A Phase Ib Randomised, Controlled, Single-blind Study to Assess the Safety, Immunogenicity of the Malaria Vaccine Candidate R21 With Matrix-M1 Adjuvant in West African Adult Volunteers
Phase: PHASE1
Conditions: Malaria
Sponsor: University of Oxford

1 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
 
A Phase Ib randomised, controlled, single-blind study to assess the safety, 
immunogenicity of the Malaria Vaccine Candidate R21 with Matrix-M1 adjuvant in 
West African adult volunteers. 
 
 
 
Study reference: 
VAC060 
 
Protocol Version: 
Final v2.1 
 
 
Date: 
14 Oct 2015 
 
OXTREC Number:  
36-15 
 
Sponsor: 
University of Oxford 
 
Funding body: 
EDCTP 
 
Author(s): 
CNRFP: Alfred B. Tiono, Issa N. Ouédraogo, Sodiomon B. 
Sirima 
 
UOXF: Egeruan Babatunde Imoukhuede, Adrian VS Hill 
 
 
 
 
 
 
 
 
 
 
 
 
 

2 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
MODIFICATION HISTORY 
List of Versions 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Version 
Date 
Author (s) 
Final v2.0 
11 June 2015 
UOXF:  Egeruan Babatunde Imoukhuede,  Adrian VS Hill 
CNRFP:  Alfred B. Tiono, Issa N. Ouédraogo, Sodiomon B. Sirima 
Final v2.1 
14 Oct 2015 
UOXF:  Egeruan Babatunde Imoukhuede,  Adrian VS Hill 
CNRFP:  Alfred B. Tiono, Issa N. Ouédraogo, Sodiomon B. Sirima 
 
 
 

3 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
PROTOCOL SIGNATURE SHEET 
 
Confidentiality Statement 
This document contains confidential information that must not be disclosed to anyone other than the Sponsor, the 
Investigator Team, members of the Independent Ethics Committee and the Burkina regulatory authority.  This 
information cannot be used for any purpose other than the evaluation or conduct of the clinical investigation without 
the prior written consent of Professor Adrian Hill. 
 
Statement of Compliance 
The trial will be conducted in compliance with the protocol, International Conference on Harmonisation Good Clinical 
Practice Guideline E6 (R1) (ICH-GCP) and the applicable regulatory requirements.   
 
Signatures 
“I have read this protocol and agree to abide by all provisions set forth therein. I agree to comply with the principles of 
the Declaration of Helsinki and the International Conference on Harmonisation Tripartite Guideline on Good Clinical 
Practice.” 
 
                                       
 
Principal Investigator: 
 _______________________________ 
 
Date: 14 Oct 2015 
 
 
 
UOXF Chief Investigator:  __
____________________________ Date: 14 Oct 2015  
 
 
Conflict of Interest 
“According to the Declaration of Helsinki, 2008, I have read this protocol, and declare no/the following (delete as 
appropriate) conflict of interest” 
 
 
 
                                           
 
Principal Investigator: 
 _______________________________ 
 
Date:  14 Oct 2015 
 
 
 
UOXF Chief Investigator:  _
______________________________ Date: 14 Oct 2015 
 
 
Details of conflict of interest (if any) 
 
 
 
 
 
 
 

4 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
CONTENTS 
 
Modification History ............................................................................................... 2 
Contents ................................................................................................................ 4 
Key Roles and General Information .......................................................................... 7 
List of abbreviations ................................................................................................ 9 
1. 
VAC060 Study synopsis ................................................................................. 10 
2. 
Background Information ............................................................................... 11 
2.1. Lifecycle of the malaria parasite .................................................................... 11 
2.2. Progress towards a malaria vaccine ............................................................... 12 
2.3. PRE-ERYTHROCYTIC STAGE OF INFECTION AS A VACCINE TARGET .................... 13 
2.3.1. 
R21 vaccine development ....................................................................... 13 
2.3.2. 
Pre-clinical studies of R21 ....................................................................... 14 
2.3.3. 
Matrix-M1 adjuvant................................................................................ 17 
3. 
Rationale ..................................................................................................... 17 
4. 
Objectives .................................................................................................... 17 
4.1. Primary Objective ......................................................................................... 17 
4.2. Secondary Objective ..................................................................................... 17 
5. 
description and justification of study design .................................................. 18 
5.1. Overview ..................................................................................................... 18 
5.2. Vaccines ...................................................................................................... 18 
5.3. Evaluation Criteria ........................................................................................ 18 
5.3.1. 
Safety endpoints..................................................................................... 18 
5.3.2. 
Immunogenicity endpoints ...................................................................... 18 
5.4. Study site ..................................................................................................... 18 
5.5. Study groups ................................................................................................ 19 
5.6. Vaccination Schedule .............................................................................. 19 
5.7. Follow-up of volunteers ................................................................................ 19 
5.7.1. 
Safety follow-up ..................................................................................... 19 
5.8. Sample size justification ................................................................................ 19 
5.9. Potential risks for volunteers ........................................................................ 19 
5.9.1. 
Phlebotomy ........................................................................................... 20 
5.9.2. 
Safety and tolerability of R21/Matrix- M1 vaccination .............................. 20 
5.10. Potential benefits for volunteers ................................................................... 20 

5 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
6. 
Inclusion and Exclusion Criteria ..................................................................... 20 
6.1. Inclusion Criteria .......................................................................................... 20 
6.2. Exclusion Criteria .......................................................................................... 21 
6.3. Withdrawal criteria ...................................................................................... 21 
7. 
Investigational Medicinal Products ................................................................ 21 
7.1. Manufacturing and presentation of vaccines .................................................. 21 
7.1.1. 
R21 vaccine ............................................................................................ 21 
7.2. Storage, dispensing and handling of Investigational Medicinal Products .......... 22 
7.2.1. 
Storage of R21/Matrix M-1 ..................................................................... 22 
7.3. Vaccination of volunteers ............................................................................. 22 
7.3.1. 
R21/Matrix-M1 vaccinations ................................................................... 22 
7.4. Concomitant medication/vaccination ............................................................ 22 
7.5. Indications for deferral of vaccine administration ........................................... 23 
7.6. Absolute contraindications to further vaccination .......................................... 23 
8. 
Study schedule and procedures ..................................................................... 23 
8.1. Identification of Study Participants ................................................................ 23 
8.1.1. 
Community sensitisation ......................................................................... 23 
8.1.2. 
Screening Visit ........................................................................................ 24 
8.1.3. 
HIV Testing ............................................................................................ 24 
8.2. Allocation of participants to Study Groups ..................................................... 24 
8.3. Enrolment .................................................................................................... 24 
8.4. STUDY VISITS AND PROCEDURES ................................................................... 25 
8.5. Laboratory Evaluations ................................................................................. 26 
8.6. Provision of care to the study participants ..................................................... 27 
8.7. Safety follow-up ........................................................................................... 29 
8.8. Immunogenicity measurements .................................................................... 29 
8.9. Data collection ............................................................................................. 29 
8.10. Study termination ........................................................................................ 30 
8.11. Definition of the Start and End of the Trial ..................................................... 30 
9. 
Assessment of scientific objectives ................................................................ 30 
9.1. Safety Evaluation ......................................................................................... 30 
9.2. Immunogenicity evaluation ........................................................................... 30 
10. 
Safety Reporting .......................................................................................... 30 
10.1. Definitions ................................................................................................... 30 
10.1.1. Adverse Event ........................................................................................ 30 

6 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
10.1.2. Adverse Drug Reaction (ADR) .................................................................. 31 
10.1.3. Serious Adverse Event (SAE) .................................................................... 31 
10.1.4. Suspected Unexpected Serious Adverse Reactions (SUSARs) ...................... 31 
10.2. Collection of Adverse Events ......................................................................... 31 
10.2.1. Local solicited Adverse Events ................................................................. 31 
10.2.2. Solicited general AEs ............................................................................... 31 
10.2.3. Unsolicited adverse events ...................................................................... 32 
10.3. Follow-up of Adverse Events ......................................................................... 32 
10.4. Reporting of Adverse Events ......................................................................... 32 
10.5. Grading the severity of adverse events .......................................................... 33 
10.5.1. Local solicited AEs ................................................................................... 33 
10.5.2. Solicited general AEs ............................................................................... 33 
10.5.3. Non Solicited AEs .................................................................................... 34 
10.5.4. Pregnancy .............................................................................................. 34 
11. 
Data Handling And Record Keeping ............................................................... 35 
11.1. Data Management ....................................................................................... 35 
11.1.1. Data Capture Methods ............................................................................ 35 
11.1.2. Types of Data ......................................................................................... 35 
11.2. Timing/Reports ............................................................................................ 35 
11.3. Archiving ..................................................................................................... 35 
11.4. Protocol Deviations ...................................................................................... 35 
12. 
Data Access and Quality Assurance ................................................................ 35 
12.1. Direct Access to Source Data/Documents ....................................................... 35 
12.2. Quality Assurance ........................................................................................ 35 
12.2.1. Modifications to the Protocol .................................................................. 35 
12.2.2. Monitoring ............................................................................................. 36 
13. 
Ethical Considerations .................................................................................. 36 
14. 
Indemnity/Compensation/Insurance ............................................................. 37 
15. 
References ................................................................................................... 38 
 
 
 

7 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
KEY ROLES AND GENERAL INFORMATION  
Trial Centre: 
Centre National de Recherche et de Formation sur le Paludisme 
(CNRFP)/ Unité de Recherche Clinique de Banfora (URC-B) 
01 BP 2208 Ouagadougou 01 
Burkina Faso 
 
 
Principal Investigator: 
Dr Alfred B. Tiono 
Centre National de Recherche et de Formation sur le Paludisme 
(CNRFP)/ Unité de Recherche Clinique de Banfora (URC-B) 
01 BP 2208 Ouagadougou 01 
Burkina Faso 
 
 
Study Physician 
Dr Sam Coulibaly 
 
 
 
Co-Investigator(s): 
Centre National de Recherche et de Formation sur le Paludisme 
(CNRFP)/ Unité de Recherche Clinique de Banfora (URC-B) 
 
 
─Dr Sodiomon B. Sirima 
─Dr Issa Nébié Ouédraogo 
─Dr Alphonse Ouédraogo 
─Dr Jean Baptiste Yaro 
─Dr Edith C. Bougouma 
─Dr Issiaka Soulama 
 
01 BP 2208 Ouagadougou 01 
Burkina Faso 
 
 
United Kingdom (Jenner Institute, Nuffield Dept. of Clinical Medicine, 
University of Oxford) 
 
─Prof Adrian Hill 
─Dr Egeruan Babatunde Imoukhuede 
─Dr Tommy Rampling 
─Dr Philip Bejon (statistician) 
─Dr Alison Lawrie 
─Dr Katie Ewer 
Centre for Clinical Vaccinology and Tropical Medicine University of 
Oxford Churchill Hospital, Oxford OX3 7LE  
 
 
Local Safety Monitor: 
Dr  KOUETA Fla 
Paediatrician National Teaching Hospital Yalgado Ouédraogo  
03 BP 7022 Ouagadougou 03 
Burkina Faso 
 
 
Data Safety and Monitoring Board: 
Chair:  Professor Geoffrey Targett 
Emeritus Professor of Immunology of Parasitic Diseases 
London School of Hygiene & Tropical Medicine 
Room 410, Keppel Street, London WC1E 7HT, UK 
 
DSMB members: 
Dr. Paul Milligan 
London School of Hygiene & Tropical Medicine 
Reader in Epidemiology and Medical Statistics 
Room 411a, Keppel Street, London WC1E 7HT, UK 
 
Professor Mahamadou Thera 

8 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
MRTC, Bamako, University of Mali 
Colline de Badalabougou 
BP E2528, Mali 
 
Dr. Bernhards Ogutu 
KEMRI 
P.O. Box 2254 - 00202 
Nairobi, Kenya 
 
Clinical Project Manager (Oxford): 
Head of Regulatory Affairs (Oxford): 
Vaccine Programme Coordinator : 
Dr Egeruan Babatunde Imoukhuede 
Dr Alison Lawrie,  
Ms Rachel Roberts 
Centre for Clinical Vaccinology and Tropical Medicine University of 
Oxford Churchill Hospital, Oxford OX3 7LJ 
 
Sponsor: 
University of Oxford 
Named contact: Heather House 
Clinical Trials and Research Governance 
Manor House 
John Radcliffe Hospital 
Oxford, OX3 9DZ 
Tel: 01865 743004 
 Fax: 01865 743002 
Email: heather.house@admin.ox.ac.uk 
 
External Monitor: 
Ceri McKenna 
Appledown Clinical Research Limited 
Orchard's End, 
Greenland's Lane, 
Prestwood, 
Gt. Missenden, 
Bucks. HP16 9QX 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

9 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
LIST OF ABBREVIATIONS 
AdCh63 
Chimpanzee Adenovirus 63 
AE 
Adverse event 
 
ALT 
Alanine aminotransferase 
AS01 
Adjuvant Systems 01 
BCG 
Bacille Calmette-Guerin 
CCVTM 
Clinical Centre for Vaccinology and Tropical Medicine 
CBF 
Clinical Biomanufacturing Facility 
ChAd63 
Chimpanzee Adenovirus 63 
CMI 
Cell-mediated immunity 
CRF 
Case report form 
CSP 
Circumsporozoite Protein 
CTL 
Cytotoxic T lymphocytes 
DSMB 
Data Safety and Monitoring Board 
EC 
Ethics Committee 
EDCTP 
European and Developing Countries Clinical Trials Partnership 
ELISA 
Enzyme-linked immunosorbant assay 
ELISPOT 
Enzyme-linked immunospot 
FBC 
Full blood count 
FP9 
Fowlpox 9 
GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
HBsAg 
Hepatitis B surface Antigen 
Hib 
Haemophilus Influenzae type b  
IB 
Investigator’s Brochure 
ICS 
Intracellular Cytokine Staining 
IDT 
Impfstoffwerk Dessau-Tornau GmbH 
IEC 
Independent Ethics Committee 
IFN-γ 
Gamma interferon 
IMP 
Investigational Medicinal Product 
IRB 
Independent Review Board 
HBV 
Hepatitis B virus 
HIV 
Human Immunodeficiency Virus 
HLA 
Human Leukocyte Antigen 
LFT 
Liver Function Test 
LSM 
Local Safety Monitor 
ME-TRAP 
Multiple epitope string with thrombospondin-related adhesion protein 
MHRA  
Medicines and Healthcare products Regulatory Agency 
MVA 
Modified vaccinia Virus Ankara 
MVVC 
Malaria Vectored Vaccines Consortium 
NHS 
National Health Service 
OXTREC 
Oxford Tropical Research Ethics Committee 
PBMC 
Peripheral blood mononuclear cells 
PCR 
Polymerase Chain Reaction 
pfu 
Plaque forming units 
PI 
Principal Investigator 
R21 
Recombinant HBsAg protein particle expressing CSP 
SAE 
Serious adverse event 
SC 
Subcutaneous 
SOP 
Standard Operating Procedures 
SUSAR 
Suspected unexpected serious adverse reaction 
vp 
 
Virus particles 
 

10 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
1. VAC060 STUDY SYNOPSIS 
 
Trial Title 
A Phase Ib randomised, controlled, single-blind study to assess the safety and 
immunogenicity of the Malaria Vaccine Candidate R21 with Matrix-M1 adjuvant in West 
African adult volunteers.  
Trial Identifier 
VAC060 
Clinical phase  
Ib 
Active ingredients of 
vaccines/products 
R21: Recombinant HBsAg protein particle expressing the central repeat and the C-
terminus of the circumsporozoite protein (CSP) 
Matrix-M1:  A saponin adjuvant 
Finished products 
R21/ Matrix-M1 
Dose(s)  
R21: 10µg and 50µg 
Matrix-M1:  50µg 
Route(s) 
R21/ Matrix-M1 i.m. to deltoid muscle 
Principal Investigator 
Dr Alfred B. Tiono  
Trial Centre 
Centre National de Recherche et Formation sur le Paludisme (CNRFP) Unité de 
Recherche Clinique, Banfora, Burkina Faso. 
Planned Trial Period 
Q3 2015 – Q2 2016 
Study Duration 
Approximately 9 months  
Participant duration 
Approximately 6 months 
Primary Objective  
 
To assess the safety and reactogenicity of three (3) doses of 10 & 50 µg of the malaria 
vaccine candidate R21 adjuvanted with Matrix-M1, given intramuscularly at 0, 1, 2 
months schedule in healthy West African adult volunteers living in a malaria-endemic 
area. 
Secondary Objective 
 
To assess the immunogenicity of three (3) doses of 10 & 50 µg of the malaria vaccine 
candidate R21 adjuvanted with Matrix-M1, given intramuscularly at 0, 1, 2 months 
schedule in healthy West African adult volunteers living in a malaria-endemic area. 
Population 
Healthy Burkinabe adults aged 18 – 45 years 
Planned Sample size 
A total of 24 volunteers will be enrolled into the groups outlined below: 
 
 
Day 
Group 1 (n=8) 
Group 2 (n=8) 
Group 3 (n = 8) 
0 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
28 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
56 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
 
 
 
 
Vaccination Schedule 
Group 1: Volunteers in group 1 will receive 10µg of R21/ Matrix-M1 at Day 0, 28 and 56 
to the deltoid muscle alternately.  The first 3 volunteers will be vaccinated in 
a staggered fashion.   
Group 2:  Volunteers in group 2 will receive 50µg of R21/ Matrix-M1 at Day 0, 28 and 56 
to the deltoid muscle alternately. The first 3 volunteers will be vaccinated in 
a staggered fashion.   
     
All vaccine administrations will be by the i.m. route 
Follow-up duration 
All volunteers will be followed up to Day 140 post-first vaccination 
Blood Sampling 
Screening, Days 0, 7, 28, 35, 56, 63, 84, 140 
Evaluation Criteria for 
Safety 
Local and systemic solicited and unsolicited adverse events  
 
Evaluation Criteria for 
Immunogenicity 
Measures of immunogenicity of R21/ Matrix-M1 
 
Antibodies to CSP and HbSAg by ELISA 
 
Ex vivo gamma-interferon T cell ELISPOT assays (Day 0 and 84 on frozen cells) 
 
ICS and flow cytometry (Day 0 and 84 on stimulated frozen whole blood)  

11 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Exploratory immunology, including RNA analysis and DNA analysis to include parasite 
and human genetic analysis. 
Study design 
Randomised, controlled, single-blind clinical trial design 
Statistical Analysis 
Observational and Descriptive 
 
2. BACKGROUND INFORMATION 
With the estimated 219 million cases of malaria worldwide in 2010 coupled with approximately 660,000 deaths 
occurring in predominantly children under the age of five years1 in Africa (where 90% of deaths are recorded), malaria 
is still the preeminent tropical infectious disease globally, with a devastating effect on human health and society. The 
enormous economic and social consequences of malaria have been well documented2. Malaria is still a potentially 
fatal hazard for travellers visiting malaria-endemic regions.  The development of an effective vaccine against malaria is 
of high priority in the context of coordinated efforts to reduce the burden of malaria and is considered necessary for 
the global eradication of malaria3. 
 
The Roll Back Malaria (RBM) Partnership (www.rollbackmalaria.org) launched in 1998 by the World Health 
Organization (WHO) has as its major goal to support the development of a vaccine against malaria as a key future 
strategy for reducing mortality from malaria. The essence of this strategy is highlighted by the limitations of other 
measures aimed at reducing the burden of malaria include the development of resistance of Anopheles mosquitoes to 
certain insecticides; the development of resistance of malaria parasites to chemotherapeutic agents4; the absence of a 
gametocidal drug suitable for mass administration5, and the risk of re-importation of malaria into geographic regions 
despite environmental elimination measures.  
 
2.1. 
Lifecycle of the malaria parasite 
The malaria lifecycle is complex with stages in both human and mosquito hosts (Figure 1).  The bite of infected female 
Anopheles mosquitoes transmits malaria sporozoites to the human host where they travel via the bloodstream to the 
liver and invade hepatocytes.  Here, during the liver stage, they mature into merozoites for 6 to 7 days. Malaria 
parasites are not detectable in the blood stream during the liver stage. The hepatocytes then rupture, releasing a large 
number of merozoites into the bloodstream signalling the onset of the blood stage.    
 
Merozoites invade erythrocytes where they multiply and after 2 days cause the erythrocyte to rupture, releasing 
progeny merozoites that in turn invade new erythrocytes. A small percentage of merozoites differentiate into 
gametocytes, which when ingested by a mosquito, unite with another gametocyte to create a zygote. The zygote 
matures and releases sporozoites which migrate to the mosquito’s salivary glands and are injected into the human 
when the mosquito feeds. Patency refers to the ability to detect parasites on examination of the peripheral blood 
during the blood stage. 
 

12 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Figure 1 Lifecycle of Malaria 
 
2.2. 
Progress towards a malaria vaccine 
The most advanced candidate malaria vaccine, RTS,S, targets antibodies against the circumsporozoite protein (CS), 
which is expressed by the sporozoite at the pre-erythrocytic stage.  RTS,S formulated with the AS01E adjuvant has now 
shown partial efficacy in Phase II6,7 and in analyses of a large Phase III trial8
.  The co-primary end point of the trial has 
recently been published6, 7. Vaccine efficacy (VE) against clinical malaria after 12 months of follow-up post dose 3 in 
the first 6000 children enrolled in the 5 to 17 month group was 55.8% (97.5% CI: 50.6, 60.4; p<0.001) and against 
severe malaria 47.3% (95% CI: 22.4, 64.2; p<0.001). In the younger age category, VE against clinical malaria after 12 
months of follow-up post dose 3 was 31.3% (p<0.0001) and VE against severe malaria was 36.6% (p=0.02).   
 
The only other vaccination approach that has demonstrated partial efficacy in humans involves using heterologous 
vectored vaccines in prime-boost sequence to induce a T cell response against the pre-erythrocytic antigen, TRAP9, 10, 
11.  T cell responses provide protection against malaria in animal models12,   in the field13-15, following irradiated 
sporozoite inoculation16-18 and following vaccination.  Immunisation of mice with irradiated sporozoites of murine 
Plasmodium provides protection against later challenge with murine malaria sporozoites12. This protective immunity 
can be transferred to non-immune mice by transferring the CD8+ T lymphocyte clones specific to pre-erythrocytic 
malaria surface antigens, the circumsporozoite protein (CS), or thrombospondin related adhesion protein (TRAP) that 
were induced by irradiated sporozoites16,17.  In the heterologous prime-boost strategy, two different vectored 
vaccines, both containing ME-TRAP, are given in sequence. This achieves an expansion of T cells reactive to TRAP, and 
to the ChAd63 vector.   Heterologous prime-boost vaccination with DNA containing ME-TRAP (DNA ME-TRAP) 
followed by recombinant MVA containing ME-TRAP (MVA ME-TRAP) led to significant reductions of an estimated 80% 
in parasite burden in the liver on human challenge with malaria infection19-21. In one study21, one of eight vaccinees 
was sterilely protected and in another19, there was a significant delay in the time to patency.  Several clinical trials 
using this heterologous prime-boost approach and several vectors (FP9, ChAd63) have been consistently carried out in 
Oxford.  Vaccination with FP9 ME-TRAP followed by MVA ME-TRAP led to an estimated 90% reduction in parasite 
burden in the liver, with two of sixteen vaccinees protected against malaria challenge20, 22. Following these promising 
findings, studies were undertaken in adults and then children in The Gambia and Kilifi23-25. Immunogenicity was lower 
than expected25, and efficacy was not seen in a study of 400 children in Kilifi district26.  From these studies it has 
emerged that T cell responses are a correlate of protection induced by these vaccination strategies, as measured by 
delay in time to patency or reduction in parasite burden in the liver on malaria challenge.20,27 Further development of 
T cell inducing vaccination in Oxford examining more immunogenic vectors in order to attain greater efficacy has led 
to preclinical and phase I and II clinical development of malaria vaccination strategies using adenoviral and MVA 
vectors in heterologous prime-boost vaccination strategies with TRAP inserts.  These clinical trials in UK and Africa 
have shown that these vaccines have been safe, tolerable and have induced adequate immune responses28, 29 that 
have been shown to be protective9.  An efficacy clinical trial in 700 children aged 5-17 months old is on-going in 
Burkina Faso and results are expected at the end of Q3 2015. 

13 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
The potential to combine these two vaccine approaches is evident.  The rest of this section provides background on 
the vaccination approach relevant to this protocol. Section 3 highlights and summarises the clinical trials relevant to 
this protocol. 
 
2.3. 
PRE-ERYTHROCYTIC STAGE OF INFECTION AS A VACCINE TARGET 
The pre-erythrocytic stage of P. falciparum infection presents an attractive target for an efficacious human vaccine, as 
sufficient reduction in the number of viable merozoites reaching the blood from the liver will prevent parasitisation of 
red blood cells and initiation of the blood stage of infection.  Anti-CS antibodies can target sporozoites, facilitating 
destruction of sporozoites prior to hepatocyte invasion. As sporozoites travel from the skin to liver within minutes, it 
may be difficult for a vaccine to achieve complete protection against P. falciparum based solely on antibodies to 
sporozoites. The liver stage of infection provides a longer window of opportunity for cell mediated immunity to 
recognize and destroy infected hepatocytes. Research suggests that, in isolation the RTS,S vaccine targeting the pre-
erythrocytic stage antigen, CS, and vaccines targeting ME-TRAP do not delay the initial emergence of parasites in to 
the blood, nor the rate of parasite multiplication in the blood, but rather reduce the size of this initial inoculum20. A 
delay to patent blood stage infection in persons receiving these vaccines reflects a reduced liver-to-blood inoculum. 
The efficacy of these pre-erythrocytic vaccine strategies can be assessed experimentally by subjecting volunteers to 
inoculation with P. falciparum sporozoites by the bite of infected mosquitoes. Complete protection against blood-
stage infection, or a delay in the time to patent blood stage infection in vaccinees compared to controls, are 
interpreted to understand vaccine-induced protection. 
 
2.3.1. R21 vaccine development 
RTS,S/AS01 vaccine, induces very strong antibody responses to the conserved central repeat of CSP, of the order of 
100 - 600 micrograms per ml, very weak mainly IL-2 containing CD4+ T cells and no CD8+ T cells to CSP.7  The most 
reproducible correlate of protection in clinical studies is with antibody levels7,8.  We propose here to test clinically a 
biosimilar of the RTS,S vaccine called R21 adjuvanted with Matrix M-1.  As a biosimilar of the RTS,S vaccine, the R21 
particle contains no sequences that are not present in RTS,S, which has been safely used in thousands of individuals.  
It is a hybrid protein of the majority of the CS protein of P. falciparum fused to the hepatitis B surface antigen (Figure 
2).  It spontaneously forms a particle just like RTS,S. However, we anticipate that R21 may be a more immunogenic 
particle than RTS,S in humans (although this has yet to be tested) for two reasons.  It induces predominantly malaria 
rather than hepatitis antibodies in pre-clinical studies probably because it has a higher proportion of malaria to 
hepatitis antigen in its composition than RTS,S.  This is made possible by expressing R21 in the better expressing yeast 
Pichia pastoris, rather than in Saccharomyces cerevisiae. Secondly, in pre-clinical studies R21 has been found to be 
exceptionally immunogenic for induction of anti-NANP antibodies, likely the key protective immune mechanism of 
RTS,S, yielding titres of a mean of 800,000 ELISA units after two immunisations which exceeds historical 
immunogenicity data obtained with the RTS,S vaccine of 150,000 ELISA units, at the UOXF laboratory.  (K Collins and A 
Hill, unpublished data).   
 
 
 
 
Figure 2:  Schematic diagram showing RTS,S and R21 fusion proteins 

14 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
2.3.2. Pre-clinical studies of R21  
Immunogenicity:  Initial pre-clinical assessment of immunogenicity was undertaken in BALB/c mice that were 
immunised intramuscularly with 0.5mg of R21 alone or in combination with an adjuvant (Alhydrogel, Abisco). Immune 
responses including antibody levels to the central NANP repeat region and antigen–specific T cell responses were 
measured three weeks after a 3-dose immunisation schedule (Figure 3).  R21 + Abisco-100, a potent saponin-based 
adjuvant used in animal studies resulted in the greatest humoral immune response at each time point in the 
vaccination schedule.  
 
 
 
Figure 3:  Pre-clinical assessment of immunogenicity with 0.5mg of R21 alone or in combination with an adjuvant 
(Alhydrogel, Abisco). 
 
The responses in all groups were boosted by a third immunisation and R21 + Abisco-100 induced the highest titres of 
NANP specific IgG and the response for this group was significantly higher than both the R21 + Alhydrogel and R21 
alone groups after the final immunisation (Figure 3.1). 
 
 
 
Figure 3.1:  Relative proportions of IgG to NANP and HBsAg after immunisation with R21 + Abisco-100 in BALB/c mice.  
 
CS-specific IFN-γ producing T cells measured after the third immunisation were only detected at a significant level in 
mice immunised with R21 + Abisco-100 (Figure 3.2). R21 alone was ineffective at inducing CS-specific T cell responses 
on its own. Further comparison to other adjuvants including a squalene-based oil-in-water emulsion (Addavax) and a 
polyionic carbomer (Carbopol) showed that Abisco-100 was the ideal adjuvant to induce high levels of humoral and 
cell-mediated immunity.  
 
NANP IgG ELISA
A - 1st immunisation
R21 alone
R21 + Alhydrogel 
R21 + Abisco
0
2
4
6
8
***
Endpoint titre (log 10)
B - 2nd immunisation
R21 alone
R21 + Alhydrogel 
R21 + Abisco
0
2
4
6
8
**
C - 3rd immunisation
R21 alone
R21 + Alhydrogel 
R21 + Abisco
0
2
4
6
8
*
*
1
2
3
4
5
6
7
1st immunisation
2nd immunisation
3rd immunisation
NANP IgG
HBsAg IgG
Endpoint titre (log 10)

15 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Figure 3.2:  CS-specific IFN-γ producing T cells measured after the third immunisation 
 
Efficacy:  Sporozoite challenge (1000 sporozoites per mouse injected intravenously) using transgenic P. berghei 
parasite were performed in BALB/c mice (Figure 3.3). R21 + adjuvant were given twice, eight weeks apart and mice 
were challenged three weeks after the second dose. Thin blood films looking for parasitaemia were performed daily 
from day 5 post-challenge. Sterile protection was defined as remaining slide negative at day 14 and significant delay in 
development of 1% parasitaemia compared to non-immunised control mice was regarded as partial efficacy.  
R21 + Abisco-100 steriley protected 100% of the challenged mice (p=< 0.0001) and R21 + Matrix M steriley protected 
87.5% (p=0.0002) and this was confirmed in a second independent challenge. (p = < 0.0001). There was no significant 
difference between the two adjuvants.  
 
Figure 3.3 (A-F): Protective efficacy elicited by saponin based ISCOM adjuvants with R21 in a transgenic sporozoite 
model. 
BALB/c mice were immunised i.m. with 0.5µg R21 + adjuvant (Abisco-100 or Matrix M), twice eight weeks apart 
(n=8/group). Mice were challenged three weeks after the final vaccination by i.v. injection of 1000 sporozoites (P. 
berghei transgenic for P. falciparum CSP) along with eight naïve mice. Two groups of adjuvant control mice 
(n=5/group) were also challenged three weeks after receiving two shots of adjuvant (Abisco-100 or Matrix M) i.m., 
eight weeks apart. Blood stage parasitemia was monitored from day 5 after challenge by thin-film blood smear, and 
time to 1% parasitemia was calculated using linear regression. The results are presented in the Kaplan-Meier survival 
graphs and survival curves were compared by Log-rank (Mantel-Cox) Test. (A) Adjuvant control = no significant 
difference, (B) R21 + Abisco-100 p<0.0001, (C) R21 + Matrix M p=0.0002, (D) R21 + Matrix M repeat p=<0.0001, (E) R21 
+ Abisco vs R21 + Matrix M = no significant difference. Blood was taken three weeks after each vaccination (Day 21 
and Day 77) for immunology and NANP specific IgG was assayed by ELISA (F), group mean responses shown and dotted 
line indicates the limit of detection. 
 
p
R21 alone
R21 + Alhydrogel 
R21 + Abisco
0
200
400
600
800
1000
SFC/106 Splenocytes

16 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
 
The durability of efficacy was assessed by undertaking sporozoite challenge in mice seven and fourteen weeks after 
immunisation. Efficacy was maintained very well at seven weeks post immunisation with 75% of mice sterilely 
protected (6/8) and this was not significantly different when compared to efficacy at three weeks post immunisation 
(p=0.4468, by Log-rank (Mantel-Cox) Test). At 14 weeks post immunisation however, sterile efficacy was reduced to 
50% (2/4) and this was 37% lower than the efficacy at three weeks. This was not significantly lower due to the small 
number in the group (p=0.0636), had there been eight mice in the group 50% sterile efficacy would have been 
significantly lower than 87.5%.  Sterile efficacy 14 weeks after immunisation is 37% lower than efficacy three weeks 
after immunisation. This reduction in protective efficacy can however be boosted to 100% if mice are challenged once 
(three weeks post immunisation) within the 14 weeks. Therefore efficacy after vaccination and one sporozoite 
infection is very durable and 100% sterile efficacy is maintained for at least 14 weeks. 
A Adjuvant only
4
5
6
7
8
9 10 11 12 13 14 15
0
20
40
60
80
100
Adjuvant only
(Abisco/Matrix M)
Naive
Time to 1% parasitaemia
Percent survival
B R21+Abisco-100
4
5
6
7
8
9 10 11 12 13 14 15
0
20
40
60
80
100
R21 + Abisco
Controls
(Abisco + naives)
Time to 1% parasitaemia
Percent survival
C R21+Matrix M
4
5
6
7
8
9 10 11 12 13 14 15
0
20
40
60
80
100
R21 + Matrix M
Controls
(Matrix M + naive)
Time to 1% parasitaemia
Percent survival
D Repeat challenge
4
5
6
7
8
9 10 11 12 13 14 15
0
20
40
60
80
100
R21 + Matrix M
Naive
Time to 1% parasitaemia
Percent survival
E R21+Abisco vs R21+Matrix M
4
5
6
7
8
9 10 11 12 13 14 15
0
20
40
60
80
100
R21+Abisco
R21+Matrix M
Time to 1% parasitaemia
Percent survival
F NANP IgG ELISA
0
1
2
3
4
5
6
7
8
R21 +Abisco
R21 + Matrix M
Day 21
Day 77
Endpoint titre (log 10)

17 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Manufacture of the clinical grade R21 particle was performed at the University of Oxford CBF (www.cbf.ox.ac.uk), with 
financial support from the UK Medical Research Council (MRC) and the EC FP7 programme. A Phase 1 CT is due to 
commence at CCVTM, University of Oxford in Q3 2015. 
 
2.3.3. Matrix-M1 adjuvant 
Matrix M™ is a saponin-based product produced using the matrix adjuvant technology patented by Isconova (now 
owned by Novovax).   Matrix-M1 is a novel adjuvant designed to stimulate both humoral and cellular immune 
responses to vaccines. Matrix-M1 is formed from selected purified saponin fractions formulated separately into 
different Matrix particles, Matrix A and Matrix C. Matrix-M1 is constituted from a mixture of Matrix A and Matrix C in 
a ratio of 85:15. 
 
Matrix M™ has previously been used to adjuvant an intranasal DNA vaccine, leading to improvements in local 
antibody responses, and increased expression of Th1 and Th2 cytokines. Matrix M™ significantly enhanced antibody 
responses to a commercial trivalent seasonal influenza vaccine. Mixed with a virosomal H9N2 avian influenza vaccine, 
Matrix-M1 induced enhanced antigen-specific humoral and CD8+ T cell response. Matrix M™ administered with an 
intramuscular H5N1 virosomal influenza vaccine induced a strong immediate and long-term humoral and cellular 
immune response and showed a dose-sparing potential. 
 
ChAd63/Matrix-M1 and MVA ME-TRAP/Matrix-M1 adjuvanted mixed vaccine combinations have been shown to be 
safe and immunogenic in the VAC048 clinical trial currently on-going in Oxford, UK.  Adverse events such as pain and 
swelling related to inflammation at the site of vaccination were observed. Systemic symptoms were also observed but 
these were generally flu-like symptoms. Adverse events were generally mild and resolved completely within 24-48hrs. 
To date, no significant safety concerns relating to the administration of the mixture of ChAd63/Matrix M-1 and MVA 
ME-TRAP/Matrix M-1 vaccine have emerged from clinical or preclinical studies. 
 
In this protocol, we plan to conduct a phase I / IIb randomised, controlled, single-blind study to assess the safety, 
immunogenicity of the malaria vaccine candidate R21 with Matrix-M1 as adjuvant, in Burkinabe adult volunteers aged 
18-45 years. 
 
3. RATIONALE 
 
In summary, the vaccination regime described here targets the pre-erythrocytic stage of the malaria parasite life cycle 
and induce specific immune responses through distinct mechanisms. The RTS,S/AS01B vaccination regime has 
demonstrated pre-erythrocytic stage immunity that has translated into sterile efficacy against experimental human P. 
falciparum infection. R21 is a bio similar of the RTS,S vaccine.  A Phase 1/IIa clinical trial of the R21/ Matrix-M1 
vaccination regime is scheduled to start in Q3 2015 at CCVTM, University of Oxford and it is expected that this trial will 
produce equally good results as obtained with the RTS,S/AS01B vaccination regime.  This will be the first time this 
vaccine will be administered to humans.  Interim results of this clinical trial will be available in late Q4 2015. 
 
Here we propose to evaluate, for the first time in an African population, the R21/ Matrix-M1 vaccination regime in a 
phase 1b safety and immunogenicity evaluation clinical trial in West African adults living in Burkina Faso. 
 
4. OBJECTIVES 
4.1. 
Primary Objective 
 
To assess the safety and reactogenicity of three (3) doses of 10 & 50 µg of the malaria vaccine candidate R21 
adjuvanted with Matrix-M1, given intramuscularly at 0, 1, 2 months schedule in healthy West African adult volunteers 
living in a malaria-endemic area. 
 
4.2. 
Secondary Objective 

18 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
To assess the immunogenicity of three (3) doses of 10 & 50 µg of the malaria vaccine candidate R21 adjuvanted with 
Matrix-M1, given intramuscularly at 0, 1, 2 months schedule in healthy West African adult volunteers living in a 
malaria-endemic area. 
 
5. DESCRIPTION AND JUSTIFICATION OF STUDY DESIGN 
5.1. 
Overview 
A randomised, controlled, single-blind clinical trial is proposed to evaluate the safety and immunogenicity of the 
malaria vaccine candidate regime of three (3) doses of R21/ Matrix-M1 compared with placebo, in healthy West 
African adult volunteers living in a malaria-endemic area.  
 
5.2. 
Vaccines 
R21: Recombinant HBsAg protein particle expressing the central repeat and the C-terminus of the circumsporozoite 
protein (CSP).   
Matrix-M1: A saponin adjuvant.  It will be administered with the R21 vaccine. 
 
5.3. 
Evaluation Criteria  
 
5.3.1. Safety endpoints 
Local and systemic solicited and unsolicited adverse events, considered possibly, probably, or definitely related to 
vaccination, occurring for 28 days after each vaccination.  Solicited adverse events will be measured up to 7 days after 
each vaccination.  Unsolicited adverse events will be measured up to 28 days after each vaccination. 
Solicited local (injection site) adverse events to be observed and recorded include pain/limitation of limb movement, 
swelling and redness/discoloration. Solicited systemic adverse events include fever, chills, headache, malaise, joint 
pains, myalgia, fatigue and nausea. 
 
SAEs occurring from first vaccination until the end of the study will be recorded 
 
 
5.3.2. Immunogenicity endpoints 
Measures of immunogenicity of R21/ Matrix-M1 will include: 
 
Antibodies to CSP and HbSAg by ELISA 
 
Ex vivo gamma-interferon T cell ELISPOT assays (Day 0 and 70 on frozen cells) 
 
ICS and flow cytometry (Day 0 and 70 on stimulated frozen whole blood)  
 
 Exploratory immunology, including RNA analysis and DNA analysis to include parasite and human genetic 
analysis. 
 
5.4. 
Study site 
The study will take place at the Banfora clinical trial site, which is located about 400 km from Ouagadougou, the 
capital city of Burkina Faso. The Unite de Recherche Clinique de Banfora (URC-B) research unit is situated within the 
complex of the Regional hospital. The trial participants will be drawn from Banfora Health demographic system that 
covers a total population of 30,000 inhabitants. From recent surveys, the bed net coverage was 80%. There is no 
implementation of Indoor Residual Spray (IRS) or Intermittent Preventive Treatment (IPT) in infants or children in the 
area. Malaria transmission is perennial with peaks during the high transmission season (June to November). The main 
malaria vectors are Anopheles gambiae and Anopheles funestus. The cumulative annual entomological rate varies 
from 55 to 400 infective bites/person/year (Moussa G et al, unpublished). P. falciparum is the main parasite present in 
more than 90% of infections. The incidence rate of uncomplicated malaria (fever and parasitaemia 5000/μL or more) 
in less than five years is 1.18 episodes/child-year at risk and around 60% of the total annual number of malaria 
episodes occurs during the high malaria transmission season41 (). The asymptomatic carriage of P. falciparum is very 
common. In 2010, during the high transmission period, the prevalence was 46.6%, 72.5% and 38.5% in 0-4 years, 5-14 
year and 15-24 years respectively. During the low transmission season, the prevalence in the same age range was 
27.2%, 60.4% and 25.5% respectively. (Tiono et al, unpublished).  Overall to date there is no evidence of the decline in 
malaria incidence in Burkina similar to what has been recently reported from other parts of sub-Saharan Africa. The 
annual malaria death toll is reaching 15,000 people and across the country’s hospitals, malaria is reportedly 
responsible for 30.7% of all hospitalization with a mortality rate of 23%.  
 

19 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
5.5. 
Study groups 
A total of 24 volunteers in 3 equal groups as outlined below: 
 
Day 0 
Group 1 (n=8) 
Group 2 (n=8) 
Group 3 (n=8) 
0 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
28 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
56 
10µg R21/ Matrix-M1 
50µg R21/ Matrix-M1 
saline 
 
 
5.6. Vaccination Schedule 
The vaccination groups are outlined below: 
 
Group 1: Volunteers in group 1 will receive 10µg of R21/ Matrix-M1 at Day 0, 28 and 56 to the deltoid muscle 
alternately.  The first 3 volunteers will be vaccinated in a staggered fashion.   
Group 2:  Volunteers in group 2 will receive 50µg of R21/ Matrix-M1 at Day 0, 28 and 56 to the deltoid muscle 
alternately. The first 3 volunteers will be vaccinated in a staggered fashion.  
 
All vaccine administrations will be by the i.m. route. 
Before vaccinations commence in this clinical trial, a DSMB review of the interim safety report for the low and high 
dose of R21 given to volunteers in the Phase I CT in Oxford will be assessed.  A DSMB review of the interim safety 
report after vaccination of the first 3 volunteers in Group 1 will be required before vaccinations commence in Group 2.    
 
The control group will undergo similar tests to the vaccinated groups including ELISA, ex-vivo and cultured ELISPOT 
and ICS  
 
5.7. 
Follow-up of volunteers 
 
5.7.1. Safety follow-up 
All volunteers will be followed up to Day 140 post-first vaccination for adverse events following vaccination. SAE’s will 
be collected throughout the duration of the study for each volunteer.   
 
Study clinicians and field workers (supervised by medical officers) will assess and record local adverse events post 
vaccination including pain, swelling and limited arm movement. In addition, systemic Adverse Events (AE’s) including 
fever, chills, headache, malaise joint pains, myalgia, fatigue and nausea will be recorded.  Each side effect will be 
classified as absent, mild, moderate or severe.  Information on AE’s will also be collected at each clinic visit by the 
study clinician. Each volunteer will be seen at home by field workers on Days 1-6 following each vaccination. Day 7 will 
be a clinic visit. The study clinicians will review reports from home visits conducted by field workers, and arrange to 
see the volunteers in person when required.  The volunteers will be asked to present themselves to the study clinic 
should they develop any illnesses during follow up.  Any emerging safety data that is considered cause for concern by 
the DSMB will be relayed to the participants during the study visits. 
 
5.8. 
Sample size justification 
 
This is an observational and descriptive safety study, where volunteers 8 volunteers each will be vaccinated with 
either 10µg of R21/ Matrix-M1 or 50µg of R21/ Matrix-M1 at Day 0, 28 and 56.  A further 8 volunteers will receive 
normal saline (placebo) as control group.  The sample size for this clinical trial balances the need to avoid exposing a 
large group to an unknown risk with the need for data from an adequate sample.  This sample size should allow 
determination of the magnitude of the outcome measures, especially of severe and serious adverse events, rather 
than aiming to obtain statistical significance for differences between groups. 
 
Specific immunogenicity to CSP and HbSAg will be assessed by a variety of immunological assays. The key 
immunogenicity assessment in this trial is whether administration of 3 doses of either 10µg of R21/ Matrix-M1 or 
50µg of R21/ Matrix-M1 induces adequate vaccine immunogenicity at the peak of the response, in comparison to a 
placebo. 
 
5.9. 
Potential risks for volunteers 

20 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
The risks of study participation are those relating to vaccination and blood sampling. Volunteers will receive three 
vaccinations with a 3-dose regimen or a placebo four weeks apart.  Vaccinations are expected to be generally well 
tolerated.  Local reactions are expected at the injection site such as pain, swelling or erythema, and less commonly 
there may be minor fever or malaise as a systemic reaction. These reactions should generally be mild and resolve 
completely after two (2) days.  
 
5.9.1. Phlebotomy 
The maximum volume of blood drawn over the study period approximately 190 mls should not compromise these 
otherwise healthy volunteers. There may be minor bruising, local tenderness or pre-syncopal symptoms associated 
with venepuncture, which will not be documented as AEs if they occur. 
 
5.9.2. Safety and tolerability of R21/Matrix- M1 vaccination 
The most frequent adverse reactions observed in previous clinical trials using the RTS,S antigen and AS01B adjuvant 
system include pain, swelling, erythema, and tenderness at the site of injection, and systemic symptoms such as low-
grade fever and short-term flu-like symptoms: fatigue, myalgia, headache, malaise. It is expected that R21 antigen (a 
biosimilar of RTS,S) with Matrix-M1 as adjuvant will produce similar or better AE profile.   
 
As with any vaccine, unexpected serious adverse events, including severe allergic reactions to the vaccine components 
may occur.  Some individuals vaccinated with adjuvant components identical to those that will be used in this study 
have reported autoimmune diseases (AID). A causal association between the adjuvant components and occurrence of 
autoimmune diseases has however not been established, as these can occur in people who get other vaccines, or no 
vaccines at all. A meta-analysis of a GSK adjuvanted vaccine showed no increased risk of AID associated to the 
adjuvant.35 
 
5.10. 
Potential benefits for volunteers  
Participants will not directly benefit from participating in this clinical trial except for adequate information on their 
health status and free medical care for the volunteers during the course of the clinical trial. 
 
In terms of protection against malaria, a biosimilar of R21 (RTS,S adjuvanted in AS01B) is the leading malaria vaccine 
candidate and has shown consistent partial efficacy in field clinical trials in Africa. We expect similar or better results 
with the R21/Matrix M-1 vaccine regime.  However, because malaria mainly affects young children, the benefits of 
participating in this study may be considered as being altruistic in nature as they would potentially benefit the wider 
society at large, if the vaccine is eventually proved to be safe and efficacious.  Volunteers will be advised that 
participating in the study does not reduce the need for continuing to use known preventive measures against malaria. 
 
6. INCLUSION AND EXCLUSION CRITERIA 
The inclusion criteria will be used at screening to identify volunteers eligible for the study, and will be checked prior to 
vaccination to confirm ongoing eligibility. Eligible volunteers will fulfil all of the inclusion criteria and none of the 
exclusion criteria.  We will continue to recruit and screen volunteers until at least 24 eligible volunteers have been 
identified, preferably until 24 eligible volunteers have been identified. If volunteers withdraw consent prior to 
receiving their first vaccination, we will replace the volunteer or screen and recruit in order to replace the number 
withdrawing consent. 
 
6.1. 
Inclusion Criteria 
The volunteer must satisfy all the following criteria to be eligible for the study: 
 
Healthy adults aged 18 to 45 years.  
 
Willingness to remain in study area for the period of the study. 
 
Able and willing (in the Investigator’s opinion) to comply with all study requirements. 
 
Women only: Must practice and show documented evidence of continuous effective contraception (e.g depo-
progesterone) or must be willing to take contraceptive measures not to become pregnant for the duration of 
the study.  Willing to have pregnancy tests at screening and vaccination time points. 
 
Agreement to refrain from blood donation during the course of the study 
 
Written informed consent to participate in the trial. 

21 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
6.2. 
Exclusion Criteria 
The volunteer may not enter the study if any of the following apply: 
 
Hb less than 10.0g/dl 
 
Receipt of an investigational product in the 30 days preceding enrolment, or planned receipt during the study 
period. 
 
Prior receipt of an investigational malaria vaccine or any other investigational vaccine likely to impact on 
interpretation of the trial data.  
 
Any confirmed or suspected immunosuppressive or immunodeficient state, including HIV infection; asplenia; 
recurrent severe infections and chronic (more than 14 days) immunosuppressant or other immune-modifying 
drugs medication (for corticosteroids, this will mean prednisone, or equivalent, ≥ 0.5 mg/kg/day) within the 
past 6 months (inhaled and topical steroids are allowed). 
 
Use of immunoglobulins or blood products within 3 months prior to enrolment.  
 
History of allergic disease or hypersensitivity reactions likely to be exacerbated by any component of the 
study vaccines. 
 
Any history of anaphylaxis post-vaccination. 
 
History of clinically significant contact dermatitis. 
 
Pregnancy, lactation or intention to become pregnant during the study. 
 
Disturbances of electrolyte balance, e.g., hypokalaemia or hypomagnesaemia 
 
History of cancer (except basal cell carcinoma of the skin and cervical carcinoma in situ). 
 
History of serious psychiatric condition that may affect participation in the study. 
 
History of splenectomy 
 
Any other serious chronic illness requiring hospital specialist supervision. 
 
HIV or Hepatitis B surface antigen seropositivity 
 
Volunteers unable to be closely followed for social, geographic or psychological reasons. 
 
Any clinically significant abnormal finding on biochemistry or haematology blood tests, urinalysis or clinical 
examination. In the event of abnormal test results, confirmatory repeat tests will be requested.  
 
Any other significant disease, disorder, or finding which may significantly increase the risk to the volunteer 
because of participation in the study, affect the ability of the volunteer to participate in the study or impair 
interpretation of the study data. 
 
6.3. 
Withdrawal criteria 
Participants may be withdrawn from the study:  
•By withdrawing consent  
•On the decision of the investigator  
•On the advice of the Data Safety Monitoring Board (DSMB)  
 
The investigator may withdraw the subject for the following reasons:  
•Any adverse event which results in the inability to comply with study procedures or affect the participant safety. 
•Ineligibility either arising during the study or retrospectively (having been overlooked during screening)  
•Significant protocol deviation. 
7. INVESTIGATIONAL MEDICINAL PRODUCTS 
 
R21 adjuvanted with Matrix-M1 is the only study vaccine for this clinical trial. 
 
7.1. 
Manufacturing and presentation of vaccines 
 
7.1.1. R21 vaccine 
R21 vaccine is manufactured under Good Manufacturing Practice (GMP) conditions by the Clinical Biomanufacturing 
Facility (CBF), University of Oxford.  Final QP certification and associated labelling will take place at the CBF, University 
of Oxford. R21 will be supplied in a sterile pharmaceutically suitable container with appropriate closure. The product 
will be stored frozen (-80°C)  
 

22 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Supply of R21 
R21 will be supplied to the clinical sites by the CBF, University of Oxford, where the vaccines were manufactured, 
vialed and labelled for trial VAC060. 
 
Supply of Matrix-M1 
Matrix-M1 was manufactured in compliance with cGMP by Apoteket Produktion & Laboratorier AB (APL) Formvägen 
5B, SE-903 03 Umeå, Sweden. Matrix- M1 is supplied as a sterile 1mg/ml solution in 2ml glass vials. 
 
7.2. 
Storage, dispensing and handling of Investigational Medicinal Products 
All study vaccines to be administered to the volunteers must be stored in a safe and locked place with no access by 
unauthorised personnel. Vaccine accountability, storage, shipment and handling will be in accordance with local SOPs.  
 
7.2.1. Storage of R21/Matrix M-1 
The R21 vaccine must be stored at the defined temperature of (-80°C). The storage conditions will be under the 
responsibility of the sponsor and then the clinical site after shipment has occurred. Any temperature deviation outside 
the defined range must be reported to the sponsor as soon as detected. Matrix–M1 will be stored between + 2 and +8 
°C. 
 
7.3. 
Vaccination of volunteers 
Each volunteer will be monitored for one hour (or longer if necessary) after each vaccination. Resuscitation (including 
intubation) equipment and medication will be available in the clinic site and a clinician trained in resuscitation will be 
present at all times during this vaccination time period.  Advanced Life Support drugs and resuscitation equipment will 
be immediately available for the management of anaphylaxis. 
 
All vaccinations will be by the intramuscular route. The study vaccines should under no circumstances be administered 
intravascularly or intradermally. For all intramuscular injections, the needle should be long enough to reach the 
muscle mass and prevent the vaccine from seeping into subcutaneous tissue, but not so long as to involve underlying 
nerves, blood vessels or bone. Vaccinators should be familiar with the anatomy of the area into which they are 
injecting vaccine. Firm pressure should be applied to the injection site (without rubbing) for at least two minutes. 
 
7.3.1. R21/Matrix-M1 vaccinations 
The R21/Matrix-M1 vaccine mixture containing either 10µg or 50µg of R21 with 50µg Matrix-M1 will be administered 
intramuscularly into the deltoid muscle starting with the left arm then alternately for the remaining two doses. 
Matrix-M1 and R21 will be mixed at the bedside immediately prior to administration according to the specific mixing 
SOP. 
 
All vaccines will be kept at room temperature between removal from the refrigerator or freezer and administration. 
If for any unforeseen reason the preferred administration site could not be used at the time of the vaccination, an 
alternative administration site will be used. The alternative site will be documented in vaccine administration records 
as well as the reasons why alternative site was used. 
 
7.4. 
Concomitant medication/vaccination 
 
At each study visit, the investigator should question the enrolled volunteer about any medication(s) taken. 
All antipyretic, analgesic and antibiotic drugs, administered at any time during the period starting with administration 
of each dose and ending 28 days after each dose of vaccine are to be recorded with generic name of the medication 
(trade names are allowed for combination drugs), medical indication, total daily dose, route of administration, start 
and end dates of treatment. 
 
Any immunoglobulin, blood products and any immune modifying drugs administered at any time during the study 
period are to be recorded with generic name of the medication (trade names are allowed for combination drugs only), 
medical indication, total daily dose, and route of administration, start and end dates of treatment.  
 
Any vaccine not foreseen in the study protocol administered during the volunteer participation in the trial is to be 
recorded with the trade name, route of administration and date(s) of administration.  

23 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Any concomitant medication administered  as prophylaxis in anticipation of reaction to the vaccination must be 
recorded in the CRF with generic name of the medication (trade names are allowed for combination drugs only), total 
daily dose, route of administration, start and end dates of treatment and coded as ‘Prophylactic’ 
 
7.5. 
Indications for deferral of vaccine administration 
The following constitute conditions for deferring administration of vaccines at vaccination time points. If any one of 
these occurs at the time scheduled for vaccination, the volunteer may be vaccinated at a later date or withdrawn, at 
the discretion of the Principal Investigator. Medical care including inpatient care if necessary will be offered.  
 
 
Acute disease at the time of vaccination. Acute disease is defined as the presence of a moderate or severe 
illness with or without fever. All vaccines can be administered to persons with a minor illness at the discretion 
of the investigators. Details of any minor illness will be recorded in the CRF. 
 
Axillary temperature of 37.5°C (99.5°F) at the time of vaccination. 
 
Receipt of any other vaccinations within one week of study vaccine 
 
7.6. 
Absolute contraindications to further vaccination 
The following AEs constitute absolute contraindications to further administration of any vaccine used in the study. If 
any of these AEs occur during the study, the subject must not receive additional doses of vaccine, but may continue 
other study procedures at the discretion of the investigator. The AE should be followed-up as per protocol 
requirement. 
 
Acute allergic reaction (significant IgE-mediated events) or anaphylaxis following the administration of 
the investigational product. 
 
Any confirmed or suspected immunosuppressive or immunodeficient condition, including human 
immunodeficiency virus (HIV) infection. 
 
8. STUDY SCHEDULE AND PROCEDURES 
Prior to the commencement of community sensitisation, the CNRFP study team will be trained on the procedures for 
community sensitisation, with the clinical trial protocol, volunteer information sheets and consent forms as training 
guides.  
 
8.1. 
Identification of Study Participants 
Community sensitisation will be undertaken to engage the community with the study and recruit volunteers for 
participation in the study. Volunteers will be assessed at screening visits to determine if they are eligible to participate 
in the study. 
8.1.1. Community sensitisation 
The CNRFP study team will hold local community meetings and explain the study to the potentially eligible adult 
volunteers. During these meetings the investigators will explain the following: the need for a vaccine; the current 
status of vaccine development (including the fact that this is likely to be a prolonged process); the study screening and 
informed consent procedure; risks of vaccination and the unproven benefits of vaccination. It will be stressed that 
these are experimental vaccine regimens and cannot be guaranteed to provide protection, and that it will therefore 
still be necessary to seek treatment for possible malaria even after vaccination and they should continue to use other 
protective measures such as bed nets. It will be explained that to aid identification, a photograph of the volunteer will 
be taken if they are eligible to be enrolled in the trial.  
 
After this meeting based on the list of adults of suitable age for participation in the trial drawn from the DSS database, 
volunteers will be asked to participate in a public lottery that is made to randomly select participants who will be 
invited for a screening visit. 
 
A set of sealed envelopes containing “YES” or “NO” will be prepared. When the proposed volunteer’s name is called 
by the investigator, the volunteer will come and randomly pick one envelope. If the envelope containing a “YES” is 
picked, the proposed volunteer’s name will be entered into the screening log sheet. If it is “NO”, the proposed 
volunteer will not be invited for screening. This method has been used in previous trials and is accepted by the 
communities as a fair way of giving the chance to each eligible volunteer at the meeting who meets the age criteria, to 

24 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
be invited for the screening visit. All proposed volunteers thus selected will be invited to the Banfora clinical trials 
centre for the screening visit. 
 
8.1.2. Screening Visit 
The Volunteer Information Sheet (VIS) will contain detailed information about the study and will be distributed to the 
proposed volunteers. The investigators will endeavour to ensure that all volunteers fully understand the risks. Any 
volunteer who appears to have less than complete understanding will be considered unable to give consent. 
Volunteers must also understand the very small chance of anaphylactic reactions and thereby the importance of 
complying with the one-hour (1hr) observation period after each vaccination. The information sheet covers these 
points in detail, and each volunteer will have the contents of the sheet explained in individual meetings. If it is 
determined by the investigator conducting the screening visit that free and informed consent is given by the volunteer 
to participate in the trial, the volunteer will be asked to complete the consent form.  
 
If unable to sign, the volunteer will be asked to thumbprint the consent form in the presence of an impartial witness 
who will be present during the screening procedures and will countersign the consent form. 
 
Fully consented volunteers will undergo the full screening procedures. This consists of medical history, physical 
examination, and blood sampling for screening tests as detailed below (Laboratory Evaluations). In summary, each 
consented volunteer will be screened for clinically significant acute or chronic disease, using both general physical 
examination and screening laboratory tests (full blood count (FBC), Hepatitis B, alanine transaminase (ALT) and 
creatinine. Likelihood of migration will be an important factor for non- enrolment. All volunteers will be informed of 
results of laboratory tests and referral to an appropriate health facility done where necessary. Volunteers will be 
informed of the need to conduct anonymous HIV testing. All volunteers will receive pre-test counselling from a trained 
counsellor who will also do post-test counselling after giving the results. HIV positivity will be an exclusion criterion. In 
order to maintain the confidentiality of those volunteers infected with HIV, we will make it clear that during screening 
one can be excluded due to a range of diseases (not just HIV) as well as abnormal laboratory results. Enrolment will be 
done within 14 days of screening.  
 
The investigator will determine whether the volunteer is eligible to participate in the study, using the findings at 
screening, including the results of the screening blood tests. Volunteers to participate in the study will fulfil all of the 
inclusion criteria, and meet none of the exclusion criteria. Volunteers enrolled will be given photo identity (ID) cards 
for ease of identification and also to serve as a reminder of the appointment dates. These will be destroyed at the end 
of the study.  The village of residence of the participant will be documented, along with the GPS coordinates of the 
homestead. 
 
8.1.3. HIV Testing 
Volunteers will have access to a trained counsellor who will do pre and post-test counselling. HIV status will be 
established using the standard rapid diagnostic kits and testing algorithm used by the Burkina Faso Ministry of Health. 
Those diagnosed positive will be referred to an appropriate health centre or District Hospital HIV clinic 
(Comprehensive Care and Research Centre) for further counselling and treatment. 
 
8.2. 
Allocation of participants to Study Groups 
Volunteers will be randomised to receive either three (3) doses of R21/ Matrix-M1 or placebo (normal saline) as 
control. Simple randomisation into the study groups will be done by an independent statistician based at the UOXF.  A 
randomisation code list will be generated by the independent statistician and its use guided by a clear Standard of 
Operating Procedure (SOP).  Allocation concealment will be employed by use of opaque sealed envelopes. As this is a 
single-blind clinical trial design, the laboratory scientists will be blinded to vaccine allocation until the end of the study.  
 
8.3. 
Enrolment 
8.4. 
PARTICIPANTS ARE CONSIDERED ENROLLED INTO THE STUDY WHEN THEY 
HAVE RECEIVED THE FIRST STUDY VACCINATION.  THIS SHOULD OCCUR ≤ 14 
DAYS AFTER THE SCREENING VISIT. THEY WILL REMAIN IN THE VACCINATING 
HEALTH FACILITY FOR 1 HOUR MINUTES POST-VACCINATION IN ORDER TO 
ENABLE MONITORING BY THE STUDY CLINICIANS. THE STUDY CLINICIANS WILL 

25 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
ASSESS AND RECORD LOCAL AS WELL AS SYSTEMIC ADVERSE EVENTS AFTER 1 
HOUR HAS ELAPSED.STUDY VISITS AND PROCEDURES 
Table 2 shows the window periods for the visits and outlines the study procedures at each visit for all study Groups. 
 
Screening: (Clinic Visit) 
Ongoing eligibility for participation will be confirmed according to the inclusion and exclusion criteria, prior to blood 
sampling.  
 
Medical history and physical examination will be performed at screening to exclude any significant medical conditions.  
 
The following lab tests will be done: FBC, ALT, urinalysis, Creatinine, rapid Hepatitis B surface antigen test, Serum β-
HCG pregnancy test (where applicable), and HIV testing 
 
Day 0 (Vaccination with R21/Matrix-M1 vaccine or placebo)  
Ongoing eligibility for participation will be confirmed according to the inclusion and exclusion criteria, prior to blood 
sampling and vaccination. 
 
Medical history, temperature monitoring, physical examination will be performed. 
 
Venepuncture for exploratory immunology (for all volunteers) and safety assessment for all participants will also be 
done prior to vaccination.  Analysis of blood samples will be performed as detailed below (Laboratory Evaluations).  
 
Volunteers are considered enrolled into the study when they receive the first study vaccination. The vaccine will be 
administered as detailed in Section 7 and according to local SOPs. The volunteer will be monitored for one hour (or 
longer if necessary) after vaccination. 
 
The CRF will be updated. 
 
Days 1-6 Field worker home visits) 
Each volunteer will be visited at home daily for 6 days post-vaccination by a field worker for assessment and recording 
of any solicited and unsolicited AEs in diary cards. If necessary the volunteer will continue to be seen regularly until 
any observed AEs have resolved or stabilised. 
 
Day 7 ±2 (Clinic visit) 
Medical history, temperature monitoring, physical examination will be performed and recorded. Analysis of blood 
samples will be performed as detailed below (Laboratory Evaluations). Any solicited/ unsolicited AEs will be reviewed 
and recorded.  
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Day 28 ±2 (Vaccination with R21/Matrix M-1 vaccine or placebo) 
Medical history, temperature monitoring, physical examination will be performed and recorded. Any solicited/ 
unsolicited AEs will be reviewed and recorded.   
 
Venepuncture for exploratory immunology and safety assessment for all participants will be done prior to vaccination.  
Analysis of blood samples will be performed as detailed below (Laboratory Evaluations).  
 
Vaccines will be administered as detailed in Section 6 and according to local SOPs. The volunteer will be monitored for 
one hour (or longer if necessary) after vaccination. 
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Days 29-34 (Field worker home visits) 
Each volunteer will be visited at home daily for 6 days post-vaccination by a field worker for assessment and recording 
of any solicited and unsolicited AEs in diary cards. If necessary the volunteer will continue to be seen regularly until 
any observed AEs have resolved or stabilised. 

26 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Day 35 ±2 (Clinic visit) 
Medical history, temperature monitoring, physical examination will be performed and recorded. Analysis of blood 
samples will be performed as detailed below (Laboratory Evaluations). Any solicited/ unsolicited AEs will be reviewed 
and recorded.   
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Days 56 ±2 (Vaccination with R21/Matrix M-1 vaccine or placebo) 
Medical history, temperature monitoring, physical examination will be performed and recorded. Any solicited/ 
unsolicited AEs will be reviewed and recorded. 
 
Venepuncture for safety assessment for all participants and exploratory immunology (for all volunteers) will be done 
prior to vaccination.  Analysis of blood samples will be performed as detailed below (Laboratory Evaluations).  
 
Vaccines will be administered as detailed in Section 6 and according to local SOPs. The volunteer will be monitored for 
one hour (or longer if necessary) after vaccination. 
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Day 57-62 (Field worker home visits) 
Each volunteer will be visited at home daily for 14 days post-vaccination by a field worker for assessment and 
recording of any solicited and unsolicited AEs in diary cards. If necessary the volunteer will continue to be seen 
regularly until any observed AEs have resolved or stabilised. 
 
Day 63 ±2 (Clinic visit) 
Medical history, temperature monitoring, physical examination will be performed and recorded. Analysis of blood 
samples will be performed as detailed below (Laboratory Evaluations). Any solicited/ unsolicited AEs will be reviewed 
and recorded.   
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Day 84 ±2 (Clinic visit) 
Medical history, temperature monitoring, physical examination will be performed and recorded.  All AE’s/SAE’s will be 
reviewed. 
 
Venepuncture for safety assessment for all participants and exploratory immunology (for all volunteers) will be done 
prior to vaccination.  Analysis of blood samples will be performed as detailed below (Laboratory Evaluations).  
 
The CRF will be updated including the records of AEs, concomitant vaccination and concomitant medications. 
 
Day 140 ±7 (Clinic visit) 
Medical history, temperature monitoring, physical examination will be performed and recorded.  All AE’s/SAE’s will be 
reviewed. 
 
Venepuncture for safety assessment for all participants and exploratory immunology (for all volunteers) will be done 
prior to vaccination.  Analysis of blood samples will be performed as detailed below (Laboratory Evaluations).  
 
The CRF will be updated including the records of AEs, concomitant vaccination. 
 
8.5. 
Laboratory Evaluations 
 
Descriptions of Blood sampling and Laboratory Evaluations 
5 mls of blood will be collected at screening (similar for all sampling groups) to test eligibility.  5 mls of blood will also 
be collected on days 0, 7, 28, 35, 56, 63, 84, and 140 for the evaluation of safety.   The following tests will be 
performed;   
• 
Full blood count  

27 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
• 
Serum ALT and creatinine  
• 
HIV antibody testing (pre-vaccination only) 
 
25 mls of blood will be collected on days 0, 28, 56, 84 and 140.  The following tests will be performed at these time 
points;  
• Antibodies to CSP and HbsAg by ELISA 
• Cellular immunology studies, including ex vivo IFNg ELISPOT and stimulation of whole blood for flow 
cytometry with intracellular cytokine staining. 
• Lymphocytes remaining after the primary immunological readout assay (IFNg ELISPOT) will be stored for 
later immunological studies. This will allow further characterisation antigen-specific T cells.  
• Plasma for antibody studies of immunity to malaria antigens.  
 
Stimulated PBMCs from the ELISPOT plate will be retained and preserved for transcriptomics analysis 
 
An aliquot of whole blood will be preserved for transcriptomics analysis 
 
8.6. 
Provision of care to the study participants 
Study contact personnel will be available 24 hours a day at the trial site clinic and at the different health facilities of 
the study population catchment areas, seven days a week, to attend to volunteers who report ill.   Volunteers 
requiring in-patient care will be admitted to the hospital where study personnel will be posted. Laboratory and 
radiological investigation will be carried out when appropriate. If necessary, volunteers requiring more specialised 
care (treatment or diagnostic procedures) will be transported to a referral hospital. Treatment for medical conditions 
will be given according to the standard treatment regimens locally. Any expenses including transport costs incurred by 
the volunteers for clinical care related to acute conditions will be borne by the trial according to the appropriate local 
arrangements. Long-term care for chronic conditions unrelated to study procedures will be delivered following local 
guidelines with no financial support from the trial. 

28 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Table 2: VAC060 Schedule of study visits and procedures for participants 
 
 
Schedule of Visits:  S = Screening; Window (days) refers to time since last visit  
 
S 
R21/ Matrix-
M1 or 
Normal 
Saline 
 
 
R21/ Matrix-
M1 or 
Normal 
Saline 
 
 
R21/ 
Matrix-M1 
or Normal 
Saline 
 
 
 
 
Attendance number 
1 
2 
3 
4 
 
5 
6 
 
7 
8 
9 
Timeline (days) 
 
0 
1-6 
7 
28 
29-34 
35 
56 
57-62 
63 
84 
140 
Window (days) 
-14 
- 
±2 
±2 
 
±2 
±2 
 
±2 
±2 
±7 
Inclusion / Exclusion criteria 
X 
X 
 
 
X 
 
 
X 
 
 
 
Informed consent 
X 
X 
 
 
 
 
 
 
 
 
 
 
Medical History 
X 
X 
 
X 
X 
 
X 
X 
 
X 
X 
X 
Physical Examination 
X 
X 
 
X 
X 
 
X 
X 
 
X 
X 
X 
Urinalysis 
X 
 
 
 
 
 
 
 
 
 
 
 
Serum Pregnancy Test (women only) 
X 
X 
 
 
X 
 
 
X 
 
 
 
Review contraindications 
X 
X 
 
 
X 
 
 
X 
 
 
 
Vaccination 
 
X 
 
 
X 
 
 
X 
 
 
 
Field Worker Home Visit/Diary Cards 
 
 
X 
 
 
X 
 
 
X 
 
 
 
Concomitant Vaccination 
 
X 
X 
X 
 
X 
X 
 
X 
X 
X 
Concomitant medication 
 
X 
X 
X 
 
X 
X 
 
X 
X 
 
AEs reviewed 
 
X 
X 
X 
 
X 
X 
 
X 
X 
X 
Diary card reviewed 
 
 
X 
 
 
X 
 
 
X 
 
 
 
Blood smear, Blood spot 
X 
 
 
 
 
 
 
 
 
 
 
HLA typing (mL) 
X 
 
 
 
 
 
 
 
 
 
 
HBV, HIV (mL) 
X 
 
 
 
 
 
 
 
 
 
 
 
Haematology (mL) 
X 
X 
 
X 
X 
 
X 
X 
 
X 
X 
X 
Biochemistry (mL) 
X 
X 
 
X 
X 
 
X 
X 
 
X 
X 
X 
Immunology 
 
X 
 
 
X 
 
 
X 
 
X 
X 
X 
Blood volume per visit (mL) 
5 
30 (25+5) 
 
5 
30 (25+5) 
 
 
30 (25+5) 
 
30 
(25+5) 
30 
(25+5) 
30 
(25+5) 

29 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
8.7. 
Safety follow-up  
Trained field workers under the supervision of the investigators will visit each enrolled volunteer for 6 consecutive 
days post-vaccination (Table 2). If necessary the volunteers will continue to be seen by the field worker on subsequent 
days for follow-up of adverse events. In the event that the field worker finds any Grade 3 solicited general or 
unsolicited symptoms, the volunteer will be brought to the vaccination centre for examination by a study clinician. 
During the field worker visits, volunteers will be asked retrospectively if any medical event that might be a SAE 
occurred since the last visit and this information will be recorded. Unreported SAEs detected in this way will be 
investigated and reported by the PI or delegate on the corresponding SAE. 
 
If a study volunteer is reported to be unwell at the time of a visit, the field worker will advise the volunteer to report 
to the trial site clinic or the nearest health facility where a study nurse will be posted and will notify this referral to the 
clinical team for follow up. In the event that the volunteer is seriously ill, the field worker will inform the PI or 
designate, and transport will be arranged, to the referral hospital (where a study physician is posted), if judged 
appropriate by the responsible clinician. 
 
In case a study volunteer is unwell and referred to the trial site clinic or health facility, a duplicate blood film will be 
obtained should the volunteer present symptoms or signs compatible with malaria (axillary temperature ≥ 37.5°C, 
history of fever within the last 24 hours,  loss of appetite, malaise, joint pains)  
 
A study clinician will review the volunteer for safety and reactogenicity assessment at the Clinic Visits, on Days 0, 7, 
28, 35, 56, 63, 84, and 140. 
 
8.8. 
Immunogenicity measurements 
These will mainly consist of measurements of humoral immunity using ELISA and cellular responses using ex vivo IFNg 
ELISPOT. In addition, flow cytometry on whole blood, and gene expression profiling will be performed.  
 
The interferon-gamma (IFNγ) enzyme-linked immunospot assay (ELISPOT) can be performed in two ways; the ex vivo 
assay that enumerates effector memory T cells and which has correlated directly with protection in two mouse 
models of malaria36; and the cultured ELISPOT that measures central memory T cell responses and correlates with 
protection in the field trial of RTS,S/AS02 in the Gambia37 and in sporozoite challenge studies of viral vector 
vaccinations in Oxford27. IFNγ secreted by T cells after interaction with infected liver cells has been shown to induce 
death of liver-stage parasites38. The ex-vivo assay will be used as the primary readout for vaccine immunogenicity in 
this study. PBMC will be stimulated with pools of 20mer peptides spanning the length of the ME-TRAP insert and 
overlapping by 10 amino acids. Additional information on T cell responses will be obtained by intracellular cytokine 
staining and flow cytometry to determine whether responding T cells are CD4+ or CD8+ and assess production of 
other cytokines such as IL-2 and TNFα . 
 
RNA analysis may also be used to examine the profile of gene expression following vaccination and during exposure as 
there is mounting evidence that gene expression profiles can predict characteristics of the immune response to 
vaccination and may possibly be used to prospectively determine vaccine efficacy39.  
 
Immune responses to vaccination may be affected by genetic factors; therefore we will  assess sequence variation in 
DNA from vaccinees by sequencing or other genotyping methods. 
 
8.9. 
Data collection  
Adverse events will be documented in individual case report forms (CRFs) for each volunteer. They will be recorded 
under two headings; local and systemic. There will be documentation of concomitant medication, concomitant 
vaccination, non-serious adverse event documentation, serious adverse event documentation and study conclusion. 
Case report forms will be kept securely.  
 
The following data will be collected for concomitant medications: medication name (generic name), dose, frequency 
and route; start and stop dates; and indication. 
Concomitant medication will be recorded according to the time period below: 
 
Antimalarial drugs, antibiotics with antimalarial activity, immune modifying drugs and blood transfusions will 
be captured for the duration of the trial.  
 
Antipyretics, analgesics, systemic antibiotics with unknown antimalarial activity will be collected from first 
vaccination until 1 month post-final vaccination. 
 
All vaccines administered, not specified in the study protocol, will be recorded for the duration of the trial.  

30 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
8.10. 
Study termination 
The study will be discontinued in the event of any of the following: 
 
New scientific information is published to indicate that volunteers in the study are being exposed to undue 
risks as a result of administration of the IMPs by any route of administration, or as a result of the follow-up 
schedule. 
 
Serious concerns about the safety of the IMPs arise as a result of one or more vaccine related SAE occurring 
in the subjects enrolled in this or any other ongoing study of the IMPs. 
 
For any other reason at the discretion of the Principal Investigator. 
 
8.11. 
Definition of the Start and End of the Trial 
The start of the trial is defined as the date of the first vaccination of the first volunteer. The end of the trial is the date 
of the last visit of the last volunteer. 
 
9. ASSESSMENT OF SCIENTIFIC OBJECTIVES 
9.1. 
Safety Evaluation 
Evaluation criteria for safety will be local and systemic solicited and unsolicited adverse events.   
 
Assessment of the safety and reactogenicity of three (3) doses of 10 & 50 µg R21/Matrix-M1 administered four (4) 
weeks apart will be undertaken by summary listing of all solicited and unsolicited local and systemic adverse events 
(including results of clinical laboratory investigations where deemed adverse events), considered possibly, probably, 
or definitely related to vaccination; and line listing of all SAEs. 
 
Safety endpoints will be summarised for groups 1 and 2. The number and percentage of volunteers in each group who 
have any local reaction will be compared (using the chi-squared test, and by calculating confidence intervals on 
differences in percentages). Similarly, the numbers who have any systemic reaction or SAE will be compared between 
groups. 
 
9.2. 
Immunogenicity evaluation 
Evaluation criteria for immunogenicity will include measures of immunogenicity of R21/Matrix-M1  
 
Antibodies to CSP and HbSAg by ELISA 
 
Ex vivo gamma-interferon T cell ELISPOT assays (Day 0 and 84 on frozen cells) 
 
ICS and flow cytometry (Day 0 and 84 on stimulated frozen whole blood)   
 
Exploratory immunology, including RNA analysis and DNA analysis to include parasite and human genetic 
analysis. 
 
Immunogenicity data will be analysed according to a detailed analytical plan.  
10. 
SAFETY REPORTING 
10.1. 
Definitions 
Definitions for the terms adverse event (or experience), adverse reaction, and unexpected adverse reaction have 
previously been agreed to by consensus of the more than 30 Collaborating Centres of the WHO International Drug 
Monitoring Centre (Uppsala, Sweden). Although those definitions can pertain to situations involving clinical 
investigations, some minor modifications are necessary, especially to accommodate the pre-approval, development 
environment. 
The following definitions, with input from the WHO Collaborative Centre, have been agreed: 
10.1.1. 
Adverse Event 
Any untoward medical occurrence in a clinical investigation subject occurring in any phase of the clinical study 
whether or not considered related to the vaccine. This includes an exacerbation of pre-existing conditions or events, 
intercurrent illnesses, or vaccine or drug interaction. Anticipated day-to-day fluctuations of pre-existing conditions 
that do not represent a clinically significant exacerbation will not be considered adverse events. Discrete episodes of 
chronic conditions occurring during a study period will be reported as adverse events in order to assess changes in 

31 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
frequency or severity. 
 
Adverse events will be documented in terms of a medical diagnosis (es). When this is not possible, the adverse event 
will be documented in terms of signs and symptoms observed by the investigator at each study visit. 
 
Pre-existing conditions or signs and/or symptoms (including any which are not recognised at study entry but are 
recognised during the study period) present in a subject prior to the start of the study will be recorded on the Medical 
History form within the subject's CRF. 
 
10.1.2. 
Adverse Drug Reaction (ADR) 
An ADR is any untoward or unintended response to a medicinal product. This means that a causal relationship 
between the study medication and an AE is at least a reasonable possibility, i.e., the relationship cannot be ruled out. 
 
10.1.3. 
Serious Adverse Event (SAE) 
A serious adverse event (experience) or reaction is any untoward medical occurrence that at any dose: 
 
results in death, 
 
is life-threatening, 
Note: The term “life-threatening” in the definition of “serious” refers to an event in which the patient was at 
risk of death at the time of the event; it does not refer to an event which hypothetically might have caused 
death if it were more severe. 
 
requires inpatient hospitalisation‡ or prolongation of existing  hospitalisation, 
 
results in persistent or significant disability/incapacity, or 
 
is a congenital anomaly/birth defect. 
Medical and scientific judgment should be exercised in deciding whether expedited reporting is appropriate in other 
situations, such as important medical events that may not be immediately life-threatening or result in death or 
hospitalisation but may jeopardise the patient or may require intervention to prevent one of the other outcomes 
listed in the definition above. These should also usually be considered serious. 
 
‡Hospitalization: In general, hospitalization signifies that the subject has been detained (usually involving at least an 
overnight stay) at the hospital or emergency ward for treatment that would not have been appropriate in the 
physician’s office or out-patient setting 
Hospitalization for either elective surgery related to a pre-existing condition which did not increase in severity or 
frequency following initiation of the study or for routine clinical procedures¶ (including hospitalization for “social” 
reasons) that are not the result of an adverse event need not be considered as adverse events and are therefore not 
serious adverse events. 
 
In addition, important medical events that may jeopardise the patient or may require intervention to prevent one of 
the other outcomes listed above should be considered serious. (Examples of such treatments are intensive treatment 
in an emergency room or at home for allergic bronchospasm; blood dyscrasias or convulsions that do not results in 
hospitalisation; or development of drug dependency or drug abuse.) 
 
10.1.4. 
Suspected Unexpected Serious Adverse Reactions (SUSARs) 
A SUSAR is a SAE that is unexpected and thought to be related to the investigational product. Administration of 
further vaccines within the trial will be suspended until a safety review is convened.   
 
10.2. 
Collection of Adverse Events 
At each post-vaccination visit all adverse events will be documented. 
 
10.2.1. 
Local solicited Adverse Events 
 
The following solicited local reactions at the injection site will be documented:  
 
swelling, 
 
Redness/discoloration 
 
Pain/limitation of limb movement.  
 
10.2.2. 
Solicited general AEs 
The solicited general AEs that will be documented include 

32 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Fever (defined as axillary temperature ≥ 37.5°C) 
 
Chills 
 
Headache 
 
Malaise 
 
Joint pains 
 
Myagia 
 
Fatigue 
 
Nausea 
The field workers will record these AEs during the home visits, according to the SOPs available at the study site.   
 
10.2.3. 
Unsolicited adverse events 
Unsolicited adverse events will only be recorded in the CRF if they occurred up to 28 days post vaccination, unless 
they meet the criteria for serious adverse event as outlined above. Serious adverse events will be collected 
throughout the study period. 
As a consistent method of soliciting adverse events, the participant will be asked a non-leading question such as: 
“Have you felt different in any way since receiving the vaccine or since the last visit?” The investigator will record only 
those adverse events having occurred within the time frames defined above. 
Adverse events already documented in the CRF, i.e. at a previous assessment and designated as ‘ongoing’ will be 
reviewed at subsequent visits, as necessary. If these have resolved, the documentation in the CRF will be completed.  
 
10.3. 
Follow-up of Adverse Events 
All AEs will be followed until resolution, stabilization of the signs or symptoms or laboratory changes occur, or until a 
non-study related causality is assigned. Clinically significant laboratory abnormalities will be followed up until they 
have returned to normal, or a satisfactory explanation has been provided.  
Volunteers who have moderate or severe on-going adverse events at the completion of the study will be advised to 
consult a physician if the event is not considered to be related to the study vaccine. A follow-up visit will be arranged 
to manage the problem and to determine the severity and duration of the event, if it is considered to be related to the 
study vaccine. If appropriate, specialist review will be arranged by CNRFP investigators. 
Any serious adverse event possibly related to the vaccine and occurring after trial termination should be reported by 
the investigator according to the procedure described below. 
 
10.4. 
Reporting of Adverse Events 
Every SAE occurring throughout the trial must be reported by telephone, email or fax to the sponsor, LSM and DSMB 
by the investigator as soon as (s)he is alerted of it and within one working day, even if the investigator considers that 
the adverse event is not related to vaccination. The investigator will then complete a SAE report form as soon as 
possible and within five working days or seven calendar days.   The contact details for reporting the SAE is 
sae@well.ox.ac.uk 
Any relevant information concerning the adverse event that becomes available after the SAE report form has been 
sent (outcome, precise description of medical history, results of the investigation, copy of hospitalisation report, etc.) 
will be forwarded to the Sponsor in a timely manner. The anonymity of the subjects shall be respected when 
forwarding this information. 
SAEs that are suspected to be related to the vaccine will be reported to the Ethics Committee within 15 calendar days 
of the site becoming aware of the event. If the event is fatal or life-threatening, the event will be reported within 7 
calendar days. 
Suspected unexpected serious adverse reactions (SUSARs) will be reported according to national regulatory 
guidelines. The sponsor pledges to inform the Authorities of any trial discontinuation and specify the reason for 
discontinuation. 
Every effort should be made by the investigator to explain each adverse event and assess its causal relationship, if any 
to the administration of the investigational product. This interpretation will be based on the type of event, the 
relationship of the event to the time of vaccine administration (the event being temporally associated with 
vaccination or reproduced on re-vaccination), and the known biology of vaccine therapy (the event having often been 
reported in literature for similar types of vaccines).  
All solicited local (injection site) reactions will be considered causally related to vaccination. 
 
The following are guidelines for assessing the causal relationship for the systemic adverse events: 
No relationship: 
 
No temporal relationship to study product; and 
 
Alternate aetiology (clinical state, environmental or other interventions); and 

33 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
 
Does not follow known pattern of response to study product 
Unlikely relationship:  
Unlikely temporal relationship to study product; and  
Alternate aetiology likely (clinical state, environmental or other interventions); and  
Does not follow known typical or plausible pattern of response to study product 
Possible relationship: 
 
Reasonable temporal relationship to study product; or 
 
Event not readily produced by clinical state, environmental or other interventions; or 
 
Similar pattern of response to that seen with other vaccines 
Probable relationship: 
 
Reasonable temporal relationship to study product; and 
 
Event not readily produced by clinical state, environment, or other interventions or 
 
Known pattern of response seen with other vaccines 
Definite relationship: 
 
Reasonable temporal relationship to study product; and 
 
Event not readily produced by clinical state, environment, or other interventions; and 
 
Known pattern of response seen with other vaccines 
 
10.5. 
Grading the severity of adverse events 
 
10.5.1. 
Local solicited AEs 
The largest diameter through the injection site of any local redness/discoloration will be recorded in millimeters. The 
largest diameter through the injection site of local swelling will be recorded in millimeters. Severity of these local 
findings will be graded using the scales below: 
 
Grading for swelling 
Grading for redness/discoloration 
Grade 
Diameter [mm] 
 
Grade 
Diameter [mm] 
0 
0 
 
0 
0 
1 
< 20 
 
1 
< 50 
2 
20 – 50 
 
2 
50 – 100 
3 
> 50 
 
3 
> 100 
 
 
The presence and severity of local pain/limitation of limb movement at the site of vaccination will be determined 
using the following scale: 
 
Grade 
Description 
0 
No pain at all 
1 
Painful on touch, no restriction in movement of limb 
2 
Painful when limb is moved 
3 
Unable to use limb due to pain 
 
 
10.5.2. 
Solicited general AEs 
 

34 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Fever 
0 
<37.5ºC 
Record axillary temperature 
1 
37.5 - 38.0ºC 
 
2 
>38.0 - 39ºC 
 
3 
>39ºC 
Chills  
0 
None 
 
1 
Chills that are easily tolerated 
 
2 
Chills that interfere with daily activity 
 
3 
Chills that prevent daily activity 
Nausea 
0 
None 
 
1 
Nausea that is easily tolerated 
 
2 
Nausea that interferes with daily activity 
 
3 
Nausea that prevents daily activity 
Headache 
0 
None 
 
1 
Headache that is easily tolerated 
 
2 
Headache that interferes with daily activity 
 
3 
Headache that prevents daily activity 
Malaise 
0 
None 
 
1 
Malaise that is easily tolerated 
 
2 
Malaise that interferes with daily activity 
 
3 
Malaise that prevents daily activity 
Myalgia 
0 
None 
 
1 
Myalgia that is easily tolerated 
 
2 
Myalgia that interferes with daily activity 
 
3 
Myalgia that prevents daily activity 
Joint pain 
0 
None 
 
1 
Joint pain that is easily tolerated 
 
2 
Joint pain that interferes with daily activity 
 
3 
Joint pain that prevents daily activity 
Fatigue 
0 
None 
 
1 
Fatigue that is easily tolerated 
 
2 
Fatigue that interferes with daily activity 
 
3 
Fatigue that prevents daily activity 
 
10.5.3. 
Non Solicited AEs 
For adverse events other than those  for which the severity scales are detailed above, AEs will be graded according to 
the DAIDS AE grading table, published in 2004.40  The DAIDS AE grading table classifies adverse events into one of four 
grades, ranging from mild to potentially life-threatening. The DAIDS AE grading table has indications for each of over 
sixty clinical parameters and forty laboratory parameters for grading adult and paediatric AEs. The table also includes 
general guidelines for estimating the grade of parameters not explicitly listed. Each grade is described broadly below: 
• Grade 1 (mild): awareness of a symptom, but the symptom is easily tolerated and causes no or minimal 
interference with usual activity. 
• Grade 2 (moderate): discomfort enough to cause greater than minimal interference with usual activity. 
• Grade 3 (severe): incapacitating; symptoms causing inability to perform usual activities; requires 
absenteeism or bed rest. 
• Grade 4 (potentially life-threatening): symptoms causing inability to perform basic self-care functions OR 
medical or operative intervention is indicated to prevent permanent impairment, persistent disability or 
death.  
 
Laboratory tests will also be graded on the DAIDS AE grading table. 
 
10.5.4. 
Pregnancy 
Participants who become pregnant during the study period must not receive additional doses of vaccine but may 
continue other study procedures to include blood draws for immunogenicity, safety and vaccine efficacy at the 
discretion of the principal investigator.  
Participants should be instructed to notify the investigator if it is determined after completion of the study that they 
became pregnant either during the study or within 30 days of the study.  

35 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
Although not considered an adverse event, pregnancy should be reported on the pregnancy report form and in the 
same way as a serious adverse event. 
A pregnancy should be followed to term, any premature terminations reported, and the health status of the mother 
and child including date of delivery and the child’s gender and weight should be reported to the sponsor. 
11. 
DATA HANDLING AND RECORD KEEPING 
11.1. 
Data Management 
Data management will be conducted at CNRFP and a copy of the anonymised database provided to the sponsor for 
archiving.  The PI will be responsible for receiving, entering, cleaning, querying and storing all data that accrues from 
the study. Responsibility for this may be delegated to the data manager at CNRFP. The data will be entered into the 
volunteers CRFs. Data will be subsequently transferred to an electronic database for analysis. 
 
If any changes to the protocol are necessary during the study a formal amendment will be presented to the sponsor 
prior to submission to the relevant ethical and regulatory agencies for approval unless to eliminate an immediate 
hazard(s) to study participant without prior ethics approval. Any unforeseen and unavoidable deviations from the 
protocol will be documented and filed in as a protocol deviation in the Trial Master File, with explanation. 
 
11.1.1. 
Data Capture Methods 
Data capture will be on paper CRFs. The CRFs will be considered source documents as healthy volunteers may not 
have hospital case-notes. 
Adverse events will be tabulated in an electronic database (OpenClinica®) for descriptive analysis. 
Immunological data will be transferred to an electronic database for analysis without any volunteer identifier apart 
from the unique volunteer number. 
 
11.1.2. 
Types of Data 
Data collected will include solicited and unsolicited adverse event data, concomitant medications, clinical laboratory 
and exploratory immunology data. Source documents will include laboratory results and the case record file 
containing the case report forms for each volunteer as the healthy volunteers participating in this study may not have 
medical notes.   
 
11.2. 
Timing/Reports 
Annual Safety Report: Due on anniversary of Regulatory Approval – sent to Regulatory and Ethical Bodies 
Annual Progress Report: Due on anniversary of Ethical Approval – sent to Ethics Committee 
 
11.3. 
Archiving 
The investigator must keep all trial documents for at least 15 years after the completion or discontinuation of the trial. 
 
11.4. 
Protocol Deviations 
Any unforeseen and unavoidable deviations from the protocol will be documented and filed in the study file with 
explanation. 
12. 
DATA ACCESS AND QUALITY ASSURANCE 
12.1. 
Direct Access to Source Data/Documents 
The PI will provide direct access to the source data documents to the Ethics Committee, to the regulatory agency, and 
to authorized representatives of the sponsor, permitting trial-related monitoring and audits. 
12.2. 
Quality Assurance 
12.2.1. 
Modifications to the Protocol 
Any amendments to the protocol that appear necessary during the course of the trial must be discussed by the 
investigator and sponsor concurrently unless to eliminate an immediate hazard(s) to study participants. If agreement 
is reached concerning the need for a substantial amendment, it will be produced in writing by the sponsor and/or the 
investigator and will be made a formal part of the protocol. Any substantial amendment requires Ethics Committee 
approval, but non-substantial amendments do not. 

36 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
All substantial amendments must also be communicated to Regulatory Authorities, if appropriate. 
An administrative or non-substantial change to the protocol is one that modifies administrative and logistical aspects 
of a protocol but does not affect the subjects’ safety, the objectives of the trial and its progress. An administrative 
change does not require Ethics Committee approval. However, the Ethics committee must be notified whenever an 
administrative or non-substantial change is made. 
The investigator is responsible for ensuring that substantial amendments to an approved trial, during the period for 
which Ethics Committee approval has already been given, are not initiated without Ethics Committee review and 
approval except to eliminate apparent immediate hazards to the subject. 
 
12.2.2. 
Monitoring 
Initiation Visit 
An initiation visit will be performed before the inclusion of the first volunteer into the study. The Monitor will verify 
and document that the material to be used during the trial has been received and that the investigational team has 
been properly informed about the trial and regulatory requirements. 
 
Interim Monitoring Visits 
The Monitor will carry out regular follow-up visits. The investigator commits to being available for these visits and to 
allow the monitoring staff direct access to volunteer medical files, if existing, and CRFs. The Monitor is committed to 
professional secrecy. 
During the visits, the Monitor may: 
 
 
Carry out a quality control of clinical trial progress: in respect of protocol and operating guidelines, data 
collection, signature of consent forms, completion of documents, SAE, sample and product management, 
cold chain monitoring 
 
Inspect the CRFs, TMF and correspondent correction sheets 
 
The Monitor will discuss any problem with the investigator and define with him the actions to be taken. 
 
 
Close-out Visit 
A close-out visit will be performed at the end of the trial. Its goals are to make sure that: 
 
The centre has all the documents necessary for archiving 
 
All unused material has been recovered 
 
All vaccines have been accounted for 
13. 
ETHICAL CONSIDERATIONS 
Ethical Review 
Before the inclusion of the first participant in the study, the protocol must be approved by Ethical Review Committees 
in Burkina and Oxford (OXTREC). 
 
Informed Consent 
All volunteers are expected to sign an informed consent document before any study procedures begin.   
 
The written information is provided in French only and the field workers will interpret the written information in a 
language the volunteers understand. The field workers involved in the informed consent discussion are trained on the 
study and the information sheet and consent form, and are trained to discuss the trial in the local languages the 
volunteers understand (Gouin, Karaboro, Dioula). The language of the consent process is documented on the consent 
form. If the volunteer is not able to read and write in French, an adult witness, impartial of the trial, will be present 
through the whole consent process and sign and date the consent form.  
 
The volunteer  should give written/thumb printed informed consent before  included in the trial, after having been 
informed of the nature of the trial, the potential risks and their obligations. Informed consent forms will be provided 
in duplicate (original kept by the investigator, one copy kept by the volunteer). 
 
Confidentiality 

37 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
All data and information generated by the site as part of the study (other than a subject’s medical records) will be kept 
confidential by the investigator and other site staff. This information and data will not be used by the investigator or 
other site personnel for any purpose other than conducting the study. 
 
Inducement 
There may be a perception amongst volunteers of benefit from physical examination, laboratory screening in the 
current study, in addition to free health care provided during the study period for non-vaccine related medical 
problems. We will also offer compensation for transport expenses for all study subjects. 
 
We do not feel these benefits are excessive, and believe it would be unreasonable to request the cooperation of a 
population in regular employment without offering compensation for time.  
14. 
INDEMNITY/COMPENSATION/INSURANCE 
Indemnity 
Compensation for any injury caused by taking part in this study will be in accordance with the guidelines of the 
Association of the British Pharmaceutical Industry (ABPI). Broadly speaking the ABPI guidelines recommend that ‘the 
sponsor’, without legal commitment, should compensate participants without them having to prove that the sponsor 
is at fault. This applies in cases where it is likely that such injury results from giving any new drug or any other 
procedure carried out in accordance with the CTP for the study. ‘The sponsor’ will not compensate participants where 
such injury results from any procedure carried out which is not in accordance with the CTP for the study. Participants’ 
right at law to claim compensation for injury where negligence can be proven is not affected. In this instance the 
University of Oxford is the Research Sponsor Institution. 
 
Compensation 
Participants enrolled in the study will be offered compensation for transport expenses.   
 
Insurance 
Oxford University Investigators participating in this trial will receive insurance coverage from the University clinical 
trials insurance policy.  The University has a specialist insurance policy in place: - Newline Underwriting Management 
Ltd, at Lloyd’s of London – which would operate in the event of any participant suffering harm as a result of their 
involvement in the research. 
 
 

38 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
15. 
REFERENCES 
 
1. World Malaria Report 2012.  World Health Organisation   http://www.who.int/malaria/publications 
2. Sachs J, Malaney P. The economic and social burden of malaria. Nature 2002;415:680-5. 
3. Das P, Horton R. Malaria elimination: worthy, challenging, and just possible. Lancet 2010; 376(9752): 1515-7 
4. White NJ. Antimalarial drug resistance. J Clin Invest 2004;113:1084-92 
5. Baird JK. Eliminating malaria--all of them.  Lancet. 2010 Dec 4; 376(9756):1883-5. doi: 10.1016/S0140-
6736(10)61494-8. Epub 2010 Oct 28. 
6. Alonso, P.L. et al. Efficacy of the RTS,S/AS02A vaccine against Plasmodium falciparum infection and disease in 
young African children: randomised controlled trial. Lancet 364, 1411-1420 (2004). 
7. Olotu, A. et al. Efficacy of RTS,S/AS01E malaria vaccine and exploratory analysis on anti-circumsporozoite 
antibody titres and protection in children aged 5-17 months in Kenya and Tanzania: a randomised controlled 
trial. The Lancet infectious diseases 11, 102-109 (2011). 
8. Agnandji, S.T. et al. First Results of Phase 3 Trial of RTS,S/AS01 Malaria Vaccine in African Children. N Engl J 
Med (2011). 
9. Ewer KJ et al. 2013. Protective CD8+ T cell Immunity to human malaria induced by chimpanzee adenovirus-
MVA immunisation. Nat Commun. 2013 Nov 28; 4: 2836.doi: 10.1038/ncomms3836 
10. McConkey SJ et al. Enhanced T-cell immunogenicity of plasmid DNA vaccines boosted by recombinant 
modified vaccinia virus Ankara in humans. Nature Medicine. 2003; 9:729-735. 
11. Webster DP et al. Enhanced T cell-mediated protection against malaria in human challenges by using the 
recombinant poxviruses FP9 and modified vaccinia virus Ankara. Proceedings of the National Academy of 
Sciences of the United States of America. 2005; 102(13): 4836-4841. 
12. Nussenzweig, R.S., et al., Protective immunity produced by the injection of x-irradiated sporozoites of 
plasmodium berghei. Nature, 1967. 216(5111): p. 160-2. 
13. Hill, A.V., et al., Common west African HLA antigens are associated with protection from severe malaria. 
Nature, 1991. 352(6336): p. 595-600. 
14. Hill, A.V., et al., Molecular analysis of the association of HLA-B53 and resistance to severe malaria. Nature, 
1992. 360(6403): p. 434-9. 
15. Todryk, S.M., et al., Correlation of memory T cell responses against TRAP with protection from clinical 
malaria, and CD4 CD25 high T cells with susceptibility in Kenyans. PLoS One, 2008. 3(4): p. e2027. 
16. Khusmith, S., M. Sedegah, and S.L. Hoffman, Complete protection against Plasmodium yoelii by adoptive 
transfer of a CD8+ cytotoxic T-cell clone recognizing sporozoite surface protein 2. Infect Immun, 1994. 62(7): 
p. 2979-83. 
17. Romero, P., et al., Cloned cytotoxic T cells recognize an epitope in the circumsporozoite protein and protect 
against malaria. Nature, 1989. 341(6240): p. 323-6. 
18. Roestenberg, M., et al., Protection against a malaria challenge by sporozoite inoculation. N Engl J Med, 2009. 
361(5): p. 468-77. 
19. McConkey SJ, Reece WH, Moorthy VS, Webster D, Dunachie S, Butcher G, et al. Enhanced T-cell 
immunogenicity of plasmid DNA vaccines boosted by recombinant modified vaccinia virus Ankara in humans. 
Nat Med 2003; 9:729-35. 
20. Bejon P, Andrews L, Andersen RF, Dunachie S,Webster D, Walther M, et al. Calculation of liver-to blood 
inocula, parasite growth rates and pre-erythrocytic vaccine efficacy, from serial quantitative polymerase 
chain reaction studies of volunteers challenged with malaria sporozoites. J Infect Dis 2005; 191:619-26. 
21. Dunachie SJ, Walther M, Epstein JE, Keating S, Berthoud T, Andrews L, et al. A DNA prime-modified vaccinia 
virus Ankara boost vaccine encoding thrombospondin-related adhesion protein but not circumsporozoite 
protein partially protects healthy malaria-naive adults against Plasmodium falciparum sporozoite challenge. 
Infect Immun 2006; 74:5933-42. 
22. Webster, D.P., et al., Enhanced T cell-mediated protection against malaria in human challenges by using the 
recombinant poxviruses FP9 and modified vaccinia virus Ankara. Proc Natl Acad Sci U S A, 2005. 102(13): p. 
4836-41. 
23. Bejon, P., et al., Safety profile of the viral vectors of attenuated fowlpox strain FP9 and modified vaccinia virus 
Ankara recombinant for either of 2 preerythrocytic malaria antigens, ME-TRAP or the circumsporozoite 
protein, in children and adults in Kenya. Clin Infect Dis, 2006. 42(8): p. 1102-10. 
24. Bejon, P., et al., Immunogenicity of the candidate malaria vaccines FP9 and modified vaccinia virus Ankara 
encoding the pre-erythrocytic antigen ME-TRAP in 1-6 year old children in a malaria endemic area. Vaccine, 
2006 

39 
VAC060_Protocol_v2.1 Final_ tracked_14 Oct 2015                                            © University of Oxford, 2015 
25. Imoukhuede EB, Berthoud T, Milligan P, K, Bojang K et al.  Safety and immunogenicity of the malaria 
candidate vaccines FP9 CS and MVA CS in adult Gambian men.  Vaccine. 2006 Oct 30; 24(42-43):6526-33. 
Epub 2006 Jun 28.   
26. Bejon, P., et al., A Phase 2b Randomised Trial of the Candidate Malaria Vaccines FP9 ME-TRAP and MVA ME-
TRAP among Children in Kenya. PLoS Clin Trials, 2006. 1(6): p. e29. 
27. Keating SM, Bejon P, Berthoud T, Vuola JM, Todryk S, Webster DP, et al. Durable human memory T cells 
quantifiable by cultured enzyme-linked immunospot assays are induced by heterologous prime boost 
immunization and correlate with protection against malaria. J Immunol 2005; 175:5675-80. 
28. O'Hara GA, Duncan CJ, Ewer KJ, Collins KA, Elias SC, Halstead FD et al.  Clinical assessment of a recombinant 
simian adenovirus ChAd63: a potent new vaccine vector.  J Infect Dis. 2012 Mar 1; 205(5):772-81. doi: 
10.1093/infdis/jir850. Epub 2012 Jan 24. 
29. Ogwang C, Afolabi M, et al.  Safety and immunogenicity of heterologous prime-boost immunisation with 
Plasmodium falciparum malaria candidate vaccines, ChAd63 ME-TRAP and MVA ME-TRAP, in healthy 
Gambian and Kenyan adults.  PLoS One. 2013;8(3):e57726.  Epub 2013 Mar 19. 
30. Osman MM, Nour BY, Sedig MF, De Bes L, Babikir AM, Mohamedani AA, Mens PF. 2010. Informed decision-
making before changing to RDT: a comparison of microscopy, rapid diagnostic test and molecular techniques 
for the diagnosis and identification of malaria parasites in Kassala, eastern Sudan. Trop Med Int Health 
15:1442-8 
31. Rantala AM, Taylor SM, Trottman PA, Luntamo M, Mbewe B, Maleta K, Kulmala T, Ashorn P, Meshnick SR. 
2010. Comparison of real-time PCR and microscopy for malaria parasite detection in Malawian pregnant 
women. Malar J 9: 269  
32. Imoukhuede EB, Andrews L, Milligan P, Berthoud T, Bojang K, Nwakanma D, Ismaili J, Buckee C, Njie F, Keita S, 
Sowe M, Lang T, Gilbert SC, Greenwood BM, Hill AV. 2007.  Low-level malaria infections detected by a 
sensitive polymerase chain reaction assay and use of this technique in the evaluation of malaria vaccines in 
an endemic area.  Am J Trop Med Hyg 76: 486-93 
33. Bojang KA, Milligan PJ, Pinder M, Vigneron L, Alloueche A, Kester KE, Ballou WR, Conway DJ, Reece WH, 
Gothard P, Yamuah L, Delchambre M, Voss G, Greenwood BM, Hill A, McAdam KP, Tornieporth N, Cohen JD, 
Doherty T. 2001.  Efficacy of RTS,S/AS02 malaria vaccine against Plasmodium falciparum infection in semi-
immune adult men in The Gambia.  Lancet 358: 1927-34 
34. Kester KE, Cummings JF, Ockenhouse CF, et al.  Randomized, double-blind, phase 2a trial of falciparum 
malaria vaccines RTS,S/AS01B and RTS,S/AS02A in malaria naïve adults; safety, efficacy and immunogenic 
associates of protection.  J Infect Dis 2009 Aug 1 200(3):337-46 doi 10.1086/600120.  
35. Verstraeten T, Descamps D, David MP, et al. Analysis of adverse events of potential autoimmune aetiology in 
a large integrated safety database of AS04 adjuvanted vaccines. Vaccine 2008;26:6630-8 
36. Plebanski, M., et al., Protection from Plasmodium berghei infection by priming and boosting T cells to a single 
class I-restricted epitope with recombinant carriers suitable for human use. Eur J Immunol, 1998. 28(12): p. 
4345-55. 
37. Reece, W.H., et al., A CD4(+) T-cell immune response to a conserved epitope in the circumsporozoite protein 
correlates with protection from natural Plasmodium falciparum infection and disease. Nat Med, 2004. 10(4): 
p. 406-10 
38. Schofield, L., et al., Gamma interferon, CD8+ T cells and antibodies required for immunity to malaria 
sporozoites. Nature, 1987. 330(6149): p. 664-6. 
39. Querec TD et al. Systems biology approach predicts immunogenicity of the yellow fever vaccine in humans, 
Nature Immunology. 2009; 10(1): 116-125 
40. Division of AIDS Table for Grading the Severity of Adult and Paediatric Adverse Events, Version 1.0, December 
2004, 
(Clarification 
August 
2009). 
[Internet] 
Accessed 
at 
http://rsc.techres.com/Document/safetyandpharmacovigilance/Table_for_Grading_Severity_of_Adult_Pedia
tric_Adverse_Events.pdf, 19 October 2012. 
41. Tiono AB, Kangoye DT, Rehman AM, Kargougou DG, Kaboré Y, Diarra A, Ouedraogo E, Nébié I, Ouédraogo A, 
Okech B, Milligan P, Sirima SB. 
Malaria incidence in children in South-west burkina faso: comparison of active and passive case detection 
methods. PLoS One. 2014 Jan 24;9(1):e86936. doi: 10.1371/journal.pone.0086936. eCollection 2014.