A Phase Ia Study to Assess Safety and Immunogenicity of the Plasmodium Vivax Malaria Vaccine Candidate Pvs25-IMX313 in Matrix-M1 Adjuvant in Healthy Adults Living in the UK
Phase: EARLY_PHASE1
Conditions: Malaria, Vivax
Sponsor: University of Oxford

 
 
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IRAS Project ID: 302239                                                                                         
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A Phase Ia study to assess safety and immunogenicity of the Plasmodium vivax malaria vaccine 
candidate Pvs25-IMX313 in Matrix-M1 adjuvant in healthy adults living in the UK 
 
Study Reference: VAC084 
Protocol Version: 2.0 
Date: 24 Mar 2022 
 
 
IRAS Project ID:  302239 
REC reference: 21/FT/0159 
EudraCT Number:  2021-003550-23 
NCT number: NCT05270265 
 
 
 
Chief Investigator: Angela M Minassian 
Sponsor: University of Oxford 
 
 
 
 
 
 
 
 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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MODIFICATION HISTORY 
Version Date 
Author(s) 
Modifications 
1.0 
17 Dec 2021 
Cyndi Goh 
Angela Minassian 
Sumi Biswas 
Created 
2.0 
24 Mar 2022 
Cyndi Goh 
Angela Minassian 
Rationale for dose selection included 
for clarity 
Langugage regarding gender and 
contraception changed to be more 
inclusive and for clarity 
Wording relating to group holding rules 
amended for clarity 
Wording relating to first volunteers 
amended for clarity 
Typographical errors corrected 
Removal of reference to form on p5 of 
GP screening letter 
 
 
 
 
 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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A Phase Ia study to assess safety and immunogenicity of the Plasmodium vivax malaria vaccine 
candidate Pvs25-IMX313 in Matrix-M1 adjuvant in healthy adults living in the UK 
Study Code:  VAC084 
Chief Investigator 
 
 
 
 
 
 
 
 
 
Investigator 
 
 
 
 
 
 
 
 
Trial sites 
 
 
 
 
Dr Angela M Minassian 
Centre for Clinical Vaccinology and Tropical Medicine 
Jenner Institute  
University of Oxford 
Churchill Hospital 
Old Road  
Headington 
Oxford, OX3 7LE 
Email: angela.minassian@ndm.ox.ac.uk 
 
Prof Sumi Biswas  
Jenner Institute  
University of Oxford 
Old Road Campus Research Building 
Roosevelt Drive  
Headington  
Oxford, OX3 7DQ 
Email: sumi.biswas@ndm.ox.ac.uk 
 
Centre for Clinical Vaccinology & Tropical Medicine 
Jenner Institute  
University of Oxford 
Churchill Hospital  
Old Road 
Headington 
Oxford, OX3 7LE 
PI at Site: Dr Angela M Minassian 
 
Sponsoring Institution 
 
 
University of Oxford 
Research Governance, Ethics and Assurance Team 
Research Services 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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University of Oxford 
Joint Research Office 
Boundary Brook House 
Churchill Drive 
Headington, OX3 7GB 
Tel: 01865 616480  
Email: ctrg@admin.ox.ac.uk 
 
External Monitor  
 
 
 
 
 
Appledown Clinical Research Ltd. 
Orchard End 
Greenlands Lane 
Prestwood 
Bucks, HP16 9QX 
Tel: 01494 867676/07941 271635 
Email: ceri.mckenna@appledown.net 
 
Local Safety Monitor  
 
Prof Brian Angus 
Centre for Clinical Vaccinology and Tropical Medicine 
Churchill Hospital  
Old Road 
Headington 
Oxford, OX3 7LE 
Tel: 01865 220289 
Email: brian.angus@ndm.ox.ac.uk 
 
Funder 
European Commission (Horizon 2020) 
Rue de la Loi / Wetstraat 170B- 
1049 Brussels 
Belgium 
 
 
 
 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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Confidentiality Statement 
This document contains confidential information that must not be disclosed to anyone other 
than the Sponsor, the Investigator Team, and members of the Independent Ethics Committee.  
This information cannot be used for any purpose other than the evaluation or conduct of the 
clinical investigation without the prior written consent of Dr Angela M Minassian. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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Investigator Agreement 
“I have read this protocol and agree to abide by all provisions set forth therein. 
I agree to comply with the principles of the International Conference on Harmonisation Tripartite 
Guideline on Good Clinical Practice.”   
Dr Angela Minassian 
 
[redacted for public release] 
 
24 Mar 2022 
Chief Investigator 
Dr Angela M Minassian 
 
 
Investigator Signature 
 
Date  
 
Conflict of Interest 
1. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare 
no/the following (delete as appropriate) conflict of interest” 
Dr Angela Minassian 
 
[redacted for public release] 
 
24 Mar 2022 
Chief Investigator 
Dr Angela M Minassian 
 
 
Investigator Signature 
 
Date  
 
Details:____ n/a ________ _________________________________________________________ 
 
2.  “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare no 
conflict of interest” 
 
Prof Sumi Biswas 
 
 
[redacted for public release] 
 
    
 
24 Mar 2022 
Co-investigator 
Prof Sumi Biswas 
 
Investigator Signature 
 
 
Date  
 
Details:_____n/a________________________________________________________________ 
 
 
 
 
 
 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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Table of Contents 
Modification History ................................................................................................................ 2 
1 Synopsis .............................................................................................................................. 11 
2 Abbreviations ...................................................................................................................... 13 
3 Background & Rationale ...................................................................................................... 15 
3.1 The need for a vaccine against Plasmodium vivax malaria ..........................................................15 
3.2 Lifecycle of the P. vivax malaria parasite ...................................................................................16 
3.3 Challenges and directions in P. vivax vaccine development ........................................................18 
3.4 Rationale for a transmission blocking vaccine against P. vivax malaria .......................................21 
3.5 Pvs25 as an antigen...................................................................................................................21 
3.6 Clinical trials of Pvs25-based vaccines ........................................................................................22 
3.7 IMX313 .....................................................................................................................................22 
4 Pvs25-IMX313/Matrix-M1 Vaccine development.................................................................. 24 
4.1 Characteristics of Pvs25-IMX313/Matrix-M1 ..............................................................................24 
4.2 Vaccine development strategy ..................................................................................................24 
4.3 Pvs25-IMX313 immunogenicity and ex vivo efficacy in pre-clinical studies ..................................24 
4.4 Clinical experience with Pvs25-IMX313/Matrix-M1 ....................................................................27 
5 Investigational Products ...................................................................................................... 29 
5.1 Pvs25-IMX313 ...........................................................................................................................29 
5.1.1 Vaccine supply .............................................................................................................................................29 
5.1.2 Vaccine formulation and packaging .............................................................................................................29 
5.1.3 Storage of Vaccines ......................................................................................................................................29 
5.2 Matrix-M1 ................................................................................................................................29 
5.2.1 Matrix-M1 supply .........................................................................................................................................29 
5.2.2 Matrix-M1 formulation and packaging ........................................................................................................29 
5.2.3 Matrix-M1 Storage and Handling .................................................................................................................29 
5.3 Administration of Pvs25-IMX313 in Matrix-M1 adjuvant ............................................................29 
6 Objectives and endpoints ..................................................................................................... 31 
6.1 Primary Objectives ....................................................................................................................31 
6.1.1 Primary Outcome Measures ........................................................................................................................31 
6.2 Secondary Objectives ................................................................................................................31 
6.2.1 
Secondary Immunological Outcome Measures ......................................................................................31 
7 Description and justification of study design ........................................................................ 32 
7.1 
Study rationale ...................................................................................................................32 
7.2  
Rationale for selected doses ...............................................................................................32 
7.3  
Study Overview ..................................................................................................................32 
7.4 
Study groups ......................................................................................................................33 
7.4.1 First volunteers ............................................................................................................................................33 
7.4.2 Dose escalation ............................................................................................................................................33 
7.4.3 Group 1 ........................................................................................................................................................34 

 
 
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7.4.4 Group 2 ........................................................................................................................................................34 
7.4.5 Group 3 ........................................................................................................................................................34 
7.5 
Duration of study ...............................................................................................................35 
7.5.1 Definition of the start and end of the trial ...................................................................................................35 
7.5.2 Duration of volunteer participation .............................................................................................................35 
7.6 
Potential risks for volunteers ..............................................................................................35 
7.6.1 
Phlebotomy ............................................................................................................................................35 
7.6.2 Vaccinations .................................................................................................................................................35 
7.7 
Potential benefits for volunteers ........................................................................................35 
8 Recruitment and withdrawal for trial volunteers .................................................................. 36 
8.1 
Informed consent ...............................................................................................................36 
8.2  
Inclusion and exclusion criteria ...........................................................................................37 
8.2.1 
Inclusion criteria .....................................................................................................................................37 
8.2.2 
Exclusion criteria .....................................................................................................................................37 
8.2.3 
Prevention of ‘Over Volunteering’ ..........................................................................................................38 
8.2.4 Vaccination and re-vaccination exclusion criteria .......................................................................................38 
8.2.5 Concomitant medications ............................................................................................................................39 
8.3 
Withdrawal of volunteers ...................................................................................................39 
8.4 
Pregnancy ..........................................................................................................................40 
9. Treatment of trial volunteers .............................................................................................. 41 
9.1 
Trial sites ...........................................................................................................................41 
9.2 
Study procedures ...............................................................................................................41 
9.2.1 
Observations ...........................................................................................................................................41 
9.2.2 
Blood tests ..............................................................................................................................................41 
9.2.3 
 Urinalysis ................................................................................................................................................42 
9.2.4 Vaccinations .................................................................................................................................................42 
9.2.5 Management of post-vaccination fevers .....................................................................................................42 
9.2.6 E-diary ..........................................................................................................................................................43 
9.2.7 Photographs .................................................................................................................................................43 
9.3 
Study visits .........................................................................................................................43 
9.3.1 
Screening visits .......................................................................................................................................43 
9.3.2 Enrolment and first vaccination (V1) with Pvs25-IMX313 in Matrix-M1 adjuvant (all groups) ...................44 
9.3.3 Reviews post-vaccination on days 1, 7 and 14 (all groups) ..........................................................................44 
9.3.4 Second vaccination (V2) with Pvs25-IMX313 in Matrix-M1 adjuvant on day 28 (all groups) ......................45 
9.3.5 Reviews post-vaccination on days 29, 35 and 42 (all groups) ......................................................................45 
9.3.6 Third vaccination (V3) with Pvs25-IMX313 in Matrix-M1 adjuvant on day 56 (all groups) .........................45 
9.3.7 Reviews post-vaccination on days 57, 63, 70 and 84 ...................................................................................45 
9.3.8 Long-term follow up post-vaccination on days 140 and 240 .......................................................................46 
10 Assessment of Safety ......................................................................................................... 48 
10.1 Definitions ..............................................................................................................................48 
10.1.1 Adverse Event (AE) .....................................................................................................................................48 
10.1.2 Adverse Reaction (AR)................................................................................................................................48 
10.1.3 Serious Adverse Event (SAE) ......................................................................................................................48 
10.1.4 Serious Adverse Reaction (SAR) .................................................................................................................49 
10.1.5 Suspected Unexpected Serious Adverse Reaction (SUSAR) .......................................................................49 
10.1.6 Foreseeable Adverse Reactions .................................................................................................................49 
10.2 Causality Assessment ..............................................................................................................49 
10.3 Reporting Procedures for all AEs ..............................................................................................50 

 
 
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IRAS Project ID: 302239                                                                                         
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10.4 Reporting Procedures for SAEs .....................................................................................................................52 
10.4.1 Reporting Procedures for SUSARs .........................................................................................52 
10.5 Development Safety Update Report ........................................................................................52 
10.6 AEs of Special Interest .............................................................................................................53 
10.7 Procedures to be followed in the event of abnormal findings ...................................................53 
10.8 Local Safety Monitor ...............................................................................................................53 
10.9 Safety Stopping/Holding Rules ................................................................................................53 
10.9.1 Group holding rules (applies to Groups 1 and 2/3) ...................................................................................53 
10.9.2 Individual stopping rules ............................................................................................................................54 
10.10 Contraception and pregnancy ................................................................................................55 
11 Statistics ............................................................................................................................ 56 
11.1  
Sample size ........................................................................................................................56 
12 Quality control and quality assurance procedures .............................................................. 57 
12.1  
Investigator procedures ......................................................................................................57 
12.2 
Monitoring .........................................................................................................................57 
12.3 
Modification to protocol .....................................................................................................57 
12.4 
Protocol deviation ..............................................................................................................57 
12.5 
Audit & inspection ..............................................................................................................57 
12.6 
Serious breaches ................................................................................................................58 
12.7 
Publication policy ...............................................................................................................58 
12.8 
Intellectual Property ...........................................................................................................58 
13 Ethics ................................................................................................................................. 59 
13.1 
Declaration of Helsinki .......................................................................................................59 
13.2 
Good Clinical Practice .........................................................................................................59 
13.3 
Approvals ...........................................................................................................................59 
13.5 
Volunteer confidentiality ....................................................................................................59 
14 Data handling and record keeping ..................................................................................... 60 
14.1 
Data handling .....................................................................................................................60 
14.2 
Record keeping ..................................................................................................................60 
14.3 
Source data and electronic case report forms (eCRFs) ..........................................................60 
14.4 
Data protection ..................................................................................................................60 
14.5 
Storage and use of data ......................................................................................................60 
15 Financing and insurance .................................................................................................... 62 
15.1 
Financing ...........................................................................................................................62 
15.2 
Insurance ...........................................................................................................................62 
15.3 
Compensation ....................................................................................................................62 
16 Appendices ........................................................................................................................ 63 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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17 References ......................................................................................................................... 64 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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1 SYNOPSIS 
Title 
A Phase Ia study to assess safety and immunogenicity of the Plasmodium 
vivax malaria vaccine candidate Pvs25-IMX313 in Matrix-M1 adjuvant in 
healthy adults living in the UK 
Trial Centre 
Clinical Centre for Vaccinology and Tropical Medicine, Churchill Hospital, 
Old Road, Headington, Oxford, OX3 7LE, UK  
Trial Identifier 
VAC084    
Clinical Phase  
Phase Ia 
Design 
Open label, single-site, first-in-human, Phase Ia study 
Population 
Healthy adults aged 18 – 45 years 
Sample Size 
Total: 24-30 volunteers 
Group 1: 8-10 volunteers receiving three doses of 10 µg Pvs25-IMX313 in  
50 µg Matrix-M1 on days 0, 28 and 56 
Group 2: 8-10 volunteers receiving three doses of 50 µg Pvs25-IMX313 in  
50 µg Matrix-M1 on days 0, 28 and 56 
Group 3: 8-10 volunteers receiving two doses of 50 µg Pvs25-IMX313 in 50 
µg Matrix-M1 on days 0 and 28, followed by one dose of 10 µg Pvs25-IMX313 
in 50 µg Matrix-M1 on day 56 
Follow-up 
duration 
Participants will be followed for approximately 8 months 
Planned trial 
period  
Approximately 11 months from first vaccination of the first volunteer 
Primary Objective To assess safety and tolerability of the Pvs25-IMX313/Matrix-M1 vaccine in 
healthy adult volunteers 
Secondary 
Objectives 
 
 
To assess the humoral and cellular immunogenicity of the Pvs25-
IMX313/Matrix-M1 vaccine, when administered to healthy adult volunteers 
in the various regimes 
To assess ex vivo efficacy of the Pvs25-IMX313/Matrix-M1 vaccine using 
membrane feeding assays, when administered to healthy adult volunteers 

 
 
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IRAS Project ID: 302239                                                                                         
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Investigational 
Products 
1. Pvs25-IMX313 - recombinant protein-nanoparticle malaria vaccine 
candidate 
2. Matrix-M1 – saponin based vaccine adjuvant  
Form 
1. Pvs25-IMX313: liquid, stored between -65oC and -90oC  
2. Matrix-M1: liquid, stored at +2oC to +8oC 
Dose 
Group 1:  
a. Pvs25-IMX313 – 10 µg, administered in a three-dose schedule 
on days 0, 28 and 56 
b. Matrix-M1 – 50 µg, administered with each Pvs25-IMX313 dose 
on days 0, 28 and 56 
Group 2: 
a. Pvs25-IMX313 – 50 µg, administered in a three-dose schedule 
on days 0, 28 and 56 
b. Matrix-M1 – 50 µg, administered with each Pvs25-IMX313 dose 
on days 0, 28 and 56 
Group 3:  
a. Pvs25-IMX313 – 50 µg, administered in a two-dose schedule on 
days 0 and 28 followed by 10 µg administered in a one-dose 
schedule on day 56 
b. Matrix-M1 – 50 µg, administered with each vfs25-IMX313 dose 
on days 0, 28 and 56 
Route 
Intramuscular injection (IM) in the deltoid region of the arm 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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2 ABBREVIATIONS 
AE 
 
 
Adverse event 
AR 
 
 
Adverse reaction 
ß-hCG  
 
Beta human chorionic gonadotropin 
CCVTM 
 
Centre for Clinical Vaccinology and Tropical Medicine 
ChAd63 
 
Chimpanzee adenovirus 63 
CHMI  
 
Controlled human malaria infection 
CI 
 
 
Chief investigator 
COVID-19 
 
Coronavirus disease 2019 
CRF 
 
 
Case report form  
DARC  
 
Duffy antigen receptor for chemokines 
DMFA  
 
Direct membrane feeding assay 
DNA 
 
 
Deoxyribonucleic acid 
DSUR  
 
Development safety update report 
ELISA  
 
Enzyme-linked immunosorbent assay 
ELISPOT 
 
Enzyme-linked immunospot assay 
FDA 
 
 
United States Food and Drug Administration 
G6PD  
 
Glucose-6-phosphate dehydrogenase 
GCP 
 
 
Good clinical practice 
GLA-SE  
 
Glucopyranosyl lipid adjuvant-stable emulsion 
GMP  
 
Good manufacturing practice 
GP 
 
 
General practitioner 
GSK 
 
 
GlaxoSmithKline 
HBsAg  
 
Hepatitis B surface antigen 
HCV 
 
 
Hepatitis C virus 
HIV 
 
 
Human immunodeficiency virus 
HTLV  
 
Human T-lymphotropic virus 
HLA 
 
 
Human leukocyte antigen 
ICH 
 
 
International Conference on Harmonisation 
IgG 
 
 
Immunoglobulin G 
IM 
 
 
Intramuscular 
IMP 
 
 
Investigational medicinal product 
kDa 
 
 
Kilodaltons  
LSM 
 
 
Local safety monitor 

 
 
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mAb 
 
 
Monoclonal antibody 
MFA 
 
 
Membrane feeding assay 
MHRA  
 
Medicines and Healthcare products Regulatory Agency 
MVA  
 
Modified vaccinia Ankara 
P. falciparum  
Plasmodium falciparum 
P. vivax 
 
Plasmodium viax 
PCR 
 
 
Polymerase chain reaction 
PHE 
 
 
Public Health England 
PI 
 
 
Principal investigator 
PIS 
 
 
Participant information sheet 
PPE 
 
 
Personal protective equipment 
PvCS 
 
 
Plasmodium vivax circumsporozoite 
PvCSP  
 
Plasmodium vivax circumsporozoite protein 
PvDBPII 
 
Plasmodium vivax Duffy-binding protein II 
QP 
 
 
Qualified person 
RAS 
 
 
Radiation-attenuated sporozoites 
RBM 
 
 
Roll back malaria 
REC 
 
 
Research Ethics Committee 
RGEA  
 
Research Governance, Ethics and Assurance Team 
RNA 
 
 
Ribonucleic acid 
SAE 
 
 
Serious adverse event 
SAR 
 
 
Serious adverse reaction 
SDEA  
 
Safety Data Exchange Agreement 
SMFA  
 
Standard membrane feeding assay 
SOP 
 
 
Standard operating procedure 
SUSAR  
 
Suspected unexpected serious adverse reaction 
TBV 
 
 
Transmission blocking vaccine 
TOPS  
 
The overvolunteering prevention system 
TMF 
 
 
Trial master file 
TRA 
 
 
Transmission reducing activity 
UOXF  
 
University of Oxford 
VLP 
 
 
Virus-like particle 
VMP001 
 
Vivax malaria protein 1 
WHO  
 
World Health Organisation 

 
 
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IRAS Project ID: 302239                                                                                         
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3 BACKGROUND & RATIONALE 
3.1 The need for a vaccine against Plasmodium vivax malaria  
Malaria is a potentially life-threatening parasitic infection and a major public health concern. 
Globally, 3.4 billion people are thought to be at risk of infection [1, 2], and in 2019 a reported 229 
million cases and 409 thousand deaths occurred from this disease [3]. Children under five years of 
age are the most severely affected, with 67% of all malaria-related deaths occurring in this age 
group around the world in 2019 [3].  
Of the five main species known to infect humans, Plasmodium vivax (P. vivax) is the most 
widespread geographically and the second largest contributor to symptomatic malaria worldwide, 
after Plasmodium falciparum (P. falciparum). There is endemic transmission of P. vivax throughout 
most of the tropics including Africa, Asia, the South Pacific, and Central and South America; an 
estimated 82% of cases comes from just four high-burden countries: India, Pakistan, Ethiopia and 
Sudan [4]. Approximately 2.5 billion individuals, or one-third of the world population are at risk of 
infection from P. vivax and the global burden of symptomatic P. vivax infection was estimated to 
be between 13.7 and 15 million cases in 2017 [4]. 
Since 2000, following concerted worldwide efforts, there have been huge gains in the global fight 
against all-cause malaria, leading to a reduction in overall mortality from 736,000 in 2000 to 
409,000 in 2019 [3]. However, this has been primarily due to a reduction in the incidence of P. 
falciparum in many areas. In fact, in regions where both parasites co-exist, there has been a steady 
increase in the proportion of P. vivax cases [5-8]. 
Moreover, there has been a prevailing attitude that P. vivax malaria is a relatively benign disease. 
This is in fact not the case as, unlike P. falciparum, P. vivax can form a dormant liver stage that can 
activate even years after the initial infection [9]. This means it is a difficult and recurrent disease 
to fully treat. In addition, the greatest risk of mortality from P. vivax is in vulnerable populations 
such as young infants and pregnant women suffering from recurrent episodes of malaria, with a 
cumulative risk of severe anaemia and associated comorbidities [10]. In patients with severe 
symptomatic disease, mortality remains unacceptably high, with estimates between 5 to 25% [2, 
11-14]. 
The socio-economic impact of this infection is not known, but in terms of overall global cost due 
to lost productivity, this is estimated at US$ 1.4 to 4.0 billion per year [15]. Those affected by P. 
vivax malaria are typically poor with inadequate access to affordable healthcare and with little 
financial reserve, perpetuating the cycle of poverty [16]. 
Clinical cases not only result from primary infection but also relapses, which occur weeks to years 
after primary infection. Relapses are due to the ability of the P. vivax parasite to remain dormant 
in the liver in the hynozoite stage, a lifecycle stage not seen with P. falciparum malaria [15]. 
Standard schizonticidal regimes are not effective against hypnozoites. Thus, radical cure of 
dormant parasites requires therapy with the 8-aminoquinolone primaquine, currently the only 
licensed hynozoiticidal anti-malaria, although tafenoquine, a new drug of the same class, has 
recently been approved by the United States Food and Drug Administration (FDA). However, both 
8-aminoquinalones carry a significant risk of causing severe haemolytic anaemia in individuals who 
are deficient in the glucose-6-phosphate dehydrogenase (G6PD) enzyme, an inherited X-lined red 
blood cell enzyme disorder common in tropical and sub-tropical areas [17]. More recently, the 
importance of the cytochrome P450 enzyme, CYP2D6, in metabolism of primaquine to the active 
metabolite has been recognised [18, 19]. A common polymorphism in the CYP2D6 gene for the 
cytochrome P450 enzyme results in poor conversion of primaquine to the active form, resulting in 

 
 
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IRAS Project ID: 302239                                                                                         
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higher treatment failure rates. It is estimated that these two factors combined may make nearly 
40% of the population at risk of P. vivax infection ineligible for primaquine therapy [17]. 
Despite nearly 20 years of advances, the WHO reports that the progress in decline of global 
mortality from all-cause malaria has slowed in recent years. Globally, the malaria mortality rate 
(i.e. deaths per 100,000 population at risk) was reported to be approximately 25 in the year 2000, 
halving to 12 in 2015, but only improving to 10 in 2019 [20]. Furthermore, current strategies are 
complicated by the rise in insecticide-resistant Anopheles mosquitoes and antimalarial resistant 
parasite strains in South East Asia and China, underlining the need for new tools for malaria control 
[21].  
The Roll Back Malaria (RBM) Partnership was launched in 1998 by the WHO, the United Nations 
Children’s Fund, the United Nations Development Programme and the World Bank. A major goal 
of The RBM Partnership is to support the development of vaccines against malaria as a key future 
strategy for reducing mortality from malaria. The WHO Global Vaccine Research Forum has set out 
a strategic framework for malaria vaccine development in the Malaria Vaccine Technology 
Roadmap, defining the goals for global malaria vaccine development community [22]. The 
Roadmap calls for the development of vaccines against both P. falciparum and P. vivax by 2030 to 
achieve two key objectives: protective efficacy of at least 75 percent against clinical malaria and 
reduction of parasite transmission to substantially reduce incidence of human malaria infection, 
enabling elimination in multiple settings.  
Efforts to develop interventions against P. vivax have lagged far behind those for P. falciparum, 
with the majority of malaria research and funding focused on the latter. Unique biological and 
epidemiological characteristics of P. vivax infection mean that control strategies that have been 
primarily targeted against P. falciparum are ineffective. For example, increasing resistance to 
chloroquine [23] and the unsuitability of primaquine in clinically vulnerable groups (e.g. pregnant 
women, infants <6 months of age and glucose-6-phosphate dehydrogenase (G6PD) deficient 
individuals) contribute to poor outcomes from P. vivax infection. An effective vaccine strategy 
against P. vivax would be particularly valuable since the dormant parasite makes pharmacological 
elimination a challenge. However, there are critical bottlenecks in the vaccine development 
process against this P. vivax parasite such as difficulty in long-term culture of P. vivax and lack of 
easy access to a safe controlled human malaria infection (CHMI) model to provide evaluation of 
vaccine efficacy in humans. 
3.2 Lifecycle of the P. vivax malaria parasite 
In the exoerythrocytic stage of human infection, the bite of an infected female Anopheles mosquito 
transmits malaria sporozoites to the human host where they travel via the bloodstream to the liver 
and invade hepatocytes (liver-stage). Here, they mature into merozoites contained within liver 
schizonts, rupturing after 6 to 30 days and causing the release of thousands of merozoites into the 
bloodstream. In parallel, some infected hepatocytes may form hypnozoites, a quiescent stage in 
the liver, resulting in dormant disease with a potential to reactivate after many months, a 
phenomenon unique to P. vivax (and Plasmodium ovale) infection.  
Merozoites then invade reticulocytes (younger, slightly larger erythrocytes) where they multiply 
and after 48 hours cause the reticulocyte to rupture, releasing progeny merozoites that in turn 
invade new reticulocytes (blood-stage). It is the blood-stage of infection that is associated with 
clinical disease and potentially severe or fatal complications. The liver stage is asymptomatic.  
During the blood-stage infection, a small percentage of parasites undergo an alternative 
development pathway, differentiating into sexual male and female gametocyte forms. Over 48 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
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hours, the intra-erythrocytic gametocytes progress through four developmental forms (stages I–
IV) before reaching maturity, as stage V gametocytes [24-27]. The earlier stages of sexual 
development are largely absent from peripheral blood and are thought to sequester 
extravascularly in the bone marrow, before returning to the circulation as mature Stage V 
gametocytes [28]. The mature gametocytes are infectious to the mosquito vector, permitting 
transmission from human to the Anopheline host.  
Following ingestion by the mosquito at the blood meal, environmental shifts, including changes in 
temperature and pH, as well as exposure to xanthurenic acid, trigger gametogenesis [29]. The 
female intra-erythrocytic sexual forms emerge as macrogametes, whilst male gametocytes 
undergo three rounds of deoxyribonucleic acid (DNA) replication before exflagellation to release 
8 motile microgametes [24].  Extracellular macro and microgametes will unite within the 
mosquito’s midgut to undergo fertilisation, forming zygotes, which go on to become motile 
ookinetes that traverse the midgut epithelium. At the basal lamina, following meiotic division, the 
ookinete further develops into the oocyst. Multiple further rounds of division follow, eventually 
culminating in rupture of the oocyst and release of sporozoites. The sporozoites subsequently 
migrate to the mosquito’s salivary glands, where they will be injected into the human host at the 
next blood meal [24-26]. This lifecycle is summarised in Figure 1. 
 
Figure 1: Lifecycle of P. vivax malaria. Figure reproduced from the Centers for Disease Control 
and Prevention website [https://www.cdc.gov/malaria/about/biology/index.html]. 

 
 
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IRAS Project ID: 302239                                                                                         
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3.3 Challenges and directions in P. vivax vaccine development 
The unique characteristics of P. vivax pose specific challenges in studying its biology and designing 
strategies to control it. For example, the lack of a method for continuous culture of P. vivax blood-
stages makes it difficult to perform in vitro growth inhibition assays to identify synergistic 
combinations of blood-stage antigens that can be targeted to inhibit P. vivax blood-stage growth 
with high efficiency. Several groups have recently succeeded in establishing short-term P. vivax 
culture for invasion assays using enriched reticulocytes from cord blood [30], but such methods 
are still dependent on access to fresh P. vivax isolates from malaria patients, limiting the routine 
use of such assays to endemic regions. In the absence of a P. vivax blood-stage culture system, 
production of infected mosquitoes for sporozoite or transmission-stage studies also requires 
access to P. vivax patients in endemic areas.  
Currently, the two most clinically advanced P. vivax candidates are both subunit vaccines based on 
P. vivax circumsporozoite protein (PvCSP), an analogue of the P. falciparum pre-erythrocytic 
target, which have both reached Phase IIa efficacy testing in mosquito-bite delivered sprorozoite 
P. vivax CHMI. The vivax malaria protein 1 (VMP001) vaccine, formulated in the GSK Adjuvant 
System AS01B, was shown to be well-tolerated and immunogenic in healthy US volunteers [31]. 
However, following CHMI at 14 days after the third immunization, the vaccine failed to induce any 
sterile protection, although a small but significant delay in parasitaemia was seen in 59% (16/27) 
volunteers compared to non-vaccinated controls. A Phase IIa trial assessing efficacy of P. vivax 
circumsporozoite protein (PvCS) formulated in Motanide ISA-51 adjuvant in mosquito-bite 
challenge is also now complete, with results published as a pre-print [32]. This study showed that 
most volunteers developed PvCS specific antibody and T-cell responses. Following CHMI at 30 days 
after the third immunization, sterile protection was observed in five of 11 naïve (42%) and four of 
11 semi-immune (36%) volunteers by showing no parasitaemia during the 60 days follow-up.  
To date, repeated exposure to radiation-attenuate sporozoites (RAS) delivered by mosquito bite 
has been the only ‘immunisation’ strategy to demonstrate any protective efficacy in P. vivax CHMI. 
Protection following exposure to irradiated sporozoites was first demonstrated in two volunteers 
in the 1970’s in Maryland, USA [33] but this result was not replicated in larger numbers until a 
recent study in Cali, Colombia [34]. In the latter study, twelve Duffy-positive participants were 
exposed to seven ‘immunisations’, receiving a median of 434 infective bites (range 362-497) from 
irradiated, infected mosquitoes over 56 weeks. Volunteers were then treated with chloroquine 
and primaquine to clear any plasmodium infections that may have developed during the 
immunization phase before challenge at 8 weeks after the last RAS exposure. ‘Immunised’ 
participants underwent CHMI alongside two controls, who were Duffy-positive individuals exposed 
to non-irradiated, non-infected mosquitoes and seven controls who were Duffy-negative 
individuals who had received the same immunization regime. Sterile efficacy was demonstrated in 
42% (5/12) Duffy-positive participants, all of which were female. Given the apparent high-dose 
exposure required to afford this level of protection, and the current lack of availability of 
cryopreserved, purified P. vivax sporozoites, this approach may be limited as a vaccination 
strategy. However, transcriptomic and anti-parasite antibody profiling may offer important 
insights into correlates of protection and new pre-erythrocytic vaccine targets.  
Apart from the viral-vectored Plasmodium vivax Duffy-binding Protein II (PvDBPII) vaccine 
candidates [35], the only other P. vivax blood-stage vaccine currently in active clinical development 
is the recombinant protein PvDBPII vaccine. This vaccine targets the cysteine-rich, region II of 
PvDBP, which serves as the functional receptor-binding domain for the essential interaction for 
reticulocyte invasion with Duffy antigen receptor for chemokines (DARC). A Phase I trial has 

 
 
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IRAS Project ID: 302239                                                                                         
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demonstrated that the PvDBPII vaccine, formulated with the glucopyranosyl lipid adjuvant-stable 
emulsion (GLA-SE), is safe and induces functional strain-transcending antibodies, which inhibit the 
PvDBP-DARC interaction in vitro [36]. 
All vaccines currently in clinical development, and almost all those in pre-clinical stages, target 
individual stages of the parasite’s life cycle and are based on single antigens (Table 1) [37]. 
However, the development of an effective vaccine will likely require combination of multiple 
antigens that provide synergy to achieve high efficacy. For example, achieving high rates of blood-
stage growth inhibition may require targeting a combination of key blood-stage antigens involved 
in reticulocyte invasion. In addition, a combination of blood-stage antigens with liver-stage 
antigens may be needed to achieve high efficacy, and inclusion of antigens from sexual and 
mosquito-stages may be needed to inhibit transmission. It is therefore necessary to initiate efforts 
to combine antigens both within and across developmental stages to achieve synergy and attain 
the goal of developing a vaccine for P. vivax malaria with high efficacy.

 
 
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IRAS Project ID: 302239                                                                                         
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Vaccine candidate  
Development phase  
Lifecycle stage  
Antigen  Delivery system  
ChAd63-PvTRAP/MVA-PvTRAP [38] 
Pre-clinical  
Liver-stage  
PvTRAP  
Prime-boost, viral vectors  
CSV-S,S [39, 40]  
Pre-clinical  
Liver-stage  
PvCSP  
HBsAg fusion-AS01B  
PvCSP-LSP [41-43] 
Phase I 
Liver-stage  
PvCSP  
Synthetic peptides-Montanide ISA 720  
VMP001 [44] 
Phase I/IIa 
Liver-stage  
PvCSP  
Rec. protein-AS01B  
ChAd64-PvAMA1/MVA-PvAMA1 [45] 
Pre-clinical  
Blood-stage  
PvAMA1  Rec. protein-adjuvant  
PvDBPII-DEKnull [46] 
Pre-clinical  
Blood-stage  
PvDBP  
Rec. protein  
PvMSP119 [47, 48] 
Pre-clinical  
Blood-stage  
PvMSP1  Rec. protein-Montanide ISA720  
PvDBPII [49, 50] 
Phase I  
Blood-stage  
PvDBP  
Rec. protein-GLA-SE  
ChAd63-PvDBPII/MVA-PvDBPII [51] 
Phase I/IIa  
Blood-stage  
PvDBP  
Prime boost, viral vectors  
PvDBPII/Matrix-M1 [NCT04201431] 
Phase I/IIa 
Blood-stage 
PvDBP 
Rec. protein-adjuvant 
AnAPN1 [52] 
Pre-clinical  
Mosquito midgut Ag  AnAPN1  Rec. protein-adjuvant  
Pvs28 [53]  
Pre-clinical  
Transmission-stage  
Pvs28  
Rec. protein-adjuvant  
Pvs25H [54, 55]  
Phase Ia END  
Transmission-stage  
Pvs25  
Rec. protein-Alhydrogel; Rec. protein-Montanide ISA 51  
Table 1:    P. vivax vaccine candidates under development. Adapted from Mueller et al (2015) [37]. Rec=recombinant. 

 
 
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IRAS Project ID: 302239                                                                                         
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3.4 Rationale for a transmission blocking vaccine against P. vivax malaria 
Transmission blocking vaccines (TBVs) aim to induce antibodies against sexual-stage antigens. 
These include antigens expressed during the gametocyte stages (inside RBCs) in humans and also 
antigens that are exclusively expressed once the parasite is in the mosquito midgut (gametes, 
zygotes and ookinetes). The induced antibodies are taken up in the blood meal and act to prevent 
sexual development in the mosquito host, thus blocking onward malaria transmission. Uniquely, 
TBVs do not aim to provide protection against infection or disease in the vaccinated individual but 
instead to reduce risk of infection at the community and population level, through interruption of 
the transmission cycle. It is widely accepted that in order to eliminate and eventually eradicate 
malaria, a multi-stage malaria vaccine will be required and a TBV will be an essential component 
for this [56]. In addition to reducing transmission, a TBV can also be used to combat the spread of 
parasite strains that might arise from immune or vaccine pressure.  
The first observations of transmission blocking immunity were reported in 1958, when lower 
infectivity to mosquitoes of the avian Plasmodium spp., P. fallax and P. gallinaceum, were 
described in turkeys and chickens which had been immunised with formalin-killed parasitised 
gametocytes  [57] [57] [57]. However, it was not until almost 20 years later, in 1976, that the 
phenomenon was reproduced, and the underlying mechanism delineated as antibody mediated 
inhibition of sexual parasite development in the mosquito midgut [58]. In 1983, following 
developments in methods for in vitro culture of the sexual stages of P. falciparum, mice were 
immunised with P. falciparum gametes leading to the generation of the first transmission-blocking 
monoclonal antibodies (mAbs), which could arrest parasite development in the mosquito midgut 
[59, 60]. Through surface radioisotope labelling of gametes followed by precipitation with these 
transmission-blocking mAbs, the first list of transmission-blocking targets were identified [60].  
The gold standard ex vivo model for establishing transmission-reducing (conferring decreased 
oocyst intensity) or transmission-blocking (reduction in mosquito infection prevalence) effects, is 
the standard membrane feeding assay (SMFA), wherein mosquitoes are fed cultured gametocytes 
in the presence of whole serum or purified IgG, from either a test or control source. Mosquitoes 
are then dissected approximately one week later and oocyst development in the mosquito is 
quantified by microscopic visualisation using mercury-bromide staining and compared between 
intervention and control groups. Since P. vivax parasites cannot be cultured, direct membrane 
feeding assays are done where the source of the parasite is infected individuals in endemic regions.  
 
3.5 Pvs25 as an antigen 
Pvs25 is a member of the P25 family of cysteine-rich 25 kDa proteins. The ookinete plasma 
membrane of all Plasmodium species has approximately 10-15 proteins on its surface, of which 
two predominate: P25 and P28 [61]. Pvs25 is present in the P. vivax species, and has an important 
role for parasite survival in the mosquito midgut, as well as possible role in traversal of the midgut 
epithelium [61-63]. Based on structural analyses, the Pvs25 molecule is thought to be triangular 
and flat, and extensively expressed on the ookinete surface, forming a protective interlocking 
sheet [64].  
Pre-clinical studies in rodent models show that Pvs25 produced in a plant-based system fused to a 
modified lichenase carrier protein (Pvs25-FhCMB) produced an in vivo transmission blocking effect 
of up to 65% reduction in intensity and 54% reduction in prevalence when used with Abisco-100 
adjuvant. In addition, when this platform was combined with heterologous prime-boost 

 
 
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IRAS Project ID: 302239                                                                                         
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vaccination with viral vectors expressing Pvs25, there was a 74% reduction in intensity and 54% 
reduction in prevalence. This observation was confirmed by direct membrane feeding assays [65]. 
Additionally, studies of the P. falciparum equivalent, Pfs25, provide further insights into the 
potential of Pvs25 to be a suitable TBV candidate. Since identification as a target of very effective 
transmission-blocking mAbs in BALB/c mice, Pfs25 has been extensively tested pre-clinically as a 
TBV candidate, reliably demonstrating that vaccine-induced Pfs25-specific IgG are effective in 
blocking transmission [60, 66-68]. 
3.6 Clinical trials of Pvs25-based vaccines  
The first clinical evaluation of a P. vivax transmission-blocking vaccine was in a 2005 Phase 1a 
clinical trial of the Pvs25 antigen formulated in Alhydrogel® [54]. The Pvs25 antigen was expressed 
in Saccharomyces cerevisiae, purified, and adsorbed onto Alhydrogel®. Amongst the 10 volunteers 
in each of the three dose groups (5, 20 and 80 µg) evaluated, the vaccine was found to be well-
tolerated with no serious adverse reactions observed. However, there was only modest 
transmission reducing activity (TRA) detected (no transmission blocking activity) in the membrane 
feeding assays, with degree of reduction correlating with antibody concentration. 
This result prompted efforts to investigate enhancement of antibody responses using a more 
potent adjuvant [55]. Here, in a Phase 1a clinical trial setting, Pvs25 expressed in S. cerevisiae was 
evaluated alongside the P. falciparum homolog, Pfs25 produced in Pichia pastoris. Both antigens 
were formulated in Montanide ISA 51 adjuvant and participants received either of the two vaccine 
candidates. However, observed suspected unexpected serious adverse reactions led to early 
cessation of the clinical trial. Two volunteers developed erythema nodosum at 18 days after 
vaccination with Pvs25/ISA 51 whilst two further volunteers developed a leukaemoid reaction 
deemed possible related to vaccination. One of these was at 14 days following vaccination with 
Pfs25/ISA 51 and the other was at 120 days following vaccination with Pvs25/ISA 51. High local 
reactogenicity was also seen, including acute local tenderness and swelling as well as induration 
at approximately 2 weeks after vaccination. Given that the systemic reactions were seen with 
Pvs25, previously well tolerated with Alhydrogel®, and that no previous severe systemic reactions 
had been reported for Montanide ISA 51, the severe systemic reactogenicity was deemed to be 
related to the specific antigen/adjuvant combination. 
3.7 IMX313  
IMX313 is based on a chimeric version of the oligomerization domains from chicken complement 
inhibitor C4-binding protein (C4bp), and forms homogeneous, self-assembling heptamers of the 
antigen fused to it. IMX313 is a 55 amino acid sequence with less than 20% homology to the 
sequence of the human protein. This C4bp oligomerization domain has been shown to 
spontaneously form soluble heptameric structures (termed nanoparticle in this study) when 
expressed in E. coli, and protein antigens fused to these domains have been shown to improve 
antibody responses over the same amount of monomeric antigen. In addition, mice immunized 
with the blood-stage malaria vaccine candidate antigen MSP119 fused to IMX313 (expressed in E. 
coli) were protected against challenge with a lethal dose of P. yoelii parasites [69]. Immunisation 
of mice and non-human primates with the Mycobacterium tuberculosis antigen 85A fused to 
IMX313 in both DNA vaccines and viral vectors also demonstrated that IMX313 enhances T-cells 
responses, with consistent increases in both CD4+ and CD8+ T cell responses [70]. Fusion of 
IMX313 with Pvs25 leads to formation of immunogenic heptamers, which may be expressed in situ 
when encoded within recombinant viral vectors or as a secreted protein-nanoparticle in Pichia 
pastoris, within a single expression system. 

 
 
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Clinically, IMX313 was first assessed in the recombinant MVA85A-IMX313 tuberculosis vaccine 
candidate, administered intradermally, in a Phase Ia trial at the Jenner Institute in Oxford. In this 
first-in-human, dose escalation study, a total of 18 volunteers received MVA85A-IMX313, with 6 
receiving a dose of 1 x 107 pfu prior to enrolment of the next groups who received either MVA85A-
IMX313 or MVA85A, both at a dose of 5 x 107 pfu. There were no safety concerns and no significant 
difference in the number of adverse events (AEs) between the MVA85A-IMX313 group and the 
MVA85A group, with the majority of AEs (both local and systemic) being mild in nature, although 
there were no significant differences in immunogenicity between MVA85A and MVA85A-IMX313 
groups [70]. Subsequently, a Phase Ia study assessing the ChAd63/MVA Pfs25-IMX313 candidate 
[study number: NCT02532049] was conducted, demonstrating favourable safety and tolerability 
for the vaccine [71]. Here, the vaccine was well tolerated at all dose levels tested, with most local 
and systemic adverse events being mild in severity, no apparent increase in adverse events with 
boost vaccination and no participants withdrawn due to AEs. 

 
 
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IRAS Project ID: 302239                                                                                         
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4 PVS25-IMX313/MATRIX-M1 VACCINE DEVELOPMENT 
4.1 Characteristics of Pvs25-IMX313/Matrix-M1 
Pvs25-IMX313 is a protein-nanoparticle vaccine based on the P. vivax transmission-blocking target 
antigen Pvs25, genetically fused to the IMX313 oligomerization domain. The Pvs25 protein is based 
on the sequence from the Sal I P. vivax strain [72]. The recombinant protein-nanoparticle is 
expressed and secreted in the Pichia pastoris expression system [73]. 
Matrix-M1 is a potent, saponin-based adjuvant, comprising partially purified extracts of the bark 
of the Quillaja saponaria Molina tree, phosphatidylcholine and cholesterol, formulated as a 40nm-
sized complex. Matrix-M has been shown to efficiently activate and recruit immune cells to the 
draining lymph node, including T-cells, B-cells, NK-cells, dendritic cells and granulocytes, which 
may lead to enhanced antigen presentation [74]. Matrix-M1 has demonstrated a favourable safety 
profile, as well as the enhancement of cellular and humoral immune responses to a range of 
vaccine [74-76].  
Pvs25-IMX313 and Matrix-M1 adjuvant will be mixed immediately prior to administration for co-
administration. 
4.2 Vaccine development strategy 
The clinical development of Pvs25-IMX313 in Matrix-M1 adjuvant is aimed towards the production 
of an effective transmission-blocking malaria vaccine for individuals in malaria endemic regions. 
This vaccine would be used in a three-dose vaccination regimen. 
This first-in-human study aims to provide an initial assessment of the frequency and magnitude of 
adverse events following vaccination with Pvs25-IMX313 in the Matrix-M1 adjuvant for evaluation 
of safety, as well as immunogenicity and ex vivo efficacy via DMFA. Data generated from this study 
will inform progression to a Phase Ib trial in an endemic region to evaluate the safety and 
immunogenicity of Pvs25-IMX313/Matrix-M1 at three doses, in a dose-escalation study, to 
establish the optimal dose and schedule in an endemic setting.  
4.3 Pvs25-IMX313 immunogenicity and ex vivo efficacy in pre-clinical studies  
The addition of the IMX313 domain to the Pvs25 protein allows heptamerization of the protein. 
This heptameric form has been shown to be significantly more immunogenic than monomeric 
Pvs25 in preclinical studies involving mice. Purified IgG from immunized mice efficiently inhibits 
parasite development in mosquito stages as analysed by DMFA. 
In pre-clinical studies, when formulated with Alhydrogel, Pvs25-IMX313 constructs induced 
significantly higher antibody titres than Pvs25 alone in CD1 mice after one immunisation (Figure 
2). Two variants of Pvs25-IMX313 were compared in this study, they were different only in the 
purification tags fused at the C terminus of the construct and exhibited no obvious difference in 
the antibody response induced. Furthermore, Pvs25-IMX313 constructs were able to induce 
similar levels of antibody response compared to constructs using Pvs25 displayed on virus-like-
particles (VLPs), namely HBsAg (Figure 2).    
 
 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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Figure 2. IMX313 oligomerization enhances Pvs25 immunogenicity: CD1 mice were immunized with 1 
µg (based on total protein content) of Pvs25-IMX313C, or Pvs25-IMX313P, or Pvs25::SpyTag-SpyCatcher::HBsAg, or 
soluble Pfs25 antigen. All vaccines were formulated with Alhydrogel. Mice received 2 doses, 3 weeks apart (on day 0 
and 21), and were sacrificed three weeks after the second dose (day 41). Anti-Pvs25 IgG levels were measured by 
endpoint ELISA. Each symbol represents serum sample from an individual mouse; lines represent the median of each 
group. Kruskal-Wallis with Dunn’s multiple comparisons test was performed to compare the three groups at the 
different timepoints: * p < 0.05, ** p < 0.01, *** p < 0.001. ST = SpyTag; SC = SpyCatcher. 
 
Based on functional activity as assessed by DMFA (results were read as TRA where higher values 
are better), the quality of the anti-Pvs25 antibody response induced by immunisation with both of 
the Pvs25-IMX313 protein-nanoparticles were significantly improved in comparison to that 
induced by soluble Pvs25 (Figure 3). Furthermore, at 250 µg/mL IgG concentration, both of the 
Pvs25-IMX313 constructs demonstrated higher %TRA than the construct using Pvs25 displayed on 
VLP (Figure 3). 
Pvs25
Pvs25-IMX313C
Pvs25-IMX313P
Pvs25::ST-SC::HBsAg
Pvs25
Pvs25-IMX313C
Pvs25-IMX313P
Pvs25::ST-SC::HBsAg
3
4
5
6
7
Endpoint titre (log10)
Day 20
Day 41
<2
✱✱
✱✱

 
 
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IRAS Project ID: 302239                                                                                         
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Figure 3 Pvs25-IMX313 induce IgG with higher TRA than soluble Pvs25: CD1 mice were immunized 
with 1 µg (based on total protein content) of Pvs25-IMX313C, or Pvs25-IMX313P, or Pvs25::SpyTag-
SpyCatcher::HBsAg, or soluble Pfs25 antigen, formulated with Alhydrogel. Mice received 2 doses, 3 weeks apart (on 
day 0 and 21), and were sacrificed three weeks after the second dose (day 41). Transmission-reducing efficacy of IgG 
induced by the vaccines was tested by DMFA: total IgG was purified from the pooled serum of each group (3 weeks 
post-boost), and the purified IgG was mixed with P. vivax  gametocytes at concentrations indicated on the X-axes, and 
fed to A. stephensi mosquitoes (n=20 per test group). IgG from ovalbumin-immunized mice was used as a negative 
control (Contrl). Midguts were dissected 7 days post-feeding. Data points represent the number of oocysts in 
individual mosquitoes; and lines show the arithmetic mean. Immunization groups are indicated on the X axis. TRA = 
Transmission-reducing activity (% inhibition in mean oocyst count per mosquito). ST=SpyTag. SC = SpyCatcher. 
 
In addition, Matrix-M and Alhydrogel have been compared head-to-head for formulation with 
Pvs25-IMX313. In our preclinical studies, Matrix-M was able to induce a higher antibody response 
compare to Alhydrogel in CD1 mice (Figure 4).  
 
 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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Figure 4. IMX313 oligomerization and Matrix-M adjuvant enhance Pvs25 immunogenicity: CD-1 
mice received 3 doses of 0.1 µg Pvs25-IMX313, formulated with either Alhydrogel or Matrix-M. Injections 
at day 0, 1 month, 2 months. Data represented here are from 2 weeks post 3rd immunization. Anti-Pvs25 
IgG levels were measured by endpoint ELISA. Each symbol represents the median of the group. Mann-
Whitney test (2-tailed); p = 0.015 
 
 4.4 Clinical experience with Pvs25-IMX313/Matrix-M1 
There is no prior clinical experience with the Pvs25-IMX313 protein in the Matrix-M adjuvant. The 
Pfs25-IMX313 insert, however, has been clinically tested in a viral vectored vaccination regimen 
ChAd63/MVA Pfs25-IMX313 in a previous Phase Ia clinical trial in Oxford, completed in May 2017 
(VAC062; NCT02532049). The Pvs25-IMX313 construct is exactly the same as the Pfs25-IMX313 
construct, except for a difference in the sequence of the 25 gene which enables the vaccine to 
target P. vivax instead of P. falciparum. 
A total of twenty-six volunteers were vaccinated with ChAd63 Pfs25-IMX313, at doses of either 5 
x 109 vp (n=5) or 5 x 1010 vp (n=21). Sixteen volunteers went on to receive a boost with MVA Pfs25-
IMX313 on day 56, of which half received a dose of 1 x 108 pfu and half a 2 x 108 pfu dose. All 
vaccinations were administered intramuscularly. The ChAd63 Pfs25-IMX313 and MVA Pfs25-
IMX313 vaccines were demonstrated to be safe and well-tolerated at all doses, with the majority 
of AEs being mild in nature. Priming vaccination with 5 × 1010 vp ChAd63 Pfs25-IMX313 followed 
by MVA Pfs25-IMX313 boost at either dose induced antigen-specific IgG responses in all 
volunteers, to levels which were significantly higher than following prime alone. However, titres 
of anti-Pfs25 IgG antibody responses were low with the peak response at 10.24 µg/mL on day 74. 
Pfs25-specific T cell responses were also detected in all volunteers, as well as Pfs25-specific B cell 
responses. The functional activity of vaccine-induced antibody responses were assessed by SMFA. 
With a median TRA of 7.2% (range -5.8% to 37.3%) in the group boosted with the lower dose of 
MVA and 25.3% (range 10.2% to 41.3%) in the higher MVA dose group, overall TRA was low, with 
serum from only 1/8 individuals boosted with the higher MVA Pfs25-IMX313 dose demonstrating 
weak but significant reduction in oocyst intensity. TRA correlated with anti-Pfs25 antibody 
concentrations (Spearman r =0.5176; P = 0.0423). Given that previous human trials have indicated 
that high levels of specific IgG are required for induction of significant transmission reducing 
activity, it is likely that lack of TRA observed relates to the low titres of anti-Pfs25 antibodies (a 
maximum of around 10µg/mL) induced by immunisation with ChAd63/MVA Pfs25-IMX313.  

 
 
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As expected, due to limited similarity between IMX313 and human C4bp, there were no detectable 
vaccine-induced auto-reactive antibody or T-cell responses to the oligomerisation domain of 
human C4bp in any of the study groups [71].  
The Pfs25-IMX313/Matrix-M1 vaccine was previously approved for testing in a clinical trial 
(VAC077; NCT: 04130282). A single volunteer was recruited to the trial and vaccinated with no 
significant adverse events. However, the trial had to be terminated due to the COVID-19 pandemic. 
A similar, larger Phase 1 study (VAC082; NCT: 04271306) is currently being conducted at the Ifakara 
Health Institute in Bagamoyo, Tanzania. This study aims to evaluate the safety, immunogenicity 
and ex vivo efficacy of Pfs25-IMX313/Matrix-M1 in healthy adults and young children. Although 
recruitment is still in the early stages, there have been no safety concerns thus far.  
Matrix-M adjuvant, in 1 or 2 formulations (named Matrix-M1 or Matrix-M2), has been 
administered to tens of thousands of individuals in clinical trials across the USA, Europe and 
Australia. In addition to trials involving vaccines to respiratory syncytial virus, influenza and SARS-
CoV-2, two malaria pre-erythrocytic candidate vaccines (ChAd63/MVA ME-TRAP and VLP R21) 
have been administered with Matrix-M1 adjuvant in four phase I and phase I/II trials conducted in 
Oxford and Burkina Faso (ClinicalTrials.gov identifiers: NCT02572388, NCT02925403, 
NCT01669512, NCT02905019). A phase III trial involving 4800 children has recently started in Africa 
with sites in Burkina Faso, Mali, Kenya and Tanzania. Available safety data demonstrates that the 
Matrix-M adjuvant is well-tolerated with acceptable short-term reactogenicity and unremarkable 
long-term safety profile with no suspected unexpected serious adverse reactions (SUSARs) in 
COVID-19 vaccine clinical trials conducted by Novavax  collaborators in subjects who received 
Matrix-M [77]. 
The profile of adverse events following vaccination with Pvs25-IMX313 may be partially predicted 
from previous studies assessing protein vaccines using the Matrix-M adjuvant. Local adverse 
events are likely to include injection site pain, erythema, swelling, itching and warmth. Forseeable 
systemic adverse events would include headache, fatigue, myalgia, arthralgia, malaise, 
feverishness, fever and nausea. 
 

 
 
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IRAS Project ID: 302239                                                                                         
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5 INVESTIGATIONAL PRODUCTS 
5.1 Pvs25-IMX313 
5.1.1 Vaccine supply 
Pvs25-IMX313 is manufactured by GenIbet (Oeiras, Portugal), where the vaccines are formulated 
and vialed. Xerimis Ltd (Reading, UK) will perform certification and QP release, labelling and 
storage of the Pvs25-IMX313 for investigational use only, and later supply to the clinical site 
(CCVTM). 
5.1.2 Vaccine formulation and packaging 
Pvs25-IMX313 protein drug product is formulated in 20 mM Tris, 150mM NaCl, pH 7.4 (adjusted 
by sodium hydroxide), at not less than 0.25 mg/ml. The final 0.2 µm filtered drug product was 
dispensed manually using an electronic dispenser preset to deliver 500 µl into type 1, 2 ml glass 
vials. The vials were sterilized and depyrogenated with a 13 mm grey bromobutyl rubber stopper 
supplied by Adelphi Healthcare Packaging (Haywards Heath, UK) and a 13 mm complete tear off, 
clear lacquered aluminium seal. The containers and closures were tested for compliance with 
defined specifications. 
5.1.3 Storage of Vaccines  
All vaccines will be stored between -65°C and –90°C. All movements of the study vaccines will be 
documented. Vaccine accountability, storage, shipment and handling will be in accordance with 
relevant local Standard Operating Procedures (SOPs) and forms. 
5.2 Matrix-M1 
5.2.1 Matrix-M1 supply 
Matrix-M1 will be supplied to Xerimis by Novavax AB (Uppsala, Sweden), a subsidiary of Novavax, 
Inc (Gaithersburg, USA), where the adjuvant is formulated and vialed. Matrix-M1 will be labelled 
for investigational use only by Xermis Ltd before supplying the adjuvant to the clinical site. 
All adjuvants will be certified for release by a qualified person (QP) at Xerimis Ltd.  
5.2.2 Matrix-M1 formulation and packaging  
Matrix-M1 is formulated at a concentration of 0.375mg/ml in PBS. The drug product is filled into 
sterile 3 mL glass vials. Matrix-M1 (85 parts Matrix A and 15 parts of Matrix C) is obtained by simply 
mixing Matrix A and C, followed by dilution in PBS, filtration though a 0.22 μm filter and filling into vials 
in a volume of 0.75 ml. Matrix-M1 is a slightly brown to colourless, slightly opalescent, non-viscous 
liquid. 
5.2.3 Matrix-M1 Storage and Handling  
Matrix-M1 is stored refrigerated at 2 to 8°C and protected from light. All movements of the 
adjuvant will be documented. Accountability, storage, shipment and handling of Matrix-M1 will be 
in accordance with relevant local SOPs and forms. 
5.3 Administration of Pvs25-IMX313 in Matrix-M1 adjuvant 
A mixture of Pvs25-IMX313 at a dose of 10 μg or 50 μg, with 50 μg of Matrix-M1 will be 
administered to all volunteers. Matrix-M1 and Pvs25-IMX313 will be mixed immediately prior to 

 
 
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administration and will be administered intramuscularly within one hour of thawing of Pvs25-
IMX313.  
All vaccines will be administered intramuscularly in the deltoid muscle of the non-dominant arm 
preferentially, according to SOP VC002 Vaccination. The vaccinating investigator will wear gloves 
and eye protection. During administration of the vaccines, Advanced Life Support drugs and 
resuscitation equipment will be immediately available for the management of anaphylaxis. On 
vaccination day, vaccines will be allowed to thaw to room temperature and administered within 1 
hour. A single vial of vaccine will be used per dose administered and any remaining vaccine not 
used will be discarded. 
 

 
 
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6 OBJECTIVES AND ENDPOINTS 
6.1 Primary Objectives  
To assess safety and tolerability of the Pvs25-IMX313/Matrix-M1 vaccine in healthy adult 
volunteers. 
6.1.1 Primary Outcome Measures 
The specific endpoints for safety and reactogenicity will be actively and passively collected data on 
adverse events. The following parameters will be assessed:  
• Occurrence of solicited local reactogenicity signs and symptoms for 7 days following each 
vaccination  
• Occurrence of solicited systemic reactogenicity signs and symptoms for 7 days following 
each vaccination  
• Occurrence of unsolicited adverse events for 28 days following the vaccination  
• Change from baseline for safety laboratory measures for 28 days following vaccination 
• Occurrence of serious adverse events during the whole study duration  
Solicited and unsolicited AE data will be collected at each clinic visit. It will be collected from e-
diary cards, clinical review, clinical examination (including observations) and laboratory results. 
This AE data will be tabulated, detailing frequency, duration and severity of AEs. 
Haematological and biochemical laboratory values will be presented according to local grading 
scales. Serious adverse events (SAEs), adverse events (AEs) of special interest and withdrawal due 
to AE(s)/SAE(s) will be described in detail. Volunteers will be followed for approximately 8 months 
following initial dose of Pvs25-IMX313/Matrix-M1 and approximately 6 months following the third 
dose.  
6.2 Secondary Objectives 
To assess the humoral and cellular immunogenicity of the Pvs25-IMX313/Matrix-M1 vaccine, when 
administered to healthy adult volunteers. 
To assess ex vivo efficacy of the Pvs25-IMX313/Matrix-M1 vaccine, when administered to healthy 
adult volunteers. 
6.2.1 Secondary Immunological Outcome Measures 
Pvs25-specific immunogenicity will be assessed by a variety of immunological assays, with 
comparison before and after vaccination. The main outcome measures will be humoral and B cell 
responses to the Pvs25 protein – total IgG, isotypes and avidity; T cell responses to Pvs25 by ex 
vivo enzyme-linked immunospot assay (ELISpot) and flow cytometry assays.  
Ex vivo functional blocking activity of purified IgG against P. vivax will be assessed by membrane 
feeding assays .  
Other established and exploratory immunology assays may be carried out, which may include 
collaboration with other specialist laboratories within or beyond Europe. This would involve 
transfer of serum/plasma, but samples would be anonymised. Volunteers will be consented for 
this. 
The secondary timepoints will be days 1, 29, 57, 140 and 240. 

 
 
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7 DESCRIPTION AND JUSTIFICATION OF STUDY DESIGN  
7.1 
Study rationale  
As outlined above, the currently available methods for preventing and treating malaria remain 
inadequate. Although focused efforts have led to substantial reductions in morbidity and mortality 
worldwide, progress has now stalled. An effective vaccine against P. vivax would contribute 
substantially to disease control and elimination efforts. Currently, only a limited number of P. vivax 
candidate vaccines have reached clinical trials (summarised in Table 1). These include two vaccines 
targeting the pre-erythrocytic stage, both of which target the P. vivax circumsporozoite protein 
(PvCSP) [31, 42]. CHMI efficacy data for only one of these candidates, VMP001/AS01B, is currently 
published [31] and whilst the vaccine was well-tolerated and immunogenic, there was limited 
efficacy following sporozoite challenge. In addition, a blood-stage vaccine targeting region II of the 
P. vivax Duffy-binding protein (ChAd63-MVA PvDBP_RII) has been studied in a Phase 1a study [35] 
and been shown to be well-tolerated with a favourable safety profile and immunogenicity. A 
related vaccine targeting the PvDBP region but formulated in Matrix-M1 is currently being studied 
in a Phase I/IIa study [VAC079, NCT04201431]. As part of the same European Commision grant 
funding the VAC084 study, a blood-stage vaccine is also being developed with the aim that blood-
stage and TBV components will be combined later to develop a multi-stage vaccine.  
Although Pvs25-IMX313 in Matrix-M1 has not previously been studied in humans, the P. 
falciparum equivalent, Pfs25-IMX313 in Matrix-M1, is currently undergoing Phase 1 study at the 
Ifakara Health Institute, Bagamoyo, Tanzania.  
This small, first-in-human study will provide initial data on the safety profile of vaccination with 
Pvs25-IMX313 in the Matrix-M1 adjuvant, as well as immunogenicity and ex vivo efficacy via 
membrane feeding assays.  
7.2  Rationale for selected doses 
The two doses of 10 μg and 50 μg Pvs25-IMX313 in 50 μg Matrix-M were selected on the basis of 
experience with other candidate protein vaccines which have been tested at the same dose in 
humans with the same adjuvant. These include the VAC077 and VAC082 trial of Pfs25-IMX313 in 
Matrix-M1 which is structurally very similar to Pvs25-IMX313. 
Mouse studies suggest that a vaccine dose of up to 250 μg/kg is safe. In this study we are testing 
a maximum dose of 50 μg Pvs25-IMX313 in 50 μg Matrix-M. This is equivalent to a dose of 0.8 
μg/kg in humans. This means that there is a safety margin based on body weight of 250/0.8 = 312-
fold in humans as compared to mice.  
7.3  Study Overview 
This is an open-label, single-centre, non-randomised, first-in-human Phase Ia study to assess the 
safety and immunogenicity of the Pvs25-IMX313 vaccine, administered in Matrix-M1 adjuvant.  
Up to 30 healthy, malaria-naïve adults aged between 18 and 45 years will be recruited into one of 
three groups (n=8-10 per group) at the Centre for Clinical Vaccinology and Tropical Medicine 
(CCVTM), Oxford over approximately 11 months.  
All volunteers will receive three doses of Pvs25-IMX313 at 10 µg or 50 µg, in 50 µg Matrix-M1, 
administered intramuscularly and given four weeks apart. Enrolment will be staggered. Initially, a 
single volunteer from Group 1 will receive the first vaccination alone, followed by clinical review 
on the day after vaccination and further monitoring via a diary card for at least a further 24 hours. 

 
 
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Provided there are no safety concerns, a further two volunteers from Group 1 may then be 
vaccinated, at least 1 hour apart. Following review of the second and third volunteers at the 48-
hour timepoint, an internal safety review will be conducted. If no safety concerns are identified, 
the remaining volunteers in the group will be vaccinated. The staggered enrolment process will be 
repeated for Group 2 for the first three volunteers. Following this, recruitment to Groups 2 and 3 
will occur in parallel.  
Volunteers will attend regular follow-up visits after each of the three vaccinations and will self-
report adverse events in the post-vaccination period via a diary card. Blood will be taken at regular 
intervals to assess safety outcomes via laboratory adverse events, as well as the immune response 
to vaccination. Volunteers will be followed until approximately 8 months from the time of 
enrolment. 
All vaccinations and follow-up visits will be performed at the CCVTM in Oxford.  
7.4 
Study groups 
There will be three study groups (Group 1, Group 2 and Group 3), each comprising 8-10 volunteers. 
There will be staggered enrolment for Groups 1 and 2 as described below in Section 7.3.1. Any 
volunteer who is withdrawn or withdraws from the study may be replaced at the discretion of the 
Investigators.  
7.4.1 First volunteers 
The first volunteer from Group 1 to receive Pvs25-IMX313/Matrix-M1 will be enrolled and 
vaccinated alone. Providing there are no safety concerns at 48 hours after the first volunteer has 
been vaccinated, the second and third volunteers from Group 1 will be vaccinated. Following a 
further 48 hour period, an internal safety review will be conducted involving the review of 
participant e-diaries by the Chief Investigator with the Lead Clinician. If at this point there are no 
safety concerns, the rest of Group 1 may be immunised. This staggered approach will be repeated 
for the second dose. After at least eight volunteers from Group 1 have received two out of three 
doses of 10 µg Pvs25-IMX313 in 50 µg Matrix-M1 and had seven days of follow-up, there will be a 
formal safety review (see below) prior to dose escalation (i.e. recruitment to Groups 2 and 3). The 
staggered enrolment process will continue for the third vaccination in Group 1 and for Group 2 for 
the first three volunteers at each of the three doses. Following this, recruitment to Groups 2 and 
3 will occur in parallel.   
7.4.2 Dose escalation 
A minimum of data from eight volunteers recruited to Group 1 will be reviewed prior to dose 
escalation (i.e. recruitment to Groups 2 or 3). The data to be reviewed will include but is not limited 
to: physical observations, patient e-diary cards and safety bloods (full blood count, urea and 
electrolytes, liver function tests). Data up to and including the first seven days following 
vaccination will be required for each of the eight volunteers prior to dose escalation. E-diary cards 
will be reviewed on a daily basis for the first eight volunteers recruited to Group 1 for the first 
seven days following vaccination. A formal safety review will be undertaken by the LSM prior to 
dose escalation. This would take the form of a safety report incorporating the relevant safety data 
described above, which would need to be reviewed and approved by the CI and LSM prior to 
recruitment into Groups 2 and 3. 

 
 
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7.4.3 Group 1 
Eight to ten volunteers will be recruited into Group 1 (Table 2) and each volunteer will attend for 
three vaccinations. On the day of enrolment, Pvs25-IMX313 will be administered at a dose of 10 
µg in 50 µg Matrix-M1 adjuvant. After approximately 4 weeks, a second dose will be administered, 
followed by a third and final vaccination approximately 4 weeks later. Second and third 
vaccinations will be administered at the same dose of both vaccine and adjuvant as at the initial 
vaccination. Volunteers will be followed until 8 months (approximately) from the time of 
enrolment.   
7.4.4 Group 2 
Eight to ten volunteers will be recruited into Group 2 (Table 2) and each volunteer will attend for 
three vaccinations. On the day of enrolment, Pvs25-IMX313 will be administered at a dose of 50 
µg in 50 µg Matrix-M1 adjuvant. After approximately 4 weeks, a second dose will be administered, 
followed by a third and final vaccination approximately 4 weeks later. Second and third 
vaccinations will be administered at the same dose of both vaccine and adjuvant as at the initial 
vaccination. Volunteers will be followed until 8 months (approximately) from the time of 
enrolment.   
7.4.5 Group 3 
Eight to ten volunteers will be recruited into Group 3 (Table 2) and each volunteer will attend for 
three vaccinations. On the day of enrolment, Pvs25-IMX313 will be administered at a dose of 50 
µg in 50 µg Matrix-M1 adjuvant. After approximately 4 weeks, a second dose will be administered, 
followed by a third and final vaccination approximately 4 weeks later. The second vaccination will 
be at the same dose as the first vaccination whilst the third vaccination will be administered at a 
lower dose of Pvs25-IMX313 10 µg in 50 µg Matrix-M1 adjuvant. Volunteers will be followed until 
8 months (approximately) from the time of enrolment.   
 
Group 
Group 
size 
D0 
D28 
D56 
Group 1 
 
8-10 
Pvs25-IMX313 10 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 10 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 10 µg 
Matrix-M1 50 µg 
Group 2 
8-10 
Pvs25-IMX313 50 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 50 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 50 µg 
Matrix-M1 50 µg 
Group 3 
 
8-10 
Pvs25-IMX313 50 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 50 µg 
Matrix-M1 50 µg 
Pvs25-IMX313 10 µg 
Matrix-M1 50 µg 
Table 2: Vaccination protocol for study groups 1, 2 and 3 
 

 
 
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7.5 
Duration of study 
7.5.1 Definition of the start and end of the trial  
The start of the trial is defined as the date of enrolment of the first volunteer. The end of the trial 
is the date of the last follow-up of the last volunteer. 
7.5.2 Duration of volunteer participation 
Volunteers will be considered to be enrolled in the trial on the receipt of the first vaccination. Total 
duration of follow-up will be approximately 8 months from enrolment. 
7.6 
Potential risks for volunteers 
7.6.1 Phlebotomy  
The maximum estimated volume of blood drawn (972mL) over the study period should not 
compromise otherwise healthy volunteers, however, volunteers will be monitored for anaemia. 
There may be minor bruising, local tenderness, pre-syncopal symptoms or syncope (rarely) 
associated with venepuncture, which will not be documented as AEs if they occur. The actual total 
blood volume may vary depending on any requirement for repeat clinical tests, e.g. in the event 
of a clinically significant abnormal result.  
7.6.2 Vaccinations 
Potential foreseeable risks from vaccination can be partially predicted from previous studies assessing 
protein vaccines, administered with the Matrix-M1 adjuvant system, which include local and systemic 
reactions. Generally, an increase in solicited AEs is seen following administration of investigational 
vaccines with Matrix-M1 adjuvant, compared to the antigen alone or placebo, however, 
reactogenicity is transient with the vast majority of adverse events resolving within 2-7 days.  
Local adverse events are likely to include: injection site pain, erythema, swelling, itching and 
warmth. Forseeable systemic adverse events would include: headache, fatigue, myalgia, 
arthralgia, malaise, feverishness, fever and nausea. The majority of local and systemic adverse 
events are forseen to be mild to moderate in nature, however, it is anticipated that rarely, these 
may be reported as severe. Of the systemic adverse events reported as severe following receipt of 
other investigational vaccines administered with Matrix-M1 adjuvant, myalgia is the most 
common.  
As with any vaccine, Guillain-Barré syndrome or immune-mediated reactions that can lead to 
organ damage including serious allergic reactions may occur, but this should be extremely rare. 
Serious allergic reactions including anaphylaxis could also occur and for this reason volunteers will 
be observed for an hour after each vaccination and vaccinated in a clinical area where Advanced 
Life Support trained physicians, equipment and drugs are immediately available for the 
management of any SARs. 
 
7.7 
Potential benefits for volunteers 
Volunteers will not benefit directly from participation in this study. However, it is hoped that the 
information gained from this study will contribute to the development of a safe and effective P. 
vivax transmission-blocking vaccine. Volunteers will also receive information about their general 
health status. 

 
 
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8 RECRUITMENT AND WITHDRAWAL FOR TRIAL VOLUNTEERS  
Volunteers may be recruited by use of an advertisement +/- registration form formally approved 
by the ethics committee(s) and distributed or posted in the following places: 
- 
In public places, including buses and trains, with the agreement of the owner/proprietor. 
- 
In newspapers or other literature for circulation. 
- 
On a website operated by the Investigators’ clinical trials group or with the agreement of 
the owner or operator (including online recruitment through our website).  
- 
As a post on Twitter, Facebook or other similar account owned and operated by the 
Investigators’ clinical trials group. 
- 
By email distribution to a group or list only with the express agreement of the network 
administrator or with equivalent authorisation, including those who have already 
expressed an interest in taking part in any clinical trial at the Oxford Vaccine Centre. This 
will include a link to the website advertisement. 
- 
On stalls or stands at exhibitions or fairs. 
- 
Via presentations (e.g. presentations at lectures or invited seminars). 
- 
Oxford Vaccine Centre databases: We may contact individuals from databases of groups 
within the CCVTM (including the Oxford Vaccine Centre database) of previous trial 
participants who have expressed an interest in receiving information about all future 
studies for which they may be eligible. 
8.1 
Informed consent 
The information sheet will be made available to the volunteer at least 24 hours prior to the 
screening visit, thus they will have a minimum of 24 hours to consider study participation. At the 
screening visit, the volunteer will be fully informed of all aspects of the trial, the potential risks and 
their obligations. The following will be emphasised: 
• Participation in the study is entirely voluntary. 
• Refusal to participate involves no penalty or loss of medical benefits. 
• The volunteer may withdraw from the study at any time.  
• The volunteer is free to ask questions at any time to allow him or her to understand the 
purpose of the study and the procedures involved. 
• There is no direct benefit from participating. 
• The volunteer’s general practitioner (GP) will be contacted to corroborate their medical 
history. Volunteers will only be enrolled in the study if written information regarding the 
volunteer’s medical history is obtained from the GP.   
• The volunteer will be registered on the TOPS database (The Overvolunteering Prevention 
System; www.tops.org.uk).  
• With volunteer consent, blood samples will be taken and any leftover samples stored 
indefinitely for use in other, ethically approved research.  
The aims of the study and all tests to be carried out will be explained. The volunteer will be given 
the opportunity to ask about details of the trial and will then have time to consider whether or not 
to participate before completing the written consent form. In addition to the consent form, 
volunteers will be asked to complete an additional separate form regarding consent for the GP to 
release medical records, a copy of which will be sent to the GP. 
The participant must personally sign and date the latest approved version of the Informed Consent 
form before any trial specific procedures are performed. 

 
 
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Written and verbal versions of the Participant Information and Informed Consent will be presented 
to the participants detailing no less than: the exact nature of the trial; what it will involve for the 
participant; the implications and constraints of the protocol; the known side effects and any risks 
involved in taking part. It will be clearly stated that the participant is free to withdraw from the 
trial at any time for any reason without prejudice to future care, without affecting their legal rights 
and with no obligation to give the reason for withdrawal. 
The participant will be allowed the opportunity to question the Investigator, their GP or other 
independent parties to decide whether they will participate in the trial. Written Informed Consent 
will then be obtained by means of participant dated signature and dated signature of the person 
who presented and obtained the Informed Consent. 
The person who obtained the consent must be suitably qualified and experienced, and have been 
authorised to do so by the Chief/Principal Investigator. A copy of the signed Informed Consent will 
be given to the participant. The original signed form will be retained at the trial site. 
8.2  Inclusion and exclusion criteria  
8.2.1 Inclusion criteria 
The volunteer must satisfy all the following criteria to be eligible for the study: 
• Healthy adult aged 18 to 45 years.  
• Able and willing (in the Investigator’s opinion) to comply with all study requirements. 
• Willing to allow the Investigators to discuss the volunteer’s medical history with their GP. 
• Volunteers with the potential to become pregnant only: must practice continuous effective 
contraception for the duration of the study (see section 10.10). 
• Agreement to refrain from blood donation for the duration of the study. 
• Able and willing to provide written informed consent to participate in the trial. 
8.2.2 Exclusion criteria 
The volunteer may not enter the study if any of the following apply: 
• History of clinical malaria (any species). 
• Travel to a clearly malaria endemic locality during the study period or within the preceding 
six months. 
• Use of immunoglobulins or blood products (e.g., blood transfusion) in the last three 
months. 
• Receipt of any vaccine in the 30 days preceding enrolment, or planned receipt of any other 
vaccine within 30 days following each study vaccination, with the exception of COVID-19 
vaccines, which should not be received between 14 days before to 7 days after any study 
vaccination. 
• Receipt of an investigational product in the 30 days preceding enrolment, or planned 
receipt during the study period. 
• Concurrent involvement in another clinical trial or planned involvement during the study 
period. 
• Prior receipt of an investigational vaccine likely to impact on interpretation of the trial data, 
as assessed by the Investigator. 

 
 
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• Any confirmed or suspected immunosuppressive or immunodeficient state, including HIV 
infection; asplenia; recurrent, severe infections and chronic (more than 14 days) 
immunosuppressant medication within the past 6 months (inhaled and topical steroids are 
allowed). 
• History of allergic disease or reactions likely to be exacerbated by any component of the 
vaccine.  
• Any history of anaphylaxis in reaction to vaccinations. 
• Pregnancy, lactation or intention to become pregnant during the study. 
• 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. 
• Any other serious chronic illness requiring hospital specialist supervision. 
• Suspected or known current alcohol abuse as defined by an alcohol intake of greater than 
25 standard UK units every week. 
• Suspected or known injecting drug abuse in the 5 years preceding enrolment. 
• Hepatitis B surface antigen (HBsAg) detected in serum. 
• Seropositive for hepatitis C virus (antibodies to HCV) at screening (unless volunteer has 
taken part in a prior hepatitis C vaccine study with confirmed negative HCV antibodies prior 
to participation in that study, and negative HCV ribonucleic acid (RNA) PCR at screening for 
this study). 
• 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. Procedures for identifying laboratory values meeting exclusion 
criteria are shown in SOP VC027.  
• 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. 
• Inability of the study team to contact the volunteer’s GP to confirm medical history. 
8.2.3 Prevention of ‘Over Volunteering’ 
Volunteers will be excluded from the study if they are concurrently involved in another trial. In 
order to check this, volunteers will be asked to provide their National Insurance or Passport 
number (if they are not entitled to a NI number) and will be registered on a national database of 
participants in clinical trials (www.tops.org.uk). 
8.2.4 Vaccination and re-vaccination exclusion criteria 
The following events associated with vaccine immunisation constitute absolute contraindications 
to further administration of vaccine. If any of these events occur during the study, the participant 
must be withdrawn and followed until resolution of the event, as with any adverse event:  
• Anaphylactic reaction following administration of vaccine.  
• Pregnancy.  
The following adverse events constitute contraindications to administration of vaccine at that 
point in time; if any one of these adverse events occurs at the time scheduled for vaccination, the 
participant may be vaccinated at a later date, or withdrawn at the discretion of the Investigator. 
The participant must be followed until resolution of the event as with any adverse event:  

 
 
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• 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 such as diarrhoea, mild upper respiratory infection with or 
without low-grade febrile illness, i.e. temperature of ≤37.5°C/99.5°F.  
• Temperature of >37.5°C (99.5°F) at the time of vaccination.  
• Current COVID-19 infection, defined as ongoing symptoms with positive COVID-19 PCR 
swab test taken during current illness or positive COVID-19 PCR swab test within preceding 
14 days without symptoms. Vaccinations will be delayed by a minimum of 2 weeks from 
the date of the first positive COVID-19 PCR swab, as long as symptoms are improving or 
resolved. It will be at the discretion of the Investigator to withdraw a participant if they 
develop severe COVID-19 disease. 
8.2.5 Concomitant medications  
As set out by the exclusion criteria, volunteers may not enter the study if they have received any 
vaccine in the 30 days prior to enrolment or there is planned receipt of any other vaccine within 
30 days of each vaccination, any investigational medicinal product within 30 days prior to 
enrolment or if receipt is planned during the study period, or any chronic (more than 14 days) 
immunosuppressant medication within 6 months prior to enrolment or if receipt is planned at any 
time during the study period (inhaled and topical steroids are permitted). One exception is COVID-
19 vaccines, which should not be received between 14 days before to 7 days after any study 
vaccinations. 
8.3 
Withdrawal of volunteers 
In accordance with the principles of the most recent revision of the Declaration of Helsinki (2013) 
and any other applicable regulations, a volunteer has the right to withdraw from the study at any 
time and for any reason and is not obliged to give his or her reasons for doing so. In addition, the 
volunteer may withdraw/be withdrawn from further study procedures at any time in the interests 
of the volunteer’s health and wellbeing, or for any of the following reasons: 
• Administrative decision by the Investigator. 
• Ineligibility (either arising during the study or retrospectively, having been overlooked at 
screening). 
• Significant protocol deviation. 
• Volunteer non-compliance with study requirements. 
• An AE, which requires discontinuation of the study involvement or results in inability to 
continue to comply with study procedures. 
The Local Safety Monitor (LSM) may recommend withdrawal of volunteers. The reason for 
withdrawal from further study procedures will be recorded in the Case Report Form (CRF) where 
available. For all AEs, appropriate follow-up visits or medical care will be arranged, with the 
agreement of the volunteer, until the AE has resolved, stabilised or a non-trial related causality 
has been assigned.  
Any volunteer who fails to attend for two or more follow-up visits will be deemed to have 
withdrawn from the study.  
If a volunteer withdraws from the study, blood samples collected before their withdrawal from the 
trial will be used/stored unless the volunteer specifically requests otherwise. Similarly, all data 
collected up to the point of withdrawal will be stored, unless they specifically request for it to be 

 
 
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destroyed. Volunteers are free to request that their blood samples be destroyed any time during 
or after the study.  
In all cases of participant withdrawal, excepting those of complete consent withdrawal, long-term 
safety data collection, including some procedures such as safety bloods, will continue as 
appropriate if participants have received one or more vaccine doses.  
8.4 
Pregnancy 
Should a volunteer become pregnant during the trial, she will be followed up as other volunteers 
and in addition will be followed until pregnancy outcome, with the volunteer’s permission. 
Depending on the timings of pregnancy outcome relative to planned study follow-up timepoints, 
this may involve additional telephone consultations. We will not routinely perform venepuncture 
on such volunteers. 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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9. TREATMENT OF TRIAL VOLUNTEERS 
This section describes the clinical procedures for evaluating study participants, vaccination and 
follow-up after vaccination.  
9.1 
Trial sites 
Volunteers will be recruited and undergo all visits at the CCVTM, Oxford, this is inclusive of 
screening, vaccinations and follow-up visits.   
9.2 
Study procedures 
Procedures will be performed at the time points indicated in the schedule of events (Table 3). 
Additional procedures or laboratory tests may be performed, at the discretion of the 
Investigators if clinically necessary.  
9.2.1 Observations 
Pulse, blood pressure and temperature will be measured at the time points indicated in the 
schedule of events (Table 3). In addition, weight and height will be measured at screening. 
9.2.2 Blood tests 
Blood will be drawn at the time points indicated in the schedule of events (Table 3) and the 
following laboratory assays performed: 
At Oxford University Hospitals NHS Foundation Trust, using NHS standard procedures: 
• Haematology: Full blood count.  
• Biochemistry: Sodium, potassium, urea, creatinine, albumin and liver function tests. 
• Diagnostic serology: HbsAg, HCV antibodies, HIV antibodies. 
• Immunology: Human Leukocyte Antigen (HLA) typing at the day of enrolment (D0). 
At the Jenner Institute research laboratories: 
• Immunology:  
o Pvs25-specific immunogenicity will be assessed by a variety of immunological assays. 
These will include ex vivo ELISpot assays for interferon gamma and antibody ELISAs for 
measuring total IgG, isotype and avidity.  
o The functional activity of the vaccine-induced antibodies will be tested using 
membrane-feeding assays. The functional activity of the purified IgG will be tested 
against P. vivax by DMFA.  
o Other exploratory immunological assays including cytokine analysis, other antibody 
assays (plasma cell and memory B cell assays), IMX313-specific immunogenicity, human 
C4b, DNA analysis of genetic polymorphisms potentially relevant to vaccine 
immunogenicity and gene expression studies, amongst others, may be performed at 
the discretion of the Investigators. B cells, plasma and/or serum may also be analysed 
and used to produce human monoclonal antibodies. 
o Samples may also be sent to collaborating laboratories within and outside the UK 
(including outside of Europe) for other immunological assays as required. This would 
involve the transfer of serum, plasma or blood cells to these laboratories, but these 
would remain pseudo-anonymised. Informed consent for this will be obtained from 

 
 
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IRAS Project ID: 302239                                                                                         
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volunteers. Immunological assays will be conducted according to the procedures 
established in the test laboratories.  
With the volunteers’ informed consent, any leftover cells, plasma, serum (or their purified 
components) will be stored indefinitely in the Oxford Vaccine Centre Biobank for future 
immunological analysis of malaria-specific or vaccine-related responses. This may include human 
DNA and RNA analysis. If a participant elects not to permit this, all of that participant’s leftover 
samples will be discarded after the required period of storage to meet Good Clinical Practice (GCP) 
and regulatory requirements. 
9.2.3  Urinalysis 
Urine will be tested for the presence of clinically significant proteinuria, glycosuria or haematuria 
(as defined in Appendix A) at the screening visits. For volunteers with the potential to become 
pregnant (see section 10.10 for definition), urine will be tested for beta-human chorionic 
gonadotrophin (β-HCG) at screening, immediately prior to each vaccination, at days 0, 28 and 56 
and on day 84.  
9.2.4 Vaccinations  
Before each vaccination, the ongoing eligibility of the volunteer will be reviewed. The vaccine will 
be administered as described above in section 5.3. The injection site will be covered with a sterile 
dressing and the volunteer will stay in the local trial site for observation for 1 hour, in case of 
immediate adverse events. Observations will be taken 30 minutes after vaccination (+/- 5 minutes) 
and the sterile dressing removed and injection site inspected. Observations will also be taken at 
60 minutes after vaccination (+/- 10 minutes).  
An oral thermometer and tape measure will be given to each volunteer, with instructions on use, 
along with the emergency 24-hour telephone number to contact the on-call study physician if 
needed.  
9.2.5 Management of post-vaccination fevers 
Post-vaccination fevers may cause diagnostic uncertainty as concurrent COVID-19 infection is a 
possibility. If a participant develops a fever within the first 24 hours after vaccination and no other 
symptoms associated with COVID-19 disease (new onset cough, anosmia or ageusia), then post-
vaccination fever is most likely. Participant will be advised to self-isolate until they become afebrile 
for 24 hours. Participants will be requested to attend their day 1 post-vaccination study visit to 
allow collection of study samples including safety blood tests. This study visit will be conducted in 
a side room with study staff wearing appropriate level 1 Personal Protective Equipment (PPE) and 
participants will be asked to attend the visit by means other than public transport.  
If a participant develops a fever post vaccination, which is associated with either new onset cough 
or anosmia or ageusia, they will be advised to self-isolate as per Public Health England (PHE) 
guidance and directed to local community based COVID-19 testing. If participants remain febrile 
more than 48 hours post vaccination, they will be advised to continue self-isolation and directed 
to local community based COVID-19 testing. Whilst awaiting their COVID-19 PCR test results, 
participants will be advised not to attend the clinic for study visits.  
If a participant tests positive for COVID-19 or whilst awaiting their COVID-19 test result, they will 
be advised to continue self-isolating as per PHE guidance. Any study visits which are due whilst a 
participant is self-isolating, should be rescheduled to occur when self-isolation ends, if this falls 
within the visit window. If a participant remains in self-isolation during the period of the visit 

 
 
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IRAS Project ID: 302239                                                                                         
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window, then the study visit will be conducted by telephone and no blood tests will be taken. Study  
visit windows have been increased in order to accommodate these scenarios.  
 
If a participant is self-isolating for a prolonged period post-vaccination (for example because they 
are a household contact of a suspected or confirmed COVID-19 case) and hence would miss all 
three initial post vaccination visits, then it will be at the Investigator’s discretion to conduct these 
as physical visits at CCVTM to allow full assessment including blood tests. These study visits would 
be conducted in a side room with study staff wearing level 1 PPE. 
If a participant tests negative for COVID-19, they can stop self-isolating and continue with study 
visits as planned. 
9.2.6 E-diary  
Following each vaccination, volunteers will be asked to complete an e- diary. Volunteers will be 
asked about foreseeable local and systemic AEs for 7 days (solicited AEs). After this, volunteers will 
be asked to record any AE daily, for 28 days (unsolicited AEs). E-diary cards will be reviewed with 
volunteers at each post-vaccination clinic visit.  
An electronic PDF copy of the e-diary or hard copy will be issued to participants if there are 
technical issues with the e-diary or if the participant has internet/accessibility issues. 
9.2.7 Photographs 
Photographs of the injection site may be taken in the event of severe pain, redness and/or 
swelling following vaccination to enable assessment of any improvement/deterioration. Severe is 
defined as a grading of 3 in any one of the categories detailed in Table 7. Photographs will not 
include the participant’s face. 
9.3 
Study visits  
All clinical reviews and procedures will be undertaken by one of the clinical team. The procedures 
to be included in each visit are documented in Table 3. Each review is assigned a time point and a 
window period within which the review will be conducted. 
9.3.1 Screening visits  
There 
will 
be 
an 
initial 
pre-screening 
questionnaire 
that 
is 
completed 
online 
(https://www.onlinesurveys.ac.uk) to assess initial eligibility. Use of this tool has been approved 
by the University of Oxford. If the prospective participant meets initial eligibility criteria, we will 
seek consent via email to subsequently contact individuals by telephone. During this pre-screening 
telephone call with a member of the clinical team to discuss the study, we will provide the 
opportunity for interested individuals to discuss the study, eligibility criteria and study 
requirements. If individuals continue to express an interest in taking part in the trial following the 
pre-screening call and if they meet eligibility criteria, a screening visit will be scheduled.  
Screening visits may take place up to 90 days prior to enrolment. Informed consent will be gained 
at screening. If consent is given, the screening procedures indicated in the schedule of events 
(Table 3) will be undertaken.  
The potential participant’s GP will be contacted with the written permission of the participant after 
screening using the GP screening letter. This will enable the study team to ascertain any significant 
medical history and act as notification that the participant has volunteered for the study.  

 
 
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During the screening the volunteers will be asked to provide their National Insurance or passport 
number so that this can be entered on to a national database which helps prevent volunteers from 
participating in more than one clinical trial simultaneously or over-volunteering for clinical trials 
(www.tops.org.uk). 
Where more than 90 days has lapsed between the initial screening visit and date of enrolment, a 
re-screening visit will be conducted to ascertain if there have been any changes in the medical 
history or physical examination findings. Urinalysis, biochemistry and haematology blood tests 
will be repeated. Other blood tests will not be repeated routinely. Where a reply from the 
potential participant’s GP has already been received, the GP will not be re-contacted unless there 
is any need to obtain futher information where appropriate (e.g. regarding a new medical event 
or finding) and at the discretion of the Investigator. 
Hepatitis B, Hepatitis C and HIV status will be checked via blood tests at the screening visit. Some 
individuals may test positive for Hepatitis C virus due to their previous involvement in a Hepatitis 
C vaccine study. If this applies to an individual and they wish to take part in our study, with their 
consent, we will contact the team responsible for the Hepatitis C vaccine study to check the 
participant’s Hepatitis C status prior to enrolment in this malaria vaccine study. A copy of the 
consent will be held by both study teams. 
9.3.2 Enrolment and first vaccination (V1) with Pvs25-IMX313 in Matrix-M1 adjuvant (all groups) 
The eligibility of the volunteer will be reviewed at the end of the screening visit and again when all 
results from the screening visit have been considered. If eligible, a day 0 visit will be scheduled for 
the volunteer to receive the first dose of vaccine and be enrolled in the trial.  
At the day 0 visit, any new medical issues or symptoms that have arisen will be assessed. Physical 
observations, urinary β-hCG test in volunteers with the potential to become pregnant and 
venepuncture for immunology and safety bloods will be undertaken according to Table 3. The 
inclusion and exclusion criteria for the study will be reviewed. If the volunteer remains eligible and 
there are no contraindications to vaccination, the Pvs25-IMX313 vaccine, formulated in Matrix-M1 
adjuvant will be administered according to the doses described in Table 3 and as described in 
Section 5.3. The visit will be carried out as described in above Section 9.2.4.  
9.3.3 Reviews post-vaccination on days 1, 7 and 14 (all groups) 
On subsequent visits, the volunteers will be assessed for local and systemic AEs, using the diary 
card, interim history, physical examination and blood tests at the time points indicated in the 
schedule of events (Table 3). Blood will also be taken for exploratory immunology analyses.  
Volunteers will be asked to record all AEs via the diary card. E-diary cards will be reviewed with 
volunteers at the clinic visits on days 1 and 7. Volunteers will be asked directly about foreseeable 
local and systemic AEs for 7 days (solicited AEs). They will also be asked daily about any new 
symptoms from day 7 until day 28 via the diary but will not be asked directly about foreseeable 
symptoms. E-diary cards will be reviewed for details of solicited and unsolicited AEs for 28 days 
after each vaccination, and any new or previously undocumented medical issues or symptoms that 
have arisen will be assessed. Physical observations and venepuncture for immunology and safety 
bloods will be undertaken according to Table 3. At any post-vaccination visit, where valid written, 
informed consent is in place, photographs may be taken for clinical comparison. This will enable 
qualitative as well as quantitative assessment of redness and swelling at the injection site. 
Confidentiality will be maintained as described in Section 13.5. 

 
 
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9.3.4 Second vaccination (V2) with Pvs25-IMX313 in Matrix-M1 adjuvant on day 28 (all groups) 
This visit will include a follow-up visit for the first vaccination and administration of the second 
vaccine. Any new medical issues or symptoms that have arisen will be assessed. Physical 
observations, urinary β-hCG test in volunteers with the potential to become pregnant and 
venepuncture for immunology and safety bloods will be undertaken according to Table 3. The 
inclusion and exclusion criteria for the study will be reviewed. If the volunteer remains eligible and 
there are no contraindications to vaccination, the Pvs25-IMX313 vaccine, formulated in Matrix-M1 
adjuvant will be administered intramuscularly in the opposite arm to that used for the first 
vaccination and as described in sections 5.3 and 9.2.4. Dosing will be determined by group 
allocation (see Table 3). 
9.3.5 Reviews post-vaccination on days 29, 35 and 42 (all groups) 
On subsequent visits, the volunteers will be assessed for local and systemic AEs, using the diary 
card, interim history, physical examination and blood tests at the time points indicated in the 
schedule of events (Table 3). Blood will also be taken for exploratory immunology analysis.  
Volunteers will be asked to record all AEs via the diary card. E-diary cards will be reviewed with 
volunteers at the clinic visits on days 29 and 35. Volunteers will be asked directly about foreseeable 
local and systemic AEs for 7 days (solicited AEs). They will also be asked daily about any new 
symptoms from day 35 until day 56 via the diary but will not be asked directly about foreseeable 
symptoms. E-diary cards will be reviewed for details of solicited and unsolicited AEs for 28 days 
after each vaccination, and any new or undocumented medical issues or symptoms that have 
arisen will be assessed. Physical observations and venepuncture for immunology and safety bloods 
will be undertaken according to Table 3. At any post-vaccination visit, where valid written, 
informed consent is in place, photographs may be taken for clinical comparison. Confidentiality 
will be maintained as described in Section 13.5. 
9.3.6 Third vaccination (V3) with Pvs25-IMX313 in Matrix-M1 adjuvant on day 56 (all groups) 
This visit will include a follow-up visit for the second vaccination and administration of the third 
vaccine. Any new medical issues or symptoms that have arisen will be assessed. Physical 
observations, urinary β-hCG test in volunteers with the potential to become pregnant and 
venepuncture for immunology and safety bloods will be undertaken according to Table 3. The 
inclusion and exclusion criteria for the study will be reviewed. If the volunteer remains eligible and 
there are no contraindications to vaccination, the PvS25-IMX313 vaccine, formulated in Matrix-
M1 adjuvant will be administered intramuscularly in the opposite arm to that used for the second 
vaccination and as described in sections 5.3 and 9.2.4. Dosing will be determined by group 
allocation (see Table 2). 
9.3.7 Reviews post-vaccination on days 57, 63, 70 and 84  
On subsequent visits, the volunteers will be assessed for local and systemic adverse events, using 
a diary card, interim history, physical examination and blood tests at the time points indicated in 
the schedule of attendances (Table 3). Blood will also be taken for exploratory immunology 
analysis.  
Volunteers will be asked to record all AEs via the diary card and at visits on days 57 and 63, during 
which e-diary cards will be reviewed with volunteers. Volunteers will be asked directly about 
foreseeable local and systemic AEs for 7 days (solicited AEs). Volunteers will be asked to continue 
to record any new symptoms from day 63 until day 84, on a daily basis via the diary but will not be 
asked directly about foreseeable symptoms. E-diary cards will be reviewed for details of solicited 

 
 
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IRAS Project ID: 302239                                                                                         
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and unsolicited AEs for 28 days after each vaccination, and any new or undocumented medical 
issues or symptoms that have arisen will be assessed. Physical observations, procedures (including 
a urinary pregnancy test at D84) and venepuncture for immunology and safety bloods will be 
undertaken according to Table 3. At any post-vaccination visit, where valid written, informed 
consent is in place, photographs may be taken for clinical comparison. Confidentiality will be 
maintained as described in Section 13.5. 
9.3.8 Long-term follow up post-vaccination on days 140 and 240  
On days 140 and 240, volunteers will be assessed for serious adverse events, by interim history, 
and physical examination, as deemed necessary by the Investigator. Blood tests will be performed 
as indicated in the schedule of attendances for these timepoints (Table 3). Blood will also be taken 
for exploratory immunology analysis. At any post-vaccination visit, where valid written, informed 
consent is in place, photographs may be taken for clinical comparison. Confidentiality will be 
maintained as described in Section 13.5. 
 
 
 
 
 
 
 
 
 
 

 
 
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IRAS Project ID: 302239                                                                                         
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S 
V1 
  
  
  
V2 
  
  
  
V3 
  
  
  
  
  
  
Attendance number 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
Timeline (days)  
  
0 
1 
7 
14 
28 
29 
35 
42 
56 
57 
63 
70 
84 
140 
240 
Window (days) 
-90 
  
±1 
±2 
-2/+4 
-7/+14 
±1 
±2 
-2/+4 
-7/+14 
±1 
±2 
-2/+4 
-7/+14 
±14 
±14 
Inclusion/Exclusion criteria 
X 
X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Informed consent 
X 
X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Physical observations* 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Medical history/ examination 
X 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Photographs 
 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Urinalysis 
X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Urine β-hCG** 
X 
X 
  
  
  
X 
  
  
  
X 
  
  
  
X  
  
  
Review contraindications 
  
X 
  
  
  
X 
  
  
  
X 
  
  
  
  
  
  
Review AEs & medications 
  
  
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Vaccination 
  
X 
  
  
  
X 
  
  
  
X 
  
  
  
  
  
  
Diary card issued 
  
X 
  
  
  
X 
  
  
  
X 
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
X 
  
  
  
X 
  
  
  
X 
  
  
Immunology plasma (mL) 
  
30 
10 
10 
30 
80 
10 
60 
30 
30 
10 
60 
30 
80 
30 
30 
Immunology serum (mL) 
  
40 
  
  
40 
40 
  
  
40 
40 
  
  
40 
40 
40 
40 
RNA preservation (mL) 
  
3 
3 
  
  
3 
3 
  
  
3 
3 
  
  
  
  
  
Haematology (mL)# 
2 
2 
  
2 
2 
2 
  
2 
2 
2 
  
2 
2 
2 
  
  
Biochemistry (mL) ## 
3 
3 
  
3 
3 
3 
  
3 
3 
3 
  
3 
3 
3 
  
  
HLA typing (mL) 
  
4 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
HIV, HBV, HCV serology (mL) 
5 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Total Blood Volume (mL) 
10 
82 
13 
15 
75 
128 
13 
65 
75 
78 
13 
65 
75 
125 
70 
70 
Cumulative blood volume (mL) 
10 
92 
105 
120 
195 
323 
336 
401 
476 
554 
567 
632 
707 
832 
902 
972$ 
Table 3: Schedule of events  
S = screening visit; V = vaccination; (X) = If considered necessary, emphasising any acute complaints. *Physical observations will include blood pressure, heart rate and temperature, 
plus height and weight at screening only. **For volunteers with the potential to become pregnant. # Haematology will comprise full blood count. ## Biochemistry will include sodium, 
potassium, urea, creatinine, albumin, liver function tests. $ Cumulative blood volume if blood taken as per schedule and excluding any repeat safety blood tests that may be 
necessary.

 
 
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10 ASSESSMENT OF SAFETY 
Safety will be assessed by the frequency, incidence and nature of AEs and SAEs arising during the 
study. The safety profile will be assessed on an ongoing basis by the Investigators during the safety 
reporting window for each participant (from provision of consent to their last study visit). The CI 
and relevant Investigators (as per the trial delegation log) will also review safety issues and SAEs 
as they arise. 
10.1 Definitions 
10.1.1 Adverse Event (AE) 
An AE is any untoward medical occurrence in a volunteer, which may occur during or after 
administration of an Investigational Medicinal Product (IMP) and does not necessarily have a 
causal relationship with the intervention. An AE can therefore be any unfavourable and 
unintended sign (including an abnormal laboratory finding), symptom or disease temporally 
associated with the study intervention, whether or not considered related to the study 
intervention. 
Each participant-reported AE will be graded by the participant according to the table for grading 
severity of adverse events (see Table 5). Severity gradings may be reviewed and discussed with the 
participants at the clinic visits. 
10.1.2 Adverse Reaction (AR) 
An AR is any untoward or unintended response to an IMP. This means that a causal relationship 
between the IMP and an AE is at least a reasonable possibility, i.e. the relationship cannot be ruled 
out. All cases judged by the reporting medical Investigator as having a reasonable suspected causal 
relationship to an IMP (i.e. possibly, probably or definitely related to an IMP) will qualify as ARs. 
10.1.3 Serious Adverse Event (SAE) 
An SAE is an AE that results in any of the following outcomes, whether or not considered related 
to the study intervention: 
• Death. 
• Life-threatening event (i.e. the volunteer was, in the view of the Investigator, at immediate 
risk of death from the event that occurred). This does not include an AE that, if it occurred 
in a more severe form, might have caused death. 
• Persistent or significant disability or incapacity (i.e. substantial disruption of one’s ability to 
carry out normal life functions). 
• Hospitalisation or prolongation of hospitalisation, regardless of length of stay, even if it is 
a precautionary measure for continued observation. Hospitalisation (including inpatient or 
outpatient hospitalisation for an elective procedure) for a pre-existing condition that has 
not worsened unexpectedly does not constitute an SAE. 
• An important medical event (that may not cause death, be life-threatening, or require 
hospitalisation) that may, based upon appropriate medical judgment, jeopardise the 
volunteer and/or require medical or surgical intervention to prevent one of the outcomes 
listed above. Examples of such medical events include allergic reaction requiring intensive 
treatment in an emergency room or clinic, blood dyscrasias, or convulsions that do not 
result in inpatient hospitalisation. 

 
 
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• Congenital anomaly or birth defect. 
10.1.4 Serious Adverse Reaction (SAR) 
An AE (expected or unexpected) that is both serious and, in the opinion of the reporting 
Investigator or Sponsors, believed to be possibly, probably or definitely due to an IMP or any other 
study treatments, based on the information provided. 
10.1.5 Suspected Unexpected Serious Adverse Reaction (SUSAR) 
All SAEs at least possibly related to Pvs25-IMX313/Matrix-M1 will be considered unexpected and 
be reported as SUSARs. 
10.1.6 Foreseeable Adverse Reactions 
The foreseeable ARs following vaccination with Pvs25-IMX313 in Matrix-M1 adjuvant are: injection 
site pain, erythema, warmth, swelling, pruritus, myalgia, arthralgia, headache, fatigue, fever, 
feverishness, malaise and nausea. These AEs will be listed as ‘solicited AEs’ providing they occur 
within 7 days of the day of vaccination. ‘Unsolicited AEs’ are AEs other than the foreseeable ARs 
occurring within the first 7 days, or any AEs occurring after the first 7 days after vaccination. 
10.2 Causality Assessment 
For every unsolicited AE, an assessment of the relationship of the event to the administration of 
the vaccine will be undertaken by the CI or the CI-delegated clinician at the coordinating site 
(Oxford). An intervention-related AE refers to an AE for which there is a possible, probable or 
definite relationship to administration of a vaccine. An interpretation of the causal relationship of 
the intervention to the AE in question will be made, based on the type of event; the relationship 
of the event to the time of vaccine administration; and the known biology of the vaccine therapy 
(Table 4).  
Causality assessment will take place during planned safety reviews, interim analyses (e.g. if a 
holding or stopping rule is activated) and at the final safety analysis, except for SAEs, for which 
causality should be assigned by the reporting Investigator. 
 
0 
No 
Relationship 
No temporal relationship to study product and 
Alternate aetiology (clinical state, environmental or other 
interventions); and 
Does not follow known pattern of response to study product 
1 
Unlikely 
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. 

 
 
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2 
Possible 
 
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 
3 
Probable 
 
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 
4 
Definite 
 
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 
Table 4: Guidelines for assessing the relationship of vaccine administration to an AE. 
10.3 Reporting Procedures for all AEs  
All AEs occurring in the 28 days following each vaccination observed by the Investigator or reported 
by the volunteer, whether or not attributed to the study vaccine, will be recorded. Recording and 
reporting of all AEs will take place as detailed in SOP VC027. We will not record bruising due to 
blood-taking or cannulation.  All AEs that result in a volunteer’s withdrawal from the study will be 
followed up until a satisfactory resolution occurs, or until a non-study related causality is assigned 
(if the volunteer consents to this). SAEs will be collected throughout the entire trial period. For 
each participant, this means the safety reporting window per participant (from provision of study 
consent to final follow-up visit) 
Each AE will be graded by the participant according to the table for grading severity of AEs (see 
Table 5). Severity gradings may be reviewed and discussed with the participants at the clinic visits. 
Volunteers will be instructed on how to self-assess the severity of these AEs. There will also be 
space on the diary card to self-document unsolicited AEs, and whether medication was taken to 
relieve the symptoms. 
The severity of clinical AEs will be assessed by the Investigators according to the scales in Tables 5-
7. Severity grading for laboratory AEs are dependent on the Oxford University Hospitals 
laboratory’s reference and will be graded according to the scales in the site-specific table that can 
be found in the Trial Master File. These ranges will be based on FDA guidance relative to local 
laboratory reference ranges [https://www.fda.gov/media/73679/download].  
 
 
 
 
 

 
 
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Grade 
Severity 
GRADE 0 
None 
GRADE 1 
Mild: 
Transient 
or 
mild 
discomfort 
(< 
48 
hours); 
no 
medical 
intervention/therapy required 
GRADE 2 
Moderate: Mild to moderate limitation in activity – some assistance may be 
needed; no or minimal medical intervention/therapy required 
GRADE 3 
Severe: Marked limitation in activity, some assistance usually required; may 
require medical intervention/therapy  
Table 5: Severity grading criteria for AEs. 
 
Physical Observations 
Grade 1 
Grade 2 
Grade 3 
Tachycardia – beats per 
min* 
101-115 
116-130 
>130 
Hypotension (systolic) 
mm Hg 
85-89 
80-84 
<80 
Hypertension (systolic) 
mm Hg** 
141-159 
160-179 
180 
Hypertension (diastolic) 
mm Hg** 
91-99 
100-109 
110 
Fever °C 
37.6 – 38.0 
38.1-39.0 
>39.0 
Table 6: Severity grading criteria for clinically significant abnormal physical observations. All 
observations should be measured at rest. *Only applies when resting heart rate is between 60 and 
100 beats per minute. Use clinical judgement when characterising bradycardia in some healthy 
participant populations (e.g. conditioned athletes). **Systolic or diastolic hypertension may only 
be confirmed as clinically significant (and therefore an AE) if persistently present when 
observations are repeated (i.e. isolated measurements of hypertension are not clinically 
significant). 
 
Adverse Event 
Grade 
Intensity 
Pain at injection site 
1 
Pain that is easily tolerated 
2 
Pain that interferes with daily activity 
3 
Pain that prevents daily activity 

 
 
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Erythema at injection site 
1 
>3 - ≤50 mm 
2 
>50 - ≤100 mm 
3 
>100 mm 
Swelling at injection site* 
1 
>3 - ≤20 mm 
2 
>20 - ≤50 mm 
3 
>50 mm 
Table 7: Severity grading criteria for local adverse events. *Erythema or swelling ≤3mm is an 
expected consequence of skin puncture and will therefore not be considered an adverse event. 
10.4 Reporting Procedures for SAEs  
This will be conducted in accordance with the SOP OVC005 Safety Reporting. In order to comply 
with current regulations on SAE reporting to regulatory authorities, the event will be documented 
accurately and notification deadlines respected. SAEs will be reported on the SAE forms to 
members of the study team immediately after the Investigators become aware of their occurrence, 
as described in SOP OVC005. Copies of all reports will be forwarded for review to the CI as the 
Sponsor’s representatives) within 24 hours of the Investigator being aware of the suspected SAE. 
The Local Safety Monitor (LSM) will be notified of SAEs which are deemed possibly, probably or 
definitely related to study interventions; the LSM will be notified immediately (within 24 hours) of 
the Investigators’ being aware of their occurrence. SAEs will not normally be reported immediately 
to the REC unless there is a clinically important increase in occurrence rate, an unexpected 
outcome, or a new event that is likely to affect safety of trial volunteers, at the discretion of the CI 
and/or LSM. In addition to the reporting above, the Investigator shall include all SAEs in the annual 
Development Safety Update Report (DSUR) report. 
10.4.1 Reporting Procedures for SUSARs 
The CI will report all SUSARs to the MHRA and REC within required timelines (15 days for all 
SUSARs, unless life-threatening or fatal). Fatal and life-threatening SUSARs will be reported to the 
MHRA and REC no later than 7 calendar days after the Sponsor or delegate is first aware of the 
reaction. Any additional relevant information will be reported within 8 calendar days of the initial 
report. All other SUSARs will be reported within 15 calendar days of awareness. The CI will also 
inform all Investigators concerned of relevant information about SUSARs that could adversely 
affect the safety of participants.  
All SUSARs and deaths occurring during the study will be reported to the Sponsor. For all deaths, 
available autopsy reports and relevant medical reports will be made available for reporting to the 
relevant authorities. 
10.5 Development Safety Update Report 
The CI will submit (in addition to the expedited reporting above) DSURs once a year throughout 
the clinical trial, or on request, to the Competent Authority (MHRA in the UK, Ethics Committee, 
HRA, Host NHS Trust and Sponsor). 

 
 
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10.6 AEs of Special Interest 
AEs of special interest will be reported as SAEs. These are: 
• Severe hypersensitivity reactions (e.g. anaphylaxis) 
• Any new, suspected autoimmune or chronic inflammatory disease 
10.7 Procedures to be followed in the event of abnormal findings 
Eligibility for enrolment in the trial in terms of laboratory findings will be assessed as detailed in 
SOP VC027. Abnormal clinical findings from medical history, examination or blood tests will be 
assessed as to their clinical significance throughout the trial. Laboratory AEs will be assessed using 
the site-specific tables in the Trial Master File (TMF). If a test is deemed clinically significant, it may 
be repeated, to ensure it is not a single occurrence. If a test remains clinically significant, the 
volunteer will be informed and appropriate medical care arranged as appropriate and with the 
permission of the volunteer. Decisions to exclude the volunteer from enrolling in the trial or to 
withdraw a volunteer from the trial will be at the discretion of the Investigator. 
10.8 Local Safety Monitor 
The LSM will be appointed to provide real-time safety oversight. The LSM will review SAEs deemed 
possibly, probably or definitely related to study interventions. The LSM will be notified within 24 
hours of the Investigators’ being aware of their occurrence. The LSM has the power to terminate 
the study if deemed necessary following a study intervention-related SAE. At the time of writing, 
the LSM is Prof Brian Angus, a Clinical Tutor in Medicine, Honorary Consultant Physician and 
Director, Centre for Tropical Medicine at the University of Oxford. All correspondence between 
the Investigator and the LSM will be conveyed by the Investigator to the trial Sponsor on their 
request. 
The LSM may be contacted for advice and independent review by the Investigator or trial Sponsor 
in the following situations: 
• Following any SAE deemed to be possibly, probably, or definitely related to a study 
intervention 
• Any other situation where the Investigator or trial sponsor feels independent advice or 
review is important. 
The study can be put on hold upon advice of the LSM, CI, study sponsor or Ethical Committee(s), 
for any single event or combination of multiple events which, in their professional opinion, 
jeopardise the safety of the participants or the reliability of the data. If the study is placed on hold 
it may only be restarted following discussion with and approval from the LSM, the ethics 
committee(s), the trial Sponsor, the MHRA and CI. 
10.9 Safety Stopping/Holding Rules  
Safety holding rules have been developed considering the fact that this is a first-in-human study.  
10.9.1 Group holding rules (applies to Groups 1 and 2/3) 
If a holding rule is activated, then further vaccinations in the group (Group 1 and/or Group 2/3) 
will not occur until an internal safety review has been conducted and it is deemed appropriate to 
restart dosing. Note that as Groups 2 and 3 will be recruited in parallel, if a group holding rule is 
activated for one of the two groups, then further vaccinations in both groups would go on hold. 
The regulatory authority must be informed of the hold via substantial amendment and a request 

 
 
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to restart dosing with pertinent data must be submitted as a further, subsequent substantial 
amendment. The internal safety review will consider:  
• The relationship of the AE or SAE to the vaccine. 
• The relationship of the AE or SAE to the vaccine dose, or other possible causes of the event. 
• If appropriate, additional screening or laboratory testing for other volunteers to identify 
those who may develop similar symptoms, and alterations to the current Participant 
Information Sheet (PIS) are discussed. 
• New, relevant safety information from ongoing research programmes on the various 
components of the vaccine (i.e. the Matrix-M1 adjuvant). 
The local ethics committee and adjuvant system manufacturers (Novavax) will also be notified if a 
holding rule is activated or released to ensure that each Party is obliged to comply with the 
performance and activities related to the clinical trial (there is a confidential Safety Data Exchange 
Agreement, SDEA, in place). All vaccinated volunteers will be followed for safety until resolution 
or stabilisation (if determined to be chronic sequelae) of their AEs. Novaxax will be provided with 
SAE and pregnancy reports and any safety concerns causing a suspension as per the SDEA beween 
Oxford and Novavax 
The group holding rules are as follows: 
• Solicited local AEs: 
o If more than 25% of doses of a vaccine are followed by the same Grade 3 solicited local 
AE beginning within 2 days after vaccination (day of vaccination and one subsequent 
day) and persisting at Grade 3 for > 72 hrs. 
• Solicited systemic AEs:  
o If more than 25% of doses of a vaccine are followed by the same Grade 3 solicited 
systemic AE beginning within 2 days after vaccination (day of vaccination and one 
subsequent day) and persisting at Grade 3 for > 48 hrs.  
• Unsolicited AEs: 
o If more than 25% of volunteers develop the same Grade 3 unsolicited AE (including the 
same laboratory AE) that is considered possibly, probably or definitely related to 
vaccination and persists at Grade 3 for > 48 hrs.  
• A serious AE considered possibly, probably or definitely related to vaccination occurs 
10.9.2 Individual stopping rules 
In addition to the above stated group holding rules, stopping rules for individual volunteers will 
apply (i.e., indications to withdraw individuals from further vaccinations). 
• 
Local reactions: Injection site ulceration, abscess or necrosis. 
• 
Laboratory AEs: 

 
 
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o The volunteer develops a Grade 3 laboratory AE considered possibly, probably or 
definitely related within 7 days after vaccination and persisting continuously at Grade 
3 for >72hrs. 
• Solicited systemic AEs:  
o The volunteer develops a Grade 3 systemic solicited AE considered possibly, probably 
or definitely related within 2 days after vaccination (day of vaccination and one 
subsequent day) and persisting continuously at Grade 3 for >72hrs.  
• Unsolicited AEs:  
o The volunteer has a Grade 3 AE, considered possibly, probably or definitely related to 
vaccination, persisting continuously at Grade 3 for >72hrs.  
o The volunteer has an acute allergic reaction or anaphylactic shock following the 
administration of the vaccine investigational product. 
• A serious AE considered possibly, probably or definitely related to vaccination occurs 
 
In addition to these pre-defined criteria, the study can be put on hold upon advice of the LSM, CI, 
study sponsor, regulatory authority or REC, for any single event or combination of events which, 
in their professional opinion, jeopardise the safety of the volunteers or the reliability of the data. 
10.10 Contraception and pregnancy 
Volunteers with the potential to become pregnant are defined as those who are fertile and able 
to become pregnant following menarche until becoming post-menopausal, unless permanently 
sterile. A post-menopausal state is defined as no menstruation for 12 months without a known 
cause. 
Volunteers with the potential to become pregnant are required to use an effective form of 
contraception during the course of the study as this is a Phase I, first-in-human, study and there is 
currently no information about the effect of this vaccine on a foetus.  
Acceptable forms of contraception for volunteers of child-bearing potential are: 
• Established use of oral, injected or implanted hormonal methods of contraception. 
• Placement of an intrauterine device or intrauterine system. 
• Male sterilization: if the vasectomised partner is the sole partner for the participant. 
• True abstinence from sex with sperm-producing partners, when this is in line with the 
preferred and usual lifestyle of the participant (periodic abstinence and withdrawal are not 
acceptable methods of contraception). 
Pregnancy, lactation or intention to become pregnant during the study is an exclusion criteria. 
 
 

 
 
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11 STATISTICS 
11.1  Sample size 
This is an observational and descriptive safety study, where healthy adult volunteers living in the 
UK will be vaccinated with Pvs25-IMX313, in formulation with the Matrix-M1 adjuvant. Up to a 
total of 30 volunteers across three equally divided groups will be vaccinated. Each volunteer will 
receive three doses of Pvs25-IMX313, formulated in 50 µg Matrix-M1, on days 0, 28 and 56 (see 
Table 2 for exact dosing regimen depending on group allocation). This sample size should allow 
estimation and comparison of the frequency and magnitude of the outcome measures, at and 
between the three different dosing regimens. As this is the first study involving the Pvs25-IMX313 
in Matrix-M1 vaccine, there is no available data to enable more precise modelling/power 
calculations. However, it is considered that eight to ten volunteers in each group will be sufficient 
to enable detection of statistically significant differences in the primary and secondary outcome 
measures (see Section 6) between the groups.  
 
 
 
 

 
 
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12 QUALITY CONTROL AND QUALITY ASSURANCE PROCEDURES 
12.1  Investigator procedures 
Approved site-specific SOPs will be used at all clinical and laboratory sites. 
12.2 Monitoring 
Monitoring will be performed by Appledown external monitors using established procedures, 
according to the principles of Good Clinical Practice (GCP).  Following written SOPs, the monitors 
will verify that the clinical trial is conducted and data are generated, documented and reported in 
compliance with the protocol and GCP. The site will provide direct access to all trial-related source 
data/documents and reports for the purpose of auditing by the Sponsor and inspection by local 
and regulatory authorities. 
12.3 Modification to protocol 
No amendments to this protocol will be made without consultation with, and the agreement of, 
the Sponsor. Any amendments to the trial that become necessary during the course of the trial 
must be discussed by the Investigator and Sponsor concurrently. If agreement is reached 
concerning the need for an amendment, it will be produced in writing by the CI and will be made 
a formal part of the protocol following ethical and regulatory approval, as applicable.  
An administrative change to the protocol is one that modifies administrative and logistical aspects 
of a protocol but does not affect the participant’s safety, the objectives of the trial and/or its 
progress. An administrative change may not require UK ethical committee approval; however, the 
REC will be notified in the event of any such change.  
The Investigator is responsible for ensuring that changes to an approved trial, during the period 
for which ethical committee(s) approval has already been given, are not initiated without review 
and approval from the ethics committee(s) and regulatory authorities except to eliminate 
apparent immediate hazards to the participant (Urgent Safety Measures only). Protocol adherence 
is a fundamental part of the conduct of a clinical study.  Changes to the approved protocol need 
prior approval unless for urgent safety reasons. Investigators must not request a protocol waiver 
to enter a patient who does not satisfy the selection criteria. 
12.4 Protocol deviation  
Any deviations from the protocol will be documented in a protocol deviation form and filed in the 
TMF.  
12.5 Audit & inspection  
The Quality Assurance manager performs systems-based internal audits to check that trials are 
being conducted, data recorded, analysed and accurately reported according to study protocols, 
departmental SOPs and in compliance with GCP. The audit schedule includes laboratory activities. 
The internal audits will supplement the independent monitoring process and will review processes 
not covered by the independent monitor. 
The Sponsor, trial site, ethical committee(s), and authorised individuals may carry out audit to 
ensure compliance with the protocol, GCP and appropriate regulations. GCP inspections may also 
be undertaken by the regulatory authority to ensure compliance with protocol and national 
regulations. The Sponsor will assist in any inspections. 

 
 
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12.6 Serious breaches 
The Medicines for Human Use (Clinical Trials) Regulations contain a requirement for the 
notification of "serious breaches" to the MHRA within 7 days of the Sponsor becoming aware of 
the breach. 
A serious breach is defined as “A breach of GCP or the trial protocol which is likely to effect to a 
significant degree –  
(a) the safety or physical or mental integrity of the participants of the trial; or 
(b) the scientific value of the trial. 
In the event that a serious breach is suspected the Sponsor must be contacted within 1 working 
day. In collaboration with the CI the serious breach will be reviewed by the Sponsor and, if 
appropriate, the Sponsor will report it to the REC, regulatory authority and the relevant NHS host 
organisation within seven calendar days. 
12.7 Publication policy 
The Investigators will be involved in reviewing drafts of the manuscripts, abstracts, press releases 
and any other publications arising from the study.  Authorship will be determined in accordance 
with the International Committee of Medical Journal Editors guidelines and other contributors will 
be acknowledged.  
12.8 Intellectual Property 
Ownership of intellectual property generated by employees of the University vests in the 
University.  The protection and exploitation of any new intellectual property is managed by the 
University’s technology transfer office, Oxford University Innovation. 

 
 
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13 ETHICS 
13.1 Declaration of Helsinki 
The Investigator will ensure that this study is conducted according to the principles of the current 
revision of the Declaration of Helsinki 2008. 
13.2 Good Clinical Practice 
The Investigator will ensure that this study is conducted in full conformity to the Medicines for 
Human Use (Clinical Trials) Regulations 2004 (SI 2004 No. 1031) and its amendments and with the 
guidelines for GCP (CPMP/ICH/135/95) July 1996.  
13.3 Approvals 
Following Sponsor approval, a copy of the protocol, proposed informed consent form, other 
written volunteer information and the proposed advertising material will be submitted to an 
appropriate REC and the MHRA for written approval. The Investigator will submit and, where 
necessary, obtain approval from the REC and the MHRA for all subsequent amendments to the 
protocol and associated trial documents. A non-substantial amendment does not require REC 
approval; however, the REC will be notified in the event of any such change. The Investigator will 
notify all deviations from the protocol or SAEs occurring at the site to the Sponsor and will notify 
the REC(s) and regulatory authorities of these if necessary and in accordance with procedures. 
13.4 
Reporting  
The CI shall submit once a year throughout the study, or on request, an Annual Progress Report to 
the REC, HRA (where required) and Sponsor. In addition, an End of Trial notification and final report 
will be submitted to the MHRA, the REC, host organisation and Sponsor. 
13.5 Volunteer confidentiality 
The research data will be pseudoanonymised; participant data will be identified by a unique study 
number in the CRF and database. Separate confidential files containing identifiable information 
and links to the study number will be stored in secured locations. These secured locations are 
either physical premises or web servers owned by the University of Oxford or in the case of the 
eCRF on the OpenClinica database. The OpenClinica database has undergone the Third Party 
Security Assessment process and been officially approved as a processor of data for which the 
University of Oxford is the controller. Only the Sponsor, Investigators, the clinical monitor, the 
ethical committee(s) and the regulatory authorities will have access to the records.  
Photographs taken (if required, with the volunteer’s written, informed consent) will not include 
the volunteer’s face and will be identified by the volunteer’s trial specific identification number 
only. Once developed, photographs will be stored as confidential records, as above. This material 
may be shown to other professional staff, used for educational purposes, or included in a scientific 
publication.  

 
 
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14 DATA HANDLING AND RECORD KEEPING 
14.1 Data handling 
The CI will be the data custodian with responsibility for delegating the receiving, entering, cleaning, 
querying, analysing and storing all data that accrues from the study. Data will be entered into the 
volunteers’ CRFs in a paper and/or electronic format (using the OpenClinica™ database). Electronic 
data will be stored on secure servers which are outsourced by OpenClinica™. OpenClinica™ meets 
FDA part 11B standards and is a system approved by the University of Oxford. This includes safety 
data, laboratory data and outcome data. Data are entered in a web browser on personal 
computers and then transferred to the OpenClinica Database by encrypted (https) transfer. Safety 
data will also be collected through an electronic diary, which is stored on a third party server (see 
Section 13.5).  
14.2 Record keeping 
The Investigators will maintain and retain appropriate medical and research records and essential 
documents for this trial in compliance with GCP and regulatory and institutional requirements for 
the protection of confidentiality of volunteers. The CI, co-investigators, clinical research nurses and 
authorised personnel will have access to records. The Investigators will permit authorised 
representatives of the Sponsor, ethical committee(s), regulatory agencies, and the monitors to 
examine (and when required by applicable law, to copy) clinical records for the purposes of quality 
assurance reviews, audits and evaluation of the study safety and progress. 
14.3 Source data and electronic case report forms (eCRFs) 
All protocol-required information will be collected in the electronic diaries and eCRFs designed by 
the Investigator. All source documents will be filed in the participants’ notes. Source documents 
are original documents, data, and records from which the volunteer’s eCRF data are obtained. For 
this study these will include, but are not limited to: the volunteer consent form, blood results, GP 
response letters, laboratory records and correspondence. In the majority of cases, electronic 
diaries and eCRF entries will be considered source data as these are the site of the original 
recording (i.e. there is no other written or electronic record of data). In this study this will include, 
but is not limited to: medical history, medication records, vital signs, physical examination records, 
urinalysis results, blood results, AE data and details of study interventions. All source data and 
volunteer CRFs will be stored securely. 
14.4 Data protection  
The trial will comply with the UK General Data Protection Regulation (UK GDPR) and Data 
Protection Act 2018. The study protocol, documentation, data and all other information generated 
will be held in strictest confidence. No information concerning the study or the data will be 
released to any unauthorised third party, without prior written approval of the Sponsor. 
14.5 Storage and use of data 
The University of Oxford will keep identifiable information from participants collected during the 
study for a maximum of 5 years after the study has finished. This will be limited to the minimum 
personal data necessary. In addition, we will securely store the anonymised research data and any 
research documents with personal information, such as consent forms, for a maximum of 5 years 
after the end of the study. The need to store this information for longer, in relation to licensing of 
the vaccine will be reviewed every 5 years. Once the study has been completed, all documents 

 
 
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would be archived in a secure facility. Files will be confidentially destroyed if storage is no longer 
required. For effective vaccines that may be licensed, secure storage of research data may be 
required for at least 15 years after the end of the study, subject to adjustments in clinical trials 
regulations. Financial information will be stored for seven years in line with University of Oxford 
financial policy.  
 
Where specific written consent is provided, personal details may be used to contact participants 
about future related research. These details and clinical information will be held separately. Note 
that the informed consent form from this study will be retained, separately, for as long as the 
participant is on the database. If participants elect to take part in another study carried out by the 
Jenner Institute, personal information and medical information including blood test results may 
be accessed to avoid unnecessary repetition.  
 

 
 
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15 FINANCING AND INSURANCE 
15.1 Financing 
The study is funded through a grant from MultiViVax, a European Commission Horizon 2020 
funded project (733073), which funds work to develop candidate multi-stage malaria vaccines 
through innovative trial designs in malaria-endemic areas.  
15.2 Insurance 
The University has a specialist insurance policy in place which would operate in the event of any 
participant suffering harm as a result of their involvement in the research (Newline Underwriting 
Management Ltd, at Lloyd’s of London).  
15.3 Compensation 
Volunteers will be compensated for their time and for the inconvenience caused by procedures as 
below.   
- Screening visit  
 
 
 
 
 
 
 
£25  
- Subsequent visits: 
 
- Travel expenses 
 
 
 
 
 
 
£15 per visit 
- Inconvenience of blood tests: 
 
 
 
 
£10 per blood donation 
- Time required for visit: 
 
 
 
 
 
£20 per hour 
Where travel expenses are greater than £15 per visit because the volunteer lives outside the city 
of the trial site, the volunteer may be given further reimbursement to meet the cost of travel 
necessary for study visits at the Investigator’s discretion.  
 
Table 8:  Estimated compensation amounts. 
Time in trial 
(approx.) 
Maximum visits Maximum volume of 
blood taken (mls) 
Compensation 
Groups 1-3 
1 year 
16 
972 
£790 

 
 
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16 APPENDICES 
Appendix A: laboratory values for exclusion 
Laboratory parameters for inclusion/exclusion in the trial will be considered on an individual basis, 
with Investigator discretion for interpretation of results and the need for repeated or further tests. 
In general, volunteers will be excluded if a result at screening constitutes what would qualify as a 
grade 1 (or higher) laboratory adverse event, according to the laboratory adverse event tables 
(filed in the TMF), on repeat of an abnormal test result. Urinalysis at screening will be assessed as 
per the table below: 
 
URINE ANALYSIS (using MULTISTIX) 
Protein* 
2+ or Protein creatinine ratio of ≥ 50 mg/mmol 
Blood£ 
2+ on two dipstick tests  
Glucose 
1+ 
Table 9: Urinalysis assessment.  
*In the event of the dipstick testing positive for protein with ≥1+ protein urine should be sent for 
a protein creatinine ratio. 
£ In the event of urine dipstick testing positive for ≥1+ blood with, or without, protein, a repeat 
dipstick test will be carried out to confirm haematuria. In volunteers of child-bearing potential, a 
menstrual history will be taken to elicit whether the participant is currently menstruating and if 
they are, urine dipstick will be repeated after 1 - 2 weeks. If blood and/or proteinuria persist in any 
volunteer, an interpretation of the results will be undertaken by the Investigator on an individual 
basis to determine if they will be excluded from the trial, and the appropriate follow-up arranged. 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
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17 REFERENCES 
1. 
Organization, W.H. Malaria.  22/06/2021]; Available from: 
https://www.who.int/data/gho/data/themes/malaria. 
2. 
Kochar, D.K., et al., Plasmodium vivax malaria. Emerg Infect Dis, 2005. 11(1): p. 132-4. 
3. 
Organisation, W.H. World malaria report 2020. 2020  22/06/2021]; Available from: 
https://www.who.int/publications/i/item/9789240015791. 
4. 
Howes, R.E., et al., Global Epidemiology of Plasmodium vivax. Am J Trop Med Hyg, 
2016. 95(6 Suppl): p. 15-34. 
5. 
Sattabongkot, J., et al., Plasmodium vivax transmission: chances for control? Trends 
Parasitol, 2004. 20(4): p. 192-8. 
6. 
Oliveira-Ferreira, J., et al., Malaria in Brazil: an overview. Malar J, 2010. 9: p. 115. 
7. 
Rodriguez, J.C., et al., Epidemiology and control of malaria in Colombia. Mem Inst 
Oswaldo Cruz, 2011. 106 Suppl 1: p. 114-22. 
8. 
Kenangalem, E., et al., Malaria morbidity and mortality following introduction of a universal 
policy of artemisinin-based treatment for malaria in Papua, Indonesia: A longitudinal 
surveillance study. PLoS Med, 2019. 16(5): p. e1002815. 
9. 
White, N.J. and M. Imwong, Relapse. Adv Parasitol, 2012. 80: p. 113-50. 
10. 
Dini, S., et al., The risk of morbidity and mortality following recurrent malaria in Papua, 
Indonesia: a retrospective cohort study. BMC Med, 2020. 18(1): p. 28. 
11. 
Barcus, M.J., et al., Demographic risk factors for severe and fatal vivax and falciparum 
malaria among hospital admissions in northeastern Indonesian Papua. Am J Trop Med 
Hyg, 2007. 77(5): p. 984-91. 
12. 
Nurleila, S., et al., Serious and fatal illness associated with falciparum and vivax malaria 
among patients admitted to hospital at West Sumba in eastern Indonesia. Am J Trop Med 
Hyg, 2012. 87(1): p. 41-49. 
13. 
Kochar, D.K., et al., A prospective study on adult patients of severe malaria caused by 
Plasmodium falciparum, Plasmodium vivax and mixed infection from Bikaner, northwest 
India. J Vector Borne Dis, 2014. 51(3): p. 200-10. 
14. 
Alexandre, M.A., et al., Severe Plasmodium vivax malaria, Brazilian Amazon. Emerg 
Infect Dis, 2010. 16(10): p. 1611-4. 
15. 
Price, R.N., et al., Vivax malaria: neglected and not benign. Am J Trop Med Hyg, 2007. 
77(6 Suppl): p. 79-87. 
16. 
Carlton, J.M., B.J. Sina, and J.H. Adams, Why is Plasmodium vivax a neglected tropical 
disease? PLoS Negl Trop Dis, 2011. 5(6): p. e1160. 
17. 
Watson, J., et al., Implications of current therapeutic restrictions for primaquine and 
tafenoquine in the radical cure of vivax malaria. PLoS Negl Trop Dis, 2018. 12(4): p. 
e0006440. 
18. 
Pybus, B.S., et al., The metabolism of primaquine to its active metabolite is dependent on 
CYP 2D6. Malar J, 2013. 12: p. 212. 
19. 
Bennett, J.W., et al., Primaquine failure and cytochrome P-450 2D6 in Plasmodium vivax 
malaria. N Engl J Med, 2013. 369(14): p. 1381-2. 
20. 
WHO. World malaria report 2020. 2020  22/06/2021]; Available from: 
https://www.who.int/publications/i/item/9789240015791. 
21. 
Wongsrichanalai, C. and C.H. Sibley, Fighting drug-resistant Plasmodium falciparum: the 
challenge of artemisinin resistance. Clin Microbiol Infect, 2013. 19(10): p. 908-16. 

 
 
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Page 65 of 68 
 
22. 
WHO. Malaria Vaccine Roadmap.  22/06/2021]; Available from: 
https://www.who.int/immunization/topics/malaria/vaccine_roadmap. 
23. 
Price, R.N., et al., Global extent of chloroquine-resistant Plasmodium vivax: a systematic 
review and meta-analysis. Lancet Infect Dis, 2014. 14(10): p. 982-91. 
24. 
Sinden, R.E., The cell biology of sexual development in plasmodium. Parasitology, 1983. 
86 (Pt 4): p. 7-28. 
25. 
Sinden, R.E., A biologist's perspective on malaria vaccine development. Hum Vaccin, 
2010. 6(1): p. 3-11. 
26. 
Meibalan, E. and M. Marti, Biology of Malaria Transmission. Cold Spring Harb Perspect 
Med, 2017. 7(3). 
27. 
de Jong, R.M., et al., Immunity against sexual stage Plasmodium falciparum and 
Plasmodium vivax parasites. Immunol Rev, 2020. 293(1): p. 190-215. 
28. 
Obaldia, N., 3rd, et al., Bone Marrow Is a Major Parasite Reservoir in Plasmodium vivax 
Infection. mBio, 2018. 9(3). 
29. 
Billker, O., et al., Identification of xanthurenic acid as the putative inducer of malaria 
development in the mosquito. Nature, 1998. 392(6673): p. 289-92. 
30. 
Russell, B., et al., A reliable ex vivo invasion assay of human reticulocytes by Plasmodium 
vivax. Blood, 2011. 118(13): p. e74-81. 
31. 
Bennett, J.W., et al., Phase 1/2a Trial of Plasmodium vivax Malaria Vaccine Candidate 
VMP001/AS01B in Malaria-Naive Adults: Safety, Immunogenicity, and Efficacy. PLoS 
Negl Trop Dis, 2016. 10(2): p. e0004423. 
32. 
Arevalo-Herrera, M.L.-D., M; Caicedo, MA; Henao-Geraldo, J; Castellanos, A; Gaitan, X; 
Devaud, J-C; Pannatier, A; Herrera, S; Onate, J; Corradin, G; Herrera, S, Randomized 
clinical trial to assess the protective efficacy of a Plasmodium vivax CS synthetic vaccine. 
Research Square, 2021. 
33. 
Clyde, D.F., Immunity to falciparum and vivax malaria induced by irradiated sporozoites: a 
review of the University of Maryland studies, 1971-75. Bull World Health Organ, 1990. 68 
Suppl: p. 9-12. 
34. 
Arevalo-Herrera, M., et al., Protective Efficacy of Plasmodium vivax Radiation-Attenuated 
Sporozoites in Colombian Volunteers: A Randomized Controlled Trial. PLoS Negl Trop 
Dis, 2016. 10(10): p. e0005070. 
35. 
Payne, R.O., et al., Human vaccination against Plasmodium vivax Duffy-binding protein 
induces strain-transcending antibodies. JCI Insight, 2017. 2(12). 
36. 
Singh, K., et al., Malaria vaccine candidate based on Duffy-binding protein elicits strain 
transcending functional antibodies in a Phase I trial. NPJ Vaccines, 2018. 3: p. 48. 
37. 
Mueller, I., A.R. Shakri, and C.E. Chitnis, Development of vaccines for Plasmodium vivax 
malaria. Vaccine, 2015. 33(52): p. 7489-95. 
38. 
Bauza, K., et al., Efficacy of a Plasmodium vivax malaria vaccine using ChAd63 and 
modified vaccinia Ankara expressing thrombospondin-related anonymous protein as 
assessed with transgenic Plasmodium berghei parasites. Infect Immun, 2014. 82(3): p. 
1277-86. 
39. 
Vanloubbeeck, Y., et al., Comparison of the immune responses induced by soluble and 
particulate Plasmodium vivax circumsporozoite vaccine candidates formulated in AS01 in 
rhesus macaques. Vaccine, 2013. 31(52): p. 6216-24. 
40. 
Yadava, A., et al., Protective efficacy of a Plasmodium vivax circumsporozoite protein-
based vaccine in Aotus nancymaae is associated with antibodies to the repeat region. 
PLoS Negl Trop Dis, 2014. 8(10): p. e3268. 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
Page 66 of 68 
 
41. 
Herrera, S., et al., Use of long synthetic peptides to study the antigenicity and 
immunogenicity of the Plasmodium vivax circumsporozoite protein. Int J Parasitol, 2004. 
34(13-14): p. 1535-46. 
42. 
Herrera, S., et al., Phase I safety and immunogenicity trial of Plasmodium vivax CS 
derived long synthetic peptides adjuvanted with montanide ISA 720 or montanide ISA 51. 
Am J Trop Med Hyg, 2011. 84(2 Suppl): p. 12-20. 
43. 
Herrera, S., et al., Safety and elicitation of humoral and cellular responses in colombian 
malaria-naive volunteers by a Plasmodium vivax circumsporozoite protein-derived 
synthetic vaccine. Am J Trop Med Hyg, 2005. 73(5 Suppl): p. 3-9. 
44. 
Yadava, A., et al., A novel chimeric Plasmodium vivax circumsporozoite protein induces 
biologically functional antibodies that recognize both VK210 and VK247 sporozoites. 
Infect Immun, 2007. 75(3): p. 1177-85. 
45. 
Bouillet, L.E., et al., Long-term humoral and cellular immune responses elicited by a 
heterologous Plasmodium vivax apical membrane antigen 1 protein prime/adenovirus 
boost immunization protocol. Infect Immun, 2011. 79(9): p. 3642-52. 
46. 
Ntumngia, F.B. and J.H. Adams, Design and immunogenicity of a novel synthetic antigen 
based on the ligand domain of the Plasmodium vivax duffy binding protein. Clin Vaccine 
Immunol, 2012. 19(1): p. 30-6. 
47. 
Rosa, D.S., et al., Immunogenicity of a recombinant protein containing the Plasmodium 
vivax vaccine candidate MSP1(19) and two human CD4+ T-cell epitopes administered to 
non-human primates (Callithrix jacchus jacchus). Microbes Infect, 2006. 8(8): p. 2130-7. 
48. 
Devi, Y.S., et al., Immunogenicity of Plasmodium vivax combination subunit vaccine 
formulated with human compatible adjuvants in mice. Vaccine, 2007. 25(28): p. 5166-74. 
49. 
Moreno, A., et al., Preclinical assessment of the receptor-binding domain of Plasmodium 
vivax Duffy-binding protein as a vaccine candidate in rhesus macaques. Vaccine, 2008. 
26(34): p. 4338-44. 
50. 
Wiley, S.R., et al., Targeting TLRs expands the antibody repertoire in response to a 
malaria vaccine. Sci Transl Med, 2011. 3(93): p. 93ra69. 
51. 
de Cassan, S.C., et al., Preclinical Assessment of Viral Vectored and Protein Vaccines 
Targeting the Duffy-Binding Protein Region II of Plasmodium Vivax. Front Immunol, 2015. 
6: p. 348. 
52. 
Armistead, J.S., et al., Antibodies to a single, conserved epitope in Anopheles APN1 
inhibit universal transmission of Plasmodium falciparum and Plasmodium vivax malaria. 
Infect Immun, 2014. 82(2): p. 818-29. 
53. 
Hisaeda, H., et al., Antibodies to malaria vaccine candidates Pvs25 and Pvs28 completely 
block the ability of Plasmodium vivax to infect mosquitoes. Infect Immun, 2000. 68(12): p. 
6618-23. 
54. 
Malkin, E.M., et al., Phase 1 vaccine trial of Pvs25H: a transmission blocking vaccine for 
Plasmodium vivax malaria. Vaccine, 2005. 23(24): p. 3131-8. 
55. 
Wu, Y., et al., Phase 1 trial of malaria transmission blocking vaccine candidates Pfs25 
and Pvs25 formulated with montanide ISA 51. PLoS One, 2008. 3(7): p. e2636. 
56. 
mal, E.R.A.C.G.o.V., A research agenda for malaria eradication: vaccines. PLoS Med, 
2011. 8(1): p. e1000398. 
57. 
Huff, C.G., D.F. Marchbank, and T. Shiroishi, Changes in infectiousness of malarial 
gametocytes. II. Analysis of the possible causative factors. Exp Parasitol, 1958. 7(4): p. 
399-417. 
58. 
Carter, R. and D.H. Chen, Malaria transmission blocked by immunisation with gametes of 
the malaria parasite. Nature, 1976. 263(5572): p. 57-60. 

 
 
VAC084 Clinical Trial Protocol                                     V2.0                                             24 Mar 2022 
IRAS Project ID: 302239                                                                                         
Page 67 of 68 
 
59. 
Ponnudurai, T., et al., The production of mature gametocytes of Plasmodium falciparum in 
continuous cultures of different isolates infective to mosquitoes. Trans R Soc Trop Med 
Hyg, 1982. 76(2): p. 242-50. 
60. 
Rener, J., et al., Target antigens of transmission-blocking immunity on gametes of 
plasmodium falciparum. J Exp Med, 1983. 158(3): p. 976-81. 
61. 
Tomas, A.M., et al., P25 and P28 proteins of the malaria ookinete surface have multiple 
and partially redundant functions. EMBO J, 2001. 20(15): p. 3975-83. 
62. 
Miura, K., et al., Transmission-blocking activity induced by malaria vaccine candidates 
Pfs25/Pvs25 is a direct and predictable function of antibody titer. Malar J, 2007. 6: p. 107. 
63. 
Baton, L.A. and L.C. Ranford-Cartwright, Do malaria ookinete surface proteins P25 and 
P28 mediate parasite entry into mosquito midgut epithelial cells? Malar J, 2005. 4: p. 15. 
64. 
Saxena, A.K., et al., The essential mosquito-stage P25 and P28 proteins from 
Plasmodium form tile-like triangular prisms. Nat Struct Mol Biol, 2006. 13(1): p. 90-1. 
65. 
Blagborough, A.M., et al., Transmission blocking potency and immunogenicity of a plant-
produced Pvs25-based subunit vaccine against Plasmodium vivax. Vaccine, 2016. 
34(28): p. 3252-9. 
66. 
Kapulu, M.C., et al., Comparative assessment of transmission-blocking vaccine 
candidates against Plasmodium falciparum. Sci Rep, 2015. 5: p. 11193. 
67. 
Jones, R.M., et al., A novel plant-produced Pfs25 fusion subunit vaccine induces long-
lasting transmission blocking antibody responses. Hum Vaccin Immunother, 2015. 11(1): 
p. 124-32. 
68. 
Goodman, A.L., et al., A viral vectored prime-boost immunization regime targeting the 
malaria Pfs25 antigen induces transmission-blocking activity. PLoS One, 2011. 6(12): p. 
e29428. 
69. 
Ogun, S.A., et al., The oligomerization domain of C4-binding protein (C4bp) acts as an 
adjuvant, and the fusion protein comprised of the 19-kilodalton merozoite surface protein 
1 fused with the murine C4bp domain protects mice against malaria. Infect Immun, 2008. 
76(8): p. 3817-23. 
70. 
Minhinnick, A., et al., A first-in-human phase 1 trial to evaluate the safety and 
immunogenicity of the candidate tuberculosis vaccine MVA85A-IMX313, administered to 
BCG-vaccinated adults. Vaccine, 2016. 34(11): p. 1412-21. 
71. 
de Graaf, H.P., RO; Taylor, I; Miura, K; Long, CA; Elias, SC; Zaric, M; Minassian, AM; 
Silk, SE; Li, L; Poulton, ID; Baker, M; Draper, SJ, Safety and Immunogenicity of 
ChAd63/MVA Pfs25-IMX313 in a Phase I First-in-Human Trial. In submission, 2021. 
72. 
Tsuboi, T., et al., Sequence polymorphism in two novel Plasmodium vivax ookinete 
surface proteins, Pvs25 and Pvs28, that are malaria transmission-blocking vaccine 
candidates. Mol Med, 1998. 4(12): p. 772-82. 
73. 
Li, Y., et al., Enhancing immunogenicity and transmission-blocking activity of malaria 
vaccines by fusing Pfs25 to IMX313 multimerization technology. Sci Rep, 2016. 6: p. 
18848. 
74. 
Bengtsson, K.L., et al., Matrix-M adjuvant enhances antibody, cellular and protective 
immune responses of a Zaire Ebola/Makona virus glycoprotein (GP) nanoparticle vaccine 
in mice. Vaccine, 2016. 34(16): p. 1927-35. 
75. 
Cox, R.J., et al., Evaluation of a virosomal H5N1 vaccine formulated with Matrix M™ 
adjuvant in a phase I clinical trial. Vaccine, 2011. 29(45): p. 8049-59. 
76. 
Venkatraman, N., et al., Safety and immunogenicity of heterologous prime-boost 
immunization with viral-vectored malaria vaccines adjuvanted with Matrix-M. Vaccine, 
2017. 35(45): p. 6208-6217. 

 
 
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IRAS Project ID: 302239                                                                                         
Page 68 of 68 
 
77. 
Heath, P., et al., Safety and efficacy of NVX-CoV2373 Covid-19 vaccine. New England 
Journal of Medicine, 2021.