A Phase I/IIa Clinical Trial to Assess the Safety, Immunogenicity and Efficacy of the Blood-stage Plasmodium Vivax Malaria Vaccine Candidate PvDBPII in Matrix M1 in Healthy Adults Living in the UK
Phase: PHASE1
Conditions: Malaria, Vivax
Sponsor: University of Oxford

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 1 of 117 
 
 
 
 
 
 
 
 
 
A Phase I/IIa clinical trial to assess the safety, immunogenicity and efficacy of the blood-stage 
Plasmodium vivax malaria vaccine candidate PvDBPII in Matrix M1 in healthy adults living in 
the UK 
 
Study Reference: VAC079 
Protocol Version: 8.0 
Date: 21st June 2022 
 
 
IRAS Project ID: 265494 
REC reference: 19/SC/0330 
EudraCT number: 2019-002872-14 
 
 
Chief Investigator: Angela M. Minassian 
Sponsor: University of Oxford 
 
 
 
 
 
 

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MODIFICATION HISTORY 
Version 
Date 
Author(s) 
Modifications 
1.0 
17th July 2019
Thomas Rawlinson 
Yrene 
Themistocleous 
Angela Minassian 
Created 
2.0 
24th October 
2019 
Yrene 
Themistocleous 
Revision of accepted methods of contraception for 
women of child bearing potential to include highly 
effective methods only 
Clarification of requirement for female participants using 
hormonal contraceptive methods to use an additional 
form of contraceptive until the next menstrual period, 
following initiation of treatment with 
artemether/lumefantrine  
2.1 
20th January 
2020 
Yrene 
Themistocleous 
Minor corrections to schedule of visits table (clarifying 
total immunology bleed and volume of blood draw for 
haematology test at screening corrected to 4mL from 
5mL)  
3.0 
14th February 
2020 
Thomas Rawlinson 
Exclusion criteria relating to prior immunoglobulin 
exposure amended. 
Removal of thick blood film as diagnostic measure during 
CHMI  
Updates and corrections to schedule of visits and bleed 
volumes, including reduction of maximal blood draw 
Addition of timing windows for recording physical 
observations on day of challenge 
Clarifications to collection of adverse event data  
Potential for use of alternative antiemetic to cyclizine 
Change to group sizes (target range now specified) and 
removal of back up volunteers 
Clarification regarding timing of screening visits and for 
re-screening procedures for participants screened >90 
days prior to enrolment 
CRP added as an exploratory measure 
Local safety monitor takes on role previously taken by 
local safety committee 
Additional wording in the protocol and PIS to clarify the 
repeat screening for blood-borne infection at day 96 post 
challenge 

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4.0 
25th 
November 
2020 
Mimi Hou 
Change of vaccination schedule for group 1 - delay of 3rd 
vaccination due to temporary trial halt 
Addition of Group 2, to be recruited if fewer than 6 
participants complete the study in Group 1 
Addition of serum bhCG to C+28/day of malaria diagnosis
Clarification that G6PD, DARC and haemoglobinopathy 
screen are only done at NHS labs 
Extension of time window for study visits 
Changes to trial procedures to account for possibility of 
COVID-19 infection during CHMI. Addition of COVID-19 
PCR swab test prior to challenge and on day of malaria 
diagnosis. Guidance on testing for COVID-19 if fever post 
vaccination and post challenge. 
Added option of using Malarone as first line anti-malaria 
treatment 
Correction of typographical errors 
Removal of measurement of T cell responses to PvDBPII 
by ELISpot from Secondary Immunological Outcome 
Measures 
Correction of error in calculation of total blood volumes 
in schedule of attendance table 
Addition of retrospective COVID-19 serology testing for 
exploratory analysis of effects of COVID serostatus on 
vaccine immunogenicity 
5.0 
19th January 
2021 
Mimi Hou 
Clarification of exclusion criteria regarding concomitant 
vaccinations and addition of specific criteria relating to 
licensed COVID-19 vaccination 
Addition of section on administration of concomitant 
COVID-19 vaccination 
Extension of time window for 2nd and 3rd vaccinations 
Threshold at which new participants are recruited into 
Group 2 amended to if less than 8 participants complete 
study in Group 1 (previously less than 6 participants)  
6.0 
4th March 
2021 
Mimi Hou 
Addition of exclusion criteria on concomitant COVID-19 
vaccination around time of CHMI.  
Shortened time window of when COVID-19 vaccination 
can be given following malaria vaccination to aid 
scheduling of COVID-19 vaccinations. 
Updated section on Conduct of CHMI in the context of 
COVID-19 pandemic 

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Removal of specific timeframe during which baseline 
observations are taken pre-challenge 
Correction of blood volume taken for HLA and total 
blood volumes 
7.0 
22nd July 
2021 
Mimi Hou 
Addition of Group 3, comprising a subset of volunteers 
originally in Group 1, who consent to undergo a 4th 
vaccination and secondary CHMI.  
Addition of secondary objective to assess vaccine 
efficacy in Group 3. 
For Groups 2 and 3 - post-challenge follow-up visits 
changed to once a day until parasite count reaches 
>1000 genome copies/ml, then to continue twice a day 
visits until diagnosis. Latest day of treatment reduced to 
C+21. 
For Groups 2 and 3 - change of post malaria treatment 
visit from T+2 to T+3. Reduction in number of observed 
doses of antimalarial medication to two observed doses. 
For Group 2 – addition of immunology bleed at D42 
Updated compensation table 
Correction of error in lumefantrine dose in section 8.5.5  
Addition of reticulocyte count to FBC taken at C-2 visit. 
Removal of need to collect unused medications after 
completion of challenge. Correction under C+56 visit – 
no collection of any diaries occur at this timepoint. 
Correction of errors in protocol – only weight taken at C-
2 visit. No CRPs are taken during post-challenge follow-
up. 
Changed Senior Laboratory Investigator address. 
Addition of New Biochemstry Building as location for 
processing of research bloods. 
8.0 
21st June 
2022 
Mimi Hou 
Samples will now be stored long term under University 
of Oxford’s HTA license at Department of Biochemistry 
instead of Oxford Vaccine Centre Biobank. 
 
 
 
 
 

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A Phase I/IIa clinical trial to assess the safety, immunogenicity and efficacy of the blood-stage 
Plasmodium vivax malaria vaccine candidate PvDBPII in Matrix M1 in healthy adults living in 
the UK 
Study Code:  VAC079 
 
Chief Investigator 
Dr Angela M Minassian 
 
 
 
 
 
 
Senior Laboratory 
Investigator 
 
 
 
 
 
 
Trial sites 
 
 
 
 
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 Simon J Draper  
New Biochemistry Building, 
Department of Biochemistry, 
South Parks Road 
Oxford, OX1 3QU 
Email: simon.draper@bioch.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 
Clinical Trials and Research Governance (CTRG)  
Research Services 
University of Oxford 
Joint Research Office 
Boundary Brook House 

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Churchill Drive 
Headington 
OX3 7GB 
Tel: 01865 289885  
Email: ctrg@admin.ox.ac.uk 
 
External Monitor  
 
 
 
 
 
Clinical Trials and Research Governance (CTRG) 
Research Services 
University of Oxford 
Joint Research Office 
Boundary Brook House 
Churchill Drive 
Headington 
OX3 7GB 
Tel: 01865 289885 
 
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 
 
 
 
 
 
 
 
 
 
 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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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. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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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 M. Minassian 
 
 
 
 
23/06/2022 
Chief Investigator 
 
 
Investigator Signature 
 
Date  
 
Conflict of Interest 
1. “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare no 
conflict of interest” 
Dr Angela M. Minassian 
 
 
 
23/06/2022 
Chief Investigator 
 
 
Investigator Signature 
 
Date  
 
Details:_________________________________________________________________________ 
 
2.  “According to the Declaration of Helsinki, 2008, I have read this protocol, and declare no 
conflict of interest” 
Prof Simon J. Draper 
 
 
 
23/06/2022 
Senior Laboratory Investigator  
 
Investigator Signature 
 
 
Date  
 
Details:________________________________________________________________________ 
 
 
 
 
 
 

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Table of Contents 
Modification History ...................................................................................................................... 2 
1 Synopsis ................................................................................................................................... 15 
2 Abbreviations ........................................................................................................................... 17 
3 Background & Rationale ........................................................................................................... 19 
3.1 
Impact of P. vivax malaria and the need for a vaccine .................................................. 19 
3.2 
Lifecycle of the Plasmodium vivax malaria parasite ...................................................... 20 
3.3 
 Challenges and directions in P. vivax vaccine development ......................................... 21 
3.4 Rationale for a blood stage vaccine against Plasmodium vivax malaria .............................. 23 
3.5 PvDBP as an Antigen .......................................................................................................... 26 
4 PvDBPII Vaccine Development .................................................................................................. 28 
4.1 PvDBPII in pre-clinical studies ............................................................................................ 28 
4.2 PvDBPII in clinical studies ................................................................................................... 28 
4.2.1 Clinical experience with a viral-vectored PvDBPII vaccine in a Phase Ia clinical trial ..... 28 
4.2.2 Clinical experience with PvDBPII with adjuvant GLA-SE in a Phase Ia clinical trial ......... 29 
4.3 Matrix M1 .......................................................................................................................... 30 
4.4 Concomitant COVID-19 vaccinations .................................................................................. 31 
5 Controlled human malaria infection ......................................................................................... 32 
5.1 
Controlled human malaria infection studies ................................................................. 32 
5.1.1 Microbial challenge studies of human volunteers ........................................................ 32 
5.1.2 
Controlled human malaria infection with Plasmodium falciparum ........................... 32 
5.1.3  
Controlled human malaria infection with Plasmodium vivax ................................ 32 
5.1.4 
P. vivax sporozoite challenge in the modern era ...................................................... 33 
5.1.5 
P. vivax challenge for assessment of vaccine efficacy ............................................... 34 
5.1.6 
Modern blood-stage controlled human P. vivax infection ........................................ 36 
5.1.7 
Rationale for blood-stage controlled human malaria infection ................................. 37 
5.2 Cryopreserved Plasmodium vivax blood inoculum ............................................................. 38 
5.2.1 
Source and preparation of the cryopreserved inoculum ........................................... 38 
5.2.2 
Collection of P. vivax infected erythrocytes .............................................................. 38 
5.2.3 
Cryopreservation and storage of blood bank ............................................................ 39 
5.2.4 
Infectivity and characterisation of parasite stock ..................................................... 39 
5.2.5 
Testing of source patient and blood donors for blood-borne and mosquito-borne 
infections ............................................................................................................................. 39 
5.2.6 
Sterility and screening for blood-borne infections of cryopreserved blood bank ...... 41 

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5.2.7 
Determination of safety, feasibility and dose of cryopreserved P. vivax inoculum .... 41 
5.3 
Conduct of CHMI trials ................................................................................................. 46 
5.3.1 Standardisation of CHMI studies .................................................................................. 46 
5.3.2 Ethical considerations of CHMI trials ........................................................................... 46 
5.3.3 
Clinical presentation post-CHMI ............................................................................... 47 
5.3.4 
Oxford’s experience conducting CHMI trials ............................................................. 47 
5.3.5 
Conducting CHMI in the context of the COVID-19 pandemic .................................... 48 
6 Investigational products ........................................................................................................... 50 
6.1 PvDBPII .............................................................................................................................. 50 
6.2 Matrix M1 .......................................................................................................................... 50 
6.3 Storage of Vaccines ............................................................................................................ 50 
6.4 Administration of Vaccines ................................................................................................ 50 
7 Objectives and endpoints ......................................................................................................... 51 
7.1 Primary Objectives ............................................................................................................. 51 
7.1.1 Primary Safety Outcome Measures.............................................................................. 51 
7.1.2 Primary Efficacy Outcome Measures ........................................................................... 51 
7.2 Secondary Objectives ......................................................................................................... 52 
7.2.2 Secondary Immunological Outcome Measures ............................................................ 52 
7.2.3 Secondary Efficacy Outcome Measures ....................................................................... 52 
8 Description and justification of study design ............................................................................ 53 
8.1 
Study rationale ............................................................................................................ 53 
8.2  
Study Overview ............................................................................................................ 53 
8.3 
Study groups ................................................................................................................ 53 
8.3.1 Control groups during challenge .................................................................................. 54 
8.4 
Duration of study ......................................................................................................... 55 
8.4.1 
Definition of the start and end of the trial ................................................................ 55 
8.4.2 Duration of volunteer participation ............................................................................. 55 
8.5 
Potential risks for volunteers ....................................................................................... 55 
8.5.1 
Phlebotomy ............................................................................................................. 55 
8.5.2 Vaccinations ................................................................................................................ 56 
8.5.3 
Risk of Infection with Blood Borne Organisms .......................................................... 56 
8.5.4 
Plasmodium vivax infection ...................................................................................... 57 
8.5.5 
Medications dispensed to volunteers in course of trial ............................................ 57 
8.5.6 
Risk of reaction to the blood sample ........................................................................ 58 
8.6 
Potential benefits for volunteers .................................................................................. 59 

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9 Recruitment and withdrawal for trial volunteers ...................................................................... 60 
9.1 
Informed consent ........................................................................................................ 60 
9.1.1 
Informed consent ..................................................................................................... 60 
9.1.2 
Informed consent questionnaire .............................................................................. 61 
9.2  
Inclusion and exclusion criteria .................................................................................... 61 
9.2.1 
Inclusion criteria ...................................................................................................... 61 
9.2.2 
Exclusion criteria ...................................................................................................... 62 
9.2.3 
Prevention of ‘Over Volunteering’ ............................................................................ 63 
9.2.4 
Vaccination and re-vaccination exclusion criteria ..................................................... 63 
9.2.5 
Exclusion criteria on day of CHMI ............................................................................. 64 
9.2.6 
Concomitant medications ........................................................................................ 64 
9.3 
Withdrawal of volunteers ............................................................................................ 65 
9.4 
Pregnancy .................................................................................................................... 65 
10 Controlled Blood-Stage Malaria Infection Inoculum ................................................................ 67 
10.1 
Preparation of the inoculum ........................................................................................ 67 
10.2 
Administration of the inoculum ................................................................................... 67 
11 Treatment of trial volunteers .................................................................................................. 68 
11.1 
Trial sites ..................................................................................................................... 68 
11.2 
Study procedures ......................................................................................................... 68 
11.2.1 
Observations ........................................................................................................ 68 
11.2.2 
Blood tests ........................................................................................................... 68 
11.2.3 
 Urinalysis ............................................................................................................. 69 
11.2.4 
 Electrocardiogram ............................................................................................... 69 
11.2.5 
Vaccinations ......................................................................................................... 69 
11.2.6 Management of post-vaccination fevers .................................................................... 70 
11.2.7 Diary card .................................................................................................................. 71 
11.2.8 
SARS-COV-2 PCR swab .......................................................................................... 71 
11.3 
Study visits ................................................................................................................... 71 
11.3.1 
Screening visits ..................................................................................................... 71 
11.3.2 Enrolment and first vaccination with PvDBPII-Matrix M1 ........................................... 72 
11.3.3 Reviews post-vaccination on days 1, 3, 7 and 14 ........................................................ 72 
11.3.4 2nd Vaccination with PvDBPII-Matrix M1 (day 28) ..................................................... 72 
11.3.5 Reviews post-vaccination on days 29, 31, 35 and 42 .................................................. 73 
11.3.6 3rd Vaccination with PvDBPII-Matrix M1 (day 56 or approximately day 365 for Group 
1) ......................................................................................................................................... 73 

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11.3.7 Reviews post-vaccination on days 57, 59, 63, and 70 for Group 2 (or 1, 3, 7 and 14 
days post 3rd vaccination for Group 1) .................................................................................. 73 
11.3.8 Group 3 only – 4th Vaccination with PvDBPII-Matrix M1 and reviews on 1, 3, 7 and 14 
days post-vaccination ........................................................................................................... 73 
11.3.9 
Two days before CHMI (C-2 (±1)) .......................................................................... 74 
11.3.10 
Day of CHMI (C0) .................................................................................................. 74 
11.3.11 
Days 1-6 post-CHMI (C+1 – C+6) ........................................................................... 75 
11.3.12 
Days 7-28 post-CHMI (C+7 – C+28) – Group 1 ....................................................... 75 
11.3.13 
Days 7-21 post-CHMI (C+7 – C+21) – Groups 2 and Group 3 ................................. 76 
11.4 
Algorithm for initiation of treatment ........................................................................... 77 
11.4.1 Participants with fever post malaria challenge and COVID-19 testing ........................ 80 
11.5 
Malaria management ................................................................................................... 81 
11.5.1 
Malaria management – Riamet ............................................................................ 81 
11.5.2 
Malaria management – Malarone ........................................................................ 82 
11.5.3 
Malaria management – supportive medications ................................................... 82 
11.5.4 
Follow-up after commencing malaria treatment .................................................. 82 
11.5.5 
Criteria for inpatient transfer to the NHS ............................................................. 82 
11.6 
Safety measures for conduct of CHMI .......................................................................... 83 
11.6.1 
Measures to be taken if a volunteer goes missing post-CHMI ............................... 83 
11.7 
Follow-up post-treatment ............................................................................................ 84 
11.7.1 
Six days after initiation of treatment (T+6) ........................................................... 84 
11.7.2 
Nine days after initiation of treatment (T+9) ........................................................ 84 
11.7.3 Day 56 post CHMI (C+56) ........................................................................................... 84 
11.7.4 
Day 96 post CHMI (C+96) ..................................................................................... 84 
11.7.5 
Day 276 post CHMI (C+276).................................................................................. 84 
12 ASSESSMENT OF SAFETY ......................................................................................................... 97 
12.1 Definitions ....................................................................................................................... 97 
12.1.1 Adverse Event (AE) .................................................................................................... 97 
12.1.2 Adverse Reaction (AR) ............................................................................................... 97 
12.1.3 Unexpected Adverse Reaction ................................................................................... 97 
12.1.4 Serious Adverse Event (SAE) ...................................................................................... 97 
12.1.5 Serious Adverse Reaction (SAR) ................................................................................. 98 
12.1.6 Suspected Unexpected Serious Adverse Reaction (SUSAR) ........................................ 98 
12.1.7 Foreseeable Adverse Reactions ................................................................................. 98 
12.1.8 Other Foreseeable Medical Occurrences ................................................................... 98 
12.1.9 Expected Serious Adverse Events............................................................................... 98 

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12.2 Causality Assessment ....................................................................................................... 98 
12.3 Reporting Procedures for All Adverse Events ................................................................... 99 
12.4 Reporting Procedures for Serious AEs (see SOP OVC005 Safety Reporting) .................... 101 
12.4.1 Reporting Procedures for SUSARS ............................................................................ 101 
12.5 Development Safety Update Report ............................................................................... 102 
12.6 Adverse Events of Special Interest.................................................................................. 102 
12.7 Procedures to be followed in the event of abnormal findings ........................................ 102 
12.8 Local Safety Monitor ...................................................................................................... 102 
12.9 Safety stopping/holding rules......................................................................................... 102 
12.9.1 Group holding rules ................................................................................................. 103 
12.9.2 Individual stopping rules .......................................................................................... 103 
13 Statistics ............................................................................................................................... 105 
13.1  
Sample size ................................................................................................................ 105 
14 Quality control and quality assurance procedures ................................................................ 106 
14.1  
Investigator procedures ............................................................................................. 106 
14.2 
Monitoring ................................................................................................................. 106 
14.3 
Modification to protocol ............................................................................................ 106 
14.4 
 Protocol deviation ..................................................................................................... 106 
14.5  
Audit & inspection ..................................................................................................... 106 
14.6 
Serious breaches ........................................................................................................ 106 
14.7 
Trial progress ............................................................................................................. 107 
14.8 
Publication policy ....................................................................................................... 107 
14.9 
Intellectual Property .................................................................................................. 107 
15 Ethics .................................................................................................................................... 108 
15.1 
Declaration of Helsinki ............................................................................................... 108 
15.2 
ICH guidelines for good clinical practice ..................................................................... 108 
15.3 
Approvals ................................................................................................................... 108 
15.4 
Reporting ................................................................................................................... 108 
15.5 
Volunteer confidentiality ........................................................................................... 108 
16 Data handling and record keeping ........................................................................................ 109 
16.1 
Data handling............................................................................................................. 109 
16.2 
Record keeping .......................................................................................................... 109 
16.3 
Source data and electronic case report forms (eCRFs) ............................................... 109 
16.4 
Data protection .......................................................................................................... 109 
17 Financing and insurance ....................................................................................................... 110 

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17.1 
Financing ................................................................................................................... 110 
17.2 
Insurance ................................................................................................................... 110 
17.3 
Compensation............................................................................................................ 110 
18 Appendices ........................................................................................................................... 111 
19 References ........................................................................................................................... 112 
 
 
 
 

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1 SYNOPSIS 
Title 
A Phase I/IIa clinical trial to assess the safety, immunogenicity and efficacy 
of the blood-stage Plasmodium vivax malaria vaccine candidate PvDBPII in 
Matrix M1 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 
VAC079     
Clinical Phase  
Phase I/IIa 
Design 
Open label, first-in-human, Phase I/IIa, blood-stage P. vivax malaria vaccine 
trial 
Population 
Healthy adults aged 18 – 45 years 
Sample Size 
Total: 12-24 volunteers 
Up to 12 volunteers in Group 1 will receive three doses of the PvDBPII 
50ug/Matrix M1 50ug candidate vaccine at 1, 2 and 12-18 months, prior to 
blood-stage CHMI 2-4 weeks after the third vaccination.  
If fewer than 8 volunteers complete the study in Group 1, then new 
volunteers will be recruited into Group 2, to make up a total of 10 to 12 
volunteers who complete 3 vaccinations and CHMI between Groups 1 and 
2. Group 2 volunteers will receive three doses of the PvDBPII 50ug/Matrix 
M1 50ug candidate vaccine at monthly intervals, prior to blood-stage CHMI 
2-4 weeks after the third vaccination. 
Up to 6 volunteers who have completed 3 vaccinations and CHMI 
previously in Group 1, will be invited to undergo a fourth vaccination at 5 
months after the third vaccination, followed by repeat CHMI 2-4 weeks 
after the fourth vaccination. These volunteers will comprise Group 3. 
All these volunteers will undergo blood stage CHMI with Plasmodium vivax.
The de-identified data from volunteers in a parallel study (VAC069), who 
will undergo the same CHMI without prior vaccination, will be used as 
infectivity controls. 
Follow-up 
duration 
Participants in Group 1 will be followed for 9 months after the third 
vaccination, approximately 2 years in total from enrolment. 
Participants in Group 2 will be followed for 1 year from enrolment. 
Participants in Group 3 will be followed for 9 months after their final 
(fourth) vaccination, approximately up to 2.5 years in total from enrolment.

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Planned trial 
period  
2.5 years from the enrolment of the first volunteer.  
Primary Objectives
 
 
To assess the safety  of the PvDBPII-Matrix M1 vaccine in healthy 
volunteers  
To establish whether the PvDBPII-Matrix M1 vaccine can demonstrate a 
reduced parasite multiplication rate in vaccinated subjects compared to 
infectivity controls in a blood-stage controlled human malaria infection 
model 
Secondary 
Objectives 
 
 
To assess the humoral and cellular immunogenicity of the PvDBPII-Matrix 
M1 vaccine candidate. 
To assess immunological readouts for association with a reduced parasite 
multiplication rate (PMR) 
To assess the durability of any reduction in PMR in volunteers who undergo 
a fourth vaccination followed by re-challenge approximately 5 months after 
the primary challenge 
Investigational 
Products 
1. PvDBPII – a recombinant protein malaria vaccine candidate 
2. Matrix-M1 – a saponin based vaccine adjuvant 
Form 
1. PvDBPII: liquid, stored between -70oC and -90oC 
2. Matrix-M1: liquid, stored at +2oC to +8oC 
Dose 
1. PvDBPII -  50µg, administered in a three-dose schedule on days 0, 
28 and 56 or 430. Fourth dose of 50µg given at day 580 for Group 3.
2.  Matrix M1 - 50 µg, administered with each PvDBPII dose on days 0, 
28 and 56 or 430. Fourth dose of 50µg given at day 580 for Group 3.
Route 
Intramuscular injection in the deltoid region of the arm 
 
 

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2 ABBREVIATIONS 
ACT 
 
  
Artemisinin-based combination therapies 
AE 
 
 
Adverse event 
AR 
 
 
Adverse reaction 
CBF  
 
 
Clinical Bio-Manufacturing Facility 
CCVTM 
 
Centre for Clinical Vaccinology and Tropical Medicine 
CHMI  
 
Controlled human malaria infection 
CI 
 
 
Chief Investigator 
CMV 
 
 
Cytomegalovirus 
CRF 
 
 
Case Report Form or Clinical Research Facility 
CTRG  
 
Clinical Trials Research Governance 
CYP2D6 
 
Enzyme predicting Primaquine metabolism – Cytochrome P450 2D6 
DARC  
 
Duffy Antigen Receptor for Chemokines 
DFA 
 
 
Direct Feeding Assay 
DSUR  
 
Development Safety Update Report 
EBV 
 
 
Epstein-Barr Virus 
ELISA  
 
Enzyme linked immunosorbent assay 
ELISPOT 
 
Enzyme linked immunospot assay 
EPI 
 
 
Expanded Programme of Immunisation 
FDA 
 
 
United States Food and Drug Administration 
GCP 
 
 
Good Clinical Practice 
GMP  
 
Good Manufacturing Practice 
HBsAg  
 
Hepatitis B surface antigen 
HCV 
 
 
Hepatitis C virus 
HIV 
 
 
Human Immunodeficiency virus 
HTLV  
 
Human T cell Lymphotrophic virus 
HLA 
 
 
Human Leukocyte Antigen 
ICH 
 
 
International Conference on Harmonisation 
IFN-

Interferon-gamma 
IM 
 
 
Intramuscular 
IMP 
 
 
Investigational Medicinal Product 
LPS 
 
 
Lipopolysaccharide 
LSM 
 
 
Local Safety Monitor 
MFA 
 
 
Membrane Feeding Assay 
MHRA  
 
Medicines and Healthcare products Regulatory Agency 

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MRC 
 
 
Medical Research Council 
g 
 
 
Microgram 
NIHR  
 
National Institute for Health Research  
pIMD  
 
potential Immune-Mediated Diseases 
PCR 
 
 
Polymerase Chain Reaction 
PIS 
 
 
Participant Information Sheet 
QP 
 
 
Qualified Person 
qPCR  
 
Quantitative polymerase chain reaction 
RAS 
 
 
Radiation-attenuated sporozoite 
RUNMC 
 
Radboud University Nijmegen Medical Center 
REC 
 
 
Research Ethics Commitee 
SAE 
 
 
Serious Adverse Event 
SAR 
 
 
Serious Adverse Reaction 
SOP 
 
 
Standard Operating Procedure 
SUSAR  
 
Suspected Unexpected Serious Adverse Reaction 
TMF 
 
 
Trial Master File 
UOXF  
 
University of Oxford 
USMMVP 
 
United States Military Malaria Vaccine Program 
VLP 
 
 
Virus-like particle 
WHO  
 
World Health Organisation 
WTCRF 
 
Wellcome Trust Clinical Research Facility 
 

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3 BACKGROUND & RATIONALE 
3.1 
Impact of P. vivax malaria and the need for a vaccine 
Plasmodium vivax (P. vivax) is one of the seven Plasmodium species known to cause human 
malaria and accounts for the most cases of non-P. falciparum malaria worldwide. Geographically, 
it is the most widespread human malaria parasite and approximately 2.5 billion people are at risk 
of contracting the infection in endemic areas [1]. It is estimated over one third of malaria cases 
outside of the African continent are due to P. vivax and that it accounts for 74% of malaria cases 
in the Americas, 37% in South-East Asia and 31% in the Eastern Mediterranean WHO regions.  
The burden is highest in Afghanistan, Ethiopia, India, Indonesia and Pakistan, which together 
account for 82% of P. vivax cases[2]. 
P. vivax malaria has long been considered, and termed, ‘benign’ malaria, but more recently large 
case series demonstrate that P. vivax infection is associated with significant morbidity and 
mortality. Complications reported include severe anaemia, respiratory and hepatic dysfunction, 
severe thrombocytopenia and coagulopathies, such as disseminated intravascular coagulation 
(DIC) [3]. P. vivax infection has also been associated with important sequelae for maternal and 
foetal health, correlating with increased rates of low birthweight, preterm delivery, and reduced 
foetal growth, as well as maternal and foetal deaths [4].  
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 [5]. 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 [6].  
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 hypnozoite stage, a lifecycle stage not seen with P. falciparum malaria [5]. 
Standard schizonticidal regimes are not effective against hypnozoites. Radical cure of dormant 
parasites requires therapy with the 8-aminoquinolone primaquine, currently the only licensed 
hypnozoitcidal anti-malarial, although tafenoquine, a new drug of the same class, given as a 
single dose, has recently been FDA-approved for adults in the United States. However, both 8-
aminoquinolones carry a significant risk of severe haemolytic anaemia in individuals who are 
deficient in the glucose-6-phosphate dehydrogenase enzyme (G6PD), an inherited X-linked red 
blood cell enzyme disorder common in tropical and sub-tropical areas [7]. More recently, the 
importance of the cytochrome P450 enzyme, CYP2D6 in metabolism of primaquine to the active 
metabolite has been recognised [8, 9]. A common polymorphism in the CYP2D6 gene for the 
cytochrome P450 enzyme results in poor conversion of primaquine to the active form, resulting 
in 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 [7].   
Control of P. vivax is also challenging, with re-emergence in areas where it has previously been 
eradicated. P. vivax is epidemiologically and biologically different to P. falciparum and control 
methods developed for P. falciparum are less efficient for P. vivax malaria [10-13]. Control efforts 
are complicated by multiple factors including difficulty detecting asymptomatic infection, 
resistance to antimalarials, a lack of understanding of parasite biology, early gametocytogenesis 
and relapses [14, 15]. Recent calls for control and ‘eradication’ of malaria worldwide [16] have 
focused attention on this neglected disease and the need for development of an effective P. 
vivax vaccine to be used alongside current control methods [17]. 
Consequently, the revised Malaria Vaccine Technology Roadmap to 2030 [17] now recognises the 
importance of P. vivax and calls for a vaccine to achieve 75% efficacy over two years – equally 

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weighted with P. falciparum in an era of renewed political will to move towards malaria 
elimination and eradication. 
However, unlike P. falciparum, to date there has been relatively little research into this 
plasmodium species and one of the limiting factors has been the inability to culture P. vivax 
parasites in vitro over a prolonged period of time. More recently, research into P. vivax malaria 
has increased with candidate vaccines being developed and taken forward to clinical trial [18, 
19]. Yet none of these vaccines have progressed past a Phase IIa trial, and options for assessing 
the immune responses to new candidates are extremely limited.  
3.2 
Lifecycle of the Plasmodium vivax malaria parasite 
The lifecycle of P. vivax is complex, involving both the vector (the Anopheles mosquito) and the 
human host (Figure 1). Sporozoites are injected into the human host when the female Anopheles 
mosquito takes a blood meal. From here they migrate within minutes to the liver, where they 
invade hepatocytes and the schizont develops (liver-stage). The mature schizont ruptures after 
around 7 days, releasing merozoites into the blood stream, which invade reticulocytes 
preferentially (unlike P. falciparum which invades all erythrocytes). As well as developing 
schizonts in the liver, P. vivax forms hypnozoites, a dormant stage which develops into schizonts 
weeks to years later, rupturing when mature and releasing merozoites to cause blood-stage 
malaria. It is this stage of the lifecycle that is responsible for relapse.  
Once merozoites have invaded reticulocytes they are termed rings, which again mature over 48 
hours into mature schizonts which release merozoites. These merozoites go on to infect other 
reticulocytes, and the blood-stage cycle continues. A subset of rings develop into gametocytes, 
the sexual-stage of the malaria lifecycle, rather than forming schizonts. This occurs early in 
blood-stage infection with P. vivax, unlike P. falciparum. These are taken up by the mosquito in a 
blood meal. The sporogonic cycle takes place within the mosquito. The microgametocyte (male) 
exflagellates and enters the macrogametocyte (female) to form a zygote within the mosquito 
midgut. Zygotes become motile (ookinetes) and penetrate the midgut wall where they develop 
into oocysts. These rupture, release sporozoites which migrate to the mosquito salivary glands 
ready to be injected when the mosquito takes another blood meal [20]. 
The liver-stage of infection is asymptomatic. Symptoms, and subsequent complications, develop 
during the blood-stage. 
 

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Figure 1: Lifecycle of P. vivax malaria 
 
3.3 
 Challenges and directions in P. vivax vaccine development  
The unique characteristics of P. vivax pose some 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 [21], 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. In order to study hepatocyte 
invasion stages of P. falciparum as well as P. vivax, attempts have been made to establish in vitro 
liver models [22, 23]. In addition, a reliable humanised mouse model for P. vivax liver-stages is 
now available [24]. These approaches, together with the P. cynomolgi model in non-human 
primates will help to identify and test novel drugs and vaccines targeting hepatocyte stages [25]. 
Studies on expression of parasite proteins in hypnozoites will determine if these latent stages can 
also be targeted with novel drugs and vaccines.  
Currently, the two most clinically advanced P. vivax vaccine candidates are both subunit vaccines 
based on P. vivax circumsporozoite protein (PvCSP), an analogue of the P. falciparum pre-

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erythrocytic target, which have both have reached Phase IIa efficacy testing in mosquito-bite 
delivered, sporozoite 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. However, following CHMI at 14 days after the third immunisation, 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 another PvCSP based candidate (long synthetic peptide CS derivatives, formulated in 
Montanide ISA 51) in mosquito bite challenge is also now complete, with results currently 
awaited (Clinicaltrials.gov identifier NCT02083068).  
To date, repeated exposure to radiation-attenuated 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 2 
volunteers in the 1970’s in Maryland but this result was not replicated in larger numbers until a 
recent study in Cali, Colombia [26, 27]. In the Cali study, twelve Duffy positive participants were 
exposed to seven “immunisations”, receiving 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 
immunisation 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 immunisation regime. Sterile efficacy was demonstrated 
in 42% (5/12) of Duffy positive participants, interestingly, all of which were female (5/7 female 
participants vs.0/5 males) [27]. 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 but 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 PvDBPII vaccine candidates [28], the only other P. vivax blood-
stage vaccine currently in active clinical development is the recombinant protein PvDBPII 
vaccine. It 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 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 [29].  
A single P. vivax transmission-blocking candidate, the recombinant protein Pvs25H, derived 
from the Pvs25 surface antigen of P. vivax ookinetes, has reached Phase I clinical testing. When 
administered in Alhydrogel, the vaccine produced promising transmission blocking activity as 
assessed in a direct membrane feeding assay (DMFA) in some volunteers, however, overall 
induced antibody titres were low [30]. To increase immunogencity, Pvs25H was tested in 
formulation with the stronger water-in-oil adjuvant, Montanide ISA51, but due to concerns 
regarding both local and systemic reactogenicity of this formulation, including erythema 
nodosum in two individuals, the trial was subsequently halted[31].   
All vaccines currently in clinical development, and almost all those in pre-clinical stages, target 
individual stages of the parasite’s lifecycle and are based on single antigens (Table 1 [18]). 
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 

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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. 
 
3.4 Rationale for a blood stage vaccine against Plasmodium vivax malaria 
The feasibility of a blood-stage vaccine for P. vivax is supported by significant literature. Firstly, 
although the human immune system will attack almost any stage of the parasite life cycle in the 
human host, naturally-acquired immunity appears to primarily target blood-stage infections [32]. 
Naturally acquired immunity develops over time with repeated exposure, to both primary blood-
stage infections and relapse infections in the blood, and individuals with acquired immunity may 
develop asymptomatic infection [5]. This is reflected in the epidemiology of disease; in high-
transmission areas, most infections, and in particular severe infections, are seen in children 
whereas in low-transmission settings the risk of infection, and severe disease, is evenly 
distributed across age groups [33]. Naturally acquired immunity to P. vivax is complex and 
remains incompletely understood, but mechanisms of protection may include antibody mediated 
inhibition of invasion, enhancement of phagocytosis, cellular inhibition and neutralisation of 
malarial toxins produced during schizont rupture, as well as cytokine release [34].  
 
Secondly, malaria occurs at the blood-stage of plasmodium infection. The potential advantage of 
a blood-stage vaccine would be that even if only partially effective, it would still confer some 
protection, with reduced disease severity, whilst allowing natural immunity to develop [35]. 
Conceptually, therefore, blood-stage malaria vaccines have significant advantages over vaccines 
targeting other life-cycle stages; if pre-erythrocytic vaccines fail to provide sterilising immunity, 
the individual would be vulnerable to severe blood-stage disease and mosquito-stage vaccines 
would offer no protection to the individual once infected. A strong case for malaria vaccine 
development is made by modelling studies which show the substantial benefit that would arise 
from deploying a vaccine that is only partially protective [36, 37]. These models, which typically 
assume a vaccine efficacy of 50%, and a protective half-life of 3 years, predict variable impacts on 
mortality, morbidity and parasite prevalence, depending upon intensity of transmission, 
deployment of other control methods, rate of immune decay and access to healthcare [36-38]. 
They all, however, reach general agreement that even with modest levels of efficacy, a vaccine 
would facilitate reductions in parasite prevalence if used in combination with other strategies. 
This role for a vaccine is very different to the ‘magic bullet’ envisaged in the past; a vaccine 
would form one arm of an integrated programme consisting of indoor residual spraying, long-
lasting bed nets, draining of mosquito habitats and provision of artemisinin-based drugs. The 
effect of a hypothetical vaccine with ninety-five percent efficacy and a protective half-life of ten 
years has also been modelled [38], and in this scenario a single mass administration of such a 
vaccine could; eliminate malaria from low-transmission zones; eliminate malaria from moderate-
transmission zones if used in combination with other methods; and reduce prevalence to below 
five percent in high-transmission zones.  
The hypnozoite stage of P. vivax means that individuals who are infected are at risk of relapse of 
disease, and a blood-stage vaccine could, importantly, protect against this (as long as relapses 
occurred within the duration of protective blood-stage immunity provided by vaccination). 
Relapses in vivax malaria occur weeks to years after initial infection, and may arise 10-14 times 
after the primary infection in the absence of eradication therapy, occurring every 3-6 weeks (in 
tropical climates) [5]. 

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Thirdly, the P. vivax Duffy-binding protein (PvDBP) is a very promising antigen for vaccine 
development (see section 3.5). P. vivax requires interaction between the Duffy-binding protein 
and the host Duffy antigen receptor for chemokines (DARC) in order for the parasite to invade 
reticulocytes during blood-stage infection.  Populations who are Duffy antigen negative are 
protected from vivax malaria, as demonstrated by a geographical scarcity of the infections in the 
areas they inhabit (for example, much of sub-Saharan Africa). Furthermore, natural immunity has 
been found to be associated with anti-PvDBP responses [39-41].  
 
 
 
 
 
 

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Vaccine candidate 
Development Phase 
Lifecycle stage 
Antigen 
Delivery system 
VMP001 
Phase I/IIa END 
Liver-stage 
PvCSP 
Rec. protein-AS01B 
CSV-S,S 
Pre-clinical 
Liver-stage 
PvCSP 
HBsAg fusion-AS01B 
PvCSP-LSP 
Phase I END 
Liver-stage 
PvCSP 
Synthetic peptides-Montanide ISA 720 
ChAd63-PvTRAP/MVA-PvTRAP 
Pre-clinical 
Liver-stage 
PvTRAP 
Prime-boost, viral vectors 
PvDBPII 
Phase I 
Blood-stage 
PvDBP 
Rec. protein-GLA-SE 
PvDBPII-DEKnull 
Pre-clinical 
Blood-stage 
PvDBP 
Rec. protein 
ChAd63-PvDBPII/MVA-PvDBPII 
Phase I/IIa 
Blood-stage 
PvDBP 
Prime boost, viral vectors 
PvMSP119 
Pre-clinical 
Blood-stage 
PvMSP1 
Rec. protein-Montanide ISA720 
PvAMA1 
Pre-clinical 
Blood-stage 
PvAMA1
Rec. protein-adjuvant 
Pvs25H 
Phase Ia END 
Transmission-stage 
Pvs25 
Rec. protein-Alhydrogel; Rec. protein-Montanide ISA 51 
Pvs28 
Pre-clinical 
Transmission-stage 
Pvs28 
Rec. protein-adjuvant 
Pvs25-IMX313 
Pre-clinical 
Transmission-stage 
Pvs25 
Rec. protein-adjuvant  
AnAPN1 
Pre-clinical 
Mosquito midgut Ag 
AnAPN1 
Rec. protein-adjuvant 
 Table 1: P. vivax vaccine candidates under development: taken from Ref [18]. 
 
 

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3.5 PvDBP as an Antigen 
From the early 1920s until the advent of penicillin in the mid-1940s, induced malaria became the 
standard treatment of neurosyphilis. An observation made soon after the introduction of 
malariotherapy to the United States, was the refractoriness of most people of African origin to 
infection with P.vivax, as opposed to universal susceptibility of persons of other origins [42]. The 
basis for this resistance was not uncovered until the mid-1970s [43] , when it was demonstrated 
that Duffy blood group positivity was required for red blood cell invasion by P. vivax, and nearly 
all West Africans, and a majority of sub-tropical African populations, are Duffy-negative . Twenty 
years later the parasite ligand which interacts with the Duffy antigen receptor for chemokines 
(DARC), known as Duffy-binding protein (DBP), was identified. 
Plasmodium merozoite invasion of erythrocytes is a complex process. The merozoite reorients 
after an initial interaction with the erythrocyte, so that its apical end faces the surface of the cell. 
A tight junction is formed between the merozoite and erythrocyte when the micronemes and 
rhoptries of the merozoite discharge their contents. Plasmodium vivax expresses homologues of 
the Py235 rhoptry proteins, referred to as PvRBP1a, PvRBP1b, PvRBP2a and PvRBP2b which bind 
to reticulocytes and not normocytes, presumably explaining the host-cell preference of P. vivax 
[44]. DBP, belongs to a family of microneme proteins that are functionally conserved across 
Plasmodium species and are thought to trigger release of rhotry proteins, which leads to the 
formation of a tight attachment between the parasite and target host cell. The erythrocyte 
membrane subsequently encases the merozoite, creating a vacuole around the invading parasite 
[45, 46]. 
Unlike P. falciparum which utilises multiple redundant invasion pathways for human erythrocyte 
invasion [44, 47], P. vivax requires interaction with the Duffy antigen receptor for chemokines 
(DARC). Further to population level evidence of the protection of individuals lacking the Duffy 
blood group antigen, as seen across much of sub-Saharan Africa, knockout of the orthologous P. 
knowlesi DBPα gene prevents invasion of Duffy-positive erythrocytes by this highly related 
parasite in vitro [45].  
DBPs have been characterised for both P. knowlesi (PkDBP) [48] nd P. vivax (PvDBP) [49]. PvDBP 
has a molecular weight of approximately 140 kDa and a 330-amino acid cysteine-rich region 
within it, known as region II (PvDBPII), is predicted to be the domain responsible for binding to 
DARC [50]. 
It is proposed that PvDBPII dimers bind either one or two DARC ectodomains and distinct 
heterotrimeric and heterotetrameric architectures may form on binding [51, 52]. Structural 
studies using the highly related P. knowlesi DBPα protein have shown that region II of PvDBP is 
polymorphic, suggesting this region is under immune pressure. However, the receptor binding 
residues within PvDBPII are highly conserved, with polymorphic residues clustered at sites that 
are distal to the DARC binding residues [53]. In-keeping with this, induction of binding inhibitory 
antibodies against these residues does not commonly occur following natural exposure [54, 55]. 
Yet, when these antibodies have been identified, as in a prospective cohort study in Papua New 
Guinea, they have been associated with reduced risk of P. vivax infection, lower P. vivax parasite 
densities and strain-transcending protection [39].  
Over recent years, the critical nature of the PvDBP-DARC interaction with the Duffy antigen 
receptor for merozoite invasion has been challenged, with various reports of P. vivax infection in 
Duffy negative individuals in the literature [56-60]. Currently, the mechanism of invasion of Duffy 
negative erythrocytes is incompletely understood, however, alternative pathways have been 
suggested through identification of novel erythrocyte invasion genes and PvDBP gene duplication 

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in clinical isolates by whole genome sequencing[61, 62]. PvDBP gene duplication has not been 
linked to cases of Duffy negative P. vivax infection, however it has been geographically correlated 
with regions also reporting Duffy negative infection, perhaps pointing to involvement of PvDBP, 
even in Duffy negative infection [63]. The further complexities of P. vivax erythrocyte invasion in 
both Duffy positive and negative individuals will continue to be elucidated, and may reveal 
important, potentially synergistic or additive antigenic targets. However, PvDBP-DARC 
interaction likely remains an essential, non-redundant pathway for merozoite invasion in the 
large majority of cases and PvDBP continues to be the current leading target for candidate P. 
vivax blood-stage vaccine development [34, 64], likely with a greater chance of success in 
comparison to the widely tested merozoite antigen targets of P. falciparum, which unlike P. vivax 
is not limited to invasion by a single pathway. 
 

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4 PvDBPII VACCINE DEVELOPMENT 
4.1 PvDBPII in pre-clinical studies 
Pre-clinically, antibodies blocking PvDBP-DARC interaction have been reproducibly induced by 
immunization using PvDBPII–based vaccines in small animal models, including in mice and 
rabbits, as well as in non-human primates [51, 52, 65, 66], and those raised against the 
P.knowlesi PkDBPα ortholog can block RBC invasion by this parasite in vitro [53]. Following 
intravenous challenge of New World Aotus monkeys with blood-stage P. vivax, longer pre-patent 
periods and lower parasitaemias were observed in animals immunised with PvDBP-RII in 
comparison to controls. Protection was conferred following immunisation with protein vaccine in 
Freund’s adjuvant but not Montanide ISA 720 (although antibody levels induced by ISA 720 were 
surprisingly low in this study) [67]. Importantly, antibodies raised against recombinant Salvador I 
(SalI) strain PvDBPII in pre-clinical animal models can inhibit binding of heterologous polymorphic 
PvDBPII domains derived from diverse field isolates [52a]. Antibodies against PvDBPII have also 
been shown to inhibit reticulocyte invasion by diverse P. vivax field isolates [68, 69]. These 
observations suggest that antibodies produced following vaccination with PvDBPII should be able 
to block diverse strains of P. vivax.  
4.2 PvDBPII in clinical studies 
4.2.1 Clinical experience with a viral-vectored PvDBPII vaccine in a Phase Ia clinical trial 
Safety and immunogenicity of the ChAd63 and MVA viral vectored vaccines targeting PvDBP_RII  
(Salvador I strain) were assessed in the VAC051 trial (Clinicaltrials.gov identifier: NCT01816113), 
an open-label dose-escalation Phase Ia study in 24 healthy UK adults in Oxford between 2013-
2014 [28]. Vaccines were delivered by the intramuscular route in a ChAd63-MVA heterologous 
prime-boost regimen using an 8-week interval. ChAd63 PvDBP was administered to 24 healthy 
malaria-naïve volunteers at doses of 5 x 109 vp (n = 4) and 5 x 1010 vp (n = 20). 8 volunteers 
received ChAd63 PvDBP alone, with the first 4 receiving the low dose (5 x 109 vp), before 15 
volunteers went on to heterologous prime-boost regimens, receiving MVA PvDBP at doses of 
either 1 x 108 pfu (n= 7) or 2 x 108 pfu (n=8).  
ChAd63 PvDBP administered alone, and particularly when preceding MVA boost, induced strong 
PvDBP_RII –specific T cell responses, as measured by IFNγ ELISpot. IgG titres were marginal 
following the higher dose prime of ChAd63, but these were boosted to similarly high levels in all 
volunteers by both doses of MVA. PvDBP_RII –specific antibody-secreting cell and memory B cell 
responses were also observed following the MVA boost and the polyclonal immune serum 
inhibited the binding of multiple allelic variants of recombinant PvDBP_RII to N-terminal DARC. 
Data from this study confirms that the PvDBP antigen is immunogenic and induces strain-
transcending antibodies, which are binding-inhibitory in vitro, when administered as a viral-
vectored vaccine in healthy UK adults.  
ChAd63-PvDBP and MVA-PvDBP were well tolerated and demonstrated a favourable safety 
profile in malaria-naive adults. There were no serious adverse events reported during the 
VAC051 trial, and the majority of vaccine-related adverse events were mild in nature.  
The most frequently reported adverse event following intramuscular administration of ChAd63 
PvDBP at either of the above doses was injection site pain, seen in the majority of individuals. 
Less frequently, erythema, swelling, warmth and itch were also reported. The overall median 
duration of local adverse events was between 1 and 4 days. The most common vaccine-related 

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systemic adverse events were fatigue, headache, nausea, myalgia and arthralgia, with a median 
duration of 1-2 days and being mostly mild in nature.  
MVA PvDBP led to injection site pain in all fifteen volunteers. At a dose of 1 x 108 pfu, pain was 
mild in the majority of individuals but reported to be severe in 1/7 volunteers and 3/8 volunteers 
receiving the higher 2 x 108 pfu. Localised swelling was also seen, which was mild in the majority 
of individuals at both doses. Warmth, erythema and itch were seen less frequently and the 
majority of all local adverse events resolved within 4 days of vaccination. Systemically, the most 
commonly reported AEs were headache and feverishness, which were mild in the majority of 
participants. At the 2 x 108 pfu dose, there was increased frequency of moderate or severe local 
and systemic adverse events compared to the 1 x 108 pfu dose, however, no systemic adverse 
event was reported to be severe for more than 24 hours. 
 
4.2.2 Clinical experience with PvDBPII with adjuvant GLA-SE in a Phase Ia clinical trial  
Safety and immunogenicity of the PvDBPII (Salvador I strain) vaccine formulated in 
glucopyranosyl lipid adjuvant-stable emulsion (GLA-SE) adjuvant was assessed in a single centre, 
phase I, randomized, controlled, dose-escalating, single-blind study in India in 2016 
(CTRI/2016/09/007289) [29]. Three doses of the PvDBPII/GLA-SE vaccine were administered by 
the intramuscular route at 4 week intervals. The vaccine was administered to 27 healthy malaria-
naïve volunteers at doses of 10 µg (n=9), 25 µg (n=9) and 50 µg (n=9). Another 9 volunteers 
received GeneVac-B (hepatitis B vaccine from Serum Institute of India) as a control arm of the 
study. The adjuvant used was glucopyranosyl lipid adjuvant-stable emulsion (GLA-SE), a synthetic 
Toll-like receptor 4 agonist (a proprietary adjuvant from Infectious Disease Research Institute, 
Seattle, USA). 
Immunogenicity data demonstrated that all three doses of PvDBPII (10, 25 and 50 μg) elicited 
antigen-specific and receptor blocking serum antibody responses. The 50 μg PvDBPII/GLA-SE 
vaccine dose elicited the highest antibody response against PvDBPII and the most persistent 
binding-inhibitory antibodies against PvDBPII. Both Day 84 and Day 180 sera inhibited binding 
not only of the homologous PvDBPII Sal I allele but also of three other PvDBPII alleles (P, O and 
AH), which are commonly found in Papua New Guinea. 
The vaccine candidate, PvDBPII/GLA-SE, which was tested in healthy Indian male adults, was safe 
and well tolerated. No significant immediate reactogenicity was observed within the first hour 
post immunization at all three dose levels (10, 25 and 50 μg PvDBPII formulated with GLA-SE [5 
μg of GLA]). No SAE was observed and no subject withdrew or was withdrawn from the study on 
account of safety. A single mild local solicited AE (pain at injection site) after the first dose of 10 
μg PvDBPII was reported. No systemic solicited AEs were observed. There were similar incidences 
and intensity of unsolicited AEs among the subjects who received PvDBPII/GLA-SE and those who 
received GeneVac-B (control hepatitis B vaccine). No clinically significant difference was 
observed in haematology, biochemistry and urinalysis parameters post vaccination. Vital signs 
showed no significant changes throughout the study. No abnormal findings were observed 
during the post-study physical examination. There was no difference in the number of subjects 
who had biological values outside the normal ranges for different haematological and 
biochemical parameters in the PvDBPII/GLA-SE and control vaccine group. 
 

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Description 
PvDBPII 
(10µg) 
N=9 
n(%) 
PvDBPII 
(25 µg) 
N=9 
n(%) 
PvDBPII 
(50 µg) 
N=9 
n(%) 
Hepatitis B 
(20 µg) 
N=9 
n(%) 
Solicited Local Adverse Events 
Pain 
1 (11.11 )* 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Swelling 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Erythema 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Induration 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Tenderness 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Solicited Systemic Adverse Events 
Fever 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Vomiting 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Diarrhoea 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Headache 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Fatigue 
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Myalgia  
0.00 (0) 
0.00 (0) 
0.00 (0) 
0.00 (0) 
Note: N= Total number of subjects randomized, n= number of subjects assigned for individual a
nalysis; %=(n/N)100;Exact Binomial proportion and 95% CIs are calculated using Clopper-
Pearson Method used in SAS 
* Mild in severity 
Table 2; adverse events from the PvDBPII with adjuvant GLA-SE Phase Ia clinical trial [29] 
 
4.3 Matrix M1 
GLA-SE, the adjuvant used in the PvDBPII vaccine trial described above, has shown comparable 
immunogenicity to Alum in human clinical trials, and the antibody titres generated may not be 
sufficient to protect against malaria, where very high antibody titres are required. When the P. 
falciparum malaria VLP, R21, was tested in combination with Matrix M1 and compared head-to-
head against R21 in combination with GSK’s potent adjuvant AS01B, the immunogenicity in 
humans was the same (ClinicalTrials.gov identifier: NCT02572388), confirming the potency of 
Matrix-M1. We would naturally choose the best adjuvant for this proof-of-concept study, and so 
have elected to combine PvDBPRII with Matrix-M1 

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Matrix-M1 (MM) is a 40nm-sized complex containing the adjuvant-active saponin Quillaja 
saponaria, phospholipid and cholesterol. Quillaja saponins are triterpene glycoside substances 
derived from the tree Quillaja saponaria. The molecular weights of the different saponins range 
from 1800 - 2000 Da. In water, saponin in concentrations of 200-500 ppm exist as monomers; at 
higher concentrations they aggregate as micelles, with a molecular weight of approximately 
100000 Da. Saponins are surface-active compounds with a variety of applications including in 
agriculture, feed, food and beverage, mining, and veterinary vaccines, and are currently being 
investigated in human vaccine clinical trials. In aqueous solution, saponins are excellent 
adjuvants and are used in commercial veterinary vaccines, e.g. vaccines against foot-and-mouth 
disease, bovine mastitis, feline leukemia and equine influenza. An HPLC-purified fraction of the 
same saponin, called QS21, is a component of the AS01 adjuvant used in the pre-erythrocytic 
vaccine RTS,S/AS01. 
Matrix-M adjuvant, in 1 of 2 formulations (named Matrix-M1 or Matrix-M2), has been 
administered to 1669 individuals, of which 1132 have received Matrix-M1, in a total of 15 clinical 
trials in the US, Europe, and Australia. These trials include assessment of two pre-erythrocytic 
malaria candidate vaccines, ChAd63/MVA ME-TRAP, and the VLP R21, adjuvanted with Matrix-
M1, within four Phase I and Phase I/II trials conducted in Oxford and Burkina Faso 
(ClinicalTrials.gov identifiers: NCT02572388, NCT02925403, NCT01669512, NCT02905019). 
Available safety data demonstrates that the Matrix-M adjuvant is well-tolerated and there have 
been no serious unexpected adverse reactions or adverse reactions of special interest reported 
to date. 
The profile of adverse events following vaccination with PvDBPII-Matrix M1 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. 
Expected systemic adverse events would include headache, fatigue, myalgia, arthralgia, malaise, 
feverishness, fever and nausea. 
4.4 Concomitant COVID-19 vaccinations 
Trial participants may become eligible for a licensed COVID-19 vaccine during the course of the 
study. During this trial, COVID-19 vaccines should not be received within 14 days before or 7 days 
after each malaria vaccination (compared to 30 days for any other vaccines) and the time 
window for the second and third vaccinations have been extended to -7/+14 days to aid 
scheduling of trial vaccinations around any COVID-19 vaccinations if required.  
The antigen of COVID-19 vaccines differ significantly from the malaria antigen tested in this trial 
and the currently approved COVID-19 vaccines are different types of vaccines compared to the 
recombinant protein vaccine used in this trial. We therefore do not expect a significant impact on 
immunogenicity to the malaria vaccines if given outside of the time window specified. 
In regards to safety assessment following malaria vaccinations, assessment of solicited adverse 
events will not be affected by COVID-19 vaccinations if they are given a minimum of 7 days after 
trial vaccinations. Assessment of unsolicited adverse events, in particular delayed events 
occurring after 2 weeks of any trial vaccinations may be more difficult to interpret. However 
assessment of causality will be based on timing and previous experience with the PvDBPII vaccine 
in the Phase Ia trial. 
 
 
 

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5 CONTROLLED HUMAN MALARIA INFECTION  
5.1 
Controlled human malaria infection studies 
5.1.1 Microbial challenge studies of human volunteers 
The deliberate infection of human volunteers with micro-organisms has contributed uniquely to 
our understanding of the pathogenesis, immune responses and the treatment and prevention of 
numerous microbial diseases including P. falciparum malaria, influenza, cholera, typhoid and 
hepatitis [70]. A review by the UK Academy of Medical Sciences on microbial challenge studies 
recognised that such studies are desirable for providing proof of concept for prophylactic and 
therapeutic interventions and can significantly accelerate progress to Phase III studies [70, 71].  
5.1.2 Controlled human malaria infection with Plasmodium falciparum 
P. falciparum CHMI has now become established as a key tool to assess the efficacy of novel 
malaria vaccines and drugs. Following the development of protocols for the continuous culture 
of P. falciparum in 1976 [72] and the generation of mature P. falciparum gametocytes in vitro in 
1981 [73], it became possible to produce laboratory-reared infectious mosquitoes, meaning that 
CHMI trials could be performed more routinely [74]. 
The first well-documented CHMI study with laboratory-reared infectious mosquitoes was carried 
out in 1986 at the US Walter Reed Army Institute of Research (WRAIR), the US Naval Medical 
Research Institute (NMRI) and the US National Institutes of Health (NIH) [75]. The following year, 
the efficacies of the first recombinant protein and synthetic peptide P. falciparum vaccines were 
reported for experimentally infected volunteers [76, 77].  
There are now an increasing number of centres conducting CHMI worldwide, including in Africa, 
and between 1985 and 2009, a total of 1,343 volunteers were experimentally infected with P. 
falciparum [78, 79]. Established methods now include via direct intravenous injection of purified, 
aseptic, cryopreserved sporozoites, as well as blood-stage CHMI, induced by injection of 
cryopreserved parasitised erythrocytes [80]. These models, in addition to mosquito bite 
delivered sporozoite challenge have significantly contributed to the P. falciparum vaccine 
pipeline, including the most clinically advanced candidate RTS,S, which is currently being 
evaluated as part of a WHO pilot programme in Phase IV studies [81]. 
5.1.3  Controlled human malaria infection with Plasmodium vivax  
In comparison to P. falciparum, there is much less experience with P. vivax in the modern CHMI 
era. A key bottleneck has been the inability to culture P. vivax long-term in vitro. Sporozoite 
challenge with P. vivax also carries the risk of relapse weeks to years after infection if 
hypnozoites are not cleared from the liver (there is no dormant liver form in the P. falciparum 
lifecycle). However, there is an extensive history of deliberate infection with P. vivax; most 
notably in malariotherapy, which was carried out for the treatment of neurosyphilis almost a 
century ago. The Austrian psychiatrist Julius Wagner-Jauregg later received a Nobel Prize for his 
work with this treatment [82] and the practice was widely adopted as the only effective 
treatment available at the time. Malariotherapy provided a wealth of information about P. vivax 
infection, which has been reviewed previously [83]. Deliberate infection with P. vivax was also 
conducted in the USA from the 1940s to 1970s in prisoners involved in the Malaria Research 
Project at the Illinois State Penitentiary. The studies mainly examined compounds for their 
potential use as antimalarials, but also assessed the ability to immunise following exposure to P. 
vivax infected irradiated mosquitoes, followed by challenge with non-radiated infected 
Anopheles  [84, 85]. Similar studies were also carried out at the United States Penitentiary, Atlanta 

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and Maryland USA. Key discoveries of the biology of P. vivax were made during this period, 
including the association between Duffy negativity and resistance to P. vivax infection [43]. These 
experiments demonstrated that CHMI with P. vivax, as well as P. falciparum, could be 
successfully carried out, but the studies were very small.  
5.1.4 P. vivax sporozoite challenge in the modern era 
More recently, over the last few years, a handful of P. vivax challenge studies have been 
reported. The majority of these studies have been mosquito-bite delivered sporozoite challenge 
trials, conducted in Cali, Colombia, the Walter Reed Army Institute of Research (WRAIR), 
Maryland, USA [86] [70] [88] and Oxford (VAC068 study, ClinicalTrials.gov Identifier: 
NCT03377296, unpublished). 
A total of four sporozoite challenge studies have been conducted in Cali. The first trial involved 
eighteen healthy volunteers, exposed to the bites of 2-10 P. vivax infected Anopheles albimanus 
mosquitoes, of which seventeen developed infection. Authors speculated that the volunteer who 
did not develop malaria had surreptitiously taken anti-malarial medication but this was never 
confirmed [86]. There were no serious adverse events (SAEs) in this trial, but seven volunteers 
required fluid therapy due to nausea and vomiting, and five developed blurred vision lasting 2-3 
days after treatment initiation with chloroquine. The second P. vivax CHMI trial was carried out 
by the same group in Colombia, aiming to demonstrate the reproducibility of this method of 
infection using three different A. albimanus mosquito lots fed on blood from three P. vivax-
infected donors [87]. Seventeen individuals whose red blood cells were positive for the Duffy 
antigen/chemokine receptor (DARC, “Duffy positive”) and five Duffy negative controls were 
enrolled then randomly assigned to three groups (with six Duffy positive individuals in two of the 
groups and five in the third group). Following 2-4 bites from infected mosquitoes, all Duffy 
positive participants (and none of the Duffy negative participants) developed blood-stage 
malaria. A third P. vivax CHMI trial was carried out in Cali among both ‘semi-immune’ 
(previously-exposed; n=9) and malaria-naïve adult volunteers (n=7) [88]. Symptoms were 
significantly worse among the malaria-naïve subjects but there were no SAEs.  
The most recent CHMI trial from the Cali group assessed “immunisation” through repeated 
exposure to radiation-attenuated sporozoites, delivered by mosquito bite. Moderate efficacy was 
demonstrated in 42% of volunteers, who were sterilely protected following sporozoite challenge 
(five out of twelve Duffy positive participants protected). There were no reported SAEs related to 
immunisation, although one volunteer developed severe elevation of hepatic transaminases (>10 
times the upper limit of normal [x ULN]) with associated abdominal pain and vomiting following 
CHMI, with no alternative cause found. These symptoms resolved spontaneously [27]. 
The first P. vivax sporozoite challenge in Europe was recently conducted in April 2018, in Oxford 
within the VAC068 study (ClinicalTrials.gov Identifier: NCT03377296). Two malaria-naïve adult 
volunteers were exposed to five infectious bites (bites by mosquitos with >10 sporozoites as 
detected on microscopic examination of salivary glands post-feeding) by Anopheles dirus 
mosquitoes. Infection in the mosquitoes was established by direct membrane feeding on the 
blood from a P. vivax infected source patient, in Songkhla, Southern Thailand, before transport of 
the infected mosquitoes to the UK. Both volunteers were successfully infected, with parasitic 
DNA being detectable from day 9 after challenge by qPCR in both subjects. Symptoms consistent 
with malaria infection, including fatigue and subjective feverishness, chills, headache, loss of 
appetite, nausea and malaise, were reported by one volunteer from day 11 after challenge and 
on day 14 by the second volunteer. Fever was detected in both subjects on day 14. Thick film 
microscopy was positive (≥2 morphologically normal asexual parasites seen in 200 high-power 
(1000x) fields) in the asymptomatic volunteer on the evening of day 13 and at day 14 in the more 

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symptomatic volunteer, leading to diagnosis on day 14 in both volunteers. A 250mL blood 
donation was made by each volunteer, immediately prior to initiation of anti-malarial treatment, 
without complication, for cryopreservation and future use as a parasitised blood inoculum for 
blood-stage challenge. After 48 hours’ anti-malarial treatment, both thick film microscopy and 
qPCR were negative (two consecutive substantially reduced results) and both volunteers 
completed the clinical follow-up period, including a two-week course of primaquine for radical 
cure of hypnozoites, until 90 days post-challenge, without safety concerns. To-date (more than 
11 months post completion of primaquine therapy) there has been nothing to suggest relapse in 
either volunteer (email questionnaire follow-up is being continued for 5 years). 
5.1.5 P. vivax challenge for assessment of vaccine efficacy  
In only two of the P. vivax CHMI studies conducted to date has efficacy of immunisation been 
assessed, with both candidate vaccines targeting the pre-erythrocytic stage and utilising the 
mosquito-bite delivered sporozoite challenge model (Table 2 – taken from Payne et al., Trends in 
Parasitology, 2017 [89]). Only one of these studies has been published to date; the 
VMP001/AS01B vaccine was tested in healthy malaria-naïve adults at the WRAIR in the USA [90]. 
VMP001 is a soluble recombinant protein vaccine [91], encoding the P. vivax circumsporozoite 
protein (PvCSP) administered with the AS01B adjuvant (GlaxoSmithKline). The vaccine was 
administered to 30 volunteers in three cohorts (10 in each) at doses of 15 µg, 30 µg and 60 µg, 
given three times at a 4 week interval between the first and second dose; the third dose was 
given 8 (15 µg cohort), 6 (30 µg cohort) or 4 (60 µg cohort) weeks after the second. Twenty-nine 
volunteers completed the vaccination phase, with twenty-seven proceeding to sporozoite CHMI, 
delivered by mosquito bite, two weeks after final vaccination, alongside 6 malaria-naïve controls. 
Vaccine protective efficacy was 0%; all volunteers had developed thick blood film-detectable 
parasitaemia by day 13. The median pre-patent period for all immunised participants was 11.9 
days versus 10.7 days for infectivity controls. The challenge was well tolerated with no untoward 
reactogenicity following mosquito bites and no untoward SAEs in challenge and treatment 
phases. Participants were treated with standard chloroquine and primaquine therapy with rapid 
clearance of infection. However, two volunteers went on to have multiple relapses. One 
participant experienced two relapses (at weeks 8 and 18 after CHMI), while the other 
experienced three (at weeks 11, 20 and 48 after CHMI) [9]. By study completion the participants 
had been followed up for 5 years, and had not had any further relapses [90]. Exploratory 
genotyping for the cytochrome P450 (CYP) allele CYP2D6 was undertaken in 25 of the 33 
volunteers. The volunteers with relapses were found to have either an intermediate-metaboliser 
phenotype or poor-metaboliser phenotype. These phenotypes were associated with significantly 
lower levels of primaquine clearance 24 hours after dosing [9]. Results from another P. vivax CSP 
vaccine candidate, CS long synthetic peptides formulated in Montanide ISA 51, assessed in a 
mosquito-bite challenge study are currently awaited (Clinicaltrials.gov identifier NCT02083068). 

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Trial Site 
Number of volunteers 
Pre-patent period 
(days)a 
Number of infected 
mosquitoes OR inoculum 
Number of volunteers with 
patent parasitaemia 
References 
Sporozoite (mosquito-bite) CHMI studies 
 
Cali, Columbia 
18 
9 – 13 
2 - 10 
17/18 
[86] 
Cali, Columbia 
17 Duffy positive 
5 Duffy negative 
9 – 16 
2 - 4 
17/17 (Duffy positive) 
0/5 (Duffy negative) 
[87] 
Cali, Columbia 
7 malaria-naïve 
9 semi-immune 
11 – 13 
2 - 4 
16/16 
[88] 
WRAIR, USA 
27 vaccinees  
6 infectivity controls 
10 – 13 
10 – 11 
5 
 
[90] 
Cali, Columbia 
12 Duffy positive vaccinees 
2 Duffy positive controls 
5 Duffy negative controls 
12 – 13 
2 - 4 
7/12 vaccinees 
2/2 Duffy positive controls 
0/5 Duffy negative controls 
[27] 
Blood-stage CHMI studies  
 
QIMRB, Australia 
2 
8-9 
13,000 genome equivalents 
2/2 
[92] 
QIMRB, 
Australia 
6 
8-9 
31,786 (± 11,947) genome 
equivalents (= 15 ± 5 viable 
P. vivax parasites) 
6/6 
[93] 
Table 3: Overview of Published Plasmodium vivax CHMI Studies. aThe pre-patent period refers to the period before malaria diagnosis: by blood film 
(sporozoite) or qPCR (blood-stage).

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5.1.6 Modern blood-stage controlled human P. vivax infection 
To date, there have been four studies of blood-stage P. vivax CHMI, of which two are published, 
all conducted at the Queensland Institute of Medical Research (QIMR), Brisbane Australia. Both 
published studies used a cryopreserved source of parasitised erythrocytes obtained from a 
returned traveller with blood group A, Rhesus negative, who was infected with P. vivax during a 
trip to the Solomon Islands [92, 93]. The first blood-stage P. vivax CHMI in Europe took place in 
January 2019, in Oxford (VAC069) – see section 5.2.7 for a full description.  
In the first blood-stage CHMI at QIMR, Brisbane, two volunteers were inoculated with 270µL of 
reconstituted parasitised red cells, containing approximately 13,000 genome equivalents. Both 
subjects were successfully infected, with PCR becoming positive on day 8 or 9. Clinical symptoms 
consistent with malaria infection were present from day 11 in one volunteer, who reported 
myalgia, headache and malaise and day 14 in the other, with both experiencing chills and sweats 
at day 14. Treatment with artemether/lumefantrine was started at day 14 and volunteers were 
admitted for ongoing monitoring. Both volunteers experienced worsening of symptoms over the 
initial 12 hours post-treatment, including nausea and vomiting, headache, sweats and rigors in 
one volunteer. Pyrexia above 39oC was detected in both volunteers. After 24 hours of anti-
malarials, all symptoms had resolved, although non-tender clinically palpable mild splenomegaly 
was detected in one subject at day 15. By day 28, splenomegaly had resolved fully and no other 
adverse events were reported by either subject in the period since discharge. Whole genome 
sequencing was performed, demonstrating a genetic sequence most closely related to Cambodia 
strains, of the publically available strains. Gametocytaemia was measured by quantitative 
reverse-transcriptase PCR for the pvs25 transcript, a mature gametocyte marker. Pvs25 
transcripts were detectable from day 11 or 12 and peaked at day 14 in both volunteers, 
coinciding with peak quantitative parasite PCR, prior to the initiation of anti-malarial treatment 
[92].  
A second study was conducted to demonstrate reproducibility of the blood-stage controlled 
infection method and assess transmissibility of infection to Anopheles stephensi mosquitoes. Six 
volunteers were infected after receiving an inoculum containing approximately 31,786 (+/- 
11947) genome equivalents, estimated by investigators using linear regression growth modelling 
to equate to 15+/-5 viable parasites. Evolution of clinical symptoms was similar to the first study, 
with volunteers developing symptoms in keeping with parasite infection on days 11-13 (mean 
12.2). There were no serious adverse events during the course of either study, but for 4 of 6 
volunteers, liver enzymes became significantly elevated, above 5 times the upper limit of normal. 
Mild bilirubinaemia was also detected in three subjects, however, increases were transient and 
did not exceed 2 times the upper limit of normal. No clinical manifestations were associated with 
biochemical abnormalities. No other cause for this deranged liver function was found and liver 
ultrasound was normal but abnormalities subsequently resolved spontaneously without 
sequelae [93]. Transmissibility of infection was assessed by both direct feeding assay (DFA) and 
membrane feeding assay (MFA), carried out 3 days prior to the anticipated peak parasitaemia 
and initiation of treatment on day 14, as predicted from the first study. A total of 16 DFAs and 32 
MFAs were performed, with rates of successful feeding above 90% across the two methods, 
without significant difference between the two. Overall, 1801 mosquitoes were dissected for 
examination of oocysts 7-9 days after feeding, of which 1.8% (n=32) were demonstrated to be 
infected, although the rate of infectivity was five times higher for DFA when compared to MFA. 
Other atypical ovoid structures were also detected by investigators on the mosquito midgut, 
which could not be identified by PCR as either microsporidia or fungal pathogens, known to 
infect mosquitoes. For standardisation of the method, if used for drug or vaccine efficacy 

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evaluation, other methods for confirmation of oocysts will be required. The reason for low 
infectivity could not be determined, however, a possible incompatibility of the Solomon Island 
derived parasite strain and the laboratory-reared A. stephensi colony, which had originally been 
sourced from India, was cited by the authors as a potential contributory factor. While small, 
these studies demonstrate the safety, feasibility and reproducibility of a P. vivax blood-stage 
challenge model and show the potential for use as a transmission model for evaluation of 
transmission-blocking vaccine candidates, although the methodology is yet to be optimised.  
5.1.7 Rationale for blood-stage controlled human malaria infection 
Mosquito-bite CHMI most closely models natural infection and the robustness and reproducibility 
of the P. vivax mosquito-bite delivered sporozoite challenge model has recently been 
demonstrated by Herrera et al. in South America [86, 87]. More recently, the safety and feasibility 
of a sporozoite P. vivax challenge, was demonstrated for the first time in Europe in April 2018 
within the ongoing VAC068 study in Oxford (ClinicalTrials.gov Identifier: NCT03377296). Since 
cryopreserved P. vivax sporozoites are yet to be developed for human trials, mosquito-bite 
delivered CHMI will remain the mainstay for CHMI in assessment of pre-erythrocytic vaccines and 
drugs. However, for vaccines and anti-malarials targeting the erythrocytic stage, the blood-stage 
offers a number of advantages over sporozoite CHMI.  
Infection of mosquitoes requires a ready supply of blood containing gametocytes, from an infected 
patient. Without the possibility of long-term P. vivax ex-vivo culture, supply is limited to endemic 
areas. For CHMI studies conducted in non-endemic zones, this necessitates infection and 
importation of mosquitos from an appropriate area, with suitable entomological facilities and 
expertise, which may be logistically demanding, particularly in cases of vaccine efficacy studies, 
where timing of challenge may be critical.  
Blood stage CHMI also permits both better characterization and standardization of the parasite 
inoculum. Parasite density of the cryopreserved isolates can be established, increasing 
homogeneity of initial parasite inoculum density, when compared to sporozoites delivered by 
mosquito bite, and allowing the possibility of controlled variations of the administered parasite 
load. Full genome sequencing, including screening for mutations known to be associated with drug 
resistance can also be performed prior to challenge. In addition, safety data can be accrued 
through multiple uses of the same inoculum. Together, standardisation of strain and density 
permit for better comparison between candidate vaccines, when evaluated through challenge 
using the same inoculum source.  
Since parasitic load introduced via intravenous injection is smaller than that released following 
schizogony at the liver-stage, as in natural infection and sporozoite challenge, there is a longer 
period of blood-stage multiplication. This provides an extended period for assessment of parasite 
multiplication rate (PMR), before the development of clinical symptoms or diagnostic criteria 
might be reached. In addition, comparison of PMR between infectivity controls from blood-stage 
CHMI performed in Oxford within the VAC054 trial, assessing efficacy of the P. falciparum apical 
membrane antigen 1 (AMA1) vaccine candidate, FMP2.1/AS01, and historic controls challenged by 
mosquito-bite, illustrated that although mean PMR was comparable, blood-stage challenge 
produced a much narrower range of PMR (smaller standard deviation with tight 95% confidence 
intervals). This provided greater power to detect partial vaccine efficacy, making blood-stage 
challenge a more robust tool for evaluation of candidate blood-stage vaccines [94]. This was 
indeed borne out in the recent Phase IIa efficacy study of the candidate vaccine RH5.1/AS01B, 
where the blood-stage challenge model was able to reliably detect a 17% reduction in the PMR 
between vaccinees and infectivity controls, with a high degree of confidence (P<0.02) (VAC063, 
ClinicalTrials.gov Identifier: NCT02927145). 

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Significantly, in addition to advantages seen with P. falciparum blood-stage CHMI, P. vivax blood-
stage challenge provides a mode of infection which by-passes the liver stage. This eliminates the 
possibility of hypnozoite formation and therefore, risk of relapse. Equally, there is no requirement 
for CYP2D6 phenotype screening and primaquine therapy. A P. vivax blood-stage challenge is 
therefore less restrictive and confers important safety benefits over sporozoite challenge.  
 
5.2 Cryopreserved Plasmodium vivax blood inoculum 
5.2.1 Source and preparation of the cryopreserved inoculum 
The cryopreserved parasitized red blood cells that will be used were produced by Dr Angela 
Minassian, Prof Simon Draper and colleagues at the University of Oxford in April 2018, as part of 
the VAC068 study (ClinicalTrials.gov Identifier: NCT03377296), approved by the South Central - 
Oxford A Research Ethics Committee.  
5.2.2 Collection of P. vivax infected erythrocytes  
Parasitised erythrocytes were collected from two P. vivax-infected volunteers (otherwise 
healthy), who donated blood following sporozoite P. vivax CHMI, delivered by mosquito bite. 
Volunteers were infected via five infectious bites (defined as bites by mosquitoes with >10 
sporozoites as detected on microscopic examination of salivary glands post-feeding) from 
infected laboratory-reared Anopheles dirus mosquitos, under controlled conditions. Infected 
mosquitos were provided by, and imported from, The Mahidol University, Bangkok. The colony is 
maintained through feeding on rigorously screened human blood (provided by the Red Cross) to 
induce egg-stimulation, animal blood is never used. For infection of mosquitos, a source patient 
was recruited from a medical clinic in one of Thailand’s Southern endemic areas – Songkhala (~7 
hours’ drive from the Vivax Research Unit in Bangkok). Microscopic diagnosis of the source 
patient was established at both the field site and in the Bangkok reference laboratory, where PCR 
analysis confirmed P. vivax mono-infection. Molecular speciation by PCR (research-grade 
laboratory assay) was verified at the Jenner Institute Laboratories and clonality of the infection 
was established by the Wellcome Trust Sanger Institute in Cambridge, UK, on a 2mL whole blood 
sample, sent directly by courier from Thailand to Oxford. Mosquitoes were infected via direct 
membrane feeding with blood from the source patient, and infectivity was confirmed at 5-7 days 
post-feeding through oocyst count on dissection of the midgut. 
Both UK volunteers (“Donors”) were followed from days 1-6 post-challenge by telephone and 
from the evening of day 6 at twice-daily clinic visits. Blood sampling was performed at each visit 
for thick film microscopy and PCR. Clinical symptoms consistent with malarial infection were 
reported from day 11.5 post-challenge in Donor 1 and from day 14 in Donor 2, with both 
becoming febrile at day 14. Thick film positivity was defined as the detection of at least two 
morphologically normal malaria parasites seen in 200 high-power (1000x) fields. Thick film 
microscopy was positive at day 14.5 in volunteer 1 and at day 13.5 in volunteer 2, corresponding 
to 31,010 and 16,717 genome copies/mL by PCR, respectively. Blood donation was performed in 
each volunteer immediately prior to initiation of treatment on the morning or afternoon of day 
14. A 250mL blood sample was collected using aseptic technique, via a whole blood donation kit 
(Leuokotrap WB, Haemonetics Corp), containing an in-line leukodepletion filter. For 
anonymization of the blood donor, blood collected for cryopreservation was labelled with either 
“Donor 1” or “Donor 2”. Traceability of the blood donor is however maintained within a 
confidential clinical record, which may be accessed by the Chief Investigator, or by key members 
of the clinical team, to whom responsibility is delegated, if requested by the sponsor or other 

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external bodies. After blood collection, treatment with artemether/lumefantrine (Riamet®) was 
administered, followed by a two week course of primaquine. No supportive treatment or 
admission was required. Follow-up (clinic visits) until 90 days post-challenge was completed, with 
no safety concerns. Ongoing follow-up by email will continue for 5 years in order to monitor for 
relapse. 
5.2.3 Cryopreservation and storage of blood bank 
After collection, blood was maintained at 37oC and transported immediately to the Jenner 
Institute Laboratory, University of Oxford. All laboratory processing was conducted under GMP-
like conditions, under QP and QA oversight. Procedures were performed within fumigated 
microbiological safety cabinets, under precautions in compliance with Containment Level 3 Code 
of Practice and with sterile technique.  
First, the red cells were separated from plasma by centrifugation of the leukodepleted blood 
before mixing with Glycerolyte 57 (at 1:2 erythrocyte to Glycerolyte 57 volume ratio). The first 
20% of Glycerolyte 57 was added dropwise with gentle agitation, the suspension was then 
incubated for 5 minutes at room temperature before the remaining Glycerolyte 57 was added. 
The RBC-Glycerolyte mixture was aliquoted into 1.5mL cryovials and frozen at -80oC for one night 
before transfer to a dedicated liquid nitrogen tank. After 6 days, samples were transferred to 
Thermo Fisher Bishop’s Stortford temperature-monitored liquid nitrogen facility, Hertfordshire, 
UK, where samples are stored on behalf of the University of Oxford. This process was performed 
under conditions similar to those used for blood thawing in previous CHMI trials at Oxford and 
according to the Jenner Institute Laboratory SOPs. 
5.2.4 Infectivity and characterisation of parasite stock 
Confirmation of parasite density within the blood collected for cryopreservation was performed 
via microscopy and quantitative PCR on the leukodepleted blood samples. Thick film microscopy 
demonstrated 11 asexual parasites per 1µL leukodepleted blood for Donor 1, and 5 asexual 
parasites per 1µL leukodepleted blood for Donor 2. Quantitative PCR confirmed the presence of 
23,566 genome copies/mL in Donor 1, and 14,078 genome copies/mL in Donor 2. This suggested 
minimal parasitic loss through the leukodepletion process when compared to the diagnostic PCR 
values of 31,010 and 16,717 genome copies/mL, respectively. 
Viability has also been demonstrated in an ex-vivo short-term culture of thawed infected red 
blood cells in enriched McCoy 5A medium. Parasitic growth was detectable by light microscopy, 
quantitative PCR and flow cytometry through an initial 40-hour growth cycle in samples collected 
from Donor 1, with normal progression of normal morphology as seen on Giemsa stained thick 
and thin films. However, parasite growth was sub-microscopic in samples obtained from Donor 
2. Therefore, we intend to proceed with further testing of cryopreserved samples from Donor 1, 
and will use this as the challenge inoculum in this and future studies. Further testing will include 
whole genome sequencing to further characterize the parasitic strain, with particular scrutiny for 
SNVs present in the PvDBP gene and the five putative drug resistant genes pvdhfr, pvdhps, pvcrt, 
pvmdr and pvmrp.  
5.2.5 Testing of source patient and blood donors for blood-borne and mosquito-borne 
infections 
Initial testing for blood-borne infections and mosquito-borne disease was carried out on blood 
samples from the original source patient in Thailand, taken at the time of P. vivax diagnosis. 
Since Anopheles is a known vector of Wucherichia bancrofti, the main causative agent of 
lymphatic filariasis, blood was screened for filarial disease via rapid diagnostic test for IgG4 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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antibodies to the W. bancrofti Wb123 antigen (SD BIOLINE Lymphatic Filariasis IgG4) at the Thai 
field site. In addition, although there is no evidence that blood-borne infections can be 
transmitted by mosquito bite, for maximal assurance of safety, serology for HIV-1 and HIV-2, 
HTLV, Hepatitis B and C and syphilis were performed on a serum sample from the Thai source 
patient, at Oxford University Hospitals NHS Trust. Due to anecdotal reports of Japanese B 
encephalitis and Chikungunya in Anopheles species, blood from the source patient was also 
screened for both arboviral infections by PCR, as performed by the WRAIR unit, who used the 
same mosquito source for a CHMI trial that assessed the efficacy of the VMP001/AS01 P. vivax 
circumsporozoite recombinant protein vaccine[90]. As a further precautionary measure, 
although Anopheles species are not known to be vectors of Dengue, Zika or West Nile virus, on 
the independent advice of clinical Tropical Medicine specialist, Prof. Nicholas Day, Professor of 
Tropical Medicine and Director of the Mahidol Oxford Tropical Medicine Research Unit in 
Thailand, PCR for these infections was performed on the source patient’s blood. In summary, 
molecular analysis (PCR) for Chikungunya, Japanese B encephalitis, West Nile, Dengue and Zika 
viruses was performed on whole blood at the Rare Imported Pathogens Laboratory (RIPL) in the 
UK. All source patient infection screen tests were negative.  
Both UK volunteers (“donors”) were subjected to screening for blood-borne infections, in line 
with the Joint UK Blood Transfusion and Tissue Transplantation Services Professional Advisory 
Committee guidelines https://www.transfusionguidelines.org/red-book/chapter-9-microbiology-
tests-for-donors-and-donations-general-specifications-for-laboratory-test-procedures/9-1-
general-requirements. Testing comprised serological tests for HIV-1 and HIV-2, hepatitis B and C, 
syphilis (anti-treponemal Ab) and HTLV-1 and HTLV-2 at screening, and nucleic acid amplification 
tests for HIV-1 and HIV-2 and hepatitis B and C, as well as repeat serological tests for HTLV-1 and 
HTLV-2 and syphilis 7 days before sporozoite challenge. Neither volunteer had travelled to any 
area which represented a risk of exposure to any other form of blood-borne infection. All tests 
were negative. Volunteers also underwent repeat serological testing for HIV, hepatitis B and C, 
syphilis, HITLV-1 and HTLV-2, 90 days after challenge was undertaken, to ensure that no 
seroconversion from a recently-acquired infection (that may have been undetectable around the 
time of challenge) had occurred since the challenge period. All tests remained negative.  
In addition, volunteers were screened for Epstein-Barr virus (EBV) and Cytomegalovirus (CMV) 
prior to enrolment. Both volunteers were IgG seropositive for both infections. However, 
leukodepletion has been shown to be effective in preventing transmission of both EBV and CMV 
[95] and PCR testing for both infections on the inoculum was negative, therefore, EBV-CMV 
concordant serostatus would not be considered a requirement for recipients of this donated 
blood in this study. This is in line with conduct of previous P. falciparum CHMI studies at Oxford, 
which have utilised a cryopreserved source of infected erythrocytes from a donor who was 
seropositive for both EBV and CMV. Initial studies using this inoculum had initially required that 
participants in CHMI trials be seropositive for both infections. However, following the 
development of PCR assays for both infections, molecular analysis of the inoculum was negative 
for both EBV and CMV. Following a risk-assessment, QIMR, Australia, subsequently removed 
seropositivity to these viruses as a criterion for participation as a recipient of the inoculum. In a 
previous P. falciparum challenge study (VAC054) at Oxford, 63% of volunteers were seronegative 
for CMV and 11% were seronegative for EBV pre-CHMI, and no cases of seroconversion were 
recorded post-CHMI [94]. Volunteers will however be informed of the theoretical risk of 
transmission of infection and this will be included within the consent form. Serostatus will be 
tested prior to CHMI at screening for volunteers as part of this protocol, and will be re-tested 
post-challenge. Serum samples post-CHMI will also be stored, for serostatus analysis of any 
infection, should this be required or requested by the Chief Investigator.  

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5.2.6 Sterility and screening for blood-borne infections of cryopreserved blood bank 
Blood collected for cryopreservation was tested for bacterial contamination using validated GMP 
and GLP laboratory techniques. These included tests of sterility by direct inoculation, 
mycoplasma specific culture and detection and quantification of bacterial endotoxin by kinetic 
chromogenic limulus amoebocyte lysate assay. All of these assays were performed by SGS 
Vitrology, Glasgow, UK, using assays compliant with harmonised European Pharmacopeia and 
United States Pharmacopeia standards.   
A further screen for blood-borne infections was conducted on the plasma, derived directly from 
the blood bank (separated from the red cells prior to freezing down), in line with testing 
procedures performed by the NHS Blood Transfusion service. DNA PCR for HIV-1, hepatitis B, 
hepatitis C, EBV, CMV, serology for HIV-2 and serology for HTLV-1, HTLV-2, and Treponema 
pallidum (RPR) was performed on thawed plasma samples at University Hospitals Birmingham 
NHS Foundation Trust laboratory. All tests were negative.  
5.2.7 Determination of safety, feasibility and dose of cryopreserved P. vivax inoculum  
Safety and feasibility of blood-stage P. vivax challenge, using the same bank of cryopreserved P. 
vivax infected erythrocytes as will be used in this trial, was assessed for the first time by CHMI in 
the VAC069 study in January 2019 (ClinicalTrials.gov identifier: NCT03797989). This proof-of-
concept assessed feasibility of infection at three different doses of inoculum, by injection at 
three dilutions, with a view to identifying the lowest concentration producing a reliable infection 
within a practicable timeframe.  
Six healthy, malaria-naïve adults aged between 18 and 50 years were recruited into three groups, 
each comprising two volunteers. Two volunteers receive a whole vial’s worth of infected 
erythrocytes, two volunteers received one fifth of the dose of parasitised erythrocytes, at a 1:5 
dilution, and the final two volunteers were inoculated with one twentieth of the dose, at a 1:20 
dilution. Intravenous inoculation of volunteers was completed without complication, with all 
volunteers safely discharged at 1 hour post-CHMI following satisfactory observation. The overall 
safety profile was in-keeping with AEs as expected in P. vivax CHMI, and comparable to AEs as 
reported in previous P. vivax blood-stage challenge studies, as summarised below [92, 93].  
Safety data was actively collected (solicited adverse events) on the following symptoms deemed 
indicative of P. vivax infection at post-challenge clinic visits until 28 days after CHMI: subjective 
feverishness, chills, rigors, sweats, headache, nausea, vomiting, diarrhoea, myalgia, arthralgia, 
lower back pain, fatigue and general malaise. Clinic visits were once or twice daily from day of 
challenge until completion of anti-malarial therapy, depending on the number of days since 
challenge and qPCR result, followed by assessment at 6 days post treatment initiation and 28 
days post-CHMI, as set out by the study protocol. The severity of adverse events were self-
assessed by volunteers who were counselled on criteria for grading as set out in Table 13. 
Unsolicted AEs and SAEs were assessed throughout the study period. 
Prior to, or at the time of malaria diagnosis, the majority of volunteers (4/6) experienced either 
no or mild AEs, whilst 2/6 volunteers reported a constellation of moderate to severe AEs 
including feverishness, chills, sweats, headache, fatigue and malaise (Figure 3). AEs reported as 
moderate or severe in intensity were more frequent post-diagnosis, following initiation of anti-
malarial therapy and were reported by 5/6 volunteers, each experiencing 1-6 AEs graded as 
severe in intensity. However, no AE reported as severe persisted at grade 3 for more than 48 
hours, with the vast majority resolving completely within 24 hours (Table 4). The most frequent 
AEs reported as moderate-severe (grade 2-3) in intensity were feverishness, chills, malaise, 
fatigue and sweats (Figure 2). The large majority of all AEs resolved within 72 hours of anti-

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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malarial treatment, with only a single volunteer reporting any solicited AE in the period between 
two and six days after treatment of mild-moderate intensity, which completely resolved at 28 
days after challenge.  
AEs relating to physical observations were mostly mild in nature, with a single grade 3 pyrexia 
occurring (graded as per local SOP), which resolved within 24 hours (Figure 4). Of the laboratory 
AEs recorded, two were grade 3 in severity (graded as per local SOP), both relating to 
lymphocytopaenia in two volunteers at the time of diagnosis (Tables 4 and 5). These grade 3 
events were transient, resolving completely within 6 days, and the abnormality was deemed to 
be in keeping with P. vivax malaria diagnosis. Given reported transaminitis in previous P. vivax 
CHMI studies, liver function tests were monitored through the post-CHMI period on days 7 and 
14 post-challenge, on day of diagnosis, days 1 and 6 following treatment commencement and 28 
days after challenge. Moderate (grade 2) transient derangement of ALT on liver function tests 
was seen in only 2/6 volunteers, detected at 6 days post-treatment. Both volunteers were 
asymptomatic and transaminases normalised by 28 days post-CHMI (within 10 days).  
To supplement clinical assessment following initiation of anti-malarial therapy and confirm 
parasitological clearance, qPCR was performed on days 1 and 2 post-treatment. There was a 
substantial reduction in parasite genome copies/mL by qPCR in all volunteers within 48 hours of 
therapy (5/6 treated with artemether/lumefantrine (Riamet), 1/6 treated with 
atavoquone/proguanil (Malarone)) and qPCR remained negative at 6 days post-treatment 
initiation in all volunteers. AEs relating to treatment were infrequently reported, and were mild-
moderate in nature where experienced (Figure 5). No grade 3 unsolicted AE was reported and 
there have been no SAEs to date in the VAC069 study. 
Feasibility of CHMI through intravenous administration of the P. vivax inoculum was 
demonstrated by the successful infection of all volunteers, with a mean day of diagnosis of 15 
days after CHMI (range 12.5-16.5). The two volunteers receiving the highest inoculum dose were 
diagnosed at days 12.5 and 15.5 respectively, those receiving a 1:5 dilution were both diagnosed 
at day 15 and those receiving the 1:20 dilution were diagnosed at days 15.5 and 16.5. All three 
dilutions were therefore demonstrated to be reliably infective. Based on these infectivity data, 
and accounting for possible variation in number of parasitised erythrocytes between thawed 
cryopreserved vials, an inoculum dilution of 1:10 (prepared as per Jenner laboratory SOP) will be 
utilised for P. vivax blood-stage CHMI in this VAC071 study.  
Volunteers are scheduled to return to undergo a second challenge in September 2019 
(approximately) and third challenge in February 2020 (approximately) to establish safety and 
feasibility of secondary and tertiary P. vivax controlled blood-stage challenge. The final follow-up 
will be 90 days after the tertiary challenge, approximately 1 and half years from enrolment. The 
natural immune response to primary, secondary and tertiary challenge, as well as 
gametocytaemia will also be explored following primary, secondary and tertiary blood-stage P. 
vivax infection. PCR quantification of gametocytes following primary challenge, as well as in-
depth immunological analysis of the natural immune response, including human and parasite 
transcriptomics, are currently underway. 

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Figure 2: Maximal severity of solicited AEs following P. vivax blood-stage CHMI in the VAC069 
study (n=6)  
Figure 
3: Maximal severity of solicited AEs reported by all volunteers (n=6) in the VAC069 study at 0-48 
hours prior to diagnosis, on day of diagnosis and 0-48 hours post-diagnosis  

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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Volunteer 
Grade 3 AE 
Timepoint 
Persisted at grade 3 
Resolution 
6901002 
Feverishness
T+1 
24 hours 
T+2 
6901003 
Feverishness
DoD/C+12.5
48 hours 
T+2 
Chills 
T+1 
24 hours 
T+2 
Sweats 
T+1 
24 hours 
T+2 
Myalgia 
T+1 
24 hours 
T+2 
Fatigue 
T+1 
24 hours 
C+28 
Malaise 
T+1 
24 hours 
T+2 
6901006 
Feverishness
T+1 
24 hours 
T+2 
Chills 
T+1 
24 hours 
T+2 
Headache 
T+1 
24 hours 
T+2 
Fatigue 
T+1 
24 hours 
T+2 
Malaise 
T+1 
24 hours 
T+2 
6901009 
Feverishness
T+1 
24 hours 
T+2 
Rigor 
T+1 
24 hours 
T+2 
Table 4: Onset and duration of grade 3 solicited AEs reported by all volunteers (n=6) in the 
VAC069 study 
0
50
100
Pyrexia (in clinic)
Tachycardia
Bradycardia
Systolic hypertension
Systolic hypotension
Diastolic hypertension
% Volunteers
Grade 0
Grade 1
Grade 2
Grade 3
 
Figure 4: Percentage of all volunteers (n=6) with AEs in recorded physical observations 
 
Figure 5: AEs possibly relating to treatment in all volunteers (n=6) 

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Table 4: Laboratory AEs in biochemistry profile (urea and electrolytes, liver function tests) in all volunteers (n=6) in the VAC069 study 
 
Laboratory AE 
Volunteer 
Sex 
Timepoint(s) 
Result 
Grade 
Action taken 
Anaemia 
6901002 
M 
T+6 
119 
1 
Repeated and resolved at next visit (C+28) 
6901005 
F 
C+16, C+34 (C+14-C+90) 
98 (98-109) 
(1-)2 
Monitored from C+14-C+90, AE ongoing at C+90 (102 - grade 1). Referred to GP for ongoing 
medical care, with follow-up of AE ongoing. 
Low white blood cell 
count 
6901002 
M 
T+1 
3.3 
1 
Consistent with P. vivax diagnosis, repeated and resolved (6.53) at next visit (T+6) 
6901003
F
T+1
2.93
1
Consistent with P. vivax diagnosis, repeated and resolved (6.86) at next visit (T+6)
6901005 
F 
T+1 (C+7-C+90) 
1.68 (1.68-3.39) 
(1-)2 
Monitored from C+7-C+90, AE ongoing at C+90 (3.36 - grade1). Referred to GP for ongoing 
medical care, with follow-up of AE ongoing.  
6901006 
M 
T+1 (C+14-T+1) 
2.36 
2 
Monitored from C+14-T+6, resolved at T+6 (4.84) - consistent with P. vivax diagnosis 
Thrombocytopaenia 
6901003 
F 
T+1 
114 
2 
Consistent with P. vivax diagnosis, repeated and resolved (233) at next visit (T+6) 
6901005
F
T+1(DoD-T+1)
115 (115-127)
(1-)2
Consistent with P. vivax diagnosis, repeated and resolved at T+6 (219)
Neutropaenia 
6901005 
F 
C+45 (C+14-C+90) 
0.76 (0.76-1.46) 
(1-)2 
Monitored from C+14-C+90, AE ongoing at C+90 (1.26 – grade 1). Referred to GP for ongoing 
medical care, with follow-up of AE ongoing. 
Lymphocytopenia 
6901002 
M 
T+1 (DoD-T+1) 
0.72 (0.72-0.86) 
(1-)2 
Consistent with P. vivax diagnosis, repeated and resolved at T+6 (1.63) 
6901003
F
T+1
0.77
1
Consistent with P. vivax diagnosis, repeated and resolved (2.53) at T+6 
6901005 
F 
DoD (DoD-T+1) 
0.44 (0.44-0.63) 
(2-)3 
Consistent with P. vivax diagnosis, repeated at resolved (1.63) at T+6 
6901006 
M 
T+1 (C+14-T+1) 
0.3(0.3-0.83) 
(1-)3 
Consistent with P. vivax diagnosis, repeated and resolved at T+6 (2.07) 
Table 5: Laboratory AEs in full blood count in all volunteers (n=6) in the VAC069 study 
Laboratory AE 
Volunteer 
Timepoint(s) 
Result 
Grade  
Action taken 
Hyponatraemia 
6901003
C+28
134
1
No clinical concern, no action taken
6901003
C+90
134
1
No clinical concern, no action taken
Hypokalaemia 
6901002 
C+90 
3.3 
1 
No clinical concern, no action taken 
6901005 
DoD 
3.3 
1 
Repeated within 24 hours at next visit, resolved (3.7) 
6901003 
C+90 
3.3 
1 
No clinical concern, no action taken 
ALT 
6901003 
T+6 
135 
2 
Asymptomatic, repeated in 48 hours (80 - grade 1) and then resolved at C+28 (21) 
6901006
T+6
56
1
Repeated and resolved (29) at next visit (C+28),
6901007 
T+6 
80 
1 
Repeated and resolved (31) at next visit (C+28)  
6901009 
T+6 
114 
2 
Asymptomatic, repeated in 72 hours and resolved (40)  

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5.3 
Conduct of CHMI trials 
5.3.1 Standardisation of CHMI studies 
Following a collaborative consensus process involving investigators from the USMMVP, Sanaria, 
University of Maryland, University of Oxford, RUNMC, The Seattle Biomedical Research Institute 
and the KEMRI-Wellcome Kilifi Research Programme, a consensus document; “Standardization of 
Design and Conduct of P. falciparum Sporozoite Challenge Trials” was developed, and provides a 
comprehensive guide to the appropriate conduct of sporozoite CHMI studies [78]. There is no 
equivalent document for controlled blood-stage infection or for P. vivax challenge, however, this 
study will be conducted in line with the following key points.  
 All volunteers should have a medical assessment no longer than 48 hours before 
challenge, including an interim medical history, directed physical examination, pregnancy 
test for female volunteers. 
 Volunteers should be questioned about the occurrence of adverse events and use of 
medication at each follow-up visit. 
 In the event that a volunteer does not attend a scheduled follow-up visit it is imperative 
that Investigators find that volunteer as quickly as possible and assess them for patent 
parasitaemia and clinical malaria. Should the volunteer withdraw consent from further 
follow-up prior to receipt of antimalarial drugs, it may be appropriate to withdraw the 
volunteer from the trial protocol and administer a course of antimalarial chemotherapy 
under close supervision.  
 Grading and reporting of adverse events should be performed using international and 
local guidelines. It should be noted that the occurrence of a low frequency of grade 3 
severe adverse events, of short duration, and with no long-term sequelae, is not 
unexpected in CHMI studies. A minority of those challenged are known to experience 
grade 3 systemic adverse events and this fact should be included in the informed consent 
form. 
 Vital signs should be recorded at least once daily and at any subsequent visits for medical 
attention. Directed physical examination should be performed when necessary.  
 It is critical that every volunteer must receive every dose of anti-malarial therapy. In some 
settings fully directly observed treatment will be essential. Where directly observed 
treatment is not used, Investigators must follow volunteers closely to ensure compliance 
with the treatment regimen.  
 After challenge, all volunteers should be followed until they have completely finished 
anti-malaria treatment.  
 Volunteers should be evaluated at least two weeks after finishing treatment. 
 A local safety monitor and an independent safety monitoring committee should be 
established to act as independent experts in evaluating adverse events. The safety 
monitor or monitoring committee may advise the Investigators on initiating anti-malarial 
treatment for a specific volunteer or volunteer group. While safety monitoring 
committees are not a requirement for Phase I trials, they should be considered a 
requirement for CHMI trials which have an efficacy/human challenge component and 
which have major potential safety concerns. 
5.3.2 Ethical considerations of CHMI trials 
Participants in CHMI trials are healthy volunteers who do not obtain direct health benefit from 
participation. Challenge trial investigators must exercise all possible safeguards for volunteer 
safety to ensure that trial participation is of minimal risk. Investigators must also ensure that 

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maximal scientific benefit accrues from each challenge trial. Key ethical considerations agreed by 
consensus of the field are outlined in a review by Moorthy from 2011, and include; [78]   
1. Volunteer safety is the paramount consideration in conduct of CHMI trials.  
2. Adherence to both international and local guidelines with respect to ethical 
considerations and in accordance with the Declaration of Helsinki and local regulatory 
and ethics committee requirements.  
3. CHMI trials should be conducted according to ICH and/or WHO Good Clinical Practice 
Guidelines with the aim of maximising scientific benefit whilst minimising risk. 
4. The raw data (both microscopy and PCR where available) from challenge trial datasets 
should be made publicly available to facilitate scientific benefit to the community.  
5. If an unexpected SAE which is possibly related to CHMI occurs at a challenge trial centre, 
recognising legal restrictions, every effort should be made to communicate information 
on this SAE to the community of challenge trial centres within 90 days of the occurrence 
of the SAE.  
5.3.3 Clinical presentation post-CHMI 
Nearly all unvaccinated volunteers in CHMI studies develop symptoms of clinical malaria 
infection; based on data predominately from P. falciparum challenge studies approximately one-
fifth of volunteers temporarily develop symptoms graded as severe (symptoms that prevent daily 
activities), but severe or life-threatening malaria has never occurred [96]. The expected time-
course, symptoms and management of clinical malaria are outlined in Section 9.  
Routine laboratory checks generally show a moderate decrease in leukocyte and platelet 
numbers during infection, with no change in haemoglobin concentration [97]. Bleeding or 
thrombogenic complications have never been described [96, 97]. Abnormalities of liver enzymes 
have been observed, but these abnormalities have rarely resulted in clinical manifestations (just 
one volunteer with raised ALT associated with abdominal pain and vomiting in the recent RCT 
from Cali [27]) and they resolved after a few days.  
It is estimated that human malaria challenge infections have now been conducted with P. 
falciparum in over 2,650 volunteers and in over 300 volunteers with P. vivax  [98]. In 2009, safety 
concerns were raised when young volunteer suffered a cardiac event shortly after treatment for 
diagnosed malaria, following a P. falciparum challenge. This was diagnosed as probable 
myopericarditis, although ischaemia could not be ruled out. Although a definite relationship 
between the cardiac event, which resolved fully and rapidly, and the experimental malaria 
infection was not established [99], it has been generally agreed that volunteers with an increased 
risk of cardiac disease should be excluded from such trials [78]. A further case of myopericarditis 
has since been identified in a recent P. falciparum CHMI study, also at the Nijmegen, Netherlands 
centre, but in this case the individual was also diagnosed with an intercurrent rhinovirus 
infection so that the relation to malaria infection is again uncertain. There was a brief episode of 
clinical chest pain and the volunteer made a full recovery [100]. 
5.3.4 Oxford’s experience conducting CHMI trials 
The University of Oxford has been conducting CHMI studies (with P. falciparum) for the last 18 
years. To date, nearly 600 volunteers have undergone CHMI in studies conducted by the 
University of Oxford, including more than 180 unvaccinated volunteers. An analysis has shown 
that the symptoms of malaria experienced by volunteers undergoing sporozoite challenge by 
mosquito bite in Oxford are broadly similar to those experienced by volunteers challenged by 
mosquito bite at Radboud University Nijmegen Medical Center (RUNMC) and the US Military 
Malaria Vaccine Program (USMMVP) [101]. In Oxford, four volunteers amongst nearly 600 
individual challengees have required admission to hospital for observation following challenge 
(SAE related to CHMI). All of these subjects made a complete recovery. This will be only the 

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second blood-stage P. vivax challenge study conducted in Oxford, but we have based our study 
design on that of the previous P. vivax blood-stage challenge studies in Australia, combined with 
our own extensive experience of P. falciparum sporozoite challenge studies and our recent 
VAC068 P. vivax sporozoite challenge study.  
In one Phase I/IIa sporozoite challenge study assessing the efficacy of viral vectored malaria 
vaccines in Oxford (VAC039; Clinicaltrials.gov reference: NCT01142765), a volunteer who 
underwent sporozoite challenge on 1st October 2010 failed to attend his next scheduled study 
visit on 7th October 2010 [102]. The police were immediately informed and began a nationwide 
search for the individual. All volunteers had been informed at screening that the police would be 
notified should they go missing following CHMI if they had not completed a full course of an 
appropriate anti-malarial treatment. The volunteer was found in the Netherlands by the local 
police 17 days following CHMI. He then had very mild malaria symptoms. He was admitted to a 
local hospital where he received appropriate treatment for P. falciparum. He had no signs of 
severe malaria but showed an altered mental state considered unrelated to malaria with 
apparent memory loss and suicidal ideation. He was therefore transferred for in-patient 
psychiatric assessment and discharged a few days later to his GP’s care. He has subsequently 
been reportedly diagnosed as schizophrenic. 
It emerged that from 2nd October 2010, the day after CHMI, the volunteer had experienced an 
alteration in his expected behaviour following his arrest the previous evening by the police 
relating to their investigation of a serious crime, of which he was later convicted. This arrest 
appeared to trigger his leaving home and disappearance and appears relevant to the subsequent 
finding of memory loss and suicidal ideation. It emerged on subsequent investigation that the 
volunteer actually had a history of some psychiatric morbidity pre-dating his involvement in the 
study by many years, which was not disclosed at screening by the volunteer or his GP. Of note, 
the volunteer had attended 9 clinic visits prior to challenge and appeared a reliable and 
appropriate volunteer.  
This event was extensively discussed with investigators, colleagues and appropriate authorities 
and non-study related causality agreed. Given the exceptional circumstances relating to this case, 
it seems very unlikely that a similar event would happen again in the future. Management of the 
event was extensively reviewed by the trial’s sponsor, the MHRA and ethical committee who felt 
that appropriate and timely action was taken. Follow-up procedures following CHMI have been 
reviewed locally, and it has been decided that for future studies volunteers should be contacted 
daily on days 1-5 post sporozoite challenge in order to make sure they are contactable and well. 
Daily telephone contact will similarly be conducted in this study on days 1-6, before daily clinic 
visits commence on day 7.  
5.3.5 Conducting CHMI in the context of the COVID-19 pandemic 
Due to the COVID-19 pandemic this trial was temporarily halted in March 2020. The incidence of 
COVID-19 continues to change in England and some ongoing transmission is likely to remain 
when this trial restarts in 2021 [103]. This trial will only proceed when rates of COVID-19 
infections in England are low and the risk of contracting COVID-19 during malaria challenge is 
small, however the risk cannot be eliminated. There is currently no evidence for the possible 
effects of co-infection with malaria and COVID-19. As the risk of co-infection cannot be 
quantified, participants will be tested for COVID-19 prior to malaria challenge and any participant 
that tests positive for COVID-19 will not proceed with malaria challenge.  
Participants who test positive for COVID-19 will be advised to self-isolate as per Public Health 
England guidance and directed to local NHS services for further care if required. Positive COVID-
19 PCR results will be reported as a notifiable disease as per Public Health England guidance. 

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Fevers which occur following malaria challenge may pose diagnostic difficulties as concurrent 
COVID-19 infection will likely remain an ongoing possibility in England. Data from the 12 primary 
and 4 secondary CHMI with P. vivax conducted in Oxford to date, show that no participants 
developed symptoms that are consistent with malaria within the first week post challenge. 
Pyrexia or subjective feelings of feverishness occurred in only half of the participants prior to 
diagnosis and treatment of malaria. However the majority of remaining participants who were 
initially apyrexial, did subsequently develop fever after commencing malaria treatment, although 
this was short lived and resolved within 2 days. Following malaria challenge, participants first 
developed pyrexia or subjective feelings of feverishness between days 10 to 20 post challenge 
(median 14 days), with corresponding malaria qPCR levels of 100 to 9600 genome copies/mL 
(median 2700 genome copies/mL). No recorded pyrexia of >37.8°C occurred at a malaria qPCR 
below 1000 genome copies/mL in any participant. Malaria diagnosis was made in participants 
within a few days following the onset of symptoms with some participants reporting no 
symptoms prior to laboratory diagnosis. Participants were diagnosed with malaria based on a 
combination of clinical symptoms, malaria qPCR and thick film microscopy between days 12 and 
21 post challenge (median 15.5 days).  
Fevers within the first week after malaria challenge are therefore unlikely to be due to malaria. 
After the first week post challenge, malaria as the cause of fever becomes increasingly likely with 
increasing time since challenge. However, as concurrent COVID-19 infection will likely remain a 
possibility, testing for COVID-19 has been added in the post-challenge period for participants 
who become febrile. For participant safety, if a participant tests positive for COVID-19 prior to 
malaria diagnosis, they will be treated for malaria regardless of if they have reached the criteria 
for malaria diagnosis.  
There is no evidence for any detrimental effect of anti-malarial medication on the outcome of 
concurrent COVID-19 infection. This is in the context of widespread antimalarial use during the 
COVID-19 pandemic in Africa, where no safety signal has emerged. Recent randomised 
controlled trials have shown no benefit of Hydroxychloroquine in COVID-19 [104], but also no 
harm, and Chloroquine will not be used for malaria treatment in this trial. The anti-malarial 
medications Riamet and Malarone are not expected to have any effects on the outcome of 
COVID-19 disease. 
Social distancing requirements to control COVID-19 will likely remain in place to some extent in 
England. To abide with these, participants’ attendances at study clinics will be carefully planned 
to adhere to social distancing guidelines. Participant flow will be arranged so that contact with 
other people is minimised and Personal Protective Equipment (PPE - compromising apron, 
gloves, surgical mask and eye protection) will be worn by members of clinical staff for all 
participant visits. This will follow an infection control SOP that covers all PPE requirements for 
clinical trials at the Oxford Vaccine Centre in the era of the COVID-19 pandemic. Any clinic visits 
that are carried out on a participant that is self-isolating will be conducted in a separate clinical 
area with side access to avoid any contact with other volunteers. 
As the national rollout of COVID-19 vaccines proceeds, some participants may be offered a 
COVID-19 vaccine around the time of planned malaria challenge. It is unknown what effect 
concomitant COVID-19 vaccine administration around the time of CHMI may have on the 
immune response to malaria infection, which is being studied in this trial. In addition 
reactogenicity from COVID-19 vaccination may be difficult to distinguish from symptoms of 
malaria and may lead to earlier diagnosis and treatment of malaria after challenge. Therefore 
concomitant COVID-19 vaccination around the time of CHMI has been added as an exclusion 
criteria in this trial. 
 
 
 

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6 INVESTIGATIONAL PRODUCTS 
6.1 PvDBPII 
PvDBPII was manufactured under Good Manufacturing Practice (GMP) conditions by Syngene 
International, Bangalore, India, and the drug product was vialed and released by Zydus Cadila, 
Ahmedabad, India.  
PvDBPII is supplied as a liquid at a concentration of 200 μg/mL in sterile aliquots in 0.5 mL clear 
glass vials. Further details relating to batch release and manufacturing can be found in the 
PvDBPII IND.   
All vaccines will be certified for release by a qualified person (QP) at the CBF, University of 
Oxford.  
6.2 Matrix M1 
Matrix M1 is manufactured under Good Manufacturing Practice conditions by Novovax AB 
(Uppsala, Sweden). Matrix-M1 will be supplied to the clinical site by Novovax AB and the 
adjuvant will be labelled for investigational use only. 
Matrix-M1 is formulated at a concentration of 0.375mg/ml in PBS. The drug product is filled into 
sterile 3mL 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 filter 0.22 μm and 
filling into vials in a volume of 2 ml. Matrix-M1 is a colourless slightly-opalescent non-viscous 
liquid. 
Final batch certification and associated labelling takes place at the CBF, University of Oxford. 
Further details relating to batch release and manufacturing can be found in the Matrix M1 IMP-
D.   
6.3 Storage of Vaccines  
All vaccines will be stored between –70°C and –90°C. All movements of the study vaccines will be 
documented. Vaccine accountability, storage, shipment and handling will be in accordance local 
SOPs.  
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. 
6.4 Administration of Vaccines 
All vaccines will be administered intramuscularly in the deltoid muscle of the arm, 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.  
For each immunization, one vial of PvDBPII will be used per subject. One vial of adjuvant will be 
used for a maximum of two immunizations. A mixture of PvDBPII at a dose of 50 μg, with 50μg of 
Matrix-M1 will be administered to all volunteers. Matrix-M1 and PvDBPII will be mixed at the 
bedside immediately prior to administration. Any vaccine not administered within the time 
frame will be destroyed. 

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7 OBJECTIVES AND ENDPOINTS 
7.1 Primary Objectives  
To assess the safety and tolerability  of the PvDBPII vaccine formulated in Matrix M1 
To establish whether the PvDBPII-Matrix M1 vaccine can demonstrate a reduced parasite 
multiplication rate in vaccinated subjects compared to infectivity controls in a blood-stage 
controlled human malaria infection model 
7.1.1 Primary Safety 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 
the 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 
diary cards, clinical review, clinical examination (including observations) and laboratory results. 
This AE data will be tabulated and frequency, duration and severity of AEs compared to the AE 
data collected for the infectivity control group undergoing parallel CHMI recruited as part of the 
VAC069 clinical trial. Hematological and biochemical laboratory values will be presented 
according to local grading scales and tabulated by group, for comparison with the laboratory AE 
data collected for infectivity control groups undergoing CHMI in parallel.  
SAEs, AEs of special interest and withdrawal due to AE(s)/SAE(s) will be described in detail. 
The vaccinated volunteers will be followed for approximately 1 year following their first 
vaccination with PvDBPII-Matrix M1. 
7.1.2 Primary Efficacy Outcome Measures 
Quantitative PCR-derived parasite multiplication rate (PMR) will be the primary efficacy endpoint 
and a comparison of the endpoint between the volunteers vaccinated with PvDBPII-Matrix M1 
and malaria-naïve controls partaking in parallel CHMI will constitute the primary analysis for 
efficacy.  
The arithmetic mean of the three replicate PCR results obtained for each individual at each 
timepoint will be used for model-fitting. Negative individual replicates will be handled as 
specified in the Jenner Institute qPCR SOP. qPCR data points which, based upon the mean of the 
three replicates, are negative or below the limit of quantification will be handled as specified in 
the Jenner Institute qPCR SOP. PMR will be calculated using a linear model fitted to log10-
transformed qPCR data.  
The distribution of PMRs will be assessed for deviation from normality visually and using a 
d’Agostino-Pearson test. Standard ladder-of-power transformations may be applied to achieve 
normality. Equality of variance will be tested with an F test. In the event of deviation from 
normality, PMRs will be compared by two-tailed Mann-Whitney test; otherwise a two-tailed two-

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sample t-test will be used, with Welch’s correction if variances are non-equal as assessed by F-
test.  
7.2 Secondary Objectives 
To assess the humoral and cellular immunogenicity of the PvDBPII-Matrix M1 vaccine candidate. 
To assess immunological readouts for association with a reduced parasite multiplication rate 
To assess the durability of any reduction in PMR in Group 3 volunteers who undergo a fourth 
vaccination followed by re-challenge approximately 5 months after the primary challenge 
7.2.2 Secondary Immunological Outcome Measures 
PvDBPII-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 P. vivax Duffy-binding protein (PvDBPII) – total IgG, isotypes and avidity; T cell 
responses to PvDBPII by flow cytometry assays and in vitro functional PvDBPII inhibitory binding 
assays. Other established and exploratory immunology assays may be carried out, including 
through collaboration with other specialist laboratories, which may be outside of UK/Europe. 
This would involve transfer of serum/plasma, but samples would be anonymised. Volunteers will 
be consented for this. 
The relationship between PMR in vaccinated subjects and anti-PvDBPII antibody responses 
induced by the PvDBPII-Matrix M1 vaccine will also be assessed.  
 
Any other immunological analyses performed will be reported as not pre-specified in the trial 
protocol. Other analyses may be detailed in the VAC079 laboratory plan. Some assays may be 
duplicated at different sites. Some of these will involve analysis of frozen samples, and others 
analysis of fresh samples. 
7.2.3 Secondary Efficacy Outcome Measures 
Quantitative PCR-derived parasite multiplication rate (PMR) will be the secondary efficacy 
endpoint for assessing the durability of vaccine induced reduction in PMR. For this secondary 
efficacy analysis comparision of the PMR will be made between volunteers in Group 3 who 
received 4 vaccinations of PvDBPII-Matrix M1 and two control groups: malaria-naïve controls 
undergoing primary CHMI and volunteers undergoing their second CHMI in the parallel VAC069 
study. PMRs will be calculated from qPCR data using the same method as above for the primary 
efficacy analysis.  
 
 
 
 
 
 

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8 DESCRIPTION AND JUSTIFICATION OF STUDY DESIGN  
8.1 
Study rationale  
As outlined above, the available methods for preventing and treating infections with P. vivax in 
the field are inadequate. An effective vaccine against P. vivax would contribute significantly to 
disease control and elimination efforts. Currently, only two P. vivax candidate vaccines, both 
targeting the pre-erythrocytic stage, have progressed to Phase II studies. CHMI efficacy data for 
only one of these candidates, VMP001/AS01B, is currently published and whilst the vaccine was 
well-tolerated and immunogenic, there was no efficacy following sporozoite challenge.  
The PvDBPII vaccine candidate, formulated with the adjuvant GLA-SE, has been demonstrated to 
be safe, tolerable and immunogenic in a single blinded, dose-escalation Phase Ia study [29]. 
Progression of this promising candidate vaccine into this Phase IIa challenge study will be the first 
trial of efficacy of a P. vivax blood-stage vaccine candidate. Even if partially efficacious, through a 
reduced parasite multiplication rate, this could represent an important tool reducing the P. vivax 
global health burden and moves towards elimination. Assessment of efficacy in this study will 
therefore inform future progression of these novel vaccines into further clinical testing into 
larger, but more expensive and logistically demanding trials, in endemic settings.  
8.2  
Study Overview 
This is an open-label, single-centre first-in-human Phase I/ IIa P. vivax blood-stage CHMI trial to 
assess the safety, immunogenicity and efficacy of the candidate malaria vaccine PvDBPII 
formulated in adjuvant Matrix M1.  
Up to 24 healthy, malaria-naïve adults aged between 18 and 45 years will be recruited at the 
CCVTM, Oxford. Volunteers in Groups 1 and 2 will receive three doses of the PvDBPII-Matrix M1 
vaccine intramuscularly. Volunteers in Group 1 who complete 3 vaccinations and CHMI, will be 
invited back for a fouth vaccination at 5 months following their third vaccination and form a new 
study group - Group 3. 
Between two to four weeks post-boost, all volunteers will undergo P. vivax blood-stage CHMI, 
induced by injection of P. vivax infected erythrocytes. Volunteers will have blood taken at regular 
intervals following vaccination and in the post-CHMI period to assess the immune response to 
vaccination and subsequent challenge, as well as parasite growth dynamics and gametocytaemia. 
Close monitoring will continue until volunteers meet criteria for treatment or until 28 days after 
challenge in Group 1 or until 21 days in Groups 2 and 3, when treatment will be started 
empirically.  
Therapy will be with either Riamet or Malarone. As infection will be mediated via intravenous 
injection of blood-stage parasites, there will be no liver-stage infection and no hypnozoite 
formation, thereby eliminating need for radical cure with primaquine therapy.   
During each CHMI period, a matched number of malaria-naïve infectivity controls, recruited as 
part of the VAC069 study, will be challenged in parallel.  
All vaccinations, follow-up visits, CHMI and follow-up in the post-challenge period will be 
performed at the CCVTM in Oxford. Follow-up will be for 9 months after the final vaccination.  
8.3 
Study groups 
This study was originally planned with one study group, Group 1, comprising 10-12 volunteers, 
who were planned to receive the same doses of the PvDBPII-Matrix M1 vaccine at 4 weekly 
intervals and be challenged with malaria at the same time.   
However this trial was temporarily halted in March 2020 due to the COVID-19 pandemic. 12 
volunteers were enrolled at the start of 2020 and 11 volunteers had completed two vaccinations 

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of the PvDBPII-Matrix M1 vaccine and were due their third and final vaccination at the end of 
March 2020, which has been delayed. All currently enrolled volunteers who consent to continue 
in the study, will now complete their third and final vaccination at 12 to 18 months and undergo 
CHMI 2 to 4 weeks after their last vaccination. These volunteers will be followed up for 9 months 
from the time of their last vaccination, approximately 2 years from enrolment.  
If fewer than 8 volunteers from Group 1 return to complete all three vaccinations followed by 
CHMI, then new volunteers will be recruited into Group 2, to make up a total of 10 to 12 
volunteers between Groups 1 and 2 who complete three vaccinations and CHMI. Group 2 
volunteers will undergo 3 vaccinations at the original 4 weekly intervals, followed by CHMI 2 to 4 
weeks after the last vaccination. They will be followed up for 12 months from enrolment.  
Volunteers who have completed 3 vaccinations and CHMI in Group 1 will be invited at one of 
their post challenge follow-up visits to return for a fourth vaccination 5 months after their third 
vaccination, followed by secondary CHMI 2-4 weeks after the last vaccination and will form 
Group 3. These volunteers will be followed up for 9 months from the time of their last 
vaccination, approximately 2.5 years from initial enrolment. 
First volunteers: 
The first volunteer to receive a dose of PvDBPII-Matrix M1 will be vaccinated alone and then 
reviewed on the day following vaccination. Providing there are no safety concerns at the Day 1 
review, or recorded in the diary over the next 24 hours, another two volunteers may be 
immunised a minimum of 2 days after the first volunteer, at least one hour apart.  Following 
review of the second and third volunteers at the 7 day timepoint, an internal safety review will 
be conducted. If no safety concerns are identified, the remaining volunteers in the group will be 
vaccinated.  
8.3.1 Control groups during challenge  
Malaria-naïve controls for CHMI, will be recruited as part of the currently ongoing VAC069 study, 
a P. vivax challenge study to assess safety and feasibility of the blood-stage P. vivax challenge 
model, and to assess the effect of previous challenge on subsequent homologous re-challenge. 
Malaria-naïve controls recruited in the VAC069 study will undergo primary challenge in parallel 
with VAC079 volunteers. The data generated from the challenge of malaria-naïve controls within 
the VAC069 study will be utilised on a de-identified basis, for comparative analysis with challenge 
of VAC079 vaccinees for assessment of vaccine efficacy. 
 
Group 
Group size 
D0 
D28 
12-18 months 
14-28 days post 
3rd vaccination 
Group 1 
10-12 
PvDBPII 50ug 
Matrix M1 50ug 
PvDBPII 50ug 
Matrix M1 50ug 
PvDBPII 50ug 
Matrix M1 50ug 
CHMI 
Table 6a: Study Group 1 
 
Group 
Group size 
D0 
D28 
D56 
D70-84 
Group 2 
Up to 12 
PvDBPII 50ug 
Matrix M1 50ug 
PvDBPII 50ug 
Matrix M1 50ug 
PvDBPII 50ug 
Matrix M1 50ug 
CHMI 
Table 6b: Study Group 2. New participants will only to be recruited into Group 2, if fewer than 8 participants in 
Group 1 complete all 3 vaccinations and CHMI, to make up a total number of 10 to 12 participants between Groups 1 
and 2. 
 

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Group 
Group 
size 
D0 
D28 
12-18 
months 
14-28 days 
post 3rd 
vaccination 
5 months 
post 3rd 
vaccination 
14-28 days 
post 4th 
vaccination 
Group 3 
(subset of 
Group 1) 
Up to 
6 
PvDBPII 
50ug 
Matrix M1 
50ug  
PvDBPII 
50ug 
Matrix M1 
50ug 
PvDBPII 
50ug 
Matrix M1 
50ug 
CHMI 
PvDBPII 
50ug 
Matrix M1 
50ug 
Second 
CHMI 
Table 6c: Study Group 3. Subset of Group 1 volunteers who consent to return for fourth vaccination, followed by 
second CHMI– all activity in this table up to and including CHMI post 3rd vaccination is undertaken as part of Group 1. 
 
8.4 
Duration of study 
8.4.1 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. 
8.4.2 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 2 years from enrolment for Group 1 volunteers, 
extended by about 1 year due to the temporary trial halt. Group 2 volunteers will be followed up 
for 1 year from enrolment. Group 3 volunteers will be followed up for 2.5 years from initial 
enrolment. 
 
8.5 
Potential risks for volunteers 
8.5.1 Phlebotomy  
The maximum volume of blood drawn over the study period should not compromise these 
otherwise healthy volunteers. There may be minor bruising, local tenderness or pre-syncopal 
symptoms associated with venepuncture, which will not be documented as AEs if they occur. The 
actual total blood volume will relate to the timing of malaria diagnosis and treatment during 
challenge.  
Male regular blood donors may donate one unit (470mL) every 12 weeks and females every 18 
weeks. However, a recent multicentre study by NHS Blood and Transplant (in Oxford and 
Cambridge) compared outcomes in 50,000 regular blood donors who were randomised to 
different intervals between blood donations (as regularly as giving 470 mL every 8 weeks in male 
participant group and every 12 weeks in females) over 2 years. There were no significant 
differences observed in quality of life, physical activity, or cognitive function although there were 
more symptoms related to blood donation, in groups donating more frequently, including 
tiredness, breathlessness, feeling faint, dizziness and restless legs. Symptoms were most 
increased among male participants.  Lower mean haemoglobin and ferritin concentrations were 
also detected.  
In this study, participants will never donate 470mL of blood in one sitting; the maximum volume 
they will donate in a single visit is 96mL so we would not expect them to report a high frequency 
of symptoms at the time of or just after blood donation. They will be closely monitored at all 
times of blood donation, and haemoglobin will be checked regularly, as described in the study 
procedures. Any abnormal result will be re-checked and referred to the GP for further 
investigation and management, as deemed clinically appropriate by the Investigator.  

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The maximum total blood volume drawn over 1 year in this study will be 1752mL in those 
participants originally in Group 1 who undergo a fourth vaccination and re-challenge in Group 3. 
For male participants, this is 1303mL less than the 8-week interval group in the NHSBT study and 
for females, 285mL less than the 12-week interval group, over the 1 year period. Therefore, we 
are confident that this volume should not compromise our volunteers. However, the additional 
effect of malarial infection on potential anaemia is acknowledged. For this reason, the full blood 
count will be monitored throughout the study period. As an additional precaution, volunteers 
weighing less than 50Kg at screening will be excluded from participation. 
8.5.2 Vaccinations 
 
Foreseeable risks from vaccination, which include local and systemic reactions, are specific to each 
IMP and full details available in the respective IBs.  
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 vaccinated in a clinical area where Advanced Life Support trained physicians, equipment 
and drugs are immediately available for the management of any serious adverse reactions. 
Based on the safety data available from the 27 volunteers who received PvDBPII vaccine 
formulated in GLA-SE in the Phase Ia study [29], as well as data available from more than 1132 
individuals who have previously received vaccines formulated in Matrix M1, the below adverse 
event profile is expected following vaccination with PvDBPII-Matrix M1: 
Local adverse events such as injection site pain would be foreseeable to occur frequently. Less 
frequent local adverse events are likely to include erythema, swelling and bruising.  Local AEs are 
likely to be mild in nature and should resolve rapidly, although there is the possibility of 
moderate or severe arm pain in some cases.   
Among the solicited systemic complaints following Matrix M1-adjuvanted vaccines, headache, 
muscle pain, fatigue, chills, joint pain, wheezing, and eyelid swelling all occurred at higher rates 
among adjuvanted vaccinees, when compared to placebo vaccinees at an excess of ≥ 1% of 
subjects. 
 
8.5.3 Risk of Infection with Blood Borne Organisms  
This controlled blood-stage challenge will involve a blood transfusion, albeit of very small volume 
relative to standard blood transfusion. As for any transfusion, the risk of transmission of a blood 
borne infection cannot be completely eliminated. However, in order to minimise the risk, 
extensive screening for blood borne infections has been performed in the direct blood donor of 
the bank, as well as the initial source patient in Thailand, who provided blood for the mosquito 
infection by direct membrane feeding. For viral infections and syphilis, testing was either 
serological or by nucleic acid amplification methods, in accordance with the Joint UK Blood 
Transfusion and Tissue Transplantation Services Professional Advisory Committee guidelines. The 
source patient was screened for antibodies to W. bancrofti at the Thai field site.  
In addition, the blood bank has been subjected to direct screening and tested negative for 
bacterial contamination, sterility testing, endotoxin screen and mycoplasma specific culture. 
Plasma derived from the bank has also been screened for blood borne infections, as detailed in 
Section 5.2.6 and has tested negative for all blood-borne infection screens by PCR.  Further to 
this, as outlined in section 5.2.5, volunteers also underwent repeat serological testing for HIV, 
hepatitis B and C, syphilis, HITLV-1 and HTLV-2, 90 days after challenge was undertaken, to 
ensure that no seroconversion from a recently-acquired infection (that may have been 

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undetectable around the time of challenge and therefore undetectable in the bank) had occurred 
since the challenge period. All tests remained negative.  
For additional safety assessment of blood-stage CHMI by intravenous injection of this new source 
of cyropreserved, infected erythrocytes, repeat blood-borne viral screening (HIV, hepatitis B and 
C, plus EBV and CMV, if negative at screening) will also be performed for all recipients of the 
inoculum at 96 days post-challenge, assessing for any change in serostatus post-exposure. 
Since the blood donor has been resident in the UK or continental Europe in countries where 
Bovine Spongiform Encephalopathy (BSE) has been reported, there is a theoretical risk of 
transmission of variant Creutzfeldt Jacob Disease (vCJD) from the inoculum. This risk is reduced 
by leukodepletion, as is practised in UK blood donation, according to the Joint UK Blood 
Transfusion and Tissue Transplantation Services Professional Advisory Committee guidelines. 
Since universal leukodepletion was introduced in the UK in 1999, no cases of transmission of 
vCJD by transfusion have been recorded [105]. The risk of infection is further reduced by the very 
small size of the inoculum. The volume to be transfused for this study will be at least nine 
hundred times smaller than the amount received in typical blood transfusion. Serum from 
volunteers will be collected before and after challenge for storage.  
8.5.4 Plasmodium vivax infection   
Volunteers are likely to develop symptomatic malaria infection following CHMI. Symptoms and 
signs will include feverishness, fever, tachycardia, hypotension, chills, rigors, sweats, headache, 
anorexia, nausea, vomiting, diarrhoea, myalgia, arthralgia, low back pain, thrombocytopenia and 
lymphopenia [101]. Unmonitored and untreated, P. vivax infection can be serious (but rarely 
fatal) and, for this reason, volunteers will be followed up closely post-challenge and only enrolled 
in the study if they are deemed reliable and capable of complying with the intensive follow-up 
schedule. A very small proportion of volunteers in previous P. vivax blood-stage and sporozoite-
stage CHMI studies have temporarily required intravenous fluid therapy for nausea and vomiting 
prior to treatment [27, 93]. If the Investigator judges the volunteer to be unwell enough to 
require intravenous fluids or continuous medical input, they shall be transferred to the John 
Radcliffe Hospital under the care of the NHS Infectious Diseases team until they are well enough 
to be discharged home.  
With regards to the above, it is relevant to note that safety and parasitological data from 128 
malaria-naïve subjects participating in CHMI studies in Oxford and the Netherlands were 
analysed and compared to a report from the US Military Malaria Vaccine Program. The authors 
found that cohorts with a longer pre-patent period or a higher peak parasitaemia did not 
consistently show a higher frequency of adverse events [106]. However, these data were on P. 
falciparum challenge, and there is a lot less experience with P. vivax challenge.   
8.5.5 Medications dispensed to volunteers in course of trial  
(a) Treatment of Plasmodium vivax Infection 
P. vivax malaria infection will be treated with oral Riamet or oral Malarone. 
Riamet contains 20mg of artemether and 120mg lumefantrine per tablet. 6 doses of 4 tablets 
will be given; the first dose is followed by additional doses after 8, 24, 36, 48 and 60 hours. If 
there is a contraindication to Riamet, oral Malarone will be used (see Summary of Material 
Product Characteristics (SmPC) for side effects of, and contraindications to Riamet and 
Malarone). At a minimum two doses (the first and third doses) of Riamet will be directly 
observed. The first doses will be observed in clinic and the volunteers will not return home until 
they are seen to be tolerating the medication.  

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Riamet is generally well tolerated, but may cause some side effects. Common side effects include 
headache, dizziness, abdominal pain and loss of appetite, sleeping problems, palpitations, 
nausea, vomiting, diarrhoea, pruritus, skin rash, cough, muscle or joint pain and fatigue. 
Volunteers will be counselled that certain side effects, for example dizziness, may impact on the 
performance of skilled tasks such as driving.  
As Riamet may increase the QT interval, Riamet will not be administered to volunteers at risk for 
QT prolongation. Exclusion criteria for this study will include prolonged QT on baseline ECG, a 
history of long QT syndrome, a family history of congenital QT prolongation or sudden death, 
cardiac arrhythmias, severe heart disease, and a history of hypokalaemia or hypomagnesaemia. 
Volunteers will be advised to avoid grapefruit juice whilst taking Riamet as it can affect the 
bioavailability of the artemether [107]. Riamet is also contraindicated in volunteers using 
concomitant medications affecting the QT interval, and will not be given to these volunteers. 
Concomitant use of Riamet may decrease effectiveness of hormonal contraceptives.  
Women using hormonal contraceptives will be advised to use an effective additional method of 
contraception whilst on Riamet treatment, until the start of the next menstruation after 
treatment. 
Malarone contains 100 mg proguanil hydrochloride and 250 mg atovaquone per tablet. 4 tablets 
will be given once daily for 3 days. The first and second dose, out of the three doses of Malarone, 
will be directly observed in clinic. Prior to starting Malarone, volunteers will be screened for drug 
interactions and contraindications (including a serum β-hCG test in female volunteers).  
Malarone is generally well tolerated but may cause some side effects, most commonly headache, 
diarrhoea, nausea, vomiting, stomach pain, dizziness rash, fever, low mood, reduced appetite, 
cough or sleep disturbance.   
(b) Treatment of Symptoms Associated with Malaria 
Volunteers will be dispensed cyclizine and paracetamol for the treatment of symptoms 
associated with malaria unless there are contraindications to these medications. Cyclizine is 
generally well tolerated, however, there is a risk of an allergic reaction. Other side effects include 
skin rashes or itching, drowsiness, headache, dry mouth, nose or throat, blurred vision, 
palpitations, difficulty urinating, constipation, anxiety, insomnia or hallucinations. Rare side 
effects include hypotonia, seizures, dizziness, hypertension, paraesthesia, jaundice, hepatitis, 
confusion or dyskinesias. Participants may be dispensed an appropriate, licensed alternative anti-
emetic to cyclizine if they are unable to take cyclizine.  Paracetamol is generally well tolerated, 
however, it can cause an allergic reaction or rarely pancytopaenia. 
8.5.6 Risk of reaction to the blood sample  
The donor of the blood used in the inoculum is Blood Group O and Rhesus (Rh) and Kell (K) 
negative. People with Blood Group O negative blood are considered ‘universal donors’, as 
recipients of their blood are unlikely to develop red cell alloantibodies when given much larger 
volumes of blood than is envisaged here. For women of child-bearing potential, transfusion of Rh 
and Kell negative blood is also essential to prevent sensitisation and the future risk of haemolytic 
disease of the newborn. In the case of Kell sensitisation, anti-K may additionally lead to 
suppression of red cell production in the fetus. The maximum volume of blood to be used in this 
study (0.025mL – 0.1mL packed red cells) is much smaller than that given in transfusion of one 
unit packed red cells (470mL). The risk of a transfusion reaction or development of antibodies to 
donor red cells, which may make blood transfusion more difficult in the future, cannot be 
completely excluded but is considered highly unlikely, both because of the small volume of blood 
used and because the blood has been leukodepleted. Nevertheless, the volunteers will be 
monitored for this possibility in the period immediately after the administration of the malaria 
parasite dose. A baseline set of observations (respiratory rate, heart rate, temperature and blood 

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pressure) will be taken before the administration of the inoculum. Volunteers will be reviewed 
15 minutes (+/- 5 minutes) and 1 hour (+/- 20 minutes) after receiving the inoculum, with a 
repeat set of observations at each time point, before they are able to leave the CCVTM. The 
serum collected for storage (as described above) could be used to test for antibodies at a later 
date if deemed necessary. 
8.6 
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 malaria vaccine regimen. Volunteers will also receive information about their general 
health status. 

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9 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 on-line recruitment through our website).  
- 
As a post on a Twitter or Facebook or other similar account owned and operated by the 
Investigators’ clinical trials group. 
- 
By e-mail distribution to a group or list only with the express agreement of the network 
administrator or with equivalent authorisation.    
- 
By email distribution to individuals who have already expressed an interest in taking part 
in any clinical trial at the Oxford Vaccine Centre. 
- 
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. 
Volunteers enrolled in Group 1 will be asked at one of their post-primary CHMI follow up visits 
whether they would be interested in enrolment into Group 3 and undergo a fourth vaccination 
and repeat challenge. Those who are will be re-consented at or after their C+96 visit, prior to 
enrolment into Group 3, which will occur at the fourth vaccination visit. 
9.1 
Informed consent 
9.1.1 Informed consent 
The information sheet will be made available to the volunteer at least 24 hours prior to the 
screening visit. 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. However if the volunteer has 
undergone CHMI and not completed a course of appropriate antimalarial therapy then 
the volunteer will need to maintain contact with the Investigators for monitoring and 
treatment. 
 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 GP will be contacted to corroborate their medical history and confirm 
that the volunteer is eligible to take part in the study. 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).  
 Blood samples will be taken and any leftover samples stored indefinitely for use in other, 
ethically approved research.  

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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.  
9.1.2 Informed consent questionnaire 
If they do decide to participate, volunteers will be asked to complete a questionnaire testing 
their understanding of the trial [78]. This helps to ensure that individuals understand the trial 
sufficiently to give informed consent. Volunteers who answer all questions in the questionnaire 
correctly, will be asked to sign the consent form, a photocopy of this signed consent form will be 
given to them to take away and the original signed consent form will be stored in the CRF. The 
form will also be signed and dated by the Investigator. Volunteers who fail to answer all 
questions correctly on their first attempt will be allowed to re-take the questionnaire one more 
time, following further discussion with the Investigator. Provided they subsequently answer all 
questions in the quiz correctly at the second attempt they may then complete the consent form 
and be screened for the trial. Failure to answer all the questions correctly at the second attempt 
will exclude them from participation. 
9.2  
Inclusion and exclusion criteria  
9.2.1 Inclusion criteria 
The volunteer must satisfy all the following criteria to be eligible for the study: 
 Healthy adult aged 18 to 45 years.  
 Red blood cells positive for the Duffy antigen/chemokine receptor (DARC). 
 Normal serum levels of Glucose-6-phosphate dehydrogenase (G6PD). 
 Negative haemoglobinopathy screen 
 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 
General Practitioner. 
 Women only: Must practice continuous highly effective contraception* for the duration 
of the study 
 Agreement to permanently refrain from blood donation 
 Written informed consent to participate in the trial. 
 Reachable (24/7) by mobile phone during the period between CHMI and completion of all 
antimalarial treatment.  
 Willing to take a curative anti-malarial regimen following CHMI.  
 Willing to reside in Oxford for the post-challenge period, until antimalarials have been 
completed. 
 Answer all questions on the informed consent quiz correctly at first or second attempt  
* Female volunteers are required to use a highly effective form of contraception during the 
course of the study as malaria challenge could pose a serious risk to both maternal health and 
the unborn foetus. 
Highly effective forms of contraception for female volunteers include: 
 Hormonal methods, as listed below (an additional form of contraception will be required 
where artemether/lumefantrine is used as the anti-malarial treatment post CHMI, as this 
can interfere with the efficacy of hormonal contraception): 
 Established use of oral, intravaginal or transdermal combined hormonal methods 
of contraception 
 Established use of oral, injected or implanted progesterone-only hormonal 
contraception associated with inhibition of ovulation (progesterone only 

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hormonal contraception where inhibition of ovulation is not the primary mode of 
action is not considered highly effective) 
 Placement of an intrauterine device (IUD) or intrauterine system (IUS). 
 Bilateral tubal occlusion or total hysterectomy. 
 Male sterilisation, if the vasectomised partner is the sole partner for the subject. 
 True abstinence (as defined as refraining from heterosexual intercourse), when this is in 
line with the preferred and usual lifestyle of the subject (periodic abstinence and 
withdrawal are not acceptable methods of contraception).  
Barrier methods of contraception (e.g. condom or occlusive cap with spermicide) may be used in 
combination with any of the above forms of contraceptive but are not considered highly 
effective methods if used as the sole method of contraception. However, where 
artemether/lumefantrine (Riamet) is used as the anti-malarial treatment post-diagnosis 
following CHMI, female participants using hormonal contraceptives must agree to using an 
additional method of contraception (e.g. barrier methods) from the time of commencing the 
artemether/lumefantrine (Riamet) drug course until the next menstrual period. This is to 
minimise any risk of possible reduced effectiveness of hormonal contraceptives during 
concomitant antimalarial therapy, due to the weak induction of cytochrome P450 enzymes 
resulting from treatment with artemether. 
9.2.2 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 systemic antibiotics with known antimalarial activity within 30 days of CHMI (e.g. 
trimethoprim-sulfamethoxazole, doxycycline, tetracycline, clindamycin, erythromycin, 
fluoroquinolones and azithromycin). 
 Use of anti-malarials within 30 days of CHMI 
 Weight less than 50kg, as measured at the screening visit 
 Receipt of immunoglobulins within the three months prior to planned administration of 
the vaccine candidate.  
 Receipt of blood products (e.g., blood transfusion) at any time in the past. 
 Peripheral venous access unlikely to allow twice daily blood testing (as determined by the 
Investigator). 
 Receipt of an investigational product in the 30 days preceding enrolment, or planned 
receipt during the study period. 
 Receipt of any vaccine in the 30 days preceding enrolment, or planned receipt of any 
other vaccine within 30 days preceding or following each study vaccination, with the 
exception of licensed COVID-19 vaccines, which should not be received between 14 days 
before to 7 days after any study vaccination 
 Planned receipt of a COVID-19 vaccine between 2 weeks before the day of CHMI until 
completion of antimalarial treatment 
 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 or the P. vivax parasite as assessed by the Investigator.  
 Any confirmed or suspected immunosuppressive or immunodeficient state, including HIV 
infection; asplenia; recurrent, severe infections and chronic (more than 14 days) 

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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 e.g. egg products, Kathon 
 History of allergic disease or reactions likely to be exacerbated by malaria infection. 
 History of clinically significant contact dermatitis 
 Any history of anaphylaxis in reaction to vaccinations 
 Pregnancy, lactation or intention to become pregnant during the study. 
 Use of medications known to cause prolongation of the QT interval and existing 
contraindication to the use of Malarone. 
 Use of medications known to have a potentially clinically significant interaction with 
Riamet and Malarone. 
 Any clinical condition known to prolong the QT interval. 
 History of cardiac arrhythmia, including clinically relevant bradycardia. 
 Disturbances of electrolyte balance, e.g. hypokalaemia or hypomagnesaemia. 
 Family history of congenital QT prolongation or sudden death. 
 Contraindications to the use of both of the proposed anti-malarial medications; Riamet 
Malarone. 
 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 or (unless has taken 
part in a prior hepatitis C vaccine study with confirmed negative HCV antibodies prior to 
participation in that study, and negative HCV RNA PCR at screening for this study). 
 Positive family history in both 1st AND 2nd degree relatives < 50 years old for cardiac 
disease. 
 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 and 
safety to participate 
9.2.3 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). 
9.2.4 Vaccination and re-vaccination exclusion criteria 
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, 

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the subject may be vaccinated at a later date, or withdrawn at the discretion of the Investigator. 
The subject must be followed until resolution of the event as with any adverse event:  
 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 1st 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.  
The following adverse events associated with vaccine immunisation constitute absolute 
contraindications to further administration of vaccine. If any of these events occur during the 
study, the subject must be withdrawn and followed until resolution of the event, as with any 
adverse event:  
 Anaphylactic reaction following administration of vaccine  
 Pregnancy  
9.2.5 Exclusion criteria on day of CHMI 
The following constitute absolute contraindications to CHMI: 
 Acute disease, defined as moderate or severe illness with or without fever 
 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. 
 Pregnancy 
9.2.6 Concomitant medications 
As set out by the exclusion criteria, volunteers may not enter the study if they have received or 
there is planned receipt of any other vaccine within 30 days of each study vaccination, with the 
exception of licensed COVID-19 vaccines, which should not be received between 14 days before 
to 7 days after any study vaccinations. In addition volunteers who are due to receive a COVID-19 
vaccine between 2 weeks before the day of CHMI until expected completion of antimalarial 
treatment (around 2 to 3 weeks after day of challenge based on experience in previous P. vivax 
CHMI studies to date) will not be enrolled in this trial. If a volunteer receives an appointment for 
COVID-19 vaccination after they have undergone malaria challenge but before reaching malaria 
diagnosis criteria, they will be advised to delay their COVID-19 vaccination until after reaching 
malaria diagnostic criteria and completion of antimalarial treatment. If a participant does not 
wish to delay their COVID-19 vaccination, they will be treated with antimalarial treatment at that 
timepoint and withdrawn from the study but will be encouraged to complete follow-up for 
safety. 
Volunteers may also not enter the study if they have any investigational medicinal product within 
the 30 days prior to enrolment, or have received or plan to receive any immunosuppressant 
medication 6 months prior to enrolment or at any time during the study period (inhaled and 
topical steroids are permitted). Volunteers will also be excluded from partaking in CHMI, if there 
is any concomitant use of systemic antibiotics with known anti-malarial activity or anti-malarial 
medication within 30 days of CHMI, however, follow-up will continue until approximately 6 
months (C+96), according to the schedule as long as valid informed consent remains in place. 

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Following CHMI, should a volunteer require treatment with systemic antibiotics at any time prior 
to reaching criteria for diagnosis, (as described in Section 11.4), the volunteer will be withdrawn 
from the study and treatment with an appropriate, curative course of anti-malarial therapy will 
be commenced immediately. Follow-up for safety will be continued until 96 days after challenge 
(C+96) in all cases except where there is complete withdrawal of consent.  
 
9.3 
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 well-being, 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 CRF. If a volunteer withdraws 
after having completed a course of antimalarials, as much continued safety data collection as 
possible, including procedures such as safety bloods, will be continued, with agreement of the 
volunteer. 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 after CHMI but before 
reaching the criterion for malaria diagnosis, a complete, appropriate, curative course of anti-
malarial therapy must be completed. The importance of this will be emphasised to volunteers at 
screening. If a volunteer refuses to take anti-malarial therapy after malaria diagnosis, a rapid 
assessment of mental state and capacity will be undertaken, with the involvement of NHS 
psychiatric and infectious diseases services. If necessary the volunteer may be detained under 
section 4 of the UK Mental Health Act until this assessment can be carried out. 
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 destroyed. Volunteers are free to request that their blood samples be destroyed anytime 
during or after the study.  
In all cases of subject withdrawal, excepting those of complete consent withdrawal, safety data 
collection will continue until 90 days after challenge (C+90), if subjects have undergone CHMI. 
9.4 
Pregnancy 
Should a volunteer become pregnant during the trial, she will be treated with antimalarials 
immediately and will be withdrawn from the study. We will not routinely perform venepuncture 
on such volunteers, other than blood films to check that the parasitaemia has been cleared by 
the antimalarials. Additionally, if a volunteer who had been negative for CMV IgG antibodies at 

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screening should become pregnant during the study, given the theoretical risk of CMV infection 
from exposure to the inoculum, venepuncture would also be performed to check CMV 
serostatus. With the volunteer’s permission she shall be followed up until pregnancy outcome. 
The management of any volunteer found to be pregnant at any time after challenge up to the 
point of malaria treatment will be discussed with the on-call infectious diseases consultant at the 
Oxford University Hospitals NHS Foundation Trust, including advice on antimalarial drug choice. 
Should a volunteer become pregnant after receiving antimalarial treatment (but prior to the end 
of the study), they shall be withdrawn from the study as soon as we have confirmed that their 
parasitaemia has cleared. With the volunteer’s permission, she shall then be followed up until 
pregnancy outcome. 
 
 
 
 

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10 CONTROLLED BLOOD-STAGE MALARIA INFECTION INOCULUM 
10.1 
Preparation of the inoculum 
Thawing and washing of the inoculum will be done with commercial solutions for human use and 
with disposable syringes and needles according to standard operating procedures used in 
previous studies at Oxford [108, 109]. Work will be carried out in the derogated category III 
laboratory at the Jenner Institute, Old Road Campus Research Building (ORCRB) or New 
Biochemistry Building. Sample manipulations will be performed within a safety cabinet that has 
been fumigated, sterilised and dedicated for this purpose.  
All procedures relating to thawing of the blood inoculum and preparation of the syringes, will be 
performed in accordance with local standard operating procedures.   
10.2 
Administration of the inoculum 
The inoculation will take place in Oxford at the CCVTM and according to local Standard Operating 
Procedure. In brief, the inoculum will be administered by intravenous injection into an indwelling 
intravenous cannula. Parasitised red blood cells will be reconstituted in 0.9% saline, to a total 
volume of 5mL. The reconstituted inoculum will be injected via an indwelling cannula, followed 
by a saline flush. Subjects will be observed for a minimum of one hour before discharge. All 
volunteers will receive the inoculum within a maximum of 4 hours of removal from frozen 
storage. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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11 TREATMENT OF TRIAL VOLUNTEERS 
This section describes the clinical procedures for evaluating study participants and follow-up 
after CHMI.  
11.1 
Trial sites 
Volunteers will be recruited and undergo all visits at the CCVTM, Oxford, this is inclusive of 
screening, vaccinations and follow-up visits, CHMI and post-CHMI follow-up.  Following CHMI 
until completion of anti-malarial therapy, all volunteers will reside in Oxford and the surrounding 
areas. After completion of treatment, volunteers may reside outside the Oxford area but will 
attend for later follow-up visits at the CCVTM, Oxford.  
11.2 
Study procedures 
Procedures will be performed at the time points indicated in the schedule of procedures (tables 
7-15). Additional procedures or laboratory tests may be performed, at the discretion of the 
Investigators if clinically necessary (e.g. urine microscopy in the event of positive urinalysis).  
11.2.1 Observations 
Pulse, blood pressure and temperature will be measured at the time points indicated in the 
schedule of procedures (Tables 7-15). Weight and height will be measured at screening and 
weight re-measured at the C-2 visit.  
11.2.2 Blood tests 
Blood will be drawn at the time points indicated in the schedule of procedures (Tables 7-15) and 
the following laboratory assays performed: 
At Oxford University Hospitals NHS Foundation Trust, using NHS standard procedures 
 Haematology; Full blood count, to include reticulocyte count at C-2 visit. Duffy 
antigen/chemokine receptor (DARC) gene positivity (either or both Fya/Fyb positive), and 
glucose-6-phosphate dehydrogenase (G6PD) levels and a haemoglobinopathy screen will 
be performed at screening only.  
 Biochemistry; Sodium, potassium, urea, creatinine, albumin, liver function tests, C 
reactive protein (CRP), magnesium and calcium, however magnesium and calcium will 
only be measured at screening. Serum will also be tested for beta-human chorionic 
gonadotrophin (β-HCG) at C-2, C+7, C+14, C+21 and C+28/day of malaria diagnosis. C-
reactive protein (CRP) will be measured on days 0, 1, 3, 7, 56, 57, 59 and 63 as an 
exploratory measure only (CRP results will not be included in safety analysis). 
 Diagnostic serology; HBsAg, HCV antibodies, HIV antibodies, CMV antibodies and EBV 
antibodies will be performed at screening and at day 96 post CHMI (C+96). EBV and CMV 
serology will only be repeated at day 96 post CHMI if they were negative at screening 
 Immunology; Human Leukocyte Antigen (HLA) typing at enrolment (D0). 
At the University of Oxford research laboratories (at the Jenner Institute and Department of 
Biochemistry) 
 Diagnostic Tests: PCR for P. vivax DNA and gametocytes. 
 Immunology: Immunological responses to primary P. vivax infection will be assessed by a 
variety of established and exploratory assays. This may include: 
o Antibodies and B cell responses to the P. vivax Duffy-binding protein (PvDBP) – 
total IgG, isotypes and avidity; 
o T cell responses to PvDBP by ex-vivo ELISpot and flow cytometry assays; 
o Functional antibody assays against parasites in vitro;  

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o Antibody responses to whole parasites and other P. vivax antigens including red 
blood cell invasion ligands as well as the use of arrays; 
o Serum cytokine analysis. 
o B cells, plasma and/or serum may be analysed and used to produce human 
monoclonal antibodies against P. vivax malaria. 
 Genetic tests: DNA analysis of genetic polymorphisms potentially relevant to 
immunological responses and gene expression studies of the host and the parasite 
amongst others may be performed at the discretion of the Investigators. 
 COVID-19 serology: Antibody to SARS-COV-2 will be measured retrospectively at 
screening, re-screening and at the final study visit following the final CHMI for each 
participant (+/- at interim time points) for restrospective exploratory analysis of any 
association between vaccine immunogenicity and COVID-19 serostatus 
 Samples may be sent to collaborating laboratories for other immunological assays or 
studies of malaria. 
 Samples may also be sent to collaborating laboratories within and outside the UK for 
immunomonitoring and/or harmonisation of key immunological assays and/or studies of 
malaria. 
Immunological assays will be conducted according to the procedures established in the test 
laboratories. Specific details of which immunological tests will be performed at each time-point 
are detailed in the separate immuno-monitoring plan – this will also clearly state which assays 
are conducted to meet the trial’s specified objectives and which are exploratory. With the 
volunteers’ informed consent, any leftover cells, plasma, serum, whole blood (or their purified 
components) will be registered under University of Oxford HTA licence 12217 and stored 
indefinitely for future analysis of human malaria infection. This may include human DNA and RNA 
analysis. Volunteers can, however, request for their remaining blood samples to be destroyed at 
any time. 
11.2.3  Urinalysis 
Urine will be tested for the presence of clinically significant proteinuria, glycosuria or haematuria 
(as defined in Appendix A) at the screening and re-screening visits. For female volunteers only, 
urine will be tested for beta-human chorionic gonadotrophin (β-HCG) at screening, immediately 
prior to each vaccination and on the day of diagnosis, immediately prior to initiation of 
treatment.  
11.2.4  Electrocardiogram 
 An electrocardiogram will be performed at the initial screening visit for detection of any 
conduction abnormalities, including long QT syndrome. 
11.2.5 Vaccinations  
Before each vaccination, the on-going eligibility of the volunteer will be reviewed. The vaccine 
will be administered as described above in section 6.4. 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.  

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Although there is extensive safety data for the use of Matrix M1 in humans and this PvDBPII 
protein has been shown to be safe when administered with adjuvant GLA-SE in a phase 1a trial 
[29], this will be the first human trial of the combination of PvDBPII and Matrix M1. 
Because of this, in the initial round of vaccination with PvDBPII-Matrix M1, the first volunteer will 
be enrolled and vaccinated alone and reviewed on the following day. Providing there are no 
safety concerns at the day 1 review, or recorded in the diary over the next 24 hours, another two 
volunteers may be immunised, a minimum of 2 days after the first volunteer and at least one 
hour apart. After a minimum of 7 days following the enrollment of the second and third 
volunteers, and following completion of day 7 clinic visits for all three volunteers, an internal 
review of safety data for the first three volunteers will be conducted. Provided there are no 
safety concerns at internal safety review, and pending CI approval, the remaining volunteers in 
Group 1 may be enrolled and vaccinated. 
The second, third and fourth doses (in Group 3 only) of the PvDBPII-Matrix M1 vaccine will be 
administered to all volunteers on the same day, or within the specified time windows (Table 6). 
11.2.6 Management of post-vaccination fevers 
Post-vaccination fevers may cause diagnostic uncertainty and concurrent COVID-19 infection 
could be a possibility. Results from this trial to date show that post-vaccination fever does occur 
in a proportion of participants, following administration of the PvDBPII-Matrix M1 vaccine. After 
the first set of vaccinations, none of the 12 participants developed a fever; however following 
the 2nd set of vaccinations, 3 out of 11 participants became febrile and therefore fevers will be 
expected to occur in some participants following their 3rd vaccination. 
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 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 
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 Personal Protective 
Equipment (PPE). 

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If a participant tests negative for COVID-19, they can stop self-isolating and continue with study 
visits as planned. 
11.2.7 Diary card 
Following each vaccination, volunteers will be asked to complete a diary card which may be in 
paper or electronic form. Volunteers will be asked about foreseeable local and systemic AE’s for 
7 days (solicited AEs). After this, volunteers will be asked to record any AE daily, for 28 days 
(unsolicited AEs). Diary cards will be reviewed with volunteers at each post-vaccination clinic 
visit.  
11.2.8 SARS-COV-2 PCR swab 
Whilst the COVID-19 pandemic is ongoing, a combined nasopharyngeal and throat swab will be 
tested for SARS-COV-2 by PCR two days before malaria challenge (C-2) in all volunteers. 
Asymptomatic COVID-19 infection will be a contra-indication to proceeding to CHMI. All 
participants who undergo CHMI will also have a COVID-19 swab taken on the day of malaria 
diagnosis when malaria treatment is commenced. Any participant who develops symptoms, 
which may be consistent with COVID-19 disease after undergoing malaria challenge and prior to 
commencing malaria treatment, may also undergo testing for COVID-19 by PCR swab. Swabs 
from symptomatic participants will be taken by the study team in-house but in a physically 
separate location from the main clinic area with separate side access, with full infection control 
precautions (as per the PPE SOP). 
If a participant develops symptoms consistent with COVID-19 after completing malaria 
treatment, they will be referred to local community testing for COVID-19.  
 
11.3 
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 Tables 7-15. Each review is assigned a 
time point and a window period within which the review will be conducted. 
11.3.1 Screening visits  
Where possible, and where consent to contact by telephone has been provided by email, 
individuals may be phoned by a member of the clinical team to discuss the study, prior to the 
screening visit. This pre-screening telephone call will provide an opportunity for interested 
individuals to discuss the study, eligibility criteria and study requirements. Where a pre-screening 
call takes place, if individuals continue to express an interest in taking part in the trial following 
the pre-screening call, 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 
procedures (Table 7) will be undertaken, including testing for Duffy antigen/chemokine 
receptor (DARC) positivity, G6PD levels and haemoglobinopathy screen.  
The subject’s general practitioner will be contacted with the written permission of the subject 
after screening to ascertain any significant medical history and as notification that the subject 
has volunteered for the study. 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). 
Abnormal clinical findings from the medical history, physical examination, urine or blood tests at 
any point in the study will be assessed according to the scales in Appendix A and site specific 

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laboratory adverse event grading tables kept in the trial master file (ref: SOP VC027). If a test is 
deemed clinically significant it may be repeated to ensure it is not a single occurrence. If an 
abnormal finding is deemed to be clinically significant, the volunteer will be informed and 
appropriate medical care arranged with the permission of the volunteer. Exclusion of the 
volunteer from enrolling in the trial or withdrawal of a volunteer from the trial will be at the 
discretion of the Investigator. 
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, medical examination findings or urinalysis, haematology or biochemistry blood tests 
only. Other blood tests and ECG will not be repeated routinely. In addition, where a reply from 
the subject’s GP has already been received, the GP will not be re-contacted unless there is any 
need to obtain further information where appropriate (e.g. regarding a new medical event or 
finding) and at the discretion of the Investigator.  
Participants who are currently enrolled in Group 1 and consent to continue in the study, will 
attend a re-screening visit prior to their third vaccination due to the long duration of the trial 
halt.  
Participants in Group 1 who would like to enrol into Group 3, will be re-consented prior to 
enrolment into Group 3. However they will not require a re-screening visit because their last visit 
C+96 in Group 1 will be only be about 1 month from enrolment into Group 3. 
11.3.2 Enrolment and first vaccination with PvDBPII-Matrix M1 
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 female volunteers and venepuncture for immunology and 
safety bloods will be undertaken according to Table 7. The inclusion and exclusion criteria for the 
study will be reviewed. If the volunteer remains eligible and there are no contraindications to 
vaccination, the PvDBPII-Matrix M1 vaccine will be administered as described in section 6.4. 
11.3.3 Reviews post-vaccination on days 1, 3, 7 and 14  
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 7). Blood will also be taken for exploratory immunology 
analysis.  
Volunteers will be asked to record all AEs via the diary card and upon attending clinic visit on 
days 1, 3 and 7, during which 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 about any new symptoms from day 7 until day 28, on a daily basis via the diary but will 
not be asked directly about foreseeable symptoms. 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 7.  
11.3.4 2nd Vaccination with PvDBPII-Matrix M1 (day 28) 
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 female volunteers and venepuncture for immunology and 
safety bloods will be undertaken according to Table 7. The inclusion and exclusion criteria for the 

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study will be reviewed. If the volunteer remains eligible and there are no contraindications to 
vaccination, the PvDBPII-Matrix M1 vaccine will be administered intramuscularly in the opposite 
arm to that used for the first vaccination and as described in sections 6.4. 
11.3.5 Reviews post-vaccination on days 29, 31, 35 and 42 
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 7). Blood will also be taken for exploratory immunology analysis.  
Volunteers will be asked to record all AEs via the diary card and upon attending clinic visit on 
days 29, 31 and 35, during which 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 about any new symptoms from day 35 until day 42, on a daily basis via the diary but will 
not be asked directly about foreseeable symptoms. 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 7. 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 15.5.  
11.3.6 3rd Vaccination with PvDBPII-Matrix M1 (day 56 or approximately day 365 for Group 1) 
For Group 2, this visit will include a follow up visit for the second vaccination and administration 
of the third vaccine. For Group 1 the 3rd vaccination will occur between 12 to 18 months after the 
second vaccination depending on the duration of the trial halt. 
Any new medical issues or symptoms that have arisen will be assessed. Physical observations, 
urinary β HCG test in female volunteers and venepuncture for immunology and safety bloods will 
be undertaken according to Table 7. The inclusion and exclusion criteria for the study will be 
reviewed. If the volunteer remains eligible and there are no contraindications to vaccination, the 
PvDBPII-Matrix M1 vaccine will be administered intramuscularly in the opposite arm to that used 
for the second vaccination and as described in sections 6.4. 
11.3.7 Reviews post-vaccination on days 57, 59, 63, and 70 for Group 2 (or 1, 3, 7 and 14 days 
post 3rd vaccination for Group 1) 
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 7). 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, 59 and 63 for 
Group 2 (or 1, 3 and 7 days post 3rd vaccination for Group 1), during which 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 70 until day 84 for Group 2 (or from 14 to 28 days after the 3rd vaccination for Group 1), 
on a daily basis via the diary but will not be asked directly about foreseeable symptoms. 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 7.  
11.3.8 Group 3 only – 4th Vaccination with PvDBPII-Matrix M1 and reviews on 1, 3, 7 and 14 
days post-vaccination 
For Group 3 only the 4th vaccination will occur approximately 5 months after their third 
vaccination and about 1 month after the volunteers have completed their C+96 visit following 

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the first CHMI (as part of Group 1). Participants will be re-consented for enrolment into Group 3, 
if not already re-consented at a prior visit. Enrolment into Group 3 will occur at the point of the 
fourth vaccination. 
Any new medical issues or symptoms that have arisen will be assessed. Physical observations, 
urinary β HCG test in female volunteers and venepuncture for immunology and safety bloods will 
be undertaken according to Table 12. The inclusion and exclusion criteria for the study will be 
reviewed. If the volunteer remains eligible and there are no contraindications to vaccination, the 
PvDBPII-Matrix M1 vaccine will be administered intramuscularly. 
Following the 4th vaccination, volunteers will be asked to record all AEs via the diary card and 
upon attending clinic visit on days 1, 3 and 7 after vaccination, during which 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 about any new symptoms from day 7 until 
day 28 after vaccination, on a daily basis via the diary but will not be asked directly about 
foreseeable symptoms. 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 12. 
 
11.3.9 Two days before CHMI (C-2 (±1))  
This visit will include a follow up visit for the final vaccination, as well as assessment for eligibility 
for CHMI. Any new medical issues or symptoms that have arisen will be assessed. Physical 
observations, serum β-HCG test in female volunteers and venepuncture for immunology, malaria 
PCR and safety bloods will be undertaken according to Table 8 for Group 1 and Table 13 for 
Groups 2 and 3. A combined nasopharyngeal and throat swab for SARS-COV-2 PCR will also be 
taken. Results of safety bloods and the SARS-COV-2 PCR test taken at this visit must be available 
and reviewed prior to challenge. Any participant with a positive SARS-COV-2 PCR swab will not 
proceed to CHMI. 
11.3.10 
Day of CHMI (C0)  
CHMI will be administered, according local standard operating procedure (SOP), this will include 
 Interim history and examination of the injection site and any body systems felt to be 
necessary by the Investigator and verify continuing eligibility / contraindications. 
 Baseline physical observations (respiratory rate, pulse, blood pressure and temperature) 
 Intravenous access via cannulation in forearm vein, flushed with normal saline. 
 For each volunteer the inoculum must be injected within 4 hours of the inoculum being 
thawed, followed by a further normal saline flush. 
 Physical observations will be repeated at 15 minutes (+/- 5 minutes)  post-administration 
of the inoculum and again at 1 hour (+/- 20 minutes), in order to assess for immediate 
adverse reactions, with regular visual observation throughout. If the volunteer should 
show signs or symptoms of a transfusion reaction, observations will be repeated and the 
volunteer will be medically reviewed by a physician. 
 Volunteers will be observed for at least 60 minutes, however, this period may be 
extended if there are any clinical concerns.  
 If there have been no symptoms or signs indicative of a transfusion reaction, the cannula 
will be removed after 1 hour  
Before leaving CCVTM, volunteers will be informed about the unlikely possibility of a delayed 
transfusion reaction and will be provided with the 24 hour emergency mobile telephone number, 
to enable them to contact a physician in the event of concern. 

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A Medic-Alert type card will be issued to each volunteer. This will contain the study physician 
contact details and a request that the research team be contacted immediately in the event of 
illness/accident. Each subject will also be issued with an accurate oral thermometer. If the 
subject does not have their own mobile telephone they will be issued with one for the duration 
of the study, and counselled about the importance of keeping it switched on or checking the 
messages regularly. In addition full contact details for each subject will be documented, including 
home and work addresses, home and work landline telephone numbers where available and 
next-of-kin address and telephone numbers. Mobile telephone numbers will be verified prior to 
challenge to ensure the volunteers are easily contactable. Subjects must also provide the 
Investigators with the name and 24 hour telephone number of a close friend, relative or 
housemate who will be kept informed of their whereabouts for the duration of the study.  
Volunteers must be resident in Oxford / the surrounding area until they have had a least one 
negative thick blood film, and (/or, if not diagnosed on the basis of blood-film) two consecutive 
qPCR results with substantial reduction in genome copies/mL after commencing antimalarial 
treatment. If volunteers do not reach malaria diagnosis, they must remain in the Oxford area 
until they have completed anti-malarial treatment. 
Volunteers will be counselled that should they fail to return for treatment having been infected 
with P. vivax malaria they could become very unwell and potentially die. They will be informed 
that should they fail to attend a scheduled clinic visit post-challenge, their nominated contact, 
next of kin and the police may be informed and a search started.  
While not frequent practice, accommodation in Oxford for the post-challenge follow-up period 
can be arranged on a case-by-case basis for eligible volunteers who do not reside in the Oxford 
area. The costs of this will be covered by the research group.  
 
11.3.11 
Days 1-6 post-CHMI (C+1 – C+6) 
In the VAC069 study, three doses of inoculum were assessed for feasibility of infection. Two 
volunteers received the equivalent of a whole vial of infected erythrocytes, two further 
volunteers received a 1:5 dilution and two received a 1:20 dilution. All volunteers were 
completely asymptomatic in the first 6 days. Patent parasitaemia, as determined by qPCR, was 
first detected after day 7, in a volunteer who had received the highest dose. For the same 
volunteer, microscopic patency was at day 12 and diagnosis was made on the same day (C+12.5). 
For the remaining volunteers, qPCR became positive after day 8 and microscopic patency was no 
earlier than day 14. Diagnostic criteria for all remaining volunteers were reached between days 
15 and 16. Therefore, a mid-range dose of 1:10 dilution is to be used in this study. qPCR 
parasitaemia patency is not expected until at least 7 days after challenge and so volunteers will 
not need to be reviewed in clinic until day 7.  However, they will be phoned daily by the clinic 
team from day 1 to 6 to make sure they are well and contactable, and will be able to contact the 
study physicians on the 24 hour emergency telephone number if needed.  
11.3.12 
Days 7-28 post-CHMI (C+7 – C+28) – Group 1 
Volunteers will be reviewed in clinic once in the morning of day 7 after challenge until day 9, 
then twice daily from day 10 until diagnosed, or if diagnosis criteria is not met, until day 28 post-
challenge inclusive (C+10 to C+28). However, from day 21 after challenge, visits may be reduced 
to once daily at the discretion of the Investigator (as agreed by the Chief Investigator) e.g., if 
qPCR is very low or undetectable, and volunteer is completely asymptomatic (qPCR results may 
be unblinded for this purpose). 
At each follow-up visit: 

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 Physical observations will be performed. Venepuncture will be performed as per schedule 
of attendance (Table 8, 9, 10 and 11), including blood for assessing parasite growth rate, 
immune response and level of gametocytaemia.  
 Volunteers will be questioned as to whether they have: 
o Experienced any of the foreseeable symptoms of malaria. 
o Experienced any other symptoms.  
o Taken any medications including over the counter medications. 
Full physical examination will be performed if deemed necessary by the Investigators. Subjects 
will be encouraged to contact one of the Investigators on the 24 hour emergency mobile 
telephone number if they develop symptoms of malaria or concerning AEs between the regular 
clinic reviews. The severity of symptoms will be assessed using grading criteria summarized in 
Section 12 below. 
Diagnosis will be made according to criteria described in section 11.4 below. If volunteers reach 
day 28 post-challenge (C+28) without reaching diagnostic criteria, they shall be started on anti-
malarial treatment empirically. Subjects will be reviewed in clinic approximately 24 and 48 hours 
after starting anti-malarial therapy as per Table 8 below. 
If a volunteer is unwell and unable to attend the CCVTM for a visit they will be reviewed on the 
phone and visited at home by one of the Investigators if necessary. Such visits will be conducted 
according to SOP VC022: Lone Working in the Community.  
11.3.13 
Days 7-21 post-CHMI (C+7 – C+21) – Groups 2 and Group 3 
Following review of the results from the first CHMI in Group 1 volunteers, the number of visits in 
the post challenge period has been reduced to minimise the number of bleeds and burden on 
volunteers. During the first CHMI, five volunteers in Group 1 continued with twice daily visits 
from C+10 until day of diagnosis, which occurred between C+19 to C+25 for these volunteers. 
One volunteer in Group 1 reduced to once a day visits from C+21 to C+25 before being diagnosed 
at C+27.  
Review of the qPCR results and parasite multiplication rate (PMR) modelling from the CHMI in 
Group 1 and the corresponding VAC069 control volunteers has shown that sufficient data can be 
obtained for accurate PMR modelling if bleeds for qPCR continue once a day until qPCR reaches 
≥1000 genome copies/ml, instead of starting twice a day visits from C+10 onwards. Once qPCR 
levels reach ≥1000 genome copies/ml, then visits will increase to twice a day in order to not 
delay diagnosis as qPCR may reach the diagnostic threshold of ≥5000 genome copies/ml with 
malaria symptoms within about half a day. PMR modelling is also minimally affected if, in 
addition to reducing the frequency of bleeds to once daily, participants are treated at C+21, 
instead of C+28, even if they have not reached malaria diagnostic criteria at that timepoint.  
Group 2 and 3 volunteers will therefore be reviewed in clinic once a day in the mornings from 
day 7 after challenge until day 9. From day 10 after challenge onwards visits continue once a day 
until the participant’s qPCR count reaches ≥1000 genome copies/ml, at which time visits will 
increase to twice a day until diagnosed, or if diagnosis criteria are not met, until day 21 post-
challenge inclusive (C+21).  
At each follow-up visit: 
 Physical observations will be performed. Venepuncture will be performed as per schedule 
of attendance (Table 13, 14 and 15), including blood for assessing parasite growth rate, 
immune response and level of gametocytaemia.  
 Volunteers will be questioned as to whether they have: 
o Experienced any of the foreseeable symptoms of malaria. 
o Experienced any other symptoms.  

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o Taken any medications including over the counter medications. 
Full physical examination will be performed if deemed necessary by the Investigators. 
Participants will be encouraged to contact one of the Investigators on the 24 hour emergency 
mobile telephone number if they develop symptoms of malaria or concerning AEs between the 
regular clinic reviews. The severity of symptoms will be assessed using grading criteria 
summarized in Section 12 below. 
Diagnosis will be made according to criteria described in section 11.4 below. If volunteers reach 
day 21 post-challenge (C+21) without meeting diagnostic criteria, they shall be started on anti-
malarial treatment empirically. Subjects will be reviewed in clinic approximately 24 and 72 hours 
after starting anti-malarial therapy as per Table 15 below. 
If a volunteer is unwell and unable to attend the CCVTM for a visit they will be reviewed on the 
phone by a clinician. If inperson assessment is considered necessary, the participant will be 
referred to appropriate NHS services.  
 
11.4 
Algorithm for initiation of treatment  
It is now well established that nucleic acid amplification-based diagnostics have greater 
sensitivity than microscopic methods for detecting malaria infection both in clinical infection and 
in the P. falciparum challenge model. In the latter, qPCR targeting the 18S ribosome DNA target 
has been used as the diagnostic tool, reliably detecting parasitaemia earlier than thick film blood 
smear.  
The qPCR assay that we have used in numerous Oxford-led CHMI P. falciparum challenge studies 
is now qualified and highly sensitive, reliably detecting parasitaemias as low as 20 parasites per 
mL of blood (0.02 parasites per μL), long before clinical symptoms manifest. The qPCR assay has 
also performed well in an international External Quality Assurance (EQA) exercise [110]  
It is also important to note that other trial centres in the USA routinely use PCR as a diagnostic 
tool in CHMI studies. NCT020-15091 was a multi-institution, Phase I, open-label, dose-escalation 
trial with CHMI, designed to assess the safety, immunogenicity, and protective efficacy of PfSPZ 
vaccine and this used PCR as the primary diagnostic [111]. 
For P. vivax, we also use a qPCR assay targeting the pan-plasmodium 18S ribosome DNA target. 
The only other centre worldwide that routinely performs blood-stage challenge studies for P. 
vivax (QIMR, Australia), uses PCR as the sole diagnostic tool in CHMI studies, and our qPCR assay 
is based on this, utilising identical primer and probe sequences. In all other aspects, apart from 
the QIMR derived detection sequences and reaction mixture employed, the extraction of DNA 
and qPCR is identical to the existing and formally validated P.falciparum qPCR method employed 
as sole, objective, diagnostic tool in the two P.falciparum trials described below. This pan-
plasmodium qPCR is in the process of undergoing a similar formal validation and work completed 
so far, demonstrates that this is similarly fit for purpose. As examples of qualification parameters 
that contribute to validation, the assay has excellent specificity (within the context of a P.vivax 
only CHMI study), linearity, acceptable accuracy (based on reference samples of microscopically 
quantified P.vivax infected blood), precision and reproducibility (based on several inter-operator 
tests with acceptable %CV values when assessing prepared samples at the described diagnostic 
thresholds in >20 replicates). The assay has an acceptable detection and reliable quantitation 
limit of 50-100 genomes per ml (well below reproducible microscopic detection levels. While 
detection is possible down to approximately 25 genomes per ml as indicated below, it is less 
reproducible at this level). The assay is also extremely robust, with both EDTA whole blood 
samples, extracted DNA and standards remaining stable and giving reproducible qPCR scores at 
4°C storage for many days (which is beyond the intended use for real time qPCR follow up). The 

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range of the qPCR is similar to that of the previously validated P.falciparum assay, with at least a 
4 log range from 100 genomes per ml to 1,000,000 for the standards employed (actual range 
used in assays is from 25 to 1,000,000 genome copies per ml). These criteria assessments are in 
accordance with the ICH Harmonised Tripartite Guideline Part II: Validation of Analytical 
Procedures: Methodology, (European Medicines Agency, 2006). This is further supported by data 
from the lab’s participation in the UKNEQAS external quality assurance scheme since mid-2018, 
with 5 rounds of samples assayed in both P.falciparum and P.vivax assays completed to date, 
with a 100% success rate.    
To date, comparative qPCR and microscopy data are available for two blood-stage P. vivax CHMI 
studies conducted with the PvW1 blood inoculum in Oxford; VAC069A&B (n=11 inclusive of 3 
participants who underwent CHMI twice) (NCT03797989) and CHMI of VAC071 Group 1 
participants (n=3) (NCT04009096). Here, the greater sensitivity of qPCR compared to thick film 
microscopy has been consistently demonstrated with 3/14 participants diagnosed by qPCR prior 
to microscopic patency and, where infection was microscopically patent (11/14 participants), 
qPCR detected parasitaemia (>25 genome copies/mL) at least 6 days prior to thick film positivity 
(as defined as ≥2 asexual forms seen in 200 high-power fields). At the pre-specified qPCR 
thresholds for diagnosis, as described in Figure 2, 3/14 participants did have a positive thick film 
result prior to reaching diagnostic criteria (≥5,000 genome copies/mL), however, none of these 
three participants had experienced symptoms consistent with the clinical threshold for diagnosis 
at the time of thick smear positivity and were later diagnosed within 1.5 days of microscopic 
patency, without complication. 
Positive thick smear results may result from chance findings, since following blood-stage 
challenge, parasites may be seen on light microscopy at any time-point. Where this occurs, and 
premature diagnosis results, this limits the qPCR dataset available for PMR modelling. Removing 
this possibility, therefore, increases the data available for analysis of the primary outcome 
measure. In addition, use of qPCR as the sole diagnostic tool leads to greater standardisation of 
diagnosis across study groups, permitting improved inter-group as well as inter-trial comparison. 
In Oxford, we have now safely conducted two P. falciparum blood-stage CHMI studies, where 
qPCR is the sole diagnostic method (Clinicaltrials.gov identifiers NCT03906474 and 
NCT02927145). Analysis of the parasitaemia at diagnosis in one of these studies, VAC063B, 
indicates that removal of microscopy as a diagnostic measure, reduced the range over which 
diagnosis was made compared to two prior P. falciparum blood-stage CHMI studies, where 
microscopy was utilised (see Figure 7).  
As microscopy findings are not considered within the primary efficacy analysis for this study, 
microscopy for VAC071 Group 1 participants (n=3) (NCT04009096) was principally utilised as an 
additional diagnostic tool for safety reasons. The data from the two previous P. vivax blood-stage 
CHMI studies were therefore analysed to apply the revised diagnostic criteria retrospectively, to 
determine if timing of diagnosis would have been altered. Using the revised criteria, the majority 
of diagnoses would not have been delayed, although it is estimated 36% of volunteers would 
have had a small delay to diagnosis. However, since clinical criteria remain integral to the 
diagnostic algorithm, this small delay should not result in any increased risk to volunteers, and 
the ability for investigators to treat any participant in the event of clinical concern, following 
discussion with the CI, regardless of parasitaemia, will ensure participant safety. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 79 of 117 
 
Figure 7: Parasitaemia at the diagnosis timepoint in unvaccinated control volunteers in three 
prior P. falciparum blood-stage CHMI studies.  
In both the VAC054  [94] and VAC063 CHMI studies, where microscopy was used as a diagnostic 
measure alongside qPCR and clinical assessment, diagnosis was made over a wider range of 
parasitaemia compared to the VAC063B study, where thick blood smear microscopy was not 
performed. Diagnostic parasitaemia ranged between 1440-273,247 parasites/mL in the VAC054 
study, 815- 124063 parasites/mL in VAC063A and 7137- 100946 parasites/mL in the VAC063B 
CHMI. Individual and median data points are shown.  
On this basis  diagnostic criteria in VAC079 will be based on symptoms in-keeping with malarial 
infection results and real-time quantitative polymerase chain reaction (qPCR) only, i.e. thick film 
microscopy will not be used as a diagnostic measure, as summarised in the algorithm in Figure 8.  
Symptoms or signs that will be considered as indicative of symptomatic malarial infection will be 
fever >37.8oC, rigors or other severe symptoms or signs related to malarial infection, including, 
but not limited to, subjective feverishness, sweats, headache, myalgia, arthralgia, nausea or 
vomiting. 
Asexual blood-stage parasitaemia will be measured by qPCR for the pan-plasmodium 18S DNA 
gene. In the presence of symptoms or signs indicative of malarial infection, ≥5,000 genome 
copies/mL will be considered diagnostic but a PCR result ≥10,000 copies/mL will be considered 
positive, regardless of symptoms. All Investigators (except the Principal Laboratory Investigator) 
will be blinded to the results. Blinding of PCR operators to the vaccination status of a volunteer 
(as opposed to an infectivity control in the parallel VAC069 study) will be maintained by assigning 
a new unique identifier to the volunteer for PCR samples from the C-2 visit. If, as judged by the 
investigator, a volunteer develops symptoms in-keeping with malaria, the volunteer will be 
discussed with the Chief Investigator. If deemed appropriate by the Chief Investigator, the qPCR 
results for this volunteer will be unblinded by the Principal Laboratory Investigator and the 
volunteer treated only if any available qPCR result is ≥5000 genome copies/mL. At the point that 
a volunteer has a qPCR result exceeding 10,000 genome copies/mL, the Principal Laboratory 
Investigator will unblind the Chief Investigator to the result and the volunteer will be treated, 
regardless of the presence of symptoms.  
Importantly, in the unlikely event of a technical failure with the qPCR machine, we also have a 
working back-up qPCR machine that will be available for use 24/7 if required. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 80 of 117 
The Investigators are able to treat any volunteer for malaria regardless of the qPCR result if they 
are clinically concerned (and have discussed the case with the Chief Investigator), or a volunteer 
wishes to withdraw from the study. If necessary, volunteers can be discussed with the LSM 
and/or the infectious diseases consultant on call at the John Radcliffe Hospital for further 
management under the care of the infectious diseases team, inclusive of parenteral anti-malarial 
therapy if deemed appropriate.  
 
 
 
 
 
 
 
 
 
 
 
 
Figure 8: Algorithm for diagnosis and initiation of treatment. N.B. If day 28 after challenge for 
Group 1, or day 21 for Groups 2 and 3, is reached without meeting any diagnostic criteria, 
treatment will be initiated regardless of qPCR results, i.e. even if parasitaemia is undetectable. 
 
11.4.1 Participants with fever post malaria challenge and COVID-19 testing 
In the first week following CHMI, if a participant becomes unwell with a febrile illness, it is highly 
unlikely that the cause of this is malaria. If a participant develops a fever between days C+1 to 
C+6 and their symptoms are consistent with possible COVID-19 disease (fever or new onset 
persistent cough or anosmia or ageusia) the participant will be advised to self-isolate as per 
Public Health England (PHE) guidelines and directed to local community based COVID-19 testing. 
If the SARS-COV-2 PCR test is negative, the participant can stop self-isolation and continue with 
study visits as per protocol. 
From C+7 onwards, if a participant develops a fever of ≥37.8°C or other symptoms consistent 
with possible COVID-19 disease, they will be advised to inform the study team before attending 
their clinic visit and commence self-isolation as per PHE guidelines. As long as the participants’ 
symptoms are judged by the Investigator as not significant enough to require referral to 
secondary care, they will be advised to attend the study clinic where their visit will be conducted 
in a separately-accessed isolated clinic room with study staff wearing appropriate PPE. If a 
participant fulfils COVID-19 testing criteria, a SARS-COV-2 PCR swab from the nasopharynx and 
throat will be taken by the study team in addition to the study specified malaria blood tests.  
If a participant has a persistent fever or study clinicians are clinically concerned for COVID-19 
disease, then a SARS-COV-2 PCR swab can be repeated at the discretion of the Investigator, if the 
first SARS-COV-2 PCR was negative. 
qPCR 
≥5,000 
genome 
copies/mL
qPCR 
<5,000 
genome 
copies/mL
Negative 
diagnosis, 
continue 
monitoring 
Positive 
diagnosis, 
begin 
treatment 
qPCR 
<10,000 
genome 
copies/mL
qPCR 
>10,000 
genome 
copies/mL
Negative 
diagnosis, 
continue 
monitoring 
Positive 
diagnosis, 
begin 
treatment 
Symptomatic 
Asymptomatic/ 
clinical criteria for diagnosis not met 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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If a participant is self-isolating because of symptoms of possible COVID-19 disease and they are 
unable to travel to the clinic safely, then a home visit may be conducted by the study team in lieu 
of a clinic visit to allow collection of safety and malaria blood tests. 
If following malaria challenge and prior to malaria diagnosis and treatment, a participant is found 
to have a positive SARS-COV-2 PCR swab from any source, they will be commenced on malaria 
treatment, irrespective of the severity of COVID-19 disease and irrespective of malaria symptoms 
or malaria qPCR result at that time point. 
All volunteers will undergo a combined nasopharyngeal and throat swab for SARS-COV-2 PCR on 
the day of malaria diagnosis when malaria treatment is commenced, unless they have had a 
negative COVID-19 PCR swab within the preceding 24 hours. The reasoning for this is that fever 
after commencement of malaria treatment is common and if a participant develops a fever after 
starting malaria treatment, a negative COVID-19 swab will allow exclusion of concurrent COVID-
19 disease and negate the need for self-isolation and household isolation.  
 
11.5 
Malaria management 
Volunteers will be treated with a standard oral course of either artemether-lumefantrine 
(Riamet) or atavoquone/proguanil (Malarone). Both are recommended treatment of P. vivax 
infection in the UK, although use for non-falciparum malaria is an unlicensed use for both 
medications. The decision on which medication will be used, will be at the Investigator’s 
discretion and depends on factors including if there are any contraindications to either 
medication. 
If a volunteer withdraws/is withdrawn from the study after administration of CHMI but before 
reaching the criteria for malaria treatment, then a complete, appropriate, curative course of anti-
malarial therapy must be completed. The importance of this will be emphasised to volunteers at 
screening. 
11.5.1 Malaria management – Riamet 
Artemisinin-based combination therapies (ACTs) containing mefloquine, piperaquine or 
lumefantrine are now the recommended treatment for P. vivax in areas where there might be 
chloroquine resistance (WHO 2015 guidelines). Although chloroquine remains effective in most 
recent studies in Thailand, there are some signs that low level chloroquine resistance may be 
becoming established along the Thai-Myanmar border.  
Riamet® is a combination drug consisting of 20mg artemether and 120mg lumefantrine per 
tablet. The treatment regime will consist of 6 doses of total 80mg artemether/480mg 
lumefantrine (4 tablets) – to clear the blood-stage infection. The first dose, which will be directly 
observed, at treatment initiation, will be followed by additional doses after 8, 24, 36, 48 and 60 
hours (window period +/- 1 hour for each dose). The 24 hour dose will also be directly observed, 
and volunteers will record the time that intervening doses were taken at home in a medication 
diary.  
Prior to starting Riamet®, volunteers will be screened for drug interactions and contraindications. 
This includes checking for pregnancy by urine pregnancy test and serum β-hCG test in female 
volunteers and prolonged QT (on pre-challenge ECG). Volunteers will be reminded of the 
potential side effects and given the patient information sheet for Riamet® and a card outlining 
when their doses of Riamet should be taken. Volunteers will be advised to avoid grapefruit juice. 
Tablets should be taken together with a fatty meal (a light snack will provided when doses are 
observed in clinic). At least two doses for each volunteer will be directly observed in clinic.  

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11.5.2 Malaria management – Malarone  
Malarone (see SmPc for Malarone) is a combination drug consisting of proguanil hydrochloride 
and atovaquone. A treatment course of Malarone consists of 4 ‘standard tablets’ of Malarone 
(proguanil hydrochloride 100 mg, atovaquone 250 mg) once daily, orally for 3 days. The first two 
doses of Malarone will be directly observed in clinic. Prior to starting Malarone, volunteers will 
be screened for drug interactions and contraindications (including a serum β-hCG test in female 
volunteers). Volunteers will be reminded of the potential side effects of Malarone and given the 
patient information sheet for Malarone. 
11.5.3 Malaria management – supportive medications  
Provided there are no contraindications, volunteers will be provided with a course of 
paracetamol (1 g orally up to four times a day) and a course of cyclizine (50 mg orally three times 
a day) (See SmPC for Paracetamol & Cyclizine). Participants may be provided with an 
appropriate, licensed alternative anti-emetic to cyclizine if they are unable to take cyclizine.  
Volunteers will be given the patient information sheet for these medications and advised how 
frequently they can take doses. Volunteers will be issued with a medication diary card on which 
they will be asked to document all doses of medications taken post-CHMI.   
All medications used in the trial will be handled and dispensed according to SOP VC021: 
Handling, Storage and Dispensing of Non-IMP Medication.  
11.5.4 Follow-up after commencing malaria treatment 
For Group 1 volunteers, at each visit, 24 hours and 48 hours after initiation of treatment, 
procedures will be performed according to Table 11, including physical observations, assessment 
of AEs and venepuncture for blood tests including measurement of malaria qPCR. Once daily 
malaria qPCR blood tests will continue until the volunteer has two consecutive qPCR readings 
with substantial reduction in genome copies per mL. 
For Group 2 and 3 volunteers post treatment visits will occur at 24 hours and 72 hours after 
initiation of treatment. During the first CHMI, a number of volunteers in Group 1 attended for an 
additional visit on T+3 as the malaria parasite count at T+2 was still positive for some 
participants. All T+3 malaria qPCRs results were significantly reduced. In order to minimise the 
number of visits, the T+2 visit has been changed to T+3. Procedures will be performed according 
to Table 15, including physical observations, assessment of AEs and venepuncture for blood tests 
including measurement of malaria qPCR. Malaria qPCR blood tests can be repeated if qPCR 
readings are not substantially reduced by T+3. 
11.5.5 Criteria for inpatient transfer to the NHS  
If any of the following criteria are met, admission under the care of the Infectious Diseases Team, 
Oxford University Hospitals NHS Foundation Trust will be considered: 
 Failure of symptoms to improve within 48 hours of starting anti-malarial therapy. 
 Unable to tolerate oral antimalarial therapy. 
 Dehydration requiring intravenous fluid therapy. 
 Signs or symptoms suggestive of pulmonary oedema.  
 Signs or symptoms of neurological dysfunction including altered consciousness. 
 Signs, symptoms or laboratory evidence of significant renal dysfunction. 
 Unanticipated concern about subject’s home circumstances. 
 Any other significant finding which the Investigators feels warrant inpatient admission - 
this includes symptoms or signs of suspected worsening COVID-19 disease e.g. respiratory 
failure, in the unlikely event of a participant testing PCR positive for COVID-19 during the 
post-challenge period. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 83 of 117 
Ultimately, the decision regarding admission will be taken by the Investigators in conjunction 
with the Infectious Diseases Consultant on call.  
11.6 
Safety measures for conduct of CHMI 
Volunteer safety is of paramount importance. The following measures are in place to safeguard 
volunteer safety: 
 Volunteers will only be enrolled in the study if both Investigators and the volunteers’ GP 
feel this is appropriate. 
 Volunteers’ understanding of the trial information will be tested by means of a 
questionnaire at screening. This provides further confidence that fully informed consent 
has been obtained. 
 If the subject does not have their own mobile telephone they will be issued with one for 
the duration of the study and counselled about the importance of keeping it switched on 
or checking the messages regularly.  
 Before challenge, full contact details for each subject will be documented, including home 
address and mobile telephone numbers. Mobile telephone numbers will be verified prior 
to challenge to ensure the volunteers are easily contactable. Home and work landline 
telephone numbers where available and next-of-kin address and telephone numbers will 
also be documented. Subjects must also provide the Investigators with the name and 24 
hour telephone number of a close friend, relative or housemate who lives nearby and will 
be kept informed of their whereabouts for the duration of the study. 
 On the day of challenge volunteers will be provided with a medic alert card containing 
contact details for the study team and brief details of the study including the optimal 
treatment for P. vivax. 
 Volunteers will be able to contact a medically qualified member of the study team 24 
hours a day throughout the study period and will be instructed to contact the Investigator 
immediately should they manifest any signs or symptoms they perceive as serious. 
 If necessary, the study team will visit volunteers in their own homes if they are unable to 
attend clinic for review.  
 At least two doses of Riamet® for each volunteer will be observed by the study team. (For 
volunteers taking Malarone, two out of three doses will be observed). 
 Volunteers will be counselled that should they fail to return for treatment having been 
infected with P. vivax they could become very unwell and potentially die. They will be 
instructed to remain in Oxford or the immediate surrounding area for the duration of the 
intensive follow-up schedule (days 0-30 post-challenge). They will be informed that 
should they fail to attend a scheduled clinic visit post-challenge, their nominated contact, 
next of kin and the police may be informed and a search started.  
11.6.1 Measures to be taken if a volunteer goes missing post-CHMI 
In the unlikely event that a volunteer should (a) fail to attend for a scheduled clinical visit or (b) 
be un-contactable by telephone after challenge and before completion of an appropriate course 
of anti-malarial therapy, the following stakeholders will be informed; 
 All Investigators.  
 The volunteer’s nominated contact and next of kin. 
 The trial Sponsor. 
 The local safety committee. 
 The ethics committee(s). 
 Relevant hospital trust R&D departments. 
 The local police department. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 84 of 117 
 Local Accident and Emergency departments. 
Active efforts will be made to locate the volunteer by the police. While all parties will aim to 
preserve the volunteer’s confidentiality, if necessary details of the volunteer’s identity and 
participation in the study may be passed to the national media in order to help locate the missing 
individual. Volunteers will be informed of this during screening. 
11.7 
Follow-up post-treatment  
11.7.1 Six days after initiation of treatment (T+6) 
Physical observations will be performed and AEs assessed. Venepuncture will be performed 
(Table 11 for Group 1, Table 15 for Groups 2 and 3). Where applicable, medication diary and 
malaria symptom diary cards will be collected from volunteers. 
11.7.2 Nine days after initiation of treatment (T+9) 
Volunteers will be phoned at 9 days post day of diagnosis visit (treatment initiation) (as per Table 
11 for Group 1, Table 15 for Groups 2 and 3). Participants will be questioned as to whether they 
have; 
o Experienced any of the foreseeable symptoms of malaria. 
o Experienced any other symptoms.  
o Taken any medications including over the counter medications. 
11.7.3 Day 56 post CHMI (C+56) 
Physical observations will be performed and AEs assessed. Venepuncture will be performed 
(Table 11 for Group 1, Table 15 for Groups 2 and 3).  
11.7.4 Day 96 post CHMI (C+96)  
Physical observations will be performed and AEs assessed. Venepuncture will be performed 
(Table 11 for Group 1, Table 15 for Groups 2 and 3).  
11.7.5 Day 276 post CHMI (C+276)  
Physical observations will be performed and AEs assessed. Venepuncture will be performed 
(Table 11 for Group 1, Table 15 for Groups 2 and 3).

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 85 of 117 
  
S 
 V1 
 V2 
 
 
 
V3 
 
 
Attendance number 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
Timeline (days) Group 1 
 
0  
1  
3  
7  
14  
28  
29 
31 
35 
42 
approx 
365 
1 day 
post 3rd  
vacc 
3 days 
post 3rd 
vacc 
7 days 
post 3rd 
vacc 
14 days 
post 3rd 
vacc 
Timeline (days) Group 2 
 
0  
1  
3  
7  
14  
28  
29 
31 
35 
42 
56 
57 
59 
63 
70 
Window (days) 
-90 
±1 
±1 
-1/+2 
-2/+4 
-3/+5 
-7/+14 
±1 
-1/+2 
-2/+4 
-3/+5 
-7/+14 
±1 
-1/+2 
-2/+4 
-3/+5 
Inclusion/Exclusion criteria 
X 
X 
  
  
  
  
X 
 
 
 
  
X  
  
  
 
 
Informed consent 
X 
(X) 
  
  
  
  
(X) 
 
 
 
  
(X)  
  
  
 
 
ICQ 
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) 
ECG 
X 
  
  
  
  
  
  
 
 
 
  
  
  
  
 
 
Urinalysis 
X 
  
  
  
  
  
  
 
 
 
  
  
  
  
 
 
Urine β-hCG** 
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 
X 
Vaccination 
X 
X 
 
 
 
X 
 
 
Diary card issued 
X 
X 
 
 
 
X 
 
 
Diary card collected 
X 
 
 
 
X 
 
 
Immunology (mL) 
  
80 
20 
20 
40 
40 
60 
20 
20 
40 
40@ 
80 
20 
20 
50 
80 
RNA preservation (mL) 
  
3 
3 
 
 
 
3 
3 
 
 
3 
3 
 
 
 
Haematology (mL)# 
4 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Biochemistry (mL) ## 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
3 
Serum βhCG (mL)** 
  
  
  
  
  
  
 
 
 
  
  
  
  
 
 
DARC (mL) 
2 
  
  
  
  
  
  
 
 
 
  
  
  
  
 
 
HLA typing (mL) 
  
4 
  
  
  
  
  
 
 
 
  
  
  
  
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 86 of 117 
HIV, HBV, HCV, EBV, CMV 
serology (mL) 
5 
  
  
  
  
  
  
 
 
 
  
  
  
  
 
 
Group 1 - Total Blood 
Volume (mL)^ 
14 
92 
28 
25 
45 
45 
68 
28 
25 
45 
5 
88 
28 
25 
55 
85 
Group 2 - Total Blood 
Volume (mL)^ 
14 
92 
28 
25 
45 
45 
68 
28 
25 
45 
45 
88 
28 
25 
55 
85 
Group 1 - Cumulative blood 
volume (mL)^ 
14 
106 
134 
159 
204 
249 
317 
345 
370 
415 
420 
508 
536 
561 
616 
701 
Group 2 - Cumulative blood 
volume (mL)^ 
14 
106 
134 
159 
204 
249 
317 
345 
370 
415 
460 
548 
576 
601 
656 
741 
Table 7: Group 1 and 2 - Schedule of events pre-CHMI 
S = screening visit; V1, V2 and V3 = 1st, 2nd and 3rd vaccinations with PvDBPII/Matrix M1; ICQ = Informed Consent questionnaire; (X) = If considered necessary, emphasising any acute 
complaints. *Physical observations will include blood pressure, pulse and temperature, plus height and weight at screening only. **For females only. # Haematology will include full 
blood count plus G6PD level and haemoglobinopathy screen at screening visit only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin and liver function tests; 
CRP on days 0, 1, 3, 7, 56, 57, 59 and 63 only; magnesium and calcium at screening only. @D42 immunology bleed taken in Group 2 only. ^Blood volumes as given in the table are 
maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the exception of additional blood volume that may be required for 
any additional tests that may be performed if deemed clinically necessary. 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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C-2 
CHMI 
C+1-C+6 
C+7 
C+8 
C+9 
C+10
C+10.5 
C+11 
C+11.5 
C+12 
C+12.5
C+13 
C+13.5 
C+14 
C+14.5
Attendance number - Group 1 
17 
18 
19 
20 
21 
22 
23 
24 
25 
26 
27 
28 
29 
30 
31 
32 
Timeline (days) - Group 1 
-2 
0 
1-6 
7 
8 
9 
10 
10 
11 
11 
12 
12 
13 
13 
14 
14 
Window (days)
±1 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Inclusion/Exclusion criteria 
X  
X  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Informed consent 
X  
 X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Daily telephone call
  
  
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) 
Urine β-hCG**
  
 
 
 
 
 
 
 
 
 
 
 
 
 
Review contraindications 
 X 
 X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Review AEs & medications 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
Diary card issued 
 X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Medic alert card issued
X
 
 
 
 
 
 
 
 
 
 
 
 
 
Malaria challenge
 
X
 
 
 
 
 
 
 
 
 
 
 
 
 
Immunology (mL) 
80 
 
 
60 
 
 
 
  
  
  
  
  
  
  
60 
  
Serum storage (mL) 
4 
 
 
 
 
 
 
  
  
  
  
  
  
  
 
  
qPCR/GctPCR (mL) 
2 
 
 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Haematology (mL)#
 2 
  
  
2 
  
  
  
  
  
  
  
  
  
  
2 
  
Biochemistry (mL) ## 
 3 
  
  
3 
 
 
 
 
 
 
 
 
 
 
3 
 
Serum βhCG (mL)** 
 5 
  
  
5 
 
 
 
 
 
 
 
 
 
 
5 
 
SARS-COV-2 PCR swab 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Total Blood Volume (mL)^ 
96 
0 
0 
72 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
72 
2 
Cumulative blood volume (mL)^
797 
797 
797 
869 
871 
873 
875 
877 
879 
881 
883 
885 
887 
889 
961 
963 
Table 8: Group 1 - Schedule of events for day before challenge to day 14 after challenge (timeline/window are in relation to the day of challenge). (X) = If considered necessary. 
Visits continue until diagnosis; when diagnostic criteria are met, venepuncture and procedures are followed according to Table 11. *Physical observations will include blood pressure, 
pulse and temperature, plus weight at C-2 visit. **For females only. #Haematology will comprise full blood count, including reticulocyte count at C-2. ## Biochemistry will include 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 88 of 117 
sodium, potassium, urea, creatinine, albumin and liver function tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any 
visit, at the discretion of the investigator, with the exception of additional blood volume that may be required for any additional tests that may be performed if deemed clinically 
necessary. 
 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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C+15 
C+15.5 
C+16 
C+16.5 
C+17 
C+17.5 
C+18 
C+18.5 
C+19 
C+19.5 
C+20 
C+20.5 
C+21 
C+21.5 
Attendance number - Group 1 
33 
34 
35 
36 
37 
38 
39 
40 
41 
42 
43 
44 
45 
46 
Timeline (days) - Group 1 
15 
15 
16 
16 
17 
17 
18 
18 
19 
19 
20 
20 
21 
21 
Window (days)
 
  
  
  
  
  
  
  
  
  
  
  
  
Inclusion/Exclusion criteria 
 
 
 
  
  
  
  
  
  
  
  
  
  
  
Informed consent 
 
 
 
  
  
  
  
  
  
  
  
  
  
  
Daily telephone call
 
 
  
  
  
  
  
  
  
  
  
  
  
Physical observations* 
 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) 
Urine β-hCG**
  
 
 
 
 
 
 
 
 
 
 
 
Review contraindications 
  
  
  
  
  
  
  
  
  
  
  
  
  
Review AEs & medications 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
Diary card issued 
  
  
  
  
  
  
  
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Medic alert card issued
 
 
 
 
 
 
 
 
 
 
 
Immunology (mL)
 
 
 
 
 
 
 
 
 
 
 
 
60 
 
Serum storage (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
qPCR/GctPCR (mL) 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Haematology (mL)#
 
 
 
 
 
 
 
 
 
 
2 
 
Biochemistry (mL) ## 
 
 
 
 
 
 
 
 
 
 
 
 
3 
 
Serum βhCG (mL)** 
 
 
 
 
 
 
 
 
 
 
 
 
5 
 
Total Blood Volume (mL)^
2
2
2
2
2
2
2
2
2
2
2
2
72
2
Cumulative blood volume (mL)^
965 
967 
969 
971 
973 
 975 
977 
979 
981 
983 
985 
987 
1059 
1061 
Table 9: Group 1 - Schedule of events for days 15-21 after challenge (timeline/window are in relation to the day of challenge). (X) = If considered necessary. Visits continue until 
diagnosis; when diagnostic criteria are met, venepuncture and procedures are followed according to Table 11. *Physical observations will include blood pressure, pulse and 
temperature. **For females only. #Haematology will comprise full blood count only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin and liver function 
tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the exception of 
additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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C+22 
C+22.5 
C+23 
C+23.5 
C+24 
C+24.5 
C+25 
C+25.5 
C+26 
C+26.5 
C+27 
C+27.5 
Attendance number – Group 1 
47 
48 
49 
50 
51 
52 
53 
54 
55 
56 
57 
58 
Timeline (days) – Group 1 
22 
22 
23 
23 
24 
24 
25 
25 
26 
26 
27 
27 
Window (days)
 
  
  
  
  
  
  
  
  
  
  
Inclusion/Exclusion criteria 
 
 
 
  
  
  
  
  
  
  
  
  
Informed consent 
 
 
 
  
  
  
  
  
  
  
  
  
Daily telephone call
 
 
  
  
  
  
  
  
  
  
  
Physical observations* 
 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) 
Urine β-hCG**
  
 
 
 
 
 
 
 
 
 
Review contraindications 
  
  
  
  
  
  
  
  
  
  
  
Review AEs & medications 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
Diary card issued 
  
  
  
  
  
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
  
  
  
  
  
  
  
Medic alert card issued
 
 
 
 
 
 
 
 
 
Immunology (mL)
 
 
 
 
 
 
 
 
 
 
 
 
Serum storage (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
qPCR/GctPCR (mL) 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Haematology (mL)#
 
 
 
 
 
 
 
 
 
 
Biochemistry (mL) ## 
 
 
 
 
 
 
 
 
 
 
 
 
Serum βhCG (mL)** 
 
 
 
 
 
 
 
 
 
 
 
 
Total Blood Volume (mL)^
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Cumulative blood volume (mL)^
1063 
1065 
1067 
1069 
1071 
1073 
1075 
1077 
1079 
1081 
1083 
1085 
Table 10: Group 1 - Schedule of events for days 22-27.5 after challenge (timeline/window are in relation to the day of challenge). (X) = If considered necessary. Visits continue until 
diagnosis; when diagnostic criteria are met, venepuncture and procedures are followed according to Table 11. *Physical observations will include blood pressure, pulse and 
temperature. **For females only. #Haematology will comprise full blood count only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin, and liver function 
tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the exception of 
additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary.

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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DOD/C+28 (if 
undiagnosed)$+ 
T+1  
(24hr dose)^^ 
T+2  
(48hr dose)^^ 
T+6 
T+9 
C+56 
C+96@ 
C+276 
Attendance number – Group 1 
59 
60 
61 
62 
n/a 
63 
64 
65 
Timeline (days) – Group 1 
 - 
 - 
-  
- 
 
56 
96 
276 
Window (days)
±1
±2
±14
±14
±14
Telephone call 
 
 
 
  
X 
  
  
  
Physical observations*
X
X
X
X
X
X
X
Medical history/ examination 
 (X) 
 (X)  
 (X) 
 (X) 
 
 (X) 
 (X) 
 (X) 
Urine β-hCG**
X
Review contraindications 
 X 
  
  
 
  
  
  
Review AEs & medications
X
X
X
X
X
X
X
Diary card issued 
  
  
  
 
  
  
  
Diary card collected 
  
  
  
 X 
 
  
  
Medic alert card issued 
  
 
 
 
 
 
Immunology (mL) 
70 
 
 
 
 
80 
70 
70 
Serum storage (mL)
4
HIV, HBV, HCV, EBV, CMV serology (mL)+++ 
 
 
 
 
 
 
5 
 
qPCR/GctPCR (mL)
2
2
2
Haematology (mL)#
2 
  
  
2 
 
2 
2 
 
Biochemistry (mL) ## 
3 
  
  
3 
 
3 
3 
 
Serum βhCG (mL)**
5
SARS-COV-2 PCR swab
X
Total Blood Volume (mL)^ 
82 
2 
2 
5 
n/a 
85 
84 
70 
Cumulative blood volume (mL)^ 
1167 
1169 
1171 
1176 
 
1261 
1345 
1415 
Table 11: Group 1 - Schedule of events from diagnosis to 276 days after challenge (timeline/window are in relation to the day of challenge or treatment day). DOD = day of 
diagnosis; C = challenge; T = treatment; (X) = If considered necessary. *Physical observations will include blood pressure, pulse and temperature. **For females only. #Haematology 
will comprise full blood count only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin and liver function tests. $ if diagnosed at C+14.5 or C+21.5 do not take 
DOD immunology bleed (will have taken full bleed at C+14 and C+21). + immediately before drug treatment. ^^ Blood draws for qPCR will continue until the volunteer has two 
consecutive qPCR readings with substantial reduction in genome copies per mL. +++ EBV and CMV serology will only be repeated at the C+96 timepoint if they were negative at 
screening. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the exception of 
additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. @Last visit for the subset of Group 1 participants that 
move into Group 3, next visit for those who go into Group 3 will be as per Table 12. 
 

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V4 
V4-D1 
V4-D3 
V4-D7 
V4-D14 
Attendance number – Group 3 
65 
66 
67 
68 
69 
Timeline (days) – Group 3 
Approx 150 
days post 3rd 
vaccination 
1 day post 4th 
vaccination 
3 days post 4th 
vaccination 
7 days post 4th 
vaccination 
14 days post 
4th vaccination 
Window (days) 
-7/+14 
±1 
-1/+2 
-2/+4 
-3/+5 
Inclusion/Exclusion criteria 
X  
  
  
 
 
Informed consent 
(X)  
  
  
 
 
Physical observations* 
X 
X 
X 
X 
X 
Medical history/examination 
(X) 
(X) 
(X) 
(X) 
(X) 
Urine β-hCG** 
X 
 
 
Review contraindications 
X 
 
 
Review AEs & medications 
X 
X 
X 
X 
X 
Vaccination 
X 
 
 
Diary card issued 
X 
 
 
Diary card collected 
 
 
Immunology (mL) 
80 
20 
20 
50 
80 
RNA preservation (mL) 
3 
3 
 
 
 
Haematology (mL)# 
2 
2 
2 
2 
2 
Biochemistry (mL) ## 
3 
3 
3 
3 
3 
Total Blood Volume (mL)^ 
88 
28 
25 
55 
85 
Group 3 - Cumulative blood 
volume (mL)^ 
1433 
1461 
1486 
1541 
1626 
Table 12: Group 3 - 4th vaccination schedule of events 
V4 = 4th vaccinations with PvDBPII/Matrix M1, to occur approximately 1 month after C+96 visit following first CHMI (X) = If considered necessary, emphasising any acute complaints. 
*Physical observations will include blood pressure, pulse and temperature. **For females only. # Haematology will include full blood count. ## Biochemistry will include sodium, 
potassium, urea, creatinine, albumin and liver function tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the 
discretion of the investigator, with the exception of additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
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C-2 
CHMI 
C+1-
C+6 
C+7 
C+8 
C+9 
C+10 
C+10.5$ 
C+11 
C+11.5$ 
C+12 
C+12.5$ 
C+13 
C+13.5$ 
C+14 
C+14.5$ 
Attendance number – Group 2 
17 
18 
19 
20 
21 
22 
23 
24 
25 
26 
27 
28 
29 
30 
31 
32 
Attendance number – Group 3 
70 
71 
72 
73 
74 
75 
76 
77 
78 
79 
80 
81 
82 
83 
84 
85 
Timeline (days)  
-2 
0 
1-6 
7 
8 
9 
10 
10 
11 
11 
12 
12 
13 
13 
14 
14 
Window (days) 
±1 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Inclusion/Exclusion criteria 
X  
X  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Informed consent 
X  
X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Daily telephone call 
  
  
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) 
Review contraindications 
 X 
 X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Review AEs & medications 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
Diary card issued 
 X 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
  
Medic alert card issued 
X 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
Malaria challenge 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Immunology (mL) 
80 
 
 
60 
 
 
 
  
  
  
  
  
  
  
60 
  
Serum storage (mL) 
4 
 
 
 
 
 
 
  
  
  
  
  
  
  
 
  
qPCR/GctPCR (mL) 
2 
 
 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Haematology (mL)# 
2 
  
  
2 
  
  
  
  
  
  
  
  
  
  
2 
  
Biochemistry (mL) ## 
3 
  
  
3 
 
 
 
 
 
 
 
 
 
 
3 
 
Serum βhCG (mL)** 
5 
  
  
5 
 
 
 
 
 
 
 
 
 
 
5 
 
SARS-COV-2 PCR swab 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Total Blood Volume (mL)^ 
96 
0 
0 
72 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
72 
2 
Group 2 - Cumulative blood 
volume (mL)^ 
837 
837 
837 
909  
911  
913  
915  
917  
919  
921  
923  
925  
927  
929  
1001 
1003 
Group 3 - Cumulative blood 
volume (mL)^  
1722 
1722 
1722 
1794 
1796 
1798 
1800 
1802 
1804 
1806 
1808 
1810  
1812 
1814  
1886 
1888 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 94 of 117 
Table 13: Group 2 (1st CHMI) and Group 3 (2nd CHMI) - Schedule of events for day before challenge to day 14 after challenge (timeline/window are in relation to the day of 
challenge). Visits continue until diagnosis; when diagnostic criteria are met, venepuncture and procedures are followed according to Table 15.  $From day 10 post challenge onwards, 
visits will continue once a day in mornings, increasing to twice daily if the malaria qPCR count is >1000 genome copies/ml. (X) = If considered necessary. *Physical observations will 
include blood pressure, pulse and temperature, plus weight at C-2 visit. **For females only. #Haematology will comprise full blood count only. ## Biochemistry will include sodium, 
potassium, urea, creatinine, albumin and liver function tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the 
discretion of the investigator, with the exception of additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 95 of 117 
  
C+15 
C+15.5$ 
C+16 
C+16.5$ 
C+17 
C+17.5$ 
C+18 
C+18.5$ 
C+19 
C+19.5$ 
C+20 
C+20.5$ 
Attendance number - Group 2  
33 
34 
35 
36 
37 
38 
39 
40 
41 
42 
43 
44 
Attendance number - Group 3  
86 
87 
88 
89 
90 
91 
92 
93 
94 
95 
96 
97 
Timeline (days) 
15 
15 
16 
16 
17 
17 
18 
18 
19 
19 
20 
20 
Physical observations* 
 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) 
Review AEs & medications 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
 X 
Diary card issued 
  
  
  
  
  
  
  
  
  
  
  
Diary card collected 
  
  
  
  
  
  
  
  
  
  
  
  
Medic alert card issued 
  
 
 
 
 
 
 
 
 
 
Immunology (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
Serum storage (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
qPCR/GctPCR (mL) 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
2 
Haematology (mL)# 
 
 
 
 
 
 
 
 
 
 
 
 
Biochemistry (mL) ## 
 
 
 
 
 
 
 
 
 
 
 
 
Serum βhCG (mL)** 
 
 
 
 
 
 
 
 
 
 
 
 
Total Blood Volume (mL)^
2
2
2
2
2
2
2
2
2
2
2
2
Group 2 - Cumulative blood 
volume (mL)^ 
1005 
1007 
1009 
1011 
1013 
1015 
1017 
1019 
1021 
1023 
1025 
1027 
Group 3 - Cumulative blood 
volume (mL)^ 
1890 
1892 
1894 
1896 
1898 
1900 
1902 
1904 
1906 
1908 
1910 
1912 
Table 14: Group 2 (1st CHMI) and Group 3 (2nd CHMI) - Schedule of events from day 15 - 20 after challenge (timeline/window are in relation to the day of challenge). Visits continue 
until diagnosis; when diagnostic criteria are met, venepuncture and procedures are followed according to Table 15. $From day 10 post challenge onwards, visits will continue once a 
day in mornings, increasing to twice daily if the malaria qPCR count is >1000 genome copies/ml. (X) = If considered necessary. *Physical observations will include blood pressure, 
pulse and temperature. **For females only. #Haematology will comprise full blood count only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin and liver 
function tests. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the 
exception of additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. 
 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 96 of 117 
 
DOD/C+21 (if 
undiagnosed)$+ 
T+1 
(24hr dose)^^ 
T+3 
T+6 
T+9 
C+56 
C+96 
C+276 
Attendance number - Group 2 
45 
46 
47 
48 
 
49 
50 
51 
Attendance number - Group 3 
98 
99 
100 
101 
n/a 
102 
103 
104 
Timeline (days) 
-
-
-
-
56
96
276
Window (days) 
 
  
  
±1 
±2 
±14 
±14 
±14 
Telephone call
X
Physical observations* 
 X 
 X 
 X 
 X 
 
 X 
 X 
 X 
Medical history/ examination
(X)
(X)
(X)
(X)
(X)
(X)
(X)
Urine β-hCG**
 X 
  
  
 
 
 
 
 
Review contraindications
X
Review AEs & medications 
 X 
 X 
 X 
 X 
 
 X 
 X 
 X 
Diary card issued 
  
  
  
 
  
  
  
Diary card collected 
  
  
  
 X 
 
  
  
Immunology (mL) 
70 
 
 
 
 
80 
70 
70 
Serum storage (mL)
4
HIV, HBV, HCV, EBV, CMV serology (mL)+++ 
 
 
 
 
 
 
5 
 
qPCR/GctPCR (mL)
2
2
2
Haematology (mL)#
2 
  
  
2 
 
2 
2 
 
Biochemistry (mL) ## 
3 
  
  
3 
 
3 
3 
 
Serum βhCG (mL)**
5
SARS-COV-2 PCR swab
X
Total Blood Volume (mL)^ 
82 
2 
2 
5 
n/a 
85 
84 
70 
Group 2 - Cumulative blood volume (mL)^  
1109 
1111 
1113 
1118 
n/a 
1203 
1287 
1357 
Group 3 - Cumulative blood volume (mL)^ 
1994
1996
1998
2003
n/a
2088
2172
2242
Table 15: Group 2 (1st CHMI) and Group 3 (2nd CHMI) - Schedule of events from diagnosis to 276 days after challenge (timeline/window are in relation to the day of challenge or 
treatment day). DOD = day of diagnosis; C = challenge; T = treatment; (X) = If considered necessary. *Physical observations will include blood pressure, pulse and temperature. **For 
females only. #Haematology will comprise full blood count only. ## Biochemistry will include sodium, potassium, urea, creatinine, albumin and liver function tests. $ if diagnosed at 
C+14.5 do not take DOD immunology bleed (will have taken full bleed at C+14). + immediately before drug treatment. ^^ Blood draws for qPCR will continue until the volunteer has 
two consecutive qPCR readings with substantial reduction in genome copies per mL. +++ EBV and CMV serology will only be repeated at the C+96 timepoint if they were negative at 
screening. ^Blood volumes as given in the table are maximal volumes and smaller blood volumes may be taken at any visit, at the discretion of the investigator, with the exception of 
additional blood volume that may be required for any additional tests that may be performed if deemed clinically necessary. 
 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 97 of 117 
12 ASSESSMENT OF SAFETY 
Safety will be assessed by the frequency, incidence and nature of adverse events and serious 
adverse events arising during the study. The safety profile will be assessed on an on-going basis 
by the Investigators. The Chief investigator, Principal Investigator, and relevant Investigators (as 
per the trial delegation log) will also review safety issues and SAEs as they arise. 
12.1 Definitions 
12.1.1 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 adverse event will be graded by the participant according to the table 
for grading severity of adverse events (see section 12.3. Severity gradings may be reviewed and 
discussed with the participants at the clinic visits. 
12.1.2 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 adverse reactions. 
12.1.3 Unexpected Adverse Reaction  
An unexpected adverse reaction is an adverse reaction, the nature or severity of which is not 
consistent with the applicable product information. 
12.1.4 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 a serious AE. 
 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 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 98 of 117 
treatment in an emergency room or clinic, blood dyscrasias, or convulsions that do not 
result in inpatient hospitalisation. 
 Congenital anomaly or birth defect. 
12.1.5 Serious Adverse Reaction (SAR) 
An adverse event (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, including P. vivax CHMI, based on the information provided. 
12.1.6 Suspected Unexpected Serious Adverse Reaction (SUSAR) 
A serious adverse reaction, the nature and severity of which is not consistent with the 
information about the medicinal product in question set out in the IB, non-investigational 
medicinal product dossier (for P. vivax blood inoculum)  or Summary of Product Characteristics 
(SmPC). 
12.1.7 Foreseeable Adverse Reactions 
The foreseeable ARs following vaccination with PvDBPII-Matrix M1 include injection site pain, 
erythema, warmth, swelling, pruritus, myalgia, arthralgia, headache, fatigue, fever, feverishness, 
malaise and nausea. These adverse events will be listed as solicited adverse events providing 
they occur within 7 days of the day of vaccination. AEs other than these, or those AEs occurring 
outside of the 7 days after vaccination, will be listed as unsolicited adverse events. 
12.1.8 Other Foreseeable Medical Occurrences 
The following medical occurrences are foreseeable: 
 Clinical P. vivax disease resulting in one or more of fever, tachycardia, hypotension, 
feverishness, chills, rigor, sweats, headache, anorexia, nausea, vomiting, myalgia, 
arthralgia, low back pain, fatigue, lymphopenia and thrombocytopenia; 
 Clinical P. vivax disease resulting in inpatient transfer of care to the NHS;  
 Adverse reactions to Riamet®, Malarone, paracetamol or cyclizine, as detailed in the 
SmPCs for these medications. 
12.1.9 Expected Serious Adverse Events 
No serious adverse events are expected in this study. 
12.2 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 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 16). 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. 

VAC079 Clinical Trial Protocol, V8.0, 21st June 2022, University of Oxford 
 
Page 99 of 117 
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. 
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 16: Guidelines for assessing the relationship of vaccine administration to an AE. 
12.3 Reporting Procedures for All Adverse Events  
All AEs occurring in the 28 days following each vaccination observed by the Investigator or 
reported by the volunteer, whether or not attributed to study medication, will be recorded. 
Recording and reporting of all AEs will take place as detailed in SOP VC027. All AEs occurring in 
post-CHMI, until 6 days after completion of antimalarial treatment (i.e. 9 days after treatment 
initiation) will also be recorded- these will be entered directly into the eCRF. 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, the event is considered stable 
or until a non-study related causality is assigned (if the volunteer consents to this). Serious 
adverse events (SAEs) will be collected throughout the entire trial period.  
Each adverse event will be graded by the participant according to the table for grading severity 
of adverse events (see Table 17). 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 adverse events will be assessed by the Investigators according to the 
scales in Tables 17-19. 

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Severity grading for laboratory AEs are dependent on the OUH laboratory’s reference and will be 
graded according to the scales in the site-specific table that can be found in the Investigator file. 
These ranges will be based on FDA guidance relative to local laboratory reference ranges [112].  
CRP may be measured as part of the biochemistry profile on days 0, 1, 3, 7, 56, 57, 59 and 63. 
However, CRP results will be used in relation to exploratory immunological analysis only, and 
therefore, any abnormal result will not be considered an AE and grading scales for CRP are not 
listed in the site-specific table, i.e. CRP results will not be considered within the safety analysis. 
 
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 17: 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 
>38.0 
>39.0 
Table 18: 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 subject 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). 
 
 
 

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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 
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 
*erythema or swelling ≤3mm is an expected consequence of skin puncture and will therefore not be considered an 
adverse event. 
Table 19: Severity grading criteria for local adverse events. 
12.4 Reporting Procedures for Serious AEs (see SOP OVC005 Safety Reporting) 
In order to comply with current regulations on serious adverse event 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 Chief Investigator and Principal Investigators (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 Chief Investigator and/or LSM. In addition to the expedited reporting above, the 
Investigator shall include all SAEs in the annual Development Safety Update Report (DSUR) 
report. 
12.4.1 Reporting Procedures for SUSARS 
The Chief Investigator will report all SUSARs to the MHRA and REC within required timelines (15 
days for all SUSARs, unless life threatening or fatal in which case 7 days, with a final report within 
a further 8 days (total 15)). The Chief Investigator 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. 

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12.5 Development Safety Update Report 
A Development Safety Update Report (DSUR) will be submitted by the Sponsor, or delegate, to 
the competent authority and ethics committee on the anniversary of the first approval date from 
the regulatory authority for each IMP. 
12.6 Adverse Events of Special Interest 
Adverse events of special interest will be reported as SAEs. These are: 
 Severe hypersensitivity reactions (e.g. anaphylaxis) 
 Any new, suspected auto-immune disease 
12.7 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 adverse events will be 
assessed using the site-specific tables in the Investigator site file (ISF)/ 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. 
12.8 Local Safety Monitor 
An independent Local Safety Monitor (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 will be 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 Local Safety Monitor, Chief Investigator, 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 subjects 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 and Chief Investigator. 
 
12.9 Safety stopping/holding rules 
Safety holding rules have been developed considering the fact that this is a first-in-human study. 
‘Solicited adverse events’ are those listed as foreseeable adverse reactions in the protocol, 
occurring within the first 7 days after vaccination (day of vaccination and six subsequent days). 

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‘Unsolicited adverse events’ are adverse events other than the foreseeable ARs occurring within 
the first 7 days, or any AEs occurring after the first 7 days after vaccination. 
 
12.9.1 Group holding rules 
If a group holding rule is activated, then further vaccinations will not occur until an internal 
safety review has been conducted and it is deemed appropriate to restart dosing. The regulatory 
authority must be informed and a request to restart dosing with pertinent data must be 
submitted as a request for a 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 (Novovax) will also be notified if 
a holding rule is activated or released.  
All vaccinated volunteers will be followed for safety until resolution or stabilisation (if 
determined to be chronic sequelae) of their AEs. 
The holding rules are as follows: 
 Solicited local adverse events: 
o If more than 25% of doses of a vaccine are followed by the same Grade 3 solicited 
local adverse event beginning within 2 days after vaccination (day of vaccination and 
one subsequent day) and persisting at Grade 3 for > 72 hrs. 
 Solicited systemic adverse events:  
o If more than 25% of doses of a vaccine are followed by the same Grade 3 solicited 
systemic adverse event beginning within 2 days after vaccination (day of vaccination 
and one subsequent day) and persisting at Grade 3 for > 48 hrs.  
 Unsolicited adverse events: 
o If more than 25% of volunteers develop the same Grade 3 unsolicited adverse event 
(including the same laboratory adverse event) that is considered possibly, probably or 
definitely related to vaccination and persists at Grade 3 for > 48 hrs.  
 A serious adverse event considered possibly, probably or definitely related to vaccination 
occurs 
 
12.9.2 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 adverse event considered possibly, 
probably or definitely related within 7 days after vaccination and persisting 
continuously at Grade 3 for >72hrs. 
 Solicited systemic adverse events:  
o The volunteer develops a Grade 3 systemic solicited adverse event 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 adverse events:  
o The volunteer has a Grade 3 adverse event, considered possibly, probably or 
definitely related to vaccination, persisting continuously at Grade 3 for >72hrs.  
o The volunteer has a serious adverse event considered possibly, probably or definitely 
related to vaccination. 
o The volunteer has an acute allergic reaction or anaphylactic shock following the 
administration of the vaccine investigational product. 
If a volunteer has an acute illness (moderate or severe illness with or without fever) or a fever 
(oral temperature greater than 37.5°C) at the scheduled time of administration of investigational 
product, the volunteer will not receive the vaccine at that time. The vaccine may be administered 
to that volunteer at a later date within the time window specified in the protocol or they may be 
withdrawn from the study at the discretion of the Investigator.  
All vaccinated volunteers will be followed for safety until the end of their planned participation in 
the study or until resolution or stabilisation (if determined to be chronic sequelae) of their AEs, 
providing they consent to this.  
In addition to these pre-defined criteria, the study can be put on hold upon advice of the Local 
Safety Monitor (LSM), Chief Investigator, Study Sponsor, regulatory authority, Research Ethics 
Committee (REC) or Local Safety Monitor (LSM), for any single event or combination of multiple 
events which, in their professional opinion, jeopardise the safety of the volunteers or the 
reliability of the data. 
  
 

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13 STATISTICS 
13.1  Sample size 
This is an open label, single site, first-in-human, Phase I/IIa, blood-stage P. vivax malaria vaccine 
trial aiming to assess the safety, immunogenicity and efficacy of the PvDBPII-Matrix M1 vaccine 
candidate against blood-stage challenge for the first time, utilising a newly produced 
cryopreserved inoculum source. The number of volunteers undergoing vaccination followed by 
malaria challenge will be 10-12.  
Outcome measures will be assessed as described in Section 7. PMR will constitute the primary 
endpoint for the study and pooled data for PMR will be compared with PMR in 10-12 malaria-
naïve infectivity controls, recruited as part of another study, VAC069, who will undergo CHMI in 
parallel to the volunteers in VAC079. Any statistically significant change in PMR in a vaccine 
group would be considered as success, however, it is not clear what level of change in PMR 
would be likely to be significant in terms of clinical outcome. As this is only the third CHMI study 
with this new bank of Plasmodium vivax, there is insufficient data to allow accurate 
modelling/power calculations, but it is considered that ten volunteers in the vaccinated group 
will be sufficient to allow detection of statistically significant differences in PMR between 
vaccinees and controls 
 
 
 
 

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14 QUALITY CONTROL AND QUALITY ASSURANCE PROCEDURES 
14.1  Investigator procedures 
Approved site-specific SOPs will be used at all clinical and laboratory sites. 
14.2 
Monitoring 
Monitoring will be performed by CTRG using established procedures, according to ICH Good 
Clinical Practice (GCP).  Following written standard operating procedures, 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 sites 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. 
14.3 
Modification to protocol 
No substantial amendments to this protocol will be made without consultation with, and 
agreement of, the Sponsor. Any substantial amendments to the trial that appear 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 Chief Investigator and will be made a formal part of the protocol following ethical approval. 
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 subject. 
14.4 
 Protocol deviation  
Any deviations from the protocol will be documented in a protocol deviation form and filed in 
the site trial master file.  
14.5  Audit & inspection 
The QA manager performs system 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 ICH GCP. The audit schedule includes laboratory activities. The 
internal audits will supplement the external monitoring process and will review processes not 
covered by the external monitor. 
The Sponsor, trial sites, 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. 
14.6 
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 subjects of the trial; or 
(b) the scientific value of the trial. 

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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 committee, Regulatory authority and the 
relevant NHS host organisation within seven calendar days.  
14.7 
Trial progress 
The progress of the trial will be overseen by the Chief Investigator. 
14.8 
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 ICMJE guidelines and other contributors will be acknowledged. Data may also be used 
for a PhD or MD thesis.  
14.9 
Intellectual Property 
Ownership of intellectual property (IP) generated by employees of the University vests in the 
University.  The protection and exploitation of any new IP is managed by the University’s 
technology transfer office, Oxford University Innovations 
 

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15 ETHICS 
15.1 
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. 
15.2 
ICH guidelines for good clinical practice 
The Investigator will ensure that this study is conducted in full conformity to Medicine for Human 
use (Clinical Trials) Regulations 2004 (SI 2004 No. 1031) and its amendments and with the ICH 
guidelines for GCP (CPMP/ICH/135/95) July 1996.  
15.3 
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 HRA for written approval. The Investigator will submit and, where 
necessary, obtain approval from the REC and the HRA 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 deviations from the protocol or SAEs occurring at the site to the Sponsor and will notify 
the REC(s) of these if necessary in accordance with procedures. 
15.4 
Reporting  
The CI shall submit once a year throughout the study, or on request, an Annual Progress Report 
to the REC, HRA (where required), host organisation and Sponsor. In addition, an End of Study 
notification and final report will be submitted to the REC and Sponsor. 
15.5 
Volunteer confidentiality 
All data will be de-identified; volunteer data will be identified only by a unique study number in 
the CRF and database. Separate confidential files containing identifiable information will be 
stored in secured locations. Only the Sponsor representative, 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.  
Results of any COVID-19 swab tests carried out through this study will be legally required to be 
reported to Public Health England. Along with the result of the COVID-19 test, the following 
details will also reported: personal details of the participant (name, date of birth, contact details, 
ethnicity, NHS number) and date of diagnosis and symptom onset if applicable.  
 

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16 DATA HANDLING AND RECORD KEEPING 
16.1 
Data handling 
The Chief Investigator will be the data manager 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. This includes safety data, laboratory 
data and outcome data. Data are entered in a web browser on PCs 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 secure server.  
16.2 
Record keeping 
The Investigators will maintain and retain appropriate medical and research records and 
essential documents for this trial in compliance with ICH E6 GCP and regulatory and institutional 
requirements for the protection of confidentiality of volunteers. The Chief Investigator, 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. 
16.3 
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; 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, urine assessments, blood results, adverse event data and details of study 
interventions. All source data and volunteer CRFs will be stored securely. 
16.4 
Data protection  
The trial will comply with the General Data Protection Regulation (GDPR) and Data Protection Act 
2018. The study protocol, documentation, data and all other information generated will be held 
in strict confidence. No information concerning the study or the data will be released to any 
unauthorised third party, without prior written approval of the Sponsor. 
Results of any COVID-19 swab tests carried out through this study will be legally required to be 
reported to Public Health England. Along with the result of the COVID-19 test, the following 
details will also reported: personal details of the participant (name, date of birth, contact details, 
ethnicity, NHS number) and date of diagnosis and symptom onset if applicable.  

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17 FINANCING AND INSURANCE 
17.1 
Financing 
The study is funded through a grant from The Wellcome Trust.  
17.2 
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). NHS indemnity operates in respect of the clinical 
treatment that is provided. 
17.3 
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 
- Admission (Illness) Compensation                                                  
£480 total 
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 20:  Estimated compensation amounts. 
 
Time in Trial
(approx.) 
Maximum No. of 
Visits 
Maximum Volume of 
Blood Taken (mL) 
Compensation 
(up to 
approximately)
Group 1 
2 years 
65 
1415 
£3500 
Group 2 
1 year 
51 
1357 
£2890 
Group 3  
total including 
Group 1 visits 
9 months 
39 
897 
£2355 
2.5 years 
104 
2242 
£5810 

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18 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 21: 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 female volunteers, a menstrual history 
will be taken to elicit whether the subject 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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