-GP 
to be generally well-tolerated when administered to healthy adults. Injection-site 
reactions following vaccination are generally mild or moderate and self-limited. 
Thereafter, there is a predictable period of early reactogenicity, including fever and 
a flu-like syndrome, which is common after receipt of other live-virus vaccines, and 
resolves within 7 days. During this time, transient decreases in white blood cells 
and platelets have also been observed but bleeding or increased risk of infection 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
25 
 
has not been reported. The early reactogenicity is likely to be a result of viremia 
with the vaccine virus which has been reported in the majority of subjects and 
resolves within 2 weeks. Mild and self-limited oral ulcers have been observed in a 
few subjects. No rVSV has been detected in oral ulcers to date.  Vaccine virus has 
been found in saliva, urine, and skin-vesicles from those vaccinated. The largest 
proportion of adult subjects that had vaccine virus detected in saliva or urine was 
5 of 20 in 2x107 pfu recipients with positive saliva on Day 7 and this decreased to 
1 of 10 on Day 14. Person-to-person transmission has not been documented. 
Joint pain (arthralgia) has been reported in the first week after vaccination in up to 
35% of vaccinees (range 5 to 35%, blinded data). In a small proportion of subjects 
(<5% at most study sites), joint swelling (arthritis) has been observed in the 2nd to 
3rd week after vaccination.19  
In one Phase 1 clinical trial in which 25 of the 51 subjects that received the study 
vaccine were vaccinated in an open-labeled manner because they were 
deployable to the Ebola outbreak, the proportion of subjects with arthritis was 
higher (11 of 51; 22%) for reasons that have not been fully elucidated. The arthritis 
is thought to be virally-mediated, and a vast majority of cases have resolved within 
several weeks.17,18 However, several subjects have reported recurrent or 
persistent arthralgia and arthritis up to 6 months of vaccination, the longest time 
followed to date.   
Recently, arthritis was also reported in a second study being conducted at the NIH 
in the US. In this open-label study of healthcare workers called the Multicenter 
Study of the Immunogenicity of Recombinant Vesicular Stomatitis Vaccine for 
Ebola-Zaire (rVSV∆G-ZEBOV-GP) for Pre-Exposure Prophylaxis in Individuals at 
Potential Occupational Risk for Ebola Virus Exposure (PREPARE), among 9 
participants enrolled, 3 developed signs and symptoms of arthritis in the knee 10-
14 days following vaccination.  Rheumatologic evaluation (including imaging and, 
in one case, diagnostic arthocentesis) of these 3 individuals supported the 
diagnosis of what was deemed likely to be arthritis, and which took 3-4 months to 
resolve to baseline.  Of note, while the complaint of arthralgia was commonly 
reported in an  earlier phase 1 study conducted jointly by NIH and the Walter Reed 
Army Institute of Research,17 arthritis had not been seen (although observed in 
other phase I/II trials conducted elsewhere27). Also, 6 of the 9 participants reported 
moderate to severe pain in the arm during the injection of the vaccination (R. 
Davey, personal communication, April 7, 2017), a complaint that had also not been 
observed in the earlier phase 1 experience.   
In a Phase 1b dose response randomized trial of the rVSVΔG-ZEBOV-GP vaccine 
with doses ranging from 3 x 103 to 1 x 108 pfu/mL given to adults in the U.S., there 
was no difference in the percent with post-vaccination arthritis among those 
vaccinated with one of the doses of rVSVΔG-ZEBOV-GP (19/418; 4.5%) 
compared to placebo (3/94; 3.2%) (p=0.78).  The percent with arthritis among 
those given the 2 x 107 dose of rVSVΔG-ZEBOV-GP was 4.3% (2/47); for those 
given the highest dose of rVSVΔG-ZEBOV-GP studied, 1 x 108 pfu/mL, 1 of 48 
(2.1%) had arthritis post-vaccination.18 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
26 
 
In a double-blind placebo controlled multicenter Phase 3 lot consistency clinical 
trial19 of adults in the US, Canada and Spain, the incidence of arthritis within 42 
days post-vaccination was statistically significantly higher in 40/790 (5.1%) vaccine 
recipients who received a dose of 2 x 107 pfu/mL when compared to 0/133 (0%) 
placebo recipients (p-value=0.008).  Among 260 participants given a higher dose 
of rVSVΔG-ZEBOV-GP (1 x 108 pfu/mL), 11 (4.2%) developed arthritis, a 
percentage similar to those given 2 x 107 pfu/mL). The cases of arthritis in this 
clinical trial were mostly mild to moderate in intensity and generally resolved within 
a week, with some participants experiencing longer duration.     
Rash (with and without vesicles) has been observed in a few subjects mostly in 
the second week after vaccination, and vaccine virus has been detected in several 
subjects in whom vesicular fluid or skin biopsy specimens were collected. The 
reported skin lesions also appear to be consistent with a virally-mediated process.  
The safety profile of rVSVΔG-ZEBOV-GP in preliminary reports from the Ebola 
epidemic region has been generally similar to that observed outside the region.19  
Data from children, HIV-positive subjects, and pregnant women 
To date, there have been a small number of patients less than 18 years of age or 
HIV-positive individuals exposed to the rVSVΔG-ZEBOV-GP vaccine in ongoing 
trials in Africa. Preliminary open-label data in 20 adolescents between 13 and 17 
years of age and 20 children between 6 and 12 years of age in Gabon suggests a 
similar reactogenicity profile to the reactogenicity profile observed in adult subjects. 
Viremia and shedding after vaccination appear to be higher in children and 
adolescents compared to adults.  Peak viremia occurs on Day 2 and decreases by 
Day 7.  Median copies/mL detected by reverse transcriptase polymerase chain 
reaction (RT-PCR) in 2x107 pfu recipients are 530 in adults, 1,592 in adolescents 
between 13 and 17 years, 1,109 in children between 6 and 12 years of age 
(Kruskal Wallis test P<0.05).  The largest proportion of pediatric subjects that had 
vaccine virus detected in saliva or urine was 14 of 18 adolescents with positive 
saliva on Day 7 (shedding on Day 14 was not assessed in this study). 
Preliminary unblinded data from one Phase 2 trial suggests that approximately 5% 
(22 subjects out of 500 total) of the volunteers that were vaccinated with rVSVΔG-
ZEBOV-GP were HIV-positive and the safety profile appeared generally similar to 
HIV-negative participants, although the small numbers limit the conclusions that 
can be drawn at this time.  
Pregnant women were excluded from the vaccine trials to date. However, a small 
number of women have been found to be pregnant after vaccination and are being 
followed for the outcome of their pregnancies. 
Immunogenicity of rVSVΔG-ZEBOV-GP 
Anti-GP antibodies are detectable by enzyme-linked immunosorbent assay 
(ELISA) at 14 days post vaccination in a majority of vaccine recipients and to date, 
approximately 96% seroconversion has been observed by 28 days post-

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
27 
 
vaccination. Durability of the immune response has also been demonstrated up to 
12 months. As the ELISA and neutralization assays are validated, more data will 
be emerging from the larger, Phase 3 studies in the clinical development program.  
Efficacy of rVSVΔG-ZEBOV-GP 
An open-label, cluster randomized ring vaccination Phase 3 trial was conducted in 
Guinea in which clusters of contacts and contacts of contacts of confirmed cases 
of Ebola were identified and randomly assigned to receive a single 2x107 pfu dose 
of rVSV∆G-ZEBOV-GP immediately or after 21 days. Efficacy and effectiveness 
have been assessed in an interim analysis and the results were published by 
Henao-Restrepo and colleagues.20  In the publication of the interim analysis, 7,651 
subjects were included in the planned interim analysis which included 48 clusters 
(4,123 subjects) randomly assigned to immediate vaccination and 42 clusters 
(3,528 subjects) randomly assigned to delayed vaccination. In the immediate 
vaccination group, there were no cases of EVD with symptom onset at least 10 
days after randomization, whereas in the delayed vaccination group there were 16 
cases of EVD from seven clusters reported, demonstrating a vaccine efficacy of 
100% (95% confidence interval [CI]: 74.7, 100.0; p=0.0036). At the cluster level, 
with the inclusion of all eligible adults, vaccine effectiveness was 75.1% (95% CI: 
7.1, 94.2; p=0.1791). No new cases of EVD were diagnosed in vaccinees from the 
immediate or delayed groups from 6 days post-vaccination. Final analyses 
involving 9,096 participants in 98 randomized clusters have been published.  
Similar to the interim analysis results, no cases of EVD with symptom onset at 
least 10 days after randomization occurred among those randomized to immediate 
vaccination, whereas in the delayed vaccination group there were 16 cases of EVD 
from seven clusters reported, demonstrating a vaccine efficacy of 100% (95% CI: 
68.9, 100.0; p=0.0045).20 
1.1.4. 
Placebo 
The placebo vials will be sterile normal saline (sodium chloride).  
1.2. 
Rationale  
As described above, in an open-label, cluster randomized ring vaccination trial of 
adults conducted in Guinea, the Merck rVSVΔG-ZEBOV-GP vaccine prevented 
EVD events diagnosed 10 days or more after randomization among close contacts 
immediately vaccinated as compared to those eligible to be vaccinated 21 days 
later. In that trial many EVD events occurred within 10 days of randomization.  
Immunogenicity testing was not performed.   
The J&J prime boost vaccine regimen, rHAd26/MVA, has completed phase 1 
testing in adults and is currently being evaluated in phase 2 and 3 clinical trials.    
Multiple questions regarding the safety and effectiveness of EVD vaccines remain 
unanswered, including the immediacy of immune responses generated by the 
Ad26.ZEBOV and rVSVΔG-ZEBOV-GP vaccines, the durability of the immune 
response following a booster at 56 days (MVA-BN-Filo following Ad26.ZEBOV 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
28 
 
prime and repeat rVSVΔG-ZEBOV-GP following rVSVΔG-ZEBOV-GP prime), and 
the safety of the different vaccine strategies.   
During the EVD epidemic in West Africa a considerable proportion of patients were 
children and the mortality rate in those aged < 5 years was very high: 89.5% (CI 
95%: 75.9, 95.8).21  No published safety and immunogenicity data are available for 
children, but at least four clinical trials enrolling children are ongoing. 
There is a need for additional effective vaccines for adults and children in the event 
that EVD incidence increases or the current outbreak is prolonged by new chains 
of sexual transmission. There are too few cases of EVD to conduct a trial with EVD 
as the outcome. Thus, the proposed trial will evaluate the safety and 
immunogenicity of three vaccine strategies in adults and in children. 
The trial will include a placebo group and will be powered to compare each vaccine 
strategy with placebo separately for adults and children for the antibody response 
at 12 months. Consideration was given to using an active vaccine as the control in 
children but in order to provide the best safety data of each vaccine strategy, a 
“true” placebo (saline) is used as the comparator. 
The trial will be double-blind until 12 months of follow-up for each participant is 
completed.  At that time the study will be unblinded. Randomized participants will 
not know their randomized assignment and clinical staff carrying out safety 
evaluations will be blinded to the participants’ randomized assignment.  Laboratory 
staff carrying out immunogenicity testing and biochemical testing will also be 
blinded to the vaccine/placebo that the participant received.  To accomplish the 
blinding in both adults and children, participants randomized to the rVSVΔG-
ZEBOV-GP vaccine without the boost will receive placebo at 56 days while 
participants randomized to rHAd26/MVA will receive the MVA-BN-Filo boost at 56 
days and those randomized to the rVSVΔG-ZEBOV-GP vaccine with a boost will 
receive a repeat rVSVΔG-ZEBOV-GP vaccination at 56 days.  A placebo with two 
fill volumes, 0.5 mL and 1.0 mL, will be used to blind the prime vaccination.  
2. 
METHODOLOGY 
2.1. 
Study design 
After obtaining informed consent, participants will be randomized to the following 
5 groups in a 2:1:2:1:1 allocation 1) Ad26.ZEBOV (prime) (0.5 mL) followed by an 
MVA-BN-Filo boost (0.5 mL) at 56 days;  2)  placebo (prime and boost at 56 days) 
(0.5 mL); 3) rVSVΔG-ZEBOV-GP (prime) (1 mL) followed by placebo boost (1 mL) 
at 56 days; 4) rVSVΔG-ZEBOV-GP (prime) (1 mL) followed by rVSVΔG-ZEBOV-
GP boost (1 mL) at 56 days; and 5) placebo (prime and boost at 56 days) (1 mL) 
(Figure 1).  At 56 days, participants assigned to the rVSVΔG-ZEBOV-GP vaccine 
without a boost and the two placebo groups will receive a placebo vaccination.  
Those initially given the Ad26.ZEBOV vaccine will receive the MVA-BN-Filo 
vaccine at 56 days.  Those assigned to the boosted rVSVΔG-ZEBOV-GP arm will 
receive a booster vaccination with the rVSVΔG-ZEBOV-GP vaccine at 56 days. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
29 
 
In each of the groups receiving the rVSVΔG-ZEBOV-GP vaccine (prime or boost), 
the undiluted dose of the rVSVΔG-ZEBOV-GP vaccine will be used. 
The trial will be conducted at sites in Guinea, Liberia, Mali, and Sierra Leone. 
In the event of a new Ebola epidemic, the study design, including the use of 
placebos, will be re-considered. 
 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
30 
 
Figure 1:  PREVAC Study Design (Version 4.0)  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
2.2. 
Primary objectives  
Separately for adults and children: 
 
• To compare each of the three vaccine strategies with the pooled placebo group 
(3 pair-wise comparisons) for antibody response 12 months after 
randomization (prime vaccination).  
In order to facilitate Merck regulatory filings and bridging of immune responses of 
the rVSVΔG-ZEBOV-GP vaccine from this study to other studies and between 
pediatric and adult populations, the following objective will be assessed specifically 
for those in the two rVSVΔG-ZEBOV-GP vaccine groups: 
• 
To compare the rVSVΔG-ZEBOV-GP vaccine (pooled rVSVΔG-ZEBOV-GP 
groups) with the matched placebo group for antibody response 28 days after 
randomization (prime vaccination).  
In order to facilitate Janssen regulatory filings of the rHAd26/MVA vaccine, the 
following objective will be assessed specifically for those in the rHAd26/MVA 
vaccine group: 
• 
To compare the rHAd26/MVA vaccine group with the matched placebo group 
for antibody response 3 months after randomization (approximately 28 days 
after the booster vaccination).  
rVSV∆G-ZEBOV-GP  
(1 mL) 
(N = 800; 400 adults 
and 400 children) 
rVSV∆G-ZEBOV-GP  
(1 mL) 
(N = 400; 200 
adults and 200 
children)  
2,800 Eligible Participants:  1,400 Adults and 1,400 
Children  
Randomized 2:1:2:1:1 
Placebo 
(1 mL) 
(N = 400; 200 
adults and 200 
children) 
Daily contact each day for one week (children only) and clinic visits at 7, 14 and 28 days  
MVA-BN-Filo 
boost (0.5 mL) 
at 56 days  
  
Placebo  
(0.5 mL) at  
56 days  
  
Placebo  
(1 mL) at 56 
days  
  
rVSV∆G-
ZEBOV-GP  
 (1 mL) at 56 
days  
Placebo  
(1 mL) at 56 
days  
  
Daily contact each day for one week (children only) and clinic visits at 63 days, and 3, 6 and 12 months and then, if 
funding permits, annually through 5 years 
Ad26.ZEBOV  
Vaccine (0.5 mL) 
(N = 800; 400 
adults and 400 
children) 
Placebo (0.5 mL) 
(N = 400; 200 
adults and 200 
children) 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
31 
 
2.3. 
Primary hypotheses 
The null hypothesis for each of the three comparisons at 12 months is that the 
antibody response for the vaccine strategy does not differ from the pooled placebo 
arm (0.5 mL and 1.0 mL groups) 
 
The following hypothesis will only apply to the rVSV∆G-ZEBOV-GP vaccine for 
the purpose of facilitating Merck regulatory filings: 
The null hypothesis is that the antibody response for the pooled rVSVΔG-
ZEBOV-GP group does not differ from the matched placebo group (0.5 mL 
and 1.0 mL groups) at Day 28 after randomization (initial vaccination). 
The following hypothesis will only apply to the rHAd26/MVA vaccine for the 
purpose of facilitating Janssen regulatory filings: 
The null hypothesis is that the antibody responses for the rHAd26/MVA 
vaccine does not differ from the matched placebo group (0.5 mL and 1.0 mL 
groups) at Month 3 after randomization. 
2.4. 
Secondary objectives 
The following secondary objectives will be assessed.  Unless otherwise stated, 
the objectives will be addressed separately for adults and children: 
• To compare the groups given the Ad26.ZEBOV prime vaccine and the 
rVSVΔG-ZEBOV-GP prime vaccine (both rVSVΔG-ZEBOV-GP groups 
combined) each with the pooled placebo group for the antibody response 
14 days after randomization (prime vaccination) (immediacy of response). 
• To compare each of the vaccine groups versus the pooled placebo group 
for the antibody response profile using measurements at 7, 14, 28, 56, 63 
days and at 3, 6 and 12 months after randomization.  
• To compare each of the vaccine groups with the pooled placebo group for 
SAEs at 12 months (see section 4.1.3).   
• To compare the groups given the Ad26.ZEBOV prime vaccine and the 
rVSVΔG-ZEBOV-GP prime vaccine (both rVSVΔG-ZEBOV-GP groups 
combined) each with the pooled placebo group for the percent with injection 
site reactions and AEs graded for severity, including targeted symptoms, 
during the first week following randomization (including the daily contacts 
for children only). 
• To compare the groups given the Ad26.ZEBOV prime vaccine and the 
rVSVΔG-ZEBOV-GP prime vaccine (both rVSVΔG-ZEBOV-GP groups 
combined) each with the pooled placebo group for percent  reporting 
injection site reactions and AEs graded for severity, including targeted 
symptoms, following prime vaccination at the vaccination visit, and through 
7,  14, and 28 days after the prime vaccination. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
32 
 
• To compare each of the vaccine groups with the pooled placebo group for 
changes from baseline in biochemical markers and complete blood count 
(CBC) measurements at 7 days after randomization (children only).   
• To compare the rHAd26/MVA  and rVSVΔG-ZEBOV-GP  boost strategies 
with the pooled placebo group for changes from baseline in biochemical 
markers and CBC measurements at 63 days after randomization (children 
only).   
• To compare the rHAd26/MVA and rVSVΔG-ZEBOV-GP boost strategies 
with the pooled placebo for percent with injection site reactions and AEs 
graded for severity, including targeted symptoms, immediately following the 
booster vaccination and through month 3 (approximately 35 days after the 
booster vaccination). 
• To compare the three vaccine strategies versus the pooled placebo group 
for long-term antibody response at 24, 36, 48 and 60 months following 
randomization (if funding permits). 
• To compare antibody responses and safety outcomes of each of the 
vaccination strategies versus the pooled placebo group in subgroups 
defined by age, gender, country, whether the volunteer is a close contact of 
an Ebola case, the presence of laboratory abnormalities at baseline, and 
has specific co-morbidities (in particular HIV and nutritional status as 
measured by body mass index). 
• For adults and children combined, to compare antibody responses and 
safety outcomes for each of the vaccination strategies versus placebo. 
• To carry out operational research which will include ethnographic, 
participatory and/or qualitative (i.e., focus groups and individual interviews) 
studies to: 1) identify issues relevant to understanding and acceptability of 
the trial, the social issues surrounding informed consent, with the primary 
goal of informing efforts to ensure autonomous fully informed individual 
consent and assent for minors; 2) describe participants' and caregivers' 
experience in the trial, and identify barriers and develop solutions to support 
trial adherence in a culturally sensitive and ethically appropriate way; and 
3) understand prevailing representations and affects surrounding the 
epidemic (including rumors), Ebola and other vaccines, the trial and other 
relevant phenomena in order to ensure effective communication around the 
trial. 
• 
In a subsample of adults in Guinea, T cell and memory B cell responses for 
the three vaccine strategies versus placebo will be compared (see 
Appendix D). 
• In a subsample of children, to compare the rVSVΔG-ZEBOV-GP vaccine 
strategies with the pooled placebo group for shedding of rVSV-ZEBOV-GP 
RNA (see Appendix E).  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
33 
 
2.5. 
Exploratory objectives to be assessed only if and when a 
correlate of protection has been established for both 
vaccine candidates 
Separately for adults and children: 
 
• 
To compare the three vaccine strategies with one another for the correlate 
of protection response at 28 days and 12 months after randomization. For 
the comparison at 28 days, the two rVSVΔG-ZEBOV-GP groups will be 
combined. 
• To compare the group randomly assigned to the Ad26.ZEBOV prime with 
the two groups (pooled) assigned the rVSVΔG-ZEBOV-GP prime vaccine 
for the correlate of protection response 14 days after randomization 
(immediacy of response).  
• To compare the three vaccine strategies with one another for the correlate 
of protection response 3 months after randomization.   
For pooled analysis of children and adults: 
• 
To compare the three vaccine strategies with one another for the correlate 
of protection response 28 days and 12 months after randomization (prime 
vaccination) (durability of response). For the comparison at 28 days the two 
rVSVΔG-ZEBOV-GP groups will be combined. 
• To compare the Ad26.ZEBOV prime vaccination with the rVSVΔG-ZEBOV-
GP prime vaccination (2 rVSVΔG-ZEBOV-GP groups combined) for the 
correlate of protection response 14 days after randomization (immediacy of 
response). 
• To compare the group randomly assigned to the Ad26.ZEBOV prime with 
the two groups (pooled) assigned the rVSVΔG-ZEBOV-GP prime vaccine 
for the correlate of protection response 28 days after randomization.   
• To compare the three vaccine strategies with one another for the correlate 
of protection response 3 months after randomization.   
• To compare the three vaccine strategies with one another for percent of 
participants who experience SAEs through 12 months after randomization.  
• To compare the three vaccine strategies with one another for the correlate 
of protection response and safety outcomes by age. 
2.6. 
Endpoints 
The primary immunogenicity endpoint is the GP-EBOV antibody response 12 
months after randomization.  This endpoint will be used to compare the 
immunogenicity of the three vaccine strategies with placebo.  The primary analysis 
for adults and children will be performed for participants who do not have elevated 
antibody levels at baseline.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
34 
 
GP-EBOV antibody response at Day 28 after randomization.  For rVSVΔG-
ZEBOV-GP arms only, Day 28 post first vaccination will be used for regulatory 
purposes for comparison to other studies and for bridging children to adults. The 
primary analysis for adults and children will be performed for participants who do 
not have elevated antibody levels at baseline. 
GP-EBOV antibody response at Month 3 after randomization (approximately 28 
days after the booster vaccination).  For the rHAd26/MVA vaccine arm only, Month 
3 will be used for regulatory purposes. The primary analysis for adults and children 
will be performed for participants who do not have elevated antibody levels at 
baseline. 
Antibodies to the Ebola virus GP will be measured with the Filovirus Animal 
Nonclinical Group (FANG) ELISA assay if available.  Other assays may also be 
used.  If a correlate of protection is identified, stored sera will be used to measure 
the correlate and carry out comparisons of the three vaccine strategies with 
placebo and with one another.  The precise definition of antibody responders will 
be defined by the trial steering committee prior to their unblinding of the study 
results. The primary analysis will exclude participants with elevated antibody levels 
at entry. 
There are a number of secondary endpoints.  Antibody levels will be determined 
at multiple time points to assess the immediacy and long-term durability of the 
response.  SAEs through 12 months will be collected.  AEs of lesser severity, 
including injection site reactions and targeted symptoms, will be collected following 
the initial vaccination and following the booster.  In addition,  substudies will 
examine T cell and memory B cell changes and viral shedding.    
2.7. 
Sample size 
Sample size was established for the 5-arm study to provide power to compare 
safety and immunogenicity separately for adults (N=1,400) and children (N=1,400) 
(see Figure 1),  Under Versions 2.0 and 3.0 of the PREVAC protocol 1,700 to 1,800 
participants (537 in Version 2.0 and an estimated 1,200 in Version 3.0) are likely 
to be randomized.  Thus, the total number randomized cumulatively in Version 2.0, 
3.0 and 4.0 could be over 4,500 participants. A statistical analysis plan describes 
how data from Version 2.0 and Version 3.0 will be combined with data from Version 
4.0 for selected objectives. 
Sample size for adults for Version 4.0 has been reduced from what was originally 
planned (from 3,500 to 1,400).  In part, this is because of the delays, and resulting 
costs, incurred before Version 4.0 could be implemented; in part, this is because 
a very conservative estimate of sample size was specified for adults in earlier 
versions of the PREVAC protocol; and in part, this is because of the plan to use 
data collected in Version 2.0 and 3.0 with data collected in Version 4.0 to address 
some objectives.  Even with this reduction in sample size for adults, power for 
comparing the major efficacy and safety outcomes for adults and children 
separately is excellent.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
35 
 
Sample size is greater than what is required to address the primary objectives 
because if a correlate of protection is identified, the vaccine strategies will be 
compared with one another for that correlate using an intention to treat analysis.  
Expected differences between vaccine groups may be smaller than comparisons 
with placebo and the correlate may have greater variability than the assay which 
will be used to measure antibody levels to address the primary, secondary and 
exploratory objectives. The larger sample size will also permit the exploration of 
subgroups and preserve power in the event there are more participants with 
elevated antibodies at baseline than anticipated.  
For the planned antibody comparisons of each vaccine strategy versus placebo at 
12 months, the planned sample is based on data from PREVAIL I.22  In PREVAIL 
I antibody levels measured at baseline, one week and at one month are briefly 
summarized below.  
• 4.0% of participants had elevated antibodies to the EBOV at baseline. An 
elevated baseline antibody level in PREVAIL I was defined using the 
distribution of antibody levels measured from sera collected in 2004 and 
2001 from 92 adults in Mali, a region where there had been no reports of 
ZEBOV transmission or disease during this time period. Baseline antibody 
levels were considered elevated in PREVAIL I if they were greater than 3 
standard deviations (SD) above the mean (607 enzyme-linked 
immunosorbent assay units/mL based on the FANG ELISA assay).35 
• Among participants without elevated antibodies at baseline (96%), the 
percent with an antibody response at week 1 was 2.5% for those given the 
rVSVΔG-ZEBOV-GP vaccine and 1.5% for those given placebo (p=0.36).  
By month 1 antibody levels had increased substantially and 83.7% of 
participants given the rVSVΔG-ZEBOV-GP vaccine had an antibody 
response as compared to 2.8% of placebo participants (p<0.001).  At 12 
months, 79.5% of participants given the rVSVΔG-ZEBOV-GP vaccine had 
an antibody response versus 6.8% of participants in the placebo group 
(p<0.001).  Responders during follow-up were defined as having a change 
in log-transformed antibody levels greater than 2 SD of the change in the 
placebo group at month 1. 
With type 1 error = 0.0167 (2-sided) to adjust for the three comparisons, separately 
for adults and for children in all age groups combined, and power = 0.90, even if 
the percent with a positive antibody response at 12 months is 50% in a vaccine 
group, with equal allocation, approximately 30 participants per group (60 
participants total) are needed assuming the percent in the placebo group with a 
positive antibody response is approximately 7%.  With unequal allocation as for 
the rVSVΔG-ZEBOV-GP with the booster versus placebo comparison, a total of 
63 participants (21 vaccinated with rVSVΔG-ZEBOV-GP with a boost and 42 
vaccinated with placebo), a difference of 50% versus 7% can also be detected at 
12 months with 87% power.  These sample size estimates indicate that power for 
the planned subgroup analysis by age is also appropriate.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
36 
 
The planned sample size is also adequate for the comparisons with placebo if 
more than 4% of participants are antibody positive at baseline and excluded from 
the primary analysis, and if there is some missing data 12 months. 
If a correlate of protection is identified, the vaccine strategies will be compared with 
one another at multiple time points.  For the contrast with the smallest sample size 
(200 adults or children assigned rVSVΔG-ZEBOV-GP with a boost versus 400 
vaccinated with rHAd26/MVA or rVSVΔG-ZEBOV-GP without a boost), power is 
0.83 to detect an absolute difference of 15% in the antibody response (50% versus 
35%) at the 0.0167 (2-sided) level of significance, assuming 10% of participants 
are excluded due to a positive antibody level at entry or for missing data.  If the 
percent with a positive antibody response at 12 months is 75%, power is 0.87 to 
detect a 15% lower antibody response for this planned contrast. 
For SAEs, we estimate that 5% of participants in the placebo group will experience 
an SAE by 12 months.  This estimate is based on data from PREVAIL I where after 
12 months of follow-up 11.8% of participants assigned to placebo developed an 
SAE.  In PREVAIL I most of the SAEs were due to malaria events which did not 
require hospitalization.35  Thus, with the exclusion of laboratory confirmed malaria 
events that did not require hospitalization, the 12 month percent with SAEs in this 
study is expected to be lower.  Among adults and children, considered separately, 
with type 1 error = 0.0167 (2-sided), power is 0.88 to detect an increase of 7% (to 
12%) for the rHAd26/MVA and for the rVSVΔG-ZEBOV-GP without a boost 
strategies compared to placebo assuming 400 per group.  For the rVSVΔG-
ZEBOV-GP with a boost (N=200) versus placebo (N=400) comparison for adults 
and children, power is 0.86 to detect an increase of 9% (to 14%). 
Sample size is also sufficient for comparing the vaccine strategies with the pooled 
placebo group for the antibody response at day 14 (an important secondary 
objective that address the immediacy of the immune response). For example, if 
the antibody response at 14 days is 25% in a vaccine group and 7% in the placebo 
group, fewer than 250 participants total are needed.   
A sample size re-estimation will be performed before enrolment has been 
completed to ensure there is adequate power for both adults and children to 
address the primary objectives. 
2.8. 
Study cohort 
This phase 2 study will be conducted in Guinea, Liberia, Mali, and Sierra Leone.  
As part of the public health campaign to prevent EVD, there will be widespread 
communication about the trial to encourage volunteers to go to a vaccination 
center near where they live.   
Our goal is to include 1,400 adults and 1,400 children.  The gender and age 
distributions of those enrolled will be closely monitored. The steering committee 
may request that more men or women or participants in specific age groups be 
enrolled to ensure broad generalizability of the findings. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
37 
 
Eligibility for enrolment in the trial will be assessed on a first-come first-serve basis; 
in case of oversubscription we will ensure that access is made on an equitable 
basis that does not compromise the external validity of the trial.     
2.8.1. 
Inclusion criteria 
The inclusion criteria for the study are broad reflecting the target population that 
would eventually receive efficacious vaccines.  
• Informed consent/assent 
• Age ≥ 1 year  
• Planned residency in the area of the study site for the next 12 months 
• Willingness to comply with the protocol requirements  
2.8.2. 
Exclusion criteria 
• Fever > 38º Celsius 
• History of EVD (self-report) 
• Pregnancy (a negative urine pregnancy test is required for females of 
child-bearing potential, i.e., females who have experienced menarche or 
who are aged 14 years and older) 
• Positive HIV test for participants < 18 years of age 
• Reported current breast-feeding 
• Prior vaccination against Ebola (self-report) 
• Any vaccination in the past 28 days or planned within the 28 days after 
randomization (initial vaccination) 
• In the judgement of the clinician, any clinically significant acute/chronic 
condition that would limit the ability of the participant to meet the 
requirements of the study protocol 
2.9. 
Randomization and blinding 
Eligible participants will be randomized in a 2:1:2:1:1 allocation to rHAd26/MVA 
(prime and boost), 0.5 mL of placebo (prime and boost), rVSVΔG-ZEBOV-GP 
without a boost, rVSVΔG-ZEBOV-GP with a boost, or, 1mL of placebo (prime and 
boost) (Figure 1).  Syringes will be pre-labelled with a unique SID number 
according to a centrally prepared randomization schedule.  The tear-off label will 
also include a bar code identifier.  For each vaccination center, the randomization 
schedule will be prepared using block randomization to ensure the desired 
allocation ratio for the five arms of the study for each vaccination center. 
It is possible that the person carrying out the vaccination will be able to differentiate 
syringes by the fill volume.  The rVSVΔG-ZEBOV-GP vaccine has a 1 mL fill 
volume and the rHAd26/MVA vaccines have a 0.5 mL fill volume.  The person 
carrying out the vaccination will not know whether vaccine or matched placebo is 
given.   
At the time of vaccination, a tear-off label on the syringe that includes the SID will 
be attached to the baseline CRF (or an eCRF will be used). This will be the primary 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
38 
 
link used between the vaccine administered and the PID, and PIDs associated with 
SIDs will be available to the pharmacy.  
With this approach, randomization does not occur until the participant is vaccinated 
with the prime vaccine.     
Participants randomized to the rVSVΔG-ZEBOV-GP vaccine without a boost for 
the initial vaccination will receive placebo (1.0 mL fill) at 56 days.  Those 
randomized to receive  rHAd26/MVA initially will receive the MVA-BN-Filo boost at 
56 days in a syringe with a 0.5 mL fill; and those randomized to rVSVΔG-ZEBOV-
GP with a boost will receive the rVSVΔG-ZEBOV-GP vaccine again at 56 days in 
a syringe with a 1 mL fill. Two groups will receive placebo (saline) for the prime 
and booster vaccination.  For one group, a 0.5 mL fill syringe will be prepared for 
each vaccination and for the other a 1.0 mL fill syringe will be prepared for each 
vaccination.  The syringe used at 56 days will be labeled with the volunteer’s PID.  
Study participants and clinical staff assessing the study participants for safety and 
laboratory outcomes will be fully blinded until all participants complete 12 months 
of follow-up.  The laboratories carrying out the safety and immunogenicity analyses 
will be blinded to the vaccine assignment.   Procedures will be put into place to 
break the blind if necessitated by a medical emergency. If unblinding of a 
participant for safety reasons is required, it will be documented. 
After 12 month follow-up visits have been completed for all participants, the study 
will be unblinded.  Follow-up may continue through 5 years if funding permits. 
3. 
STUDY PLAN 
Vaccination centers will be established in West Africa to consent, vaccinate and 
follow study participants. A CRF with a unique PID will be completed for each 
participant consented and screened.  The CRF may be paper or an eCRF.  The 
procedures described in this protocol are in terms of a paper CRF.  If an eCRF is 
used, the procedures described will be adapted for their use. 
At the time of vaccination, study volunteers will be given an identification card that 
identifies them as a study participant.  The identification card will include contact 
information in the event they experience vaccine side effects, or signs and 
symptoms of Ebola. Volunteers will be encouraged to contact the vaccination 
center by telephone at any time of day if they have any AEs.  If the participant 
experiences an AE requiring immediate attention, they will be given instructions on 
which hospital to go to for evaluation and treatment.  
Women of childbearing potential will be told to use adequate birth control 
measures for at least 3 months following the initial vaccination and booster 
vaccination.  Men who are sexually active with women of child-bearing potential 
will be told to use condoms for at least 3 months following the initial vaccination 
and booster vaccination.  Participants will also be instructed to avoid sharing items 
during the period of potential virus shedding. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
39 
 
Following randomization, all participants will be seen at 7, 14, 28, 56 and 63 days, 
and at 3, 6 and 12 months.  After 12 months, participants will be seen annually if 
funding permits.  In addition to these study visits, volunteers will be encouraged to 
contact the vaccination center by telephone if they experience AEs.  
3.1. 
Screening/Baseline data collection 
Prior to randomization, children must have an HIV test.  There is no specific time 
period within which the HIV test must have been performed. If the test is 
negative, they will be eligible for randomization 
Baseline data will include: 
• Informed consent type and date 
• Birth month/year and gender 
• Urine pregnancy test result for females who are of childbearing potential, 
i.e., females who have experienced menarche or who are aged 14 years 
and older 
• High risk occupation (e.g., work involved or might involve contact with living 
or dead persons with Ebola such as a nurse, physician, ambulance driver, 
burial crew member or other healthcare worker or not a healthcare worker) 
• History of past contact with a person with EVD 
• Name and contact information for participant (to be kept locally) 
• Names and contact information of 2 alternate contact people (to be kept 
locally)  
• Body temperature 
• Height and weight 
• Mid-upper arm circumference (for children 1-5 years).  
• Serum for aspartate transaminase (AST), alanine transaminase (ALT), 
creatinine, and potassium 
• Blood for CBC with differential, platelet count, hemoglobin, hematocrit, 
mean corpuscular volume (MCV), and red cell distribution width (RDW)  
• Blood for assessing HIV and syphilis serostatus 
• Stored serum for immunogenicity testing and future research 
• Stored cells for assessing T cell and memory B cell function (subsample of 
adults enrolled at a single site in Guinea) 
• Stored saliva for assessing viral shedding (subsample of children) 
• Antipyretics prescribed/offered 
3.2. 
Vaccine administration 
Following informed consent and the collection of baseline data, the study vaccines 
will be administered to participants who meet all of the eligibility criteria via 
intramuscular injection in the upper, outer aspect of the arm (deltoid region).  
Children may be vaccinated in the thigh.  Prior to injection, the area to be injected 
will be prepared using an alcohol swab. Vaccine will be administered using a 3 mL 
syringe with a sterile needle. While holding the needle at an approximate 90-
degree angle to the skin, the needle will be directed through the skin, into the 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
40 
 
muscle of the arm (or thigh).  The location of  vaccination will be recorded on the 
case report form. 
At the time of prime vaccination administration, the tear-off portion of the SID label 
on the syringe will be placed on the baseline CRF. 
Study participants/family members will also be provided information on how to 
minimize risk of infection with Ebola. 
After vaccination, participants will be watched closely for at least 30 minutes.  
Injection site reactions, targeted symptoms of any grade severity, and grade 3 and 
4 AEs that occur during this observation period will be recorded.   
Participants may be offered medication post-vaccination for fever and pain relief 
depending on each site’s policy. If participants receive or take their own medication 
for fever and pain following vaccination, it will be recorded. 
The same procedures will be followed for the MVA-BN-Filo and rVSVΔG-ZEBOV-
GP booster/placebo vaccination at 56 days.  As stated below, the window for the 
booster vaccination at 56 days is 53 to 66 days (-3,+10 days). 
3.3. 
Follow-up data collection 
Following randomization, all participants will be seen at the following visits (visit 
window):  7 (± 3 days), 14 (± 3 days), 28 (± 7 days), 56 (53 to 66 days) and 63 
days (7 ± 3 days after the booster vaccination), and at 3 (± 14 days), 6 (± 1 month), 
and 12 (± 1 month) months.  Participants may continue to be seen at 24 (± 1 
month), 36 (± 1 month), 48 (± 1 month) and 60 (± 1 month) months if funding 
permits. 
At each follow-up visit, volunteers will be asked a few questions to assess the 
development of any serious medical conditions (e.g., malaria), the possibility of 
unreported  SAEs, and for women whether they are pregnant.  Temperature will 
be recorded, and blood will be drawn for immunogenicity testing and future 
research.   
The following additional information will be collected at 7, 14, 28, and 63 days and 
at 3 months (approximately 35 days after the booster) following randomization: 
• Grade 3 and 4 AEs and targeted symptoms since vaccination 
• Injection site reactions since vaccination 
The following additional specimens and information will be obtained for children:  
1) during the first week following the prime and booster vaccination, there will be 
daily contacts with children to assess injection site reactions, targeted symptoms 
and SAEs, and to measure temperature;  2) on day 7 and 63, serum for AST, ALT, 
creatinine,  and potassium and blood for CBC with differential, platelet count, 
hemoglobin, hematocrit, MCV, and RDW will be obtained; 3) on day 28, at months 
3 and 6 and annually weight will be recorded;  4) annually height will be measured; 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
41 
 
and 5) mid-upper arm circumference (children aged 1-5 years) will be measured 
at months 3, 6, and 12, and then annually if funding permits.   
For a subsample of adults, at baseline and days 7, 14, 56, 63, 70, and month 12, 
cells for assessing  T cell and memory B cell function will be obtained. If funding 
permits, cells will also be collected at 24, 36, 48 and 60 months,  
On day 56 prior to the booster vaccination, a urine pregnancy test will be performed 
for females of child-bearing potential, i.e., females who have experienced 
menarche or who are aged 14 years and older.  If a woman is pregnant, she cannot 
receive the booster vaccination.  However, she will remain under follow-up. 
Following the booster vaccination, participants will be watched closely for at least 
30 minutes.  Injection site reactions, targeted symptoms of any grade severity, and 
grade 3 and 4 AEs that occur during this observation period will be recorded.   
After 12 months of follow-up have been completed for all participants, participants 
will be informed which vaccine they received and the results of the study. 
3.4. 
Laboratory Tests 
Safety laboratory tests will be performed by a clinical laboratory at the site.  
Safety laboratory reports will be generated and returned to the vaccination 
center(s) prior to the next scheduled visit.   
At one site peripheral blood mononuclear cells (PBMCs) will be collected for a 
subsample of adults and stored for the investigation of T cell and memory B cell 
responses. 
At one site saliva will be collected for a subsample of children for measurement of 
viral shedding. 
Immunogenicity testing will be performed using a validated FANG GP-ELISA 
assay on baseline, Day 28, Month 3 and Month 12 specimens. The participating 
pharmaceutical companies will be provided samples from these visits and have 
the primary responsibility for obtaining this testing. A non-validated FANG GP-
ELISA assay or another assay may also be used at these and other time points. 
3.5. 
Biometric Data 
Digital finger prints and iris scans may also be collected, digitized and stored in a 
secure database.  These data would be used to ensure appropriate matching of 
study participants with PIDs.  
4. 
ASSESSMENT OF SAFETY 
A manual of operations will describe procedures for reporting deaths, Ebola 
events, pregnancies, pregnancy outcomes, and SAEs, including assessments of 
severity and the relationship of the vaccine to events.  Below a brief summary is 
provided. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
42 
 
4.1. 
SPECIFICATION OF SAFETY PARAMETERS 
4.1.1. 
Definition of AE 
An AE is any untoward medical occurrence in a randomized volunteer who is 
administered the trial product (vaccine or placebo), whether or not considered 
related to the vaccination.  
4.1.2. 
Definition of an Adverse Reaction (AR) 
An AR is any untoward and unintended response related to an investigational 
medicinal product (in this case, vaccine or placebo). 
4.1.3. 
Definition of SAE 
An AE is considered "serious" if, in the view of either the investigator or sponsor, 
it results in any of the following outcomes: 
a. Death 
b. Events that are life-threatening 
Note: The term ‘life-threatening’ in the definition of ‘serious’ refers to an event 
in which the participant was at risk of death at the time of the event. It does not 
refer to an event, which hypothetically might have caused death, had it been 
more severe. 
c. Events requiring hospitalisation 
d. Events requiring prolongation of existing hospitalisation, 
Note: In general, hospitalization signifies that the participant has been 
admitted at the hospital or emergency ward for observation and/or treatment 
that would not have been appropriate in the physician’s office or in an out-
patient setting. Complications that occur during hospitalization are also 
considered as SAEs. If a complication prolongs hospitalization or fulfils any 
other serious criteria, the event will also be considered serious. When in doubt 
as to whether ‘hospitalization’ occurred or was necessary, the AE should be 
considered serious. 
Hospitalization for elective treatment of a pre-existing condition (known or 
diagnosed prior to informed consent signature) that did not worsen from 
baseline is NOT considered an SAE. 
e. Events resulting in a persistent or significant incapacity or substantial 
disruption of the ability to conduct normal life functions 
Note: The term disability means a substantial disruption of a person’s ability to 
conduct normal life functions. This definition is not intended to include 
experiences of relatively minor medical significance such as uncomplicated 
headache, nausea, vomiting, diarrhea, influenza-like illness, and accidental 
trauma (e.g. sprained ankle) which may interfere or prevent everyday life 
functions but do not constitute a substantial disruption. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
43 
 
f. 
Congenital anomaly or birth defects in the offspring of a study participant 
g. Important medical events, that may not result in death, be life-threatening, or 
require hospitalization may be considered serious when, based upon 
appropriate medical judgment, they may jeopardize the participant and may 
require medical or surgical intervention to prevent one of the outcomes listed 
in this definition.  Examples of such medical events include: 
• Any suspected transmission of an infectious agent by a medicinal product 
• Allergic bronchospasm requiring intensive treatment in an emergency 
room or at home 
• Blood dyscrasias or convulsions that do not result in inpatient 
hospitalization 
• Development of drug dependency or drug abuse. 
• Cranial nerve disorders, including paralyses/paresis (e.g. Bell’s palsy)  
• Optic neuritis 
• Multiple sclerosis 
• Transverse myelitis 
• Guillain-Barré syndrome, including Miller Fisher syndrome, Bickerstaff’s 
encephalitis and other variants 
• Acute disseminated encephalomyelitis, including site specific variants: 
e.g. non-infectious encephalitis, encephalomyelitis, myelitis, 
myeloradiculomyelitis 
• Myasthenia gravis and Lambert-Eaton myasthenic syndrome  
• Immune-mediated peripheral neuropathies and plexopathies, (including 
chronic inflammatory demyelinating polyneuropathy, multifocal motor 
neuropathy and polyneuropathies associated with monoclonal 
gammopathy). 
• Narcolepsy 
• Isolated paresthesia of >7 days duration 
Note: For this protocol, EVD events and laboratory confirmed malaria events that 
do not require hospitalization will not be considered as SAEs.  Laboratory 
diagnostic criteria for confirmation of malaria should be based on local criteria. 
4.1.4. 
Serious Adverse Reaction (SAR) 
Any AR which is also considered serious. 
4.1.5. 
Suspected Unexpected Serious Adverse Reaction (SUSAR) 
An AR that is both unexpected (not consistent with the applicable product 
information) and also meets the definition of a SAR. 
4.1.6. 
Definition of Unanticipated Problems (UP) 
An UP is any incident, experience or outcome that meets all of the following criteria: 
1. Unexpected in terms of nature, severity, or frequency in relation to: 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
44 
 
a. the 
research 
risks 
that 
are 
described 
in 
the 
ethical 
committee/institutional review board (IRB)-approved research 
protocol and informed consent document; Investigator’s Brochure or 
other study documents; and 
b. the characteristics of the population being studied; and 
 
2. Possibly, probably, or definitely related to participation in the research 
(“possibly related” means there is a reasonable possibility that the incident, 
experience, or outcome may have been caused by the procedures involved 
in the research); and 
 
3. Places study participants or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously 
known or recognized. 
 
Furthermore, a UP could be an expected event that occurs at a greater frequency 
in one of the vaccine groups than the placebo group.  
4.2. 
CLASSIFICATION OF AN AE 
4.2.1. 
Relationship to study product 
The relationship assessment is one of the criteria used when determining 
regulatory reporting requirements. 
Relationship of SAEs should be assessed by the site physician using the following 
question: 
Is there a reasonable possibility that the AE may have been caused by the study 
product (vaccine)? 
YES 
: The AE is known to occur with the study product, there is a 
reasonable possibility that the study product caused the AE, or 
there is a temporal relationship between the study product and 
event. Reasonable possibility means that there is evidence to 
suggest a causal relationship between the study product and the 
AE. 
NO 
: There is not a reasonable possibility that the administration of 
the study product caused the event, there is no temporal 
relationship between the study product and event onset, or an 
alternate etiology has been established. 
4.2.2. 
Unexpectedness 
Only SAEs will be assessed as expected or unexpected. An SAE will be 
considered unexpected if the nature, severity, or frequency of the event is not 
consistent with the risk information previously described for the vaccine. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
45 
 
An SAE is considered "unexpected" if it is not listed in the Investigator’s Brochure 
or is not listed at the specificity or severity that has been observed; or, if an 
Investigator’s Brochure is not required or available, is not consistent with the risk 
information described in the general investigational plan or elsewhere in the 
protocol, as amended.  
For example, under this definition, hepatic necrosis would be unexpected (by virtue 
of greater severity) if the Investigator’s Brochure referred only to elevated hepatic 
enzymes or hepatitis. Similarly, cerebral thromboembolism and cerebral vasculitis 
would be unexpected (by virtue of greater specificity) if the Investigator’s Brochure 
listed only cerebral vascular accidents.  
"Unexpected," as used in this definition, also refers to AEs or suspected ARs that 
are mentioned in the Investigator’s Brochure as occurring with a class of drugs or 
as anticipated from the pharmacological properties of the drug, but are not 
specifically mentioned as occurring with the particular drug under investigation. 
4.3. 
TIME PERIOD AND FREQUENCY FOR EVENT ASSESSMENT 
AND FOLLOW-UP 
4.3.1. 
Time Period and Frequency Assessment 
4.3.1.1. 
AE 
Injection site reactions, targeted symptoms of any grade severity, and unsolicited 
Grade 3 or 4 AEs will be collected and reported on CRFs as defined by the protocol 
(see Appendix A).  AEs of grade 1 or 2 severity other than injection site reactions 
and targeted symptoms will not be collected.   Grade 3 or 4 signs or symptoms 
associated with laboratory abnormalities and clinically significant grade 3 and 4 
laboratory abnormalities documented by the investigators as AEs should be 
reported on the CRFs. All other laboratory abnormalities will be summarized using 
the protocol required laboratory data that is in the central database. Laboratory 
abnormalities that are considered clinically significant should be reported as AEs 
or SAEs irrespective of the presence of symptoms.  
Sites will be notified of laboratory abnormalities identified as part of the protocol 
required testing (baseline for all participants and also 7 days after the prime and 
booster vaccination for children).  Laboratory abnormalities will be summarized for 
each DSMB review. Details for documenting laboratory abnormalities in the 
participants medical record are provided in the manual of operations. 
Injection site reactions will include redness of skin, swelling/induration, 
pain/tenderness with activity, and itching at injection site. 
The targeted symptoms will include reduced activity/somnolence, fatigue, 
vomiting, chills, abnormal sweating. skin lesions (macules, papules, purpura, 
petechiae), mouth ulcers, decreased appetite, feverishness, diarrhea, nausea, 
headache, dizziness, abdominal pain, muscle pain,  joint swelling, and joint pain.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
46 
 
For non-verbal children, subjective symptoms may not be assessed and 
irritability/fussiness, crying, and screaming will be assessed.   
Injection site reactions and symptoms will be graded as follows: Grade 1 – 
symptoms causing no or minimal interference with usual social & functional 
activities; Grade 2 - symptoms causing greater than minimal interference with 
usual social & functional activities; Grade 3 - symptoms causing inability to perform 
usual social & functional activities; and Grade 4 - symptoms causing inability to 
perform basic self-care functions OR medical or operative intervention indicated to 
prevent permanent impairment, persistent disability, or death. 
4.3.1.2. 
SAEs, EVD events, Malaria Events 
SAE and EVD events will be assessed and reported on CRFs as soon as the site 
becomes of aware of the event.  For EVD events, where possible, a sample of 
blood will be stored for future research. Malaria events will be assessed at each 
follow-up visit.  Malaria events that meet the SAE definition (e.g., result in 
hospitalization) will be reported on the SAE form. 
4.3.1.3. 
Pregnancy 
Women who report being pregnant in the first 3 months of follow-up after the 
prime vaccination will be followed for the outcome of their pregnancy.   
The medical surveillance of the women and their children will be reinforced: a 
particular attention must be given to serious pathology occurring during pregnancy.  
An SAE form should be completed if an AE during pregnancy or birth outcome 
meets the SAE definition. Congenital anomalies or birth defects should also be 
reported as SAE. 
4.3.1.4. 
UP 
UPs that are not related to the study vaccines, for example, a breach of subject 
confidentiality, will be reported to the respective Ethical/Institutional Review 
Committees in accordance with the requirements of the Committee for oversight 
of the particular site where the UP occurs.  UPs related to the study vaccines will 
be reported per AE/SAE reporting requirements in 4.3.1.1 and 4.3.1.2.  Details for 
UP reporting are provided in the manual of operations. 
4.3.2. 
Time of collecting information 
All SAE events will be collected from the signing of informed consent until the end 
of the study of the participant. 
4.3.3. 
Follow-up 
All SAE events must be followed until clinical recovery is complete and laboratory 
results have returned to normal or baseline, or until the event has stabilized. 
Follow-up should continue regardless of when this occurred in relation to the study 
conclusion. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
47 
 
4.4. 
GENERAL REPORTING, RESPONSIBILITIES AND 
TIMELINES 
Events Reported on CRFs by the investigators:  
AEs (injection site reactions, targeted symptoms of any grade of severity and 
unsolicited grade 3 and 4 AEs), UPs, SAEs, deaths, Ebola events, malaria events, 
and pregnancy outcomes will be reported on CRFs.   
Events to be notified by the investigators to the Medical Officer and to Sponsors:  
SAEs  and Ebola events will be reported within 24 hours of their knowledge of 
the event. 
The Medical Officer will assess the expectedness and relationship of SAE to the 
study vaccines.  All events will be coded using the Medical Dictionary for 
Regulatory Activities (MedDRA®).   
The plan for DSMB reporting is described in section 7.2 and in the statistical 
analysis plan. 
Events to be notified to US Food and Drug Administration (FDA):  
Only SUSARs will be notified to the FDA by the NIH within the legal timeframe: 
 7 calendar days in case of death or life-threatening events after the 
sponsor's initial receipt of the information; relevant complementary 
information should be collected and notified within 8 extra-days 
 15 calendar days for all other events after the sponsor's initial receipt of the 
information; relevant complementary information should also be collected 
and notified within 8 extra-days 
 
Events to be notified to IRB/ethics committee (EC): Each sponsor will be 
responsible for the SUSAR and other events notification to their respective ECs 
or IRBs in compliance with local regulations. 
Safety Reports: 
The Safety Reports will be sent to the sponsors for review and validation. NIH is 
responsible for the Safety Reports submission to the FDA and each sponsor is 
responsible for the Safety Reports submission to their respective Health Authority 
and IRB/EC in compliance with local regulations.  NIH will also forward Safety 
Reports to industry partners in accordance with provisions identified in the 
respective clinical trial agreements. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
48 
 
4.5. 
HALTING RULES 
4.5.1. 
Study halting rules 
The DSMB for this study will meet frequently to assess the overall safety of study 
participants.  The DSMB may pause enrolment in the event of vaccine-related 
deaths or SAEs that are considered vaccine-related. The DSMB may also pause 
enrolment or request that participants be notified if there is an increased frequency 
of unanticipated adverse effects. 
4.5.2. 
Participant halting rules 
If a participant experiences an SAR (that is attributed to initial vaccination), the 
booster vaccination will not be given.  These participants will continue to be 
followed until the end of the study.  
5. 
CLINICAL MANAGEMENT 
5.1. 
AEs and symptoms and signs of EVD 
Study participants will be given information on whom to call or see in the event 
they experience AEs and symptoms of EVD. If the participant experiences an AE 
requiring hospitalization, they will be given instructions on which hospital to go to 
for evaluation and treatment.  The management of suspected and confirmed EVD 
will be according to local standards. 
5.2. 
HIV and syphilis counseling  
Participants who test positive for HIV or syphilis at study entry will be provided with 
counselling and referred for treatment according to local standards.    
5.3. 
Study withdrawal 
Every effort should be made to follow participants for blood draws and SAEs until 
the end of the study.  Following vaccination, participants may withdraw from follow-
up in the study at any time.  They may resume participation at any time. Reasons 
for withdrawal will be recorded. 
5.4. 
Pregnancy 
Women who report being pregnant in the first 3 months of follow-up after the 
prime vaccination will be followed for the outcome of their pregnancy.  The 
outcome will be reported on a CRF. Women who report being pregnant prior to 
the booster vaccination or who have a positive pregnancy test at 56 days will not 
receive the booster vaccination at 56 days. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
49 
 
5.5. 
Childhood vaccinations 
Children will not be enrolled if they received a vaccination in the 28 days before 
randomization.  Children will be advised not to receive another vaccination in the 
28 days after the prime vaccination as well as prior to and after the booster 
vaccination unless the benefit of that vaccination is judged to be greater than the 
potential risk.  For example, if a child has an unexpected exposure that places 
them at high risk of a disease (e.g., rabies, meningitis), they may be vaccinated.  
6. 
PHARMACY PROCEDURES 
6.1. 
 
Study agents 
6.1.1. 
 Ad26.ZEBOV and MVA-BN-Filo vaccines 
The drug products, Ad26.ZEBOV and MVA-BN-Filo will be provided by Johnson & 
Johnson. Ad26.ZEBOV is supplied in single dose glass vials with an extractable 
volume of 0.5 mL at a dose of 5x1010 vp. Ad26.ZEBOV is colorless to slightly 
yellowish or brownish, clear to slightly opalescent suspension, practically free of 
particles.  
The MVA-BN®-Filo vaccine is supplied in single dose glass vials with an 
extractable volume of 0.5 mL at a dose of 1x108 infectious units. MVA-BN®-Filo is 
a milky, light yellow suspension with no visible extraneous particles.   
6.1.2. 
rVSV∆G-ZEBOV-GP 
The drug product, rVSV∆G-ZEBOV-GP, will be provided by Merck. The vaccine 
will be a sterile, aqueous, buffered solution composed of rVSV∆G-ZEBOV-GP drug 
product filled into single use vials with an extractable volume of 1 mL at a nominal 
dose of 2x107 pfu/mL. The product is a clear, colorless aqueous suspension free 
from particles.  The certificate of analysis for the lot of rVSVΔG-ZEBOV-GP to be 
used, indicated a first assay test result of 1.3x108 pfu/mL, with the geometric mean 
of available assay results of 9.4 x 107.   
In Version 2.0 of PREVAC, the rVSVΔG-ZEBOV-GP vaccine was not used.  In 
Version 3.0, the rVSVΔG-ZEBOV-GP vaccine was given after 2-fold dilution 
(approximately 5x107 pfu/mL) for prime and boost.  In Version 4.0, the undiluted 
dose (geometric mean of assays = 9.4x107 pfu/mL) is used for both the prime and 
boost vaccinations. 
6.1.3. 
Sterile Normal Saline 
The placebo is sterile normal saline (sodium chloride 0.9 percent for injection, 
United States Pharmacopeia, preservative free).   

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
50 
 
6.2. 
Study agent presentation and storage 
6.2.1. 
Study agent labels 
Clinical supplies will be affixed with a clinical label in accordance with regulatory 
requirements. 
6.2.2. 
Study agent storage 
Storage requirements as well as procedures for temperature deviations for the 
investigational vaccines are described in the Pharmacy Procedures.   
Normal saline will be stored at room temperature in accordance with the 
manufacturer label.  
If deviations in storage temperature occur from the normal allowance for the 
pharmacy freezer, the affected investigational products will be quarantined and the 
storage temperature excursion will be promptly reported to the Principal 
Investigators, the IND sponsor, and product manufacturer: Johnson & Johnson for 
Ad26.ZEBOV and MVA-BN-Filo, Merck for rVSV∆G-ZEBOV-GP.  The excursion 
will be evaluated and investigated and action will be taken to restore and maintain 
the desired temperature limits.  Pending the outcome of the investigation, the 
manufacturer will notify the Principal Investigator, sponsor, and pharmacist if 
continued clinical use of the product is acceptable. 
6.3. 
Preparation of study agents for injection 
For the initial vaccination, products will be prepared in individual syringes at the 
vaccination site pharmacies in each country.  The syringes for the 56 day 
vaccination will also be prepared by these pharmacies. 
Products will remain under the manufacturer required temperature range prior to 
preparation of syringes detailed in the Pharmacy Procedures. 
Examples of syringe labels will be included in the Pharmacy Procedures as well 
as details related to product receipt, accountability, storage (shelf-life), preparation 
and disposition. 
6.4. 
Study agent accountability 
Study pharmacist in each country will be responsible for maintaining an accurate 
record of vaccine supplies for this study, as well as documenting the destruction 
of unused syringes according to local standards for biohazardous waste disposal.  
Electronic documentation as well as paper copies may be used.  Accountability 
instructions will be included in the Pharmacy Procedures.   

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
51 
 
7. 
EVALUATION 
7.1. 
Planned data analyses 
Final analyses will be carried out according to a statistical analysis plan that is 
updated prior to unblinding.  The planned analyses below only describe those 
planned for participants enrolled under Version 4.0.  Separate analyses will be 
carried out for participants enrolled under Version 2.0 and 3.0. These analyses 
and analyses which will be pooled across protocol versions are described in the 
statistical analysis plan. 
Prior to unblinding, the analysis plan for the immunogenicity substudy 
(Appendix D) and the viral shedding substudy (Appendix E) will also be finalized.   
For the purposes of regulatory filings there is a specific primary objective for each 
participating pharmaceutical company involving a single follow-up time point and 
the use of a validated assay for immunogenicity testing.  Thus, the two companies 
may also prepare a statistical analysis plan prior to unblinding and regulatory filing 
that describes analysis plans for their primary objective (e.g., comparisons with 
matched placebo for immunogenicity due to gamma irradiation of a subset of 
specimens) as well as plans for controlling type 1 error for investigations of 
secondary endpoints and subgroups. 
The following plan outlines data analyses that will be carried out during the trial by 
the unblinded statisticians for the independent DSMB. 
Adults and children will be analyzed separately and combined.  Baseline 
characteristics and follow-up completeness will be summarized by vaccine group. 
Prior to carrying out the endpoint analyses which pool the two placebo groups, the 
two placebo groups will be compared for the immunogenicity and safety outcomes. 
The primary analyses for adult and children will be carried out at the 0.0167 (2-
sided) level of significance and will compare the three vaccine strategies with the 
pooled placebo group for the percent with a positive antibody response at 12 
months. Each pair-wise comparison will be carried out using Mantel-Haenszel chi-
square statistics which consider stratification by vaccination center.  Similar 
methods will also be used to compare groups for the immediacy of the antibody 
response at 14 days.  These analyses will exclude those participants with elevated 
antibody levels at baseline.  Participants will not be screened at study entry for 
elevated antibody levels.  These exclusions will be carried out during analysis.  
Analyses will also be carried out for all randomized participants irrespective of 
antibody level at baseline. 
Similar methods will be used to compare antibody responses of each vaccine 
strategy with the pooled placebo group at other time points, including 28 days after 
randomization and 28 days following the booster vaccination.  The two additional 
primary objectives for immune responses, pooled rVSVΔG-ZEBOV-GP groups 
versus pooled placebo groups 28 days after randomization for Merck, and  
rHAd26/MVA versus pooled placebo groups approximately 28 days following 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
52 
 
booster vaccination for Janssen, will be carried out the 0.05 (2-sided) level of 
significance.  
All comparisons (for adults, children, and for adults and children combined) of 
immune responses and safety outcomes at other time points will be carried out at 
the 0.0167 (2-sided) level of significance when there are three pair-wise 
comparisons made (e.g., each vaccine strategy versus pooled placebo) and at the 
0.025 (2-sided) level of significance when there are two pair-wise comparisons 
made (e.g., for comparisons at days 7, 14, 28 and 56 with the pooled placebo 
group in which the two rVSVΔG-ZEBOV-GP strategies are pooled).   
SAEs will be classified by system organ class according to MedDRA®. The total 
number of events and the number of participants with at least one event will be 
reported.   Mantel-Haenszel chi-square statistics will be used for each pair-wise 
comparison of the number of participants with an SAE through 12 months of follow-
up; time-to-event methods, e.g., Kaplan-Meier, Cox regression, will also be used 
to compare groups for SAEs over the 12 month follow-up period.  Analyses will 
also be carried out for SAEs judged to be related to the vaccine.  If follow-up is 
extended to 60 months, similar analyses will be performed for SAEs reported 
through 60 months.   
 
Each vaccine group will be compared with the pooled placebo group using Mantel-
Haenszel chi-square tests and time-to-event methods for diagnoses of laboratory-
confirmed malaria through 12 months of follow-up.    
 
For safety and efficacy outcomes assessed before the booster at 56 days, the two 
groups given the rVSVΔG-ZEBOV-GP prime vaccination will be combined for 
comparisons with pooled placebo group  For outcomes that are assessed after 56 
days, each of the three vaccination strategies will be compared with the pooled 
placebo group. 
 
Mantel-Haenszel chi-square tests will be used to compare each vaccine with 
placebo for the percent with grade 3 or 4 AEs  and with targeted symptoms 
reported at 7, 14, and 63 days.  Similar methods will be used to compare injection 
site reactions reported immediately following prime vaccination and at 7 and 14 
days.  Likewise, treatment differences in injection site reactions reported 
immediately following booster vaccination and through day 63 (7 days after booster 
vaccination) will be summarized with similar methods. In the event that there is 
evidence that safety outcomes differ between the two placebo groups, analyses 
will also be performed for each vaccine versus its matched placebo. 
Longitudinal mixed models will be used to compare log-transformed antibody 
levels collected at multiple follow-up time points during follow-up.  Results will be 
summarized as geometric means and geometric mean ratios. These analyses will 
be carried out among participants who did not have elevated antibody levels at 
baseline.  Baseline titer level will be included as a covariate.  These analyses will 
be supplemented with scatter plots that display fold increases in antibody titers, 
line plots, box plots and reverse cumulative distributions of antibody levels at each 
visit for each vaccine group. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
53 
 
For children, changes in biochemical and CBC test values after 7 and 63 days will 
be compared for each vaccine versus the pooled placebo group using analysis of 
covariance with the baseline laboratory level as a covariate. Laboratory test results 
will be graded for severity according to the Division of AIDS (DAIDS) AE Table and 
local laboratory norms, and the proportion with grade 3 or 4 laboratory 
abnormalities will be compared.  
For children, findings based on contacts during the 7 days following the prime and 
booster vaccination will be summarized each day.  Trends over the 7 days will be 
examined for each vaccine versus placebo comparison. 
Subgroup analyses for the primary endpoints and major secondary outcomes, 
including SAEs and other safety data, will be performed to determine whether 
vaccine versus placebo differences vary qualitatively across various baseline-
defined subgroups.  Subgroup analyses will be performed using regression 
analysis by age, gender, country, vaccination center, health care worker or job 
involving close contact with EVD cases, baseline laboratory abnormalities, HIV 
status, and nutritional status as determined by body mass index.  An overall test 
of heterogeneity will provide evidence of whether the magnitude of the vaccine 
pair-wise comparison for efficacy/safety outcomes varies across these baseline 
subgroups. 
In addition to the separate analysis for adults and children for each vaccine group 
versus placebo for antibody levels and safety outcomes, the endpoints will also be 
summarized pooling the data for adults and children.  Methods similar to those 
described above will be used after stratification on age. 
Analyses which compare each vaccine group with placebo for immunogenicity and 
safety outcomes and adjust for age, gender, vaccination center, health care worker 
or job involving close contact with EVD cases, baseline laboratory abnormalities, 
HIV status, and nutritional status as determined by body mass index will also be 
carried out with regression. 
7.2. 
Independent DSMB 
The statistical analysis plan will be reviewed with the independent DSMB prior to 
beginning the study and after any changes are made to it.  It will be updated by 
the investigators prior to unblinding the data or if a correlate of protection is found. 
The DSMB will closely monitor accumulating safety and immunogenicity data for 
adults and children in each age group (1-4, 5-11, and 12-17 years).  The DSMB 
will review data for participants enrolled under Versions 2.0, 3.0 and 4.0. 
After 70 children aged ≥12 to ≤17 years have been enrolled (20 vaccinated with 
Ad26.ZEBOV, 30 vaccinated with rVSVΔG-ZEBOV-GP, and 20 vaccinated with 
placebo) and followed for 28 days, the DSMB will be asked to consider whether 
vaccination of children ≥5 to ≤11 years can commence.  The decision to begin 
enrolling younger children will be based on the severity of AEs experienced and 
the likely relationship of AE to a vaccine, the frequency of adverse effects, and 
whether the AEs were time-limited. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
54 
 
If judged safe to enrol younger children, 70 children aged ≥5 to ≤11 years will be 
enrolled and after they have been followed for 28 days the DSMB will be asked to 
consider whether vaccination of children at least 1 year of age to ≤ 4 years can 
commence.  The decision to begin enrolling children at least 1 year of age to ≤ 4 
years will be based on AE severity, frequency and time-limitedness. 
Once the decision is made that the enrolment of children 1 to 4 years is safe, the 
goal will be to target the enrolment of the 1,400 children to achieve a similar 
distribution in the 3 age groups. 
Safety data for children will be reviewed at least monthly until the DSMB is satisfied 
that the vaccines are safe at which time less frequent reviews may be carried out.   
The DSMB will be immediately notified of SAEs considered vaccine-related.  The 
DSMB may pause enrolment in the event of vaccine-related deaths or SAEs that 
are considered vaccine-related. The DSMB may also pause enrolment or request 
that participants be notified if there is  an increased frequency of unanticipated 
adverse effects. 
In addition to monitoring safety and immunogenicity outcomes, the DSMB will also 
review the completeness of follow-up and other aspects of study conduct. 
After each meeting the DSMB will recommend continuing the study as planned, 
modifying the study, or terminating the study. 
There is no plan to terminate the trial early based on the immunogenicity findings. 
7.3. 
Steering Committee 
The trial will be conducted under the direction of a steering committee.  Members 
of the steering committee will be blinded to interim safety and immunogenicity 
results.  The steering committee will monitor the enrolment and follow-up of 
participants, including the collection of AEs and blood for immunogenicity testing.  
The steering committee may request that more men or women or participants in 
specific age groups be enrolled to ensure broad generalizability of the findings.    
The steering committee will be responsible for the planned sample size re-
estimation before enrolment is completed.  The sample size re-estimation will be 
carried out using data pooled across the vaccine strategies. The steering 
committee will also be responsible for identifying new information for participants 
that arises during the trial that may relate to the willingness of participants to 
continue participating in the trial. Members of the steering committee will meet with 
the DSMB during an open session to discuss study conduct.   

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
55 
 
8. 
PROTECTION OF HUMAN SUBJECTS AND OTHER 
ETHICAL CONSIDERATIONS 
8.1. 
 Local review of protocol and informed consent 
Prior to initiation, the protocol, the informed consent and assent forms, and 
participant information materials will be submitted to and approved by the 
INSERM IRB/ EC; and LSHTM EC; and the ECs in Guinea, Liberia, Mali, and 
Sierra Leone.  Any future amendments to the study protocol, consent, or 
participant materials will likewise be submitted and approved by these ECs.   
8.2. 
 Ethical conduct of the study 
The trial will be conducted according to the Declaration of Helsinki in its current 
version and any subsequent revisions; US regulations governing protection of 
human subjects such as 21 US Code of Federal Regulations (CFR) 50; the 
requirements of Good Clinical Practice (GCP) as defined in published guidelines; 
the US Office for Human Research Protections; and with local law and regulation, 
whichever affords greater protection of human subjects. 
A team of social anthropologists will undertake a longitudinal survey to collect the 
experiences of both participants and health professionals within the trials.  They 
will inform in real time the investigators of issues that may be expressed by the 
persons carrying out the field work. 
8.3. 
 
Informed consent of study participants 
Participating adults must sign/mark the appropriate approved informed consent 
form prior to any study-related procedures being conducted. Informed consent for 
children will be given by one of the parents or legal guardian of the child. A video 
and/or picture booklets that describe the study may be used to ensure that illiterate 
volunteers understand the study requirements and risks and benefits.  A witness 
will document that subjects who are illiterate reviewed the booklet/video and were 
provided the opportunity to ask questions about the consent information.  Details 
of consent and assent procedures for minors who are able to provide assent will 
be described in the manual of operations.  
Minors who decline participation in the study after reviewing the assent materials 
will not be enrolled even if their parent(s) or legal guardian consent to their 
participation. 
Community mobilization efforts will help identify local stakeholders to ensure that 
the informed consent process is locally acceptable and takes into account 
background health, scientific and basic literacy levels, and is effective. 
8.4. 
 
Confidentiality of study participants 
The confidentiality of all study participants will be protected in accordance with 
GCP Guidelines and national regulations.  

PREVAC 
Version 4.0 
March 10, 2018 
56 
Data generated as result of this study are anonymized and are to be available for 
inspection on request by IRBs and the regulatory health authorities, including 
external site auditors and inspectors.  For example, monitors may visit the clinical 
research sites to verify the existence of signed informed consent documents and 
the prompt and accurate recording of SAEs. The monitors may also inspect the 
clinical site regulatory files to ensure applicable guidelines are being followed.  
8.5. 
Compensation 
Participants in the study will be compensated for their time and inconvenience.  
Compensation will be provided at each regularly scheduled study visit/contact.  
The governments of each country will recommend the amount of compensation to 
be provided.  
8.6. 
Dissemination of trial results 
Participants in each study and representatives of the community will be kept 
informed of enrolment and follow-up statistics.  When the trial has been completed, 
the results will be shared with participants and community representatives. 
8.7. 
Risk/Benefit Assessment 
The sponsors evaluated the risks of the respective interventions to the benefit 
from enrollment in the study.  In accordance with US regulation 45CFR46, 
minimal risk is defined as the probability and magnitude of harm or discomfort 
anticipated in the research are not greater in and of themselves than those 
ordinarily encountered in daily life or during the performance of routine physical 
or psychological examinations or tests.  As such, consistent with the 
administration of any vaccine, approved or investigational, and considering the 
safety profile of the study vaccines described in sections 1.1.2 and 1.1.3, the 
study intervention represents greater than minimal risk to both adults and 
children.  The direct benefit to both adults and children for those randomized to 
the study vaccines result from the immunogenic response observed from the 
study vaccines in previous studies.  Evidence indicates a reservoir of Ebola 
remains in West Africa that is capable of generating sporadic outbreaks in the 
near future.23  The immunogenic response generated by these vaccines has the 
potential to protect vaccinated individuals from Ebola acquisition.  One study of 
the rVSVΔG-ZEBOV-GP vaccine provided evidence of clinical 
efficacy.33  Additionally, access to healthcare is limited in West Africa.24 During 
the 2015 PREVAIL I study, participants included in the study were provided 
laboratory testing not available through the public health system resulting in the 
identification of a number of underlying conditions that allowed individuals to 
enter the health care system for treatment.35  For PREVAC, the baseline and 
continued follow-up of volunteers will provide the opportunity for early 
identification of unrecognized illnesses and referral to the health care system for 
treatment.   

PREVAC 
Version 4.0 
March 10, 2018 
57 
The sponsors believe the research in adults is approvable as research involving 
greater than minimal risk to subjects but with a prospect of direct benefit to the 
individual subjects. In children, the investigators believe the research is 
approvable under Subpart D 45 CFR 46.405 as the risk is greater than minimal 
risk but may provide prospect of direct benefit. This request is based on evidence 
from a Guinean study reporting that children, especially those under age 5, were 
particularly vulnerable and highly susceptible to death.25 In their paper describing 
the impact of Ebola on children in Guinea, the authors found an overall case 
fatality rate of 62.9% for children between 1 month and 20 years.  For those 
between one month and five years, the case fatality was 82.9%. The WHO 
statistics for Guinea for all patients in the 2014-2016 outbreak was 67%.2 
Because 45 CFR 46.408 requires that the consent of both parents be obtained 
unless there is the potential for benefit, the sponsors have identified that there is 
a potential for benefit due to the vulnerability of children to Ebola and therefore 
request under provisions permitted under 45 CFR 46.408(b) that consent from 
one parent to enroll a child be sufficient.  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
58 
 
APPENDIX A 
Time and Events Schedule for Baseline and Regularly Scheduled 
Follow-up Visits 
 
Adults 
Children 
Screening (children must have a documented negative HIV test prior to 
randomization) 
 
X 
Baseline  
 
 
Informed consent/assent 
X 
X 
Initial vaccination 
X 
X 
Demographics a) 
X 
X 
Contact information b) 
X 
X 
Indicators of increased risk c) 
X 
X 
Height, weight and temperature 
X 
X 
Mid-upper arm circumference for children 1-5 years 
 
X 
Pregnancy test for females of child-bearing potential, i.e., females who have 
experienced menarche or who are aged 14 years and older 
X 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Blood sample for chemistries and CBC 
X 
X 
Blood sample for HIV/syphilis testing 
X 
X 
Saliva sample for assessing viral shedding (subsample) 
 
X 
Stored blood for assessing T cell and memory B cell function in a subset of 
participants in Guinea 
X 
 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs following prime vaccination 
X 
X 
HIV pre-counseling 
X 
X 
Days 1 to 6 following prime and booster vaccination 
 
 
Contact for injection site reactions, targeted symptoms of any grade severity, 
grade 3 or 4 AEs, measurement of temperature, and possible SAEs in 
children 
 
X 
Day 7 and Day 63 
 
 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs 
X 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Saliva sample for assessing viral shedding (subsample) 
 
X 
Blood sample for assessing T cell and memory B cell function in a subset of 
adults in Guinea 
X 
 
HIV and syphilis post-counseling referral 
X 
X 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
59 
 
Blood sample for chemistries and CBC 
 
X 
Temperature 
X 
X 
Day 14  
 
 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs 
X 
X 
Temperature 
X 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Saliva sample for assessing viral shedding (subsample) 
 
X 
Blood sample for assessing T cell and memory B Cell function in a subset of 
adults in Guinea 
X 
 
Day 28 
 
 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs 
X 
X 
Temperature 
X 
X 
Weight 
 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Saliva sample for assessing viral shedding (subsample) 
 
X 
Day 56  
 
 
Booster vaccination 
X 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Pregnancy test for females  of childbearing potential, i.e., females who have 
experienced menarche or who are aged 14 years and older 
X 
X 
Temperature 
X 
X 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs following booster vaccination 
X 
X 
Saliva sample for assessing viral shedding (subsample) 
 
X 
Blood sample for assessing T cell and memory B cell function in a subset of 
adults in Guinea 
X 
 
Day 70 
 
 
Blood sample for assessing T cell and memory B cell function in a subset of 
adults in Guinea 
X 
 
Months 3 and 6 
 
 
Injection site reactions, targeted symptoms of any grade severity, and grade 3 
or 4 AEs (Month 3 only) 
X 
X 
Temperature 
X 
X 
Weight 
 
X 
Mid-upper arm circumference for children 1-5 years 
 
X 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
60 
 
Blood sample for immunogenicity testing and future research 
X 
X 
Saliva sample for assessing viral shedding (subsample at Month 3 only) 
 
X 
Months 12, 24, 36, 48, and 60 
 
 
Temperature 
X 
X 
Weight 
 
X 
Height 
 
X 
Mid-upper arm circumference for children 1-5 years 
 
X 
Blood sample for immunogenicity testing and future research 
X 
X 
Blood sample for assessing T cell and memory B cell function in a subset of 
adults in Guinea 
X 
 
SAEs throughout follow-up 
X 
X 
EVD events (reported as soon as aware) throughout follow-up and 
where possible a stored blood sample for future research 
X 
X 
Death (reported as soon as aware) throughout follow-up 
X 
X 
Pregnancy outcome for women who become pregnant in the first three 
months of follow-up after prime vaccination  
X 
X 
a) 
Birth month and year, gender 
b) 
Contact information for self and 2 contacts who will know how to locate the volunteer 
c) 
Role as health care worker, contact with persons known to have EVD  
 
 
  
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
61 
 
APPENDIX B 
Reference Documents on PREVAC Study Website 
The PREVAC website will maintain updated links to the following documents 
referenced in the PREVAC protocol and to other information pertinent to the 
study: 
PREVAC Procedures 
 
Pharmacy Manual of Operations  
 
Investigator’s Brochure for each vaccine 
 
The Division of AIDS (DAIDS) Table for Grading the Severity of Adult and 
Pediatric Adverse Events, Version 2.0, November 2014. 
 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
62 
 
APPENDIX C 
Site Investigators and Ethics Review Boards 
Site Investigators: 
Abdoul Habib Beavogui 
Guinea (Landreah and Maferinyah) 
Alseny Modet Camara 
Guinea (Landreah) 
Mamadou Saliou Diallo 
Guinea (Maferinyah) 
Mark Kieh 
Liberia 
Stephen B. Kennedy 
Liberia 
Bailah Leigh 
Sierra Leone 
Mohamed H. Samai  
Sierra Leone 
Gibrilla Fadlu Deen 
Sierra Leone 
Alie Wurie 
Sierra Leone 
Samba Sow 
Mali Center for Vaccine Development 
Milli Tapia 
Mali Center for Vaccine Development 
Seydou Doumbia 
Mali University Clinical Research Center 
Mahamadou Diakite 
Mali University Clinical Research Center 
 
Ethics Review Boards: 
INSERM IRB (IRB00003888 – FWA00005831) 
Paris, France 
 
LSHTM Research EC 
London, United Kingdom 
 
National Research Ethics Board (NREB) 
Monrovia, Liberia 
 
Comité National d’Ethique pour la Recherche en Santé (CNERS) 
Conakry, Guinea 
 
Sierra Leone Ethics and Scientific Review Committee (SLESRC) 
Freetown, Sierra Leone 
 
Faculty of Medicine, Pharmacy, and Odonto-Stomatology (FMPOS) and IRB of 
the University of Maryland, Mali 
 
Regulatory Sponsor in Liberia: 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
63 
 
Project PREVAIL will be the Regulatory Sponsor In Liberia to the Liberian 
Medicines and Health Products Regulatory Authority (LMHRA). 
INSERM has entered into an institutional authorization agreement with the NIH to 
allow the NIH to rely upon the ethical review of the INSERM EC.  The INSERM 
EC will conduct initial and annual continuing review of the research in 
accordance with the requirements identified in the Federal-Wide Assurance 
granted by the Office for Human Research Protections of the US Department of 
Health and Human Services. 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
64 
 
APPENDIX D 
PREVAC Immunological Substudy Protocol 
(To Be Performed in Guinea Only)  
Mondor Immunomonitoring Center, MIC (C. Lacabaratz)/Vaccine Research 
Institute, VRI (Head: Pr. Y. Lévy) 
1. Rationale 
A variety of vaccine platforms against Ebola have been generated in mice, guinea 
pigs and NHPs. Different types of vaccines against EBOV have been proposed in 
clinical trials, including in majority recombinant vaccines expressing the EBOV 
surface GP. But question regarding their efficiency remains unanswered.  
Among these vaccines, the Ad26.ZEBOV vaccine is used in prime boost strategy 
in combination with MVA-BN-Filo in phase I study: EBOVAC1 trial is actually 
conducted in the United Kingdom (UK), the US and African countries to assess the 
safety and immunogenicity of the candidate prime-boost regimen in healthy 
volunteers. Enrolment of the first human trial started in December 2015, 
preliminary post-prime safety and immunogenicity data are expected in March 
2015 and the full safety and humoral/cellular immunogenicity data in May/June 
2015. Phase II trials (EBOVAC2) are actually ongoing in healthy volunteers in 
Europe (France and UK) and non-epidemic African countries. 
Alternatively, some data from an open-label, cluster randomized ring vaccination 
trial conducted in Guinea, showed that the rVSVΔG-ZEBOV-GP vaccine 
prevented EVD events occurring at least 10 days after randomization in close 
contacts who were immediately vaccinated, but duration of the vaccine effect was 
not assessed. Two other phase 1 dose-escalation trials using rVSVΔG-ZEBOV-
GP vaccine were conducted in US. No short-term safety concerns were identified 
after a single administration of the rVSVΔG-ZEBOV-GP vaccine, and anti-Ebola 
humoral responses (titers of antibodies against ZEBOV GP assessed by ELISA) 
were identified in all the volunteers 28 days after vaccination. 
Up to now, very few data on the quality of cellular immune responses induced by 
the vaccines and no data on their middle/long-term duration are available. 
Moreover, no comparison of immunogenicity profiles between different vaccine 
strategies with and without a booster has been assessed.  
2. Objectives 
The objective of the immunological sub-study is to analyze the immunogenicity of 
the 2 prophylactic vaccine candidates (rVSVΔG-ZEBOV-GP from Merck and 
rHAd26/MVA from Janssen) used in the PREVAC study in a population at risk of 
EBOV infection. We will focus on T cell responses induced by the vaccines in adult 
population and on their persistence until 1 year after vaccination and more 
(middle/long term). 
 
This hypothesis will be addressed with both the diluted (Version 3.0) and undiluted 
(Version 4.0) rVSVΔG-ZEBOV-GP vaccine. 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
65 
 
 
3. Endpoints 
In addition to the analysis of humoral response (neutralizing antibodies and 
antibodies mediating antibody-dependent cellular cytotoxicity), analysis of the 
immunogenicity of the study vaccine regimens will include the characterization of 
specific cellular response. The quality, the precocity as well as the durability of the 
T cell responses induced by the different vaccines will be raised, and we can 
propose the following assays. 
The primary endpoints will focus on quantitative and qualitative aspects of specific 
T cell response. 
- The frequency and functional capacity of EBOV-specific T cells will be determined 
using interferon gamma (IFNγ) enzyme-linked immunospot (ELISpot) assay after 
stimulation of PBMCs with EBOV-specific peptides (15-mer peptide pools, to be 
determined) before (d0), 14 days after each vaccination (d14 and d70) and at week 
48. 
A positive IFNγ ELISpot response to a given peptide pool will be defined by a mean 
spot forming unit (SFU) across replicate wells ≥ 55 SFU/106 cells and ≥ 4-fold the 
negative control. A response to a peptide pool after vaccination will be considered 
positive if not already positive at baseline and specific IFNγ ELISPOT response 
will be defined by a positive response to at least one of the peptide pools. 
The number and the frequency (with 95% one-sided confidence interval) of 
subjects with an EBOV-specific IFNγ ELISpot response, as well as the magnitude 
(intensity) of this response will be calculated for each arm.  
- Activation of cluster of differentiation (CD) 4+ and CD8+ T cell subsets and their 
cytokine expression patterns will be determined by flow cytometry after EBOV-
specific stimulation (including, but not limited to, IFNγ, interleukin (IL) 2, and tumor 
necrosis factor alpha (TNF-alpha) using intracellular cytokine staining (ICS) before 
(d0), 14 days after each vaccination (d14 and d70) and at week 48. The 
polyfunctionality of specific CD4+ and CD8+ T cells (capacity to produce more than 
1 cytokine) will be analyzed as well. 
The secondary endpoints will include:  
- Phenotypic analysis of B and T cell subsets before (d0), 14 days after vaccination 
(d17 and 70) and w48. The frequency of B (naive, memory, switched, plasma cells) 
and T cell subsets (naive, memory, effector, cytotoxic capacity and T follicular 
helper [Tfh] cells) will be assessed by flow cytometry combining a panel of different 
markers: e.g. CD19, CD21, CD27, CD38, immunoglobulin (Ig) D, IgM and CD3, 
CD4, CD8, CD45RA, CC-chemokine receptor 7 (CCR7), CD27, Perforin, 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
66 
 
Granzyme B, CXCR5, programmed cell death protein 1 (PD-1), respectively for B 
and T cell panels. 
- Measurement of serum cytokine/chemokine levels (inflammation/activation 
markers) before (d0) and after vaccination (d7 and d63) using Luminex technology 
allowing detection up to 30 analytes in a small sample volume. 
- Ex vivo gene expression profile in whole blood before and 3h after each 
vaccination (d0, d56), d7 and d63 to evaluate gene abundance by microarray in 
order to find the components of the vaccine signature and to compare them 
between Ebola vaccines. Highly sensitive and quantitative assays were developed 
to process samples including quality controls at each step of processing, from 
sampling to the results report. The technology used is based on the BeadChip 
Illumina platform which consists of the direct hybridization assay that offers the 
highest level of multiplexing for whole-genome expression profiling available, with 
the most up-to-date expression content and high-throughput processing. Each 
array on the HumanHT-12 v4 Expression BeadChip targets more than 34,000 
annotated genes. 
4. Sub-study eligibility criteria 
 
Adult participants from Guinea 
Participation approval in immunological sub-study (additional blood draws, 
specific consent)  
 
5. Sub-study components and procedures 
 
For the evaluation of the different immunogenicity endpoints, samples will be 
collected at the time points and in volumes as indicated in the Table below:  
 
Blood volumes (ml) 
Type of 
tube 
Assays 
d0 
Prime 
d7 
d14 
d56 
Boos
t 
d63 
d70 
M12-60 
EDTA 
(PBMC) 
IFNg Elispot 
12 
  
12 
  
  
12 
   12 
ICS 
12 
  
12 
  
  
12 
   12 
B / T 
phenotype 
6 
 
6 
  
 
6 
    6 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
67 
 
Dry 
tube 
(serum) 
Luminex 
4 
4 
  
  
4 
  
  
Tempu
s (RNA) Gene profile 
6* 
3 
  
6* 
3 
  
  
  
Blood total 
vol.  
40 
7 
30 
6 
 7 
 30 
   30 
*: 3 ml before and 3 hours after vaccine injection 
 
Number of tubes 
 
Type of 
tube 
d0 
Prime 
d7 
d14 
d56 
Boos
t 
d63 
d70 
M12-60 
EDTA  
7 ml tube  
4 
 
4 
  
 
4 
    4 
Dry  
5 ml tube  
1 
1 
  
  
1 
  
  
Tempus 
2* 
1 
  
2* 
1 
  
  
*: 1 tube before and 3 hours after vaccine injection 
 
6. Sample size calculation 
 
According to methodologist’s calculation respecting allocation ratio of 
randomization 2:1:2:1:1, we plan to enroll up to 230 participants in the substudy 
under Versions 2.0, 3.0, and 4.0 (in total). 
 
Separate analyses will be carried out for participants enrolled under Versions 3.0 
and 4.0. 
 
7. Sites 
 
Whole blood samples for this sub-study will be collected at Conakry (Guinea) site 
on 3 different types of tubes (tubes without anticoagulant, EDTA and TempusTM 
Blood RNA tubes), and then:  

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
68 
 
- sent immediately to the immunological sites (2 "selected" sites in Conakry for 
PBMC isolation:  Landréah and Institut National de Santé Publique)  
- processed at the immunological sites according to current versions of approved 
Standard Operating Procedure, 
- and stored according to a specific procedure before shipping to Mondor 
Immunomonitoring Platform, Créteil, France.  
Different Standard Operating Procedures will contain all details regarding the 
collection, handling, labeling, and shipment of blood samples to the laboratory in 
charge of immunomonitoring. 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
69 
 
APPENDIX E 
PREVAC SALIVA SAMPLE SUBSTUDY 
(To Be Performed in Liberia Only) 
Rationale 
This substudy of viral shedding of rVSV∆G-ZEBOV-GP is being conducted to 
estimate the proportion of children who have detectable vaccine virus by RT-PCR 
and to quantify vaccine virus levels shed after a prime and a boost vaccine dose. 
In Phase 1 studies that collected specimens to monitor shedding of rVSV∆G-
ZEBOV-GP, the largest proportion of adult subjects that had vaccine virus detected 
in saliva or urine was 5/20 in 2x107 pfu North American recipients with positive 
saliva on Day 7. This decreased to 1 of 10 on Day 14.29  Preliminary open-label 
data in African adults, adolescents between 13 and 17 years of age and children 
between 6 and 12 years of age suggests that shedding after vaccination is higher 
in children and adolescents compared to adults.  While no shedding was detected 
in adults, 14/18 adolescents and 7/20 children had positive saliva on Day 7.   
Shedding in children beyond Day 7 has not been assessed to date and will be 
assessed in this substudy. 
Objective 
The objective of the saliva substudy is to estimate the proportion of children who 
shed vaccine virus and to quantify the rVSVΔG-ZEBOV-GP vaccine shed in 
children (participants aged < 18 years) after the prime and boost vaccinations.  For 
estimates on days 7, 14, 28, and 56 (prior to booster), the two rVSVΔG-ZEBOV-
GP groups will be pooled.  On day 63 and at 3 months (after the booster), the two 
rVSVΔG-ZEBOV-GP groups will be considered separately. Our hypothesis is that 
viral shedding will be greatest 7 days after vaccination (prime or booster) and will 
decline afterwards. 
 
This hypothesis will be addressed with both the diluted (Version 3.0) and undiluted 
(Version 4.0) rVSVΔG-ZEBOV-GP vaccine.  
 
Procedures and Measurements 
At a single vaccination center, approximately 0.5-1.0 mL of saliva will be collected 
from children for whom informed consent is obtained at baseline (prior to prime 
vaccination), days 7, 14, 28, 56 (prior to booster), and 63 (7 days following 
booster), and at 3 months (approximately 35 days after booster).  The saliva will 
be divided into at least 2 x 200 µL aliquots in RNA microfuge tubes.  600 µL of 
Trizol LS or equivalent transport media will be added to each aliquot, the tubes will 
be sealed, mixed and stored at -70° C or lower, prior to shipment for quantitative 
RT-PCR (RT-qPCR) testing to determine levels of virus present in the samples 
(threshold value will be determined based on assay characteristics).  Testing will 
be performed on all substudy participants blinded to vaccine group.  
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
70 
 
Statistical Considerations 
We plan to enroll approximately 140 children in the substudy in each of Versions 
3.0 and 4.0 (280 total).  For both Version 3.0 and 4.0, efforts will be made to enroll 
1/3 of the total number of children in each age group (1-4, 5-11, and 12-17 years). 
 
With 140 children enrolled from the 5 randomized groups, there will be 
approximately 60 participants who are given the rVSVΔG-ZEBOV-GP vaccine as 
prime and 40 participants in the pooled placebo group. Approximately 40 
participants who are enrolled in the substudy would receive the rHAd26 /MVA 
vaccine,  The 95% confidence interval half-widths are presented for several 
percentages for 60 children receiving the rVSVΔG-ZEBOV-GP prime vaccine: 
10% ±8%, 30% ±12% and 50% ±13%.  
 
There will be approximately 20 participants  who are given the rVSVΔG-ZEBOV-
GP prime vaccine and booster vaccine at 56 days.  40 participants will receive a 
placebo booster. The 95% confidence interval half-widths are presented for 
several percentages receiving the rVSVΔG-ZEBOV-GP prime and booster vaccine 
for 20 children: 10% ±13%, 25% ±19%, and 30% ±20%.  
 
Counts and percentages of children with vaccine virus detected in saliva samples 
as well as mean, and medians (25th, 75th percentiles) for virus levels (copies/mL) 
will be presented for days 7, 14, 28, 56 (prior to the booster vaccination) and for 
day 63 and 3 months.  For the first four time points, the rVSVΔG-ZEBOV-GP prime 
vaccine with and without the rVSVΔG-ZEBOV-GP boost will be pooled; for the last 
2 time points (after the booster vaccination), these groups will be considered 
separately. 
 
Separate analyses will be carried out for participants enrolled under Versions 3.0 
and 4.0.    
 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
71 
 
APPENDIX F 
NIH Roles in the Conduct of PREVAC 
This appendix is required by NIH Office of Human Subjects Research Protections 
in order to establish an institutional reliance agreement for NIH to rely upon the 
INSERM Ethical Review Committee. 
NIH roles in the conduct of PREVAC are described below: 
• Dr. Lane is a member of the tripartite executive committee for the trial 
along with Yves Levy of INSERM and Peter Piot of LSHTM.  The INSERM 
representative chairs the executive committee. 
• PREVAC will be conducted in Guinea, Mali, Sierra Leone, and Liberia with 
the respective partnership of INSERM, LSHTM, and NIAID.  Trial 
execution in Liberia will be conducted under the auspices of Project 
PREVAIL. 
• NIAID resources will be involved in supporting the clinical trial 
infrastructure in Liberia and Mali.  
• NIAID staff will be on site to oversee the proper conduct of the research in 
Liberia and Mali but the actual execution of the research will be done by 
Liberians and Malians (e.g., preparing vaccines, administering consent, 
obtaining lab samples, vaccine administration, assessing vaccine 
reactions, completing case report forms, follow-up visits). 
• NIAID staff will be involved in overseeing the clinical lab as well as the 
research lab at the Liberian Institute of Biomedical Research.  
• NIAID funds support a contract with Leidos Biomedical who has hired staff 
in Liberia and also sub-contracted additional staff for the conduct of the 
study.  
• NIAID funds support a contract with Leidos Biomedical who provides sub-
contract funding to INSERM for the conduct of the study in Guinea.  
Specific functions in which NIAID collaborates with the other partners 
include: 
o Trial governance 
o Protocol development 
o Coordination with industry partners for provision of test articles 
o Data and safety management board support 
o Site development 
o Laboratory management 
o Staff training 
o Clinical trial monitoring 
• NIAID is the regulatory sponsor of record for the IND to the US FDA 
through the DCR OCRPRO. 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
72 
 
REFERENCES 
 
1 World Health Organization. Ebola Situation Report.  30 March 2016; 
http://apps.who.int/ebola/current-situation/ebola-situation-report-30-march-
2016.  Accessed 9 April 2016. 
 
2 World Health Organization. Ebola virus disease. 2018; 
http://www.who.int/mediacentre/factsheets/fs103/en/. Accessed March 10, 
2018. 
 
3 Feldmann H, Geisbert TW. Ebola haemorrhagic fever. Lancet 2011;377:849-
862. 
 
4 Chertow DS, Kleine C, Edwards JK, et al. Ebola virus disease in West Africa--
clinical manifestations and management.  N Engl J Med 2014;371:2054-2057. 
 
5 Bah EI, Lamah MC, Fletcher T, et al.  Clinical presentation of patients with 
Ebola virus disease in Conakry, Guinea.  N Engl J Med 2015; 372:40-47. 
 
6 Stanley DA, Honko AN, Asiedu C, et al.  Chimpanzee adenovirus vaccine 
generates acute and durable protective immunity against ebolavirus challenge.  
Nature Medicine 2014; 20:1126-1129. 
 
7 Investigator’s Brochure for MVA-BN-Filo.  
8 Verheust C, Goossens M, Pauwels K, Breyer D.  Biosafety aspects of modified 
vaccinia virus Ankara (MVA)-based vectors used for gene therapy or 
vaccination.  Vaccine 2012; 30:2623-2632. 
 
9 Investigator’s Brochure for rVSVΔG-ZEBOV-GP.   
10 Jones SM, Feldmann H, Stroher U, et al. Live attenuated recombinant vaccine 
protects nonhuman primates against Ebola and Marburg viruses. Nature 
Medicine 2005; 11:786-790. 
 
11 Geisbert TW, Daddario-Dicaprio KM, Geisbert JB, et al. Vesicular stomatitis 
virus-based vaccines protect nonhuman primates against aerosol challenge 
with Ebola and Marburg viruses. Vaccine 2008; 26:6894-6900. 
 
12 Qiu X, Fernando L, Alimonti JB, et al. Mucosal immunization of cynomolgus 
macaques with the VSVDeltaG/ZEBOVGP vaccine stimulates strong ebola 
GP-specific immune responses. PloS One 2009; 4:e5547. 
 
13 Feldmann H, Jones SM, Daddario-DiCaprio KM, et al. Effective post-exposure 
treatment of Ebola infection. PLoS Pathogens 2007; 3:e2. 
doi:10.1371/journal.ppat.0030002. 
 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
73 
 
 
14 Cobleigh MA, Bradfield C, Liu Y, Mehta A, Robek MD. The immune response 
to a vesicular stomatitis virus vaccine vector is independent of particulate 
antigen secretion and protein turnover rate. J Virology 2012; 86:4253-4261. 
15 Johnson JE, Nasar F, Coleman JW,  et al. Neurovirulence properties of 
recombinant vesicular stomatitis virus vectors in non-human primates. 
Virology. 2007;360: 36-49. 
16 Mire CE, Miller AD, Carville A, Westmoreland SV, Geisbert JB, Mansfield KG, 
et al. Recombinant vesicular stomatitis virus vaccine vectors expressing 
filovirus glycoproteins lack neurovirulence in nonhuman primates. PLoS Negl 
Trop Dis. 2012;6:e1567 
17 Regules JA, Beigel JH, Paolino KM, et al.  A recombinant vesicular stomatitis 
virus Ebola vaccine. N Engl J Med 2017; 376:330-341.  
 
18 Heppner DG, Kemp TL, Martin BK, et al.  Safety and immunogenicity of the 
rVSV∆G-ZEBOV-GP Ebola virus vaccine candidate in healthy adults: a phase 
1b randomized, multicenter, double-blind, placebo-controlled, dose-response 
study.  Lancet Infect Dis 2017; 17:854-866. 
   
19 Halperin SA, Arribas JR, Rupp R, et al.  Six-month safety data of recombinant 
vesicular stomatitis virus-Zaire Ebola virus envelope glycoprotein vaccine in a 
phase 3 double-blind, placebo-controlled randomized study in healthy adults.   
J Infect Dis 2017; doi: 10.1093/infdis/jix189. 
  
20 Henao_Restrepo AM, Camacho A, Longini IM, et al.  Efficacy and 
effectiveness of rVSV-vectored vaccine in preventing Ebola virus disease: final 
results from the Guinea ring vaccination, open-label,cluster-randomised trial 
(Ebola Ça Suffit!) Lancet 2017; 389:505-518. 
 
21 WHO Ebola Response Team.  Ebola virus among children in West Africa. N 
Engl J Med (Letter to Editor) 2015; 372:1274-1277. 
 
22 Kennedy SB, Bolay F, Kieh M, et al. Phase 2 placebo-controlled trial of 2 
vaccines to prevent Ebola in Liberia N Engl J Med 2017; 377:1438-1447. 
 
23  WHO Ebola Response Team, Agua-Agum J, Allegranzi B, et al. After Ebola in 
West Africa--Unpredictable Risks, Preventable Epidemics. N Engl J Med. 
2016;375:587-596. 
24 G. B. D. 2015 Healthcare Access and Quality Collaborators. Healthcare 
Access and Quality Index based on mortality from causes amenable to 
personal health care in 195 countries and territories, 1990-2015: a novel 
 

PREVAC 
 
Version 4.0 
  
 
March 10, 2018 
 
74 
 
 
analysis from the Global Burden of Disease Study 2015. Lancet. 
2017;390:231-266. 
25 Cherif MS, Koonrungsesomboon N, Kasse D, et al. Ebola virus disease in 
children during the 2014-2015 epidemic in Guinea: a nationwide cohort study. 
Eur J Pediatr. 2017;176:791-796.