A Clinical Study to Assess the Safety and Feasibility of Controlled Blood-stage Plasmodium Vivax Human Malaria Infection Through Experimental Inoculation of Cryopreserved Infected Erythrocytes in Healthy Thai Adults
Phase: NA
Conditions: Plasmodium Vivax Infection
Sponsor: University of Oxford

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 1 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
PART A: PROJECT INFORMATION 
A1. TITLE OF THE STUDY IN THAI AND ENGLISH 
English:   A clinical study to assess the safety and feasibility of controlled blood-stage Plasmodium 
vivax human malaria infection through experimental inoculation of cryopreserved infected 
erythrocytes in healthy Thai adults 
Study Code:  MAL21001 
Trial Registry Name: ClinicalTrials.gov; ClinicalTrials.gov ID: NCT05071079 
A2. RESEARCH PROPOSAL VERSION DATE:  
v.7.0, dated 3 January 2024  
Type of submission     Initial review (first time submission) 
   Revision according to TMEC suggestions No… 
   Amendment No. ….4… 
A3. PRINCIPAL INVESTIGATOR NAME: Professor Nicholas Day and Research Professor            
Dr. Jetsumon Sattabongkot Prachumsri 
 
 
 
 Faculty Staff (Go to A4.1) 
 
 Student, ID………………..…  (Please check the following, and go to A4.2)  
 
 Research for Thesis 
   Research for Thematic Paper 
 
  M.Sc. (Trop. Med.)  
 
  M.C.T.M. 
 
 
  Ph.D. (Trop. Med.)  
 
  M.C.T.M. (T.P.) 
 
 
  Ph.D. (Clin. Trop. Med.) 
 
  Other, specify …………… 
 
 
 Other  
Mahidol-Oxford Tropical Medicine Research Unit (MORU) Staff  
                                         (Please specify, and go to A4.1) 
A4. LIST NAME, AFFILIATION AND CONTACT DETAILS OF ALL INVESTIGATORS 
A4.1 For Faculty staff and other 
Name 
Position 
Contact address 
E-mail address 
Professor Nicholas 
Day 
Co-Principal 
Investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU) 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
nickd@tropmedres.ac 
Research Professor 
Dr. Jetsumon 
Sattabongkot 
Prachumsri 
 
Co-Principal 
Investigator 
Mahidol Vivax Research Unit 
(MVRU) 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road  
jetsumon.pra@mahidol.ac
.th 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 2 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Name 
Position 
Contact address 
E-mail address 
Bangkok 10400 Thailand 
Assist. Professor 
Borimas 
Hanboonkunupakarn  
 
Accountable 
Investigator 
(FTM Thai 
Staff) and  
Site-
Principal 
Investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU)  
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
borimas@tropmedres.ac 
 
Professor Sasithon 
Pukrittayakamee 
 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU)  
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
yon@tropmedres.ac 
Assist. Professor 
Podjanee Jittamala 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU)  
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
podjanee@tropmedres.ac 
Assoc. Professor 
Kittiyod 
Poovorawan 
 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU) Faculty of Tropical 
Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
kittiyod.poo@mahidol.ac.
th 
Professor Sir Nick 
White 
 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU) Faculty of Tropical 
Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
nickwdt@tropmedres.ac 
Professor Arjen 
Dondorp 
 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU) Faculty of Tropical 
Medicine 
arjen@tropmedres.ac 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 3 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Name 
Position 
Contact address 
E-mail address 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
Dr. Angela M 
Minassian 
 
Co-
investigator 
Centre for Clinical 
Vaccinology and Tropical 
Medicine, 
Jenner Institute, University 
of Oxford, 
Churchill Hospital, Old 
Road, Headington, 
Oxford, OX3 7LE 
angela.minassian@ndm.o
x.ac.uk 
Professor Philip 
Bejon 
 
Co-
investigator 
Kemri-Wellcome Trust 
research program 
Kilifi, Kenya 
pbejon@kemri-
wellcome.org 
Professor Mallika 
Imwong 
Co-
investigator 
Molecular Tropical Medicine 
and Genetics 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
mallika.imw@mahidol.ac.
th 
Assist. Prof. Wang 
Nguitragool 
Co-
investigator 
Molecular Tropical Medicine 
and Genetics 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
Wang.ngu@mahidol.edu 
Professor Kesinee 
Chotivanich 
 
Co-
investigator 
Mahidol-Oxford Tropical 
Medicine Research Unit 
(MORU) Faculty of Tropical 
Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
nok@tropmedres.ac 
Dr. Wanlapa 
Roobsoong 
 
Co-
investigator 
Mahidol Vivax Research Unit 
(MVRU) 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
wanlapa.ros@mahidol.edu 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 4 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Name 
Position 
Contact address 
E-mail address 
Assist. Professor 
Patchara Sriwichai  
Co-
investigator 
The Department of Medical 
Entomology 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
patchara.sri@mahidol.ac.t
h 
Assoc. Professor 
Pornsawan 
Leaungwutiwong 
 
Co-
investigator 
Tropical Medicine Diagnostic 
Reference laboratory 
Department of Microbiology 
and Immunology 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
 
pornsawan.lea@mahidol.a
c.th 
A4.2 For students:   Not applicable 
Name 
Position 
Contact address 
E-mail address 
 
Principal Investigator 
 
 
 
Accountable Investigator 
(Student/Advisor) 
 
 
 
Advisor 
 
 
 
Co-advisor 
 
 
 
Co-advisor 
 
 
A5. RESPONSIBILITY OF PI 
  Whole study  
 
    Partial responsibility  
A6. NUMBER OF YOUR ONGOING RESEARCH PROJECT(S) 
1) Professor Nicholas Day:  5 studies 
2) Research Professor Dr. Jetsumon Sattabongkot Prachumsri:  8 studies 
A7. CONTACT PERSON  
1. Professor Nicholas Day (Co-Principal Investigator) 
Mahidol-Oxford Tropical Medicine Research Unit (MORU) 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 5 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Intervention study 
Bangkok 10400 Thailand  
Email: nickd@tropmedres.ac 
2. Research Professor Dr. Jetsumon Sattabongkot Prachumsri (Co-Principal Investigator) 
Mahidol Vivax Research Unit (MVRU) 
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
Tel. 02 306 9187 
Email: jetsumon.pra@mahidol.ac.th 
Mobile phone: 08 1611 4241 
3. Assist. Professor Borimas Hanboonkunupakarn (Accountable Investigator and Site-Principal 
Investigator) 
Mahidol-Oxford Tropical Medicine Research Unit (MORU)  
Faculty of Tropical Medicine 
Mahidol University 
420/6 Rajvithi Road 
Bangkok 10400 Thailand 
Email: borimas@tropmedres.ac 
Mobile phone: 08 6970 5705  
A8. NATURE OF THE STUDY- STUDY INVOLVING SPECIMEN COLLECTION 
  Clinical trial phase…-…/  
  Bioequivalence/ pharmacokinetic drug study 
  Prospective epidemiological research 
  Laboratory study 
A9. IS THE PROJECTS A SINGLE CENTER OR MULTI-CENTER? 
  Single center 
  Multicenter (within Thailand)  
 Please specify the study sites ……………………………………………........ 
  Multicenter (International) 
 Please specify the study sites …………………………………........................ 
 
 
 
 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 6 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
A10. PROJECT SUMMARY IN THAI AND ENGLISH 
A10.1 Full protocol summary in English (not more than 1,000 words per language) 
This project is the second part of a 5-year research program entitled "Malaria Infection Study in 
Thailand (MIST)" and known as MIST2. MIST2 primary objectives are to assess the safety and 
feasibility of blood-stage controlled human P. vivax malaria infection (CHMI) in healthy adult 
Thai volunteers through experimental injection of cryopreserved P. vivax infected erythrocytes, 
and to choose the optimal inoculation dose for future P. vivax CHMI studies. In this study, 
blood-stage CHMI will be conducted in 8 volunteers per inoculum stock who will be infected 
with P. vivax by experimental injection with cryopreserved P. vivax infected erythrocytes, 
which were collected in MIST1. As there will be 6 stocks of inocula from 6 volunteers in the 
MIST1 study, which have different quantity and stage of parasites. The total number of 
volunteers would be up to 48. The volunteers will be monitored closely as in-patients in the 
Hospital for Tropical Diseases, and will be treated according to the Research Proposal 
Submission Form. 
 
A10.2 Part of study conducted by PI  
  Whole Study  
  Part of the Study 
A11. SOURCE(S) OF FUNDING/ SPONSOR(S) AND BUDGET (Information required for 
review and consideration) 
  Funded by:    
Wellcome Trust  
 
 Budget amount:  
2,000,000 THB 
 
  Expecting fund from: ………………… 
Budget amount: …………………………………...………….……………. 
A12.  DECLARE CONFLICT OF INTEREST  
The Co-Principal Investigators and the Investigators declare they have no conflict of interest. 
PART B: DETAILS OF THE STUDY (Describe only the responsibilities of the PI) 
B1. BACKGROUND AND RATIONALE 
The P. vivax malaria problem 
Plasmodium vivax (P. vivax) is one of the five Plasmodium species to cause human malaria and 
accounts for the most cases of non- falciparum malaria worldwide.  It is now the most widespread 
human malarial infection in endemic areas outside Africa.  The main reasons for the wide 
geographical distribution of P. vivax are likely to be related to the following aspects of P. vivax 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 7 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
biology: (1) its ability to relapse from dormant liver stage (hypnozoite), (2) its high transmission 
potential due to early production of gametocytes, high infectivity to mosquitoes and shorter 
development cycle in the vector - host compared to other plasmodia (1, 2). 
P. vivax has been considered as causing ‘benign’ malaria, but recent large case series have 
demonstrated that P. vivax infection is associated with significant morbidity and mortality (3). 
Clinical cases are not only due to primary infection following the infected mosquito bite but also 
relapses from the hypnozoite stage, which occur weeks to years after primary infection (4). Standard 
schizonticidal regimens are not effective against the hypnozoite. Radical cure of the hypnozoite stage 
requires therapy with the 8-aminoquinolines primaquine and tafenoquine. However, both 
medications carry a significant risk of severe haemolytic anemia in individuals with glucose-6-
phosphate dehydrogenase deficiency (G6PD deficiency) (5). There is no cheap, reliable point-of-
care G6PD test, so many P. vivax patients do not currently receive adequate anti-hypnozoite 
treatment. More recently, the importance of the cytochrome P450 enzyme CYP2D6 in the 
metabolism of primaquine to the active metabolite has been recognized (6) (7). A common 
polymorphism in the CYP2D6 gene for the cytochrome P450 enzyme results in poor conversion of 
primaquine to the active form, resulting in treatment failures. It is estimated that these two factors 
combined may make nearly 40% of the population at risk of P. vivax infection potentially ineligible 
for primaquine therapy (5). 
Recent calls for control and ‘eradication’ of malaria worldwide 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. Consequently, the revised Malaria Vaccine Technology Roadmap to 2030 
now recognizes the importance of P. vivax and calls for development of a vaccine to achieve 75% 
efficacy over two years – equally weighted with P. falciparum in an era of renewed political will to 
move towards malaria elimination and eradication (8) (9). More recently, research into vivax malaria 
has increased with candidate vaccines being developed and taken forward to clinical trial (10) (11).  
 
Challenges and directions in P. vivax vaccine development.  
P. vivax malaria vaccine approach 
Most of the P. vivax malaria vaccine candidates that are currently under development target 
individual stages of the P. vivax parasite’s life cycle. Several highly abundant parasite proteins have 
been identified as targets of natural immunity and in the recent years the list of possible candidates 
has expanded.  
Subunit vaccines for P. vivax have been developed based on these candidate antigens. There are 
three major approaches for the development of malaria vaccines that correspond to the three stages 
of the parasite’s life cycle as follow:  
 Pre-erythocytic antigens; vaccine using these antigens aim to prevent initial infection by 
targeting the infective stage (sporozoites) that were introduced by the mosquito bite. 
Alternatively, these vaccines can target antigens expressed by liver stage parasites to prevent 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 8 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
the emergence of merozoites into the blood stream e.g. P. vivax circumsporozoite protein 
(PVCSP), P. vivax thrombospondin related adhesive protein (PvTRAP) 
 Blood stage antigens; since all of the symptoms of malaria occur during this stage, the 
majority of vaccines targeting antigens expressed during the blood stage are designed 
primarily to prevent disease or morbidity and mortality associated with the disease when 
parasites are in red blood cell (RBC). One approach is to target merozoite antigens to 
prevent red blood cell invasion and reduce the density and prevalence of parasites in the 
infected host RBC e.g. receptor-binding region II of P. vivax Duffy binding protein 
(PvDBPII), P. vivax merozoite surface proteins (MSP 1), P. vivax apical merozoite antigen 
1 (PvAMA1), P. vivax reticulocyte binding protein/ homolog (PvRBP) 
 Transmission blocking antigens; the transmission blocking vaccines target antigens 
expressed during parasite lifecycle stages in the mosquito (e.g. gametocyte or oocyst 
antigens). Transmission blocking vaccines aim to block malaria transmission from 
mosquitoes to humans by preventing the malaria parasite from developing in the 
mosquito. Although these vaccines would not directly prevent infection or disease, they 
would assist elimination efforts through preventing the onward transmission of infections 
e. g. parasite proteins expressed on surface of zygote (Pvs25) and ookinete (Pvs28), alanyl 
aminopeptidase 1 (AnAPN1), a highly conserved midgut surface antigen of Anopheles 
mosquitoes that is essential for ookinete invasion and development). (Table 1, Figure 1) 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 9 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Figure 1. Malaria vaccine leading candidate antigens
 
 
Ref. Barry and Arnott et al. Frontiers in Immunology, 2014 (359) 
 
 
 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 10 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Table 1. P. vivax malaria vaccine candidates under development 
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-DEKnull 
Pre-clinical 
Blood-stage 
PvDBP 
Rec. protein 
PvDBPII 
Phase I 
Blood-stage 
PvDBP 
Rec. protein-GLA-SE 
PvDBPII 
Phase I/IIa 
Blood-stage 
PvDBP 
Rec. protein- Matrix-M1 
ChAd63-
PvDBPII/MVA-
PvDBPII 
Phase I/IIa 
Blood-stage 
PvDBP 
Prime boost, viral vectors 
PvMSP119 
Pre-clinical 
Blood-stage 
PvMSP1 
Rec. protein-Montanide 
ISA720 
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 
 
Why controlled human challenge trial?  
Conventional vaccine development is complex. The average development timeline is between 8 and 
18.5 years, estimated costs are $200 million to $900 million, and the probability of success of less 
than 10% (12, 13). Vaccine clinical trials aim to answer questions related to the immune response, 
immune correlates of protection, protective efficacy, therapeutic window, and target populations. It 
is expected that most vaccines will fail because of poor “immune correlates of infection” and 
“protective efficacy”. Increasing recognition of this critical bottleneck in conventional vaccine 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 11 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
development has led to demand for a proof-of-concept clinical trial: the controlled human 
challenge trial. 
 
In July 2014, the World Health Organization (WHO) held a consultation on Clinical Evaluation of 
Vaccines: regulatory expectations and controlled human challenge trials were considered important 
issues for facilitating vaccine development (14). Controlled human challenge trials are thus a prime 
example of ways in which clinical development of vaccines can be accelerated. Controlled human 
challenge trials in malaria initially raised ethical debate, but have now gained acceptance through 
demonstrating their potential and the fact that they have been safely performed in >3,000 volunteers 
all over the world (15). 
 
The development of a safe and reproducible P. vivax-controlled challenge model in humans could 
greatly accelerate clinical development of P. vivax vaccines, through selecting efficacious candidates 
for further clinical testing in more expensive and logistically challenging studies in the field.  
 
P. vivax human challenged study in Thailand 
This study is part of the long term project "Malaria Infection Study in Thailand (MIST)" funded by 
Wellcome Trust to develop the feasibility of P. vivax controlled human challenge trial in an endemic 
area. The following section provides an overview of the projects in this program, which comprises 
multiple studies to be conducted conduct over the next 5 years in the Faculty of Tropical Medicine, 
Mahidol University, Thailand. 
 
Most vivax malaria challenge studies performed so far have been in non-endemic settings such as in 
the US, UK, and Australia. However, the findings in those settings may not be extrapolatable to the 
target population for future vaccine deployment, which is more heterogeneous both in terms of vivax 
immunity and genetic background. The best volunteer population in which to test new vaccines is 
the eventual target population for vaccine deployment. To date, the MIST 1 is the only CHMI to be 
performed in a P. vivax endemic population in Asia.   
 
P. vivax human challenge studies conducted in Thailand would allow us to understand the 
immunological correlates of protection in an endemic setting, thereby informing the development of 
new P. vivax vaccine candidates and rapid testing of the protective efficacy of candidate vaccines in 
the at-risk population in which they will be deployed. Advancing the development of such methods 
needs renewed emphasis on understanding the biology, pathogenesis and transmission of P. vivax.  
 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 12 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
The overall objectives of the MIST program 
 
 
 Proof of feasibility and safety study and providing P. vivax banked infected blood inocula 
for future P. vivax blood stage human challenge studies.  
 
 Test the protective efficacy of pre-erythrocytic, blood stage and (ultimately) transmission 
blocking P. vivax vaccine candidates in the target population. 
 
 Characterise correlates of P. vivax immune protection in the target population. 
 
 Test the efficacy of new drug candidates under development. 
 
 
o To achieve these major aims written above, the programme will compose of 4 to 7 studies. 
Each subsequent study will be informed by the previous studies in the series and each study 
will be covered by separate protocols with separate Ethics Committee submissions and 
approvals.   
 
o The MIST2 study (covered by this current protocol): Blood stage inoculum human 
challenge in volunteers from target population (using banked blood from MIST1) to find the 
proper dose for the subsequent studies assessing blood and transmission stage vaccine 
candidates.  
 
The MIST2 study 
 
During mid- year 2020, the Faculty of Tropical Medicine Ethics Committee (FTM-EC) and the 
Oxford Tropical Research Ethics Committee (OxTREC) approved the protocol “A clinical study to 
assess the feasibility of a controlled human Plasmodium vivax malaria infection model through 
experimental sporozoite infection in Thai adults (MIST1)”, MAL19002 (FTM-EC #TMEC19-067, 
OxTREC # 43-19 The study was registered to the Clinicaltrials.gov, # NCT04083508. 
 
The MIST1 study aimed to assess the safety and feasibility of controlled human P. vivax malaria 
infection following infection by the natural route of delivery – mosquito bite. Another major 
objective of the study was to provide a source of P. vivax infected blood to use in future vaccine 
efficacy trials of a blood- and transmission-stage vaccines.  
 
Up to date, sporozoite inoculation by mosquito bite has been done in four healthy subjects 
successfully in the Hospital for Tropical Diseases and blood banks were prepared for the challenge 
stocks to be used in MIST2 and further blood-stage vaccine trials. 
 
This MIST2 protocol is a blood stage inoculum dose finding study. It is a blood-stage P. vivax human 
challenge study with the primary aim of assessing the safety and feasibility of a challenge model 
using banks of cryopreserved P. vivax infected erythrocytes for the first time (using the inocula 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 13 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
produced from MIST1) to identify the dose of the inocula to be used in future studies. Four different 
doses of inoculum will be assessed, by injection at four dilutions of each banked blood, with a view 
to identifying the lowest concentration producing a reliable infection within a practicable timeframe.  
This study will serve as the basis for a challenge model for future P. vivax candidate vaccine efficacy 
studies. 
 
Blood stage P. vivax vaccine experience 
 
There have been 8 studies of blood-stage P. vivax human challenge, of which 5 were published. Five 
studies were conducted at the Queensland Institute of Medical Research (QIMR), Brisbane, 
Australia, and 3 studies have been recently conducted by the University of Oxford, UK. The details 
of the currently unpublished Oxford studies are as follows: 
In 2018, Oxford University undertook the first proof-of-concept studies in Europe of P. vivax human 
challenge (ClinicalTrials.gov: NCT03377296). Two healthy UK adult volunteers were safely 
infected with a clonal strain of P. vivax by the bite of infected mosquitoes transported from Thailand. 
And 250 mL blood was obtained from both volunteers at a prespecified parasitaemia/clinical 
threshold, leading to the production of a P. vivax-parasitized erythrocytes (pRBC) for use in future 
blood-stage human challenge studies as summarized below.  
VAC 069 A-F (NCT03797989) was the first blood-stage human challenge study using pRBCs from 
a sporozoite inoculation study by mosquito bite. The safety and feasibility of blood-stage infection 
at 3 different dilutions (whole vial or a neat dose, 1:5 dilution and 1:20 dilation) of thawed inoculum 
were tested in 6 malaria-naïve healthy UK adult volunteers (2 subjects x1 dilution strength) 
(VAC069A). Intravenous inoculation of volunteers was completed without complication. All 
volunteers were diagnosed within 12.5-16.5 days of infection and all had detectable gametocytaemia 
by qPCR within 1-3 days of peak parasitaemia. After antimalarial treatment with 
artemether/lumefantrine or atavoquone/proguanil, qPCR was negative by 6 days post-treatment 
initiation in all volunteers. The full study will be reported soon (Minassian et al., in preparation).  
The majority of volunteers (4/6) experienced either no or mild AEs, whilst 2/6 volunteers reported 
malaria related clinical signs and symptoms. The majority of the AE symptoms resolved within 24 
hours after the treatment initiation. Of the laboratory AEs recorded, transient lymphocytopenia and 
derangement of alanine transaminase (ALT) was seen in 2 volunteers. Both volunteers were 
asymptomatic.  
 
Based on these infectivity data, and accounting for possible variation in the number of pRBC 
between thawed cryopreserved vials, an inoculum dilution of 1:10 was selected for future P. vivax 
blood-stage human challenge studies. The inoculum dilution of 1:10 was further tested in 2 malaria-
naïve healthy UK adult volunteers (VAC 069 B) and showed qPCR positivity on day 9. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 14 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
The VAC 069 protocol was also designed to include re-infection (re-challenge blood -stage human 
control trial) and immunity studies (VAC 069 C - F). All re-challenge blood-stage human challenges 
were performed in parallel with leading candidate blood-stage vaccine efficacy studies  
B2. STUDY OBJECTIVES AND ENDPOINTS 
Primary objectives: 
1. To assess the safety and feasibility of blood-stage controlled human P. vivax malaria 
infection in healthy adult Thai volunteers through experimental injection of cryopreserved 
P. vivax infected erythrocytes at different doses 
2. To choose the optimal inoculation dose for future P. vivax CHMI studies. 
Primary endpoints: 
1. Safety and feasibility of primary P. vivax blood-stage CHMI, as measured by (S)AE 
occurrences and successful infection (development of detectable persistent parasitaemia by 
thick blood film +/- clinical symptoms) 
2. Choosing the optimal inoculation dose for future P. vivax CHMI studies, which will be the 
lowest concentration that produces a reliable infection within a comparable timeframe as 
compare to the highest concentration.  
 
Secondary objectives: 
1. To assess the immune response to P. vivax infection in volunteers, through experimental 
injection of P. vivax infected erythrocytes 
2. To assess gametocytaemia during P. vivax infection in volunteers, through experimental 
injection of P. vivax infected erythrocytes 
3. To assess transmission of gametocytes from infected volunteers to Anopheles mosquitoes 
Secondary endpoints: 
1. Immune response to experimental P. vivax infection through blood-stage challenge, as 
measured by antibody, cytokines, B cell and T cell responses 
2. Gametocytaemia, as measured by qPCR in experimental P. vivax infection through blood-
stage challenge 
3. Transmissibility of gametocytes from the infected volunteer to Anopheles mosquito vector 
B3. STUDY SITE(S) 
Clinical Therapeutics Unit (FTMCTU), Hospital for Tropical Diseases, Mahidol Vivax Research 
Unit (MVRU), Medical Entomology Department, Tropical Medicine Diagnostic Reference 
Laboratory (TMDR), and Malaria Laboratory, Faculty of Tropical Medicine, Mahidol University, 
Bangkok, Thailand  
B4. STUDY TIMELINES: 
1. There will be an up to 14 days screening process prior to the malaria challenge day.   
2. The duration that individual volunteers will be in the study is 1 year from starting anti-
malarial treatment until the end of the follow-up  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 15 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
3. After completion of the last volunteer’s last visit, there will be an estimated 3 months for 
database lock and an estimated 3 months to analyse the data.  
4. The total duration of this study is approximately 5 year and 9 months starting after EC 
approvals. 
B5. STUDY POPULATION AND SAMPLE                                                                                   
 
B5.1 Target population  
Forty-eight healthy Thai adults, aged from 20 to 55 years with at least an undergraduate degree, 
will be recruited at the FTMCTU at the Hospital for Tropical Diseases, Faculty of Tropical 
Medicine, Mahidol University, Bangkok, Thailand. More volunteers would be required if there 
are more stocks of inoculum in the future. 
 
B5.2 Subject selection criteria  
B5.2.1 Subject/inclusion criteria  
The volunteer must meet all of the following criteria to be eligible for the study: 
1. Healthy Thai adult aged 20 to 55 years with weight at least 50 kg. 
2. Red blood cells positive for the Duffy antigen/chemokine receptor (DARC) 
3. Women only: Must practice continuous effective contraception for the duration of study 
period until 3 months post-challenge. 
4. COVID-19 vaccination at least two doses of COVID-19 vaccines approved by WHO.   
5. Agreement to refrain from blood donation during the course of the study and for 1 year after the 
initiation of antimalarial treatment.  
6. Willing to be admitted in the Hospital for Tropical Diseases for clinical monitoring, until 
antimalarial treatment is completed and their symptoms are settling.  
7. Willing to take a curative antimalarial treatment following CHMI. 
8. Willing to reside in Bangkok and its vicinity for 2 months after malarial treatment initiation. 
9. Able to read and write in Thai. 
10. Provide written informed consent to participate in the trial 
11. Answer all questions on the informed consent quiz correctly 
12. Educational level: has at least an undergraduate degree 
  
 B5.2.2 Subject/exclusion criteria  
The volunteer MUST NOT enter the study if any of the following apply: 
1. Positive malaria qPCR OR malaria film  
2. Presence of any medical condition (either physical or psychological) which in the judgment 
of the investigator would place the participant at undue risk or interfere with the results of 
the study (e.g. serious underlying cardiac, renal, hepatic or neurological disease; severe 
malnutrition; congenital defects or febrile condition) 
3. Presence of chronic disease or chronically use of medication 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 16 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
4. Use of systemic antibiotics with known antimalarial activity in the 30 days before 
challenge (e.g. trimethoprim-sulfamethoxazole, doxycycline, tetracycline, clindamycin, 
erythromycin, fluoroquinolones and azithromycin)  
5. Use of immunoglobulins or blood products (e.g. blood transfusion) at any time in the 1 year 
preceding enrolment 
6. Receipt of an investigational product, any vaccine in the 30 days preceding enrolment (D0), 
or planned receipt during the study period 
7. 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. 
8. Any confirmed, or suspected immunosuppressive, or immunodeficient state, including HIV 
infection, asplenia, history of splenectomy, recurrent, severe infections, and chronic 
infection 
9. Immunosuppressant medication within the past 6 months preceding enrolment (D0) (inhaled 
and topical steroids are allowed) 
10. History of allergic disease or reactions likely to be exacerbated by malaria infection 
11. Female participant who is pregnant as evidenced by positive beta-human chorionic 
gonadotropin (β-HCG) test, lactating, or planning pregnancy during the course of the study  
12. Contraindications to the use of antimalarial treatment (e.g. chloroquine, atovaquone / 
proguanil or dihydroartemisinin/piperaquine)  
13. Use of medications known to have a potentially clinically significant interaction with the 
antimalarial drug that will be used in this study (chloroquine, atovaquone / proguanil or 
dihydroartemisinin/piperaquine) 
14. Known existing positive family history in both 1st AND 2nd degree relatives < 50 years old 
for cardiac disease 
15. History of cardiac arrhythmia, including clinically relevant bradycardia 
16. Family history of congenital QT prolongation or sudden death 
17. Any clinical condition, including using medications, known to prolong the QT interval. 
18. Screening electrocardiogram (ECG) demonstrates a QTc interval ≥ 450 ms.  
19. Suspected or known or history of alcohol abuse  
20. Suspected or known or history of drug abuse.  
21. Concurrently participating in another clinical study, at any time during the study period 
22. Haemoglobin < 11 g/dL 
23. Positive hepatitis B surface antigen or seropositive for hepatitis C virus 
24. Finding on safety laboratory values as defined below: 
 Abnormal AST (AST > 40 U/L for male, and > 32 U/L for female [upper normal range]) 
 Abnormal ALT (ALT > 41 U/L for male, and > 33 U/L for female [upper normal range]) 
 Abnormal serum creatinine (Scr) (Creatinine [Cr] > 1.17 mg/dL for male, and > 0.95 mg/dL 
for female [upper normal range])  
 Abnormalities corrected calcium and magnesium blood levels  
25. Blood group Rhesus negative 
26. Blood incompatibility to the inoculum  
27. Positive for COVID-19 diagnosed by RT-PCR 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 17 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
B5.3 Sample size calculation 
In order to find the optimum dose of each blood-stage inoculum (the lowest concentration that the 
volunteers reliably develop infection, i.e., 2/2, within the comparable time frame as compare to the 
highest concentration and within 21 days), one blood-stage inoculum stock will be tested in 4 
different doses: whole dose inoculum (neat vial), 1:5, 1:10, and 1:20 dilution. Each dose will be 
inoculated into two volunteers resulting in 8 volunteers. We plan to collect six stocks of blood-stage 
inoculum from MIST1, four already obtained and two will be collected,  48 volunteers will be 
recruited: 8 to be challenged with each inoculum stock.   
B5.4 Recruitment Methods 
Volunteers will be screened and recruited at the FTMCTU, Faculty of Tropical Medicine, Mahidol 
University in Bangkok following the Recruitment Process for Healthy Volunteer Standard Operating 
Procedure (SOP).  
In brief, the volunteers may be recruited by use of an advertisement approved by the ethics 
committee and distributed or posted in the following places: 
 On a MIST website operated by the study group   
 In public places with the agreement of the owner/proprietor 
 Via presentations (e.g., presentations at lectures or invited seminars) 
In addition, we may contact individuals who have participated in previous clinical trials from the 
FTMCTU’s databases. These volunteers will have expressed an interest in receiving information 
about all future studies in FTMCTU for which they may be eligible. 
B5.5 Withdrawal/discontinuation criteria  
In accordance with the principles of the current revision of the Declaration of Helsinki (updated 
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). 
 Volunteer non-compliance with study requirements. 
 An AE which requires discontinuation of the study involvement or which results in inability 
to continue to comply with study procedures. 
 Current COVID-19 infection during a participant’s admission in the hospital 
The medical monitors may recommend the withdrawal of volunteers. The reason for withdrawal 
from further study procedures will be recorded in the Case Record Form (CRF). If a volunteer 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 18 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
withdraws after having completed a course of antimalarials, as much long-term safety data collection 
as possible, including procedures, such as safety bloods, will continue to be collected, with the 
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, stabilized, or a non-trial-
related causality has been assigned. 
If a volunteer withdraws from the study after the challenge but before reaching the criterion for 
malaria diagnosis, a complete, appropriate, curative course of antimalarial therapy must be 
completed with standard chloroquine treatment. The importance of this will be emphasized to 
volunteers at screening. If a volunteer refuses to take antimalarial therapy after malaria diagnosis, a 
rapid assessment of mental state and capacity will be undertaken, with the involvement of psychiatric 
and infectious diseases specialists. If necessary and if in accordance with the law in Thailand, the 
volunteer may be detained for appropriate medical management. 
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 volunteer withdrawal, except those of complete consent withdrawal, safety data 
collection will continue if volunteers have already undergone the challenge process.  
B5.6 Multi-site research 
- Not applicable 
B6. RESEARCH METHODOLOGY 
B6.1 Details of study design 
Summary of trial design 
Controlled Plasmodium vivax human malaria infection through experimental inoculation of 
cryopreserved infected erythrocytes in healthy Thai adults, with a randomized double-blind design. 
Overview 
This blood-stage P. vivax human challenge study conducted for the first time in Asia has two 
primary aims: (1) to assess the safety and feasibility of the challenge model using the bank of 
cryopreserved P. vivax infected erythrocyte inocula prepared during the MIST1 study, and (2) to 
determine the optimal inoculation dose for future P. vivax CHMI studies. This will be assessed by 
intravenously injecting the volunteers with four different doses of inoculum: whole dose and 1:5, 
1:10, and 1:20 dilutions. If safe and feasible, this study will serve as the basis for a challenge model 
for future P. vivax candidate vaccine efficacy studies. The secondary objectives are to determine the 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 19 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
immune response to P. vivax infection, the resulting gametocytaemia, and transmissibility of the 
gametocytes from the infected volunteers.   
Healthy, Thai adults aged between 20 and 55 years will be recruited and randomized at the Faculty 
of Tropical Medicine, Mahidol University in Bangkok.  
CHMI will be induced by injection of P. vivax infected erythrocytes and all follow-up in the post-
challenge period will be performed at the Faculty of Tropical Medicine, Mahidol University in 
Bangkok. 
Volunteers will have blood taken at regular intervals post-CHMI to assess the immune response to 
P. vivax infection, as well as parasite growth dynamics and gametocytaemia. We will also assess 
transmission of P. vivax gametocytes from the infected volunteers to mosquitos using a membrane 
feeding assay (MFA).  
Close monitoring will continue until volunteers meet criteria for treatment or until 21 days after 
challenge, when treatment will be started empirically. 
Therapy will be with a standard course of chloroquine where not contraindicated. As infection will 
be induced via intravenous injection of blood-stage parasites, there will be no liver-stage infection 
and no hypnozoite formation, thereby eliminating the need for radical cure with primaquine therapy. 
Follow-up at study site will be up to 1 year after antimalarial treatment initiated. 
B6.2 Details of procedures for specimen/data collection 
B6.2.1. 
Screening and eligibility assessment 
B6.2.1.1. Screening visit 
The screening will aim to recruit 48 healthy volunteers.  
Screening visits may take place up to 14 days prior to enrolment. Informed consent will be conducted 
at screening. If consent is given, the screening procedures will be undertaken, including testing for 
Duffy antigen/chemokine receptor (DARC) positivity, blood diseases, blood biochemistry, blood-
borne infections, and serum pregnancy test. The full list of laboratory tests and the schedules of 
procedures is listed in Table 4. 
Abnormal clinical findings from the medial history, physical examination, or laboratory tests at any 
point in the study will be assessed according to the scales in APPENDIX C: Safety terms and 
administrative information. If a test is deemed clinically significant it may be repeated to ensure it 
is not a single or spurious 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.  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 20 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Cross matching will be performed to check blood compatibility between volunteer and the blood 
inoculum. 
Exclusion of the volunteer from enrolling in the trial or withdrawal of a volunteer from the trial will 
be at the discretion of the Investigators. 
B6.2.1.2. Day before CHMI (Admission day; D -1) 
Any new medical issues or symptoms that have arisen will be assessed. All eligible volunteers will 
have physical examination/observations will be performed during this visit. Serum β-HCG (woman), 
malaria diagnosis, complete blood count (CBC), biochemistry, and COVID-19 testing (RT-PCR will be 
tested during this visit. In addition, COVID-19 serology test will also be done to look at participants’ 
immunity from the current/previous COVID-19 infection (asymptomatic) and/or prior immunization. 
G6PD test and malaria immunological profiles will be tested for baseline information only and will 
not be used as exclusion criteria. CMV IgM/IgG antibody, EBV VCA IgM/IgG antibody, and anti 
EBNA and EA complex antibody will be tested. Results of blood taken at this visit must be available 
and reviewed prior to challenge. 
B6.2.1.3. Day of CHMI (D0) 
Half of the number of challenged volunteers must also be available during the period of the malaria 
challenge (during the whole inoculation process) for back-up, in case one of the volunteers in the 
batch withdraws at the last minute or any unexpected event occurs. This means that confirmation of 
whether or not a back-up volunteer will be needed for the challenge will not be made until the 
challenge (the whole inoculation process) has been successfully completed. 
CHMI will be administered according to 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 verification of eligibility / contraindications. 
 Physical observations (including respiratory rate, pulse rate, blood pressure and temperature), 
with recordings to be repeated if this is longer than 60 minutes before administration of the 
parasitized erythrocyte inoculum. 
 Intravenous access via cannulation in a 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 post-administration of inoculum and again 
at 1 hour, in order to assess for immediate adverse reactions, with regular visual observation 
throughout, in line with The British Society for Haematology Guideline for the administration 
of blood products (23). If the volunteer shows 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. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 21 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 If there have been no symptoms or signs indicative of a transfusion reaction, the cannula will 
be removed after 1 hour. 
B6.2.2. 
Days 1-28 post CHMI (D1 to D28) 
This section describes the clinical procedures for evaluating study volunteers and management after 
challenge.  
B6.2.2.1 Day 1 post challenge to malaria qPCR show positive (D qPCR+)  
The volunteers will be assessed once daily from day 1 post challenge until malaria qPCR becomes 
positive (D qPCR+).  The assessment includes a clinical well-being check, and the volunteers will be 
questioned whether they have experienced any malaria-related symptoms (such as fever, chills, 
sweating, malaise). The once daily general vital signs and physical examination will be done. Blood 
will be drawn once daily for monitoring blood parasitaemia (malaria blood film, qPCR, and 
gametocyte qPCR) and membrane feeding to assess the transmissibility of the gametocyte. 
Immunological profile and CBC will be performed on day 4 and DqPCR+.  If volunteer present with a 
clinical feature(s) of COVID-19 at the discretion of the trial physician, and the malaria blood film 
and malaria qPCR are both negative, COVID-19 testing will be performed. The complete list of 
laboratory procedures is shown in Table 5. 
 
B6.2.2.2 Day after qPCR+ (Day After qPCR+) to day of reach treatment criteria (D reach treatment criteria) 
The clinical well-being will be checked. Blood will be drawn twice daily for monitoring blood 
parasitemia (malaria blood film, qPCR, and gametocyte qPCR) and membrane feeding to assess the 
transmissibility of the gametocyte. Malaria immunology, CBC, and blood biochemistry will be 
assessed on D reach treatment criteria. In addition, blood collection will be performed to test for CMV 
IgM/IgG antibody, EBV VCA IgM/IgG antibody, and anti EBNA and EA complex antibody at 2-4 
weeks after the inoculation.  
Volunteers will be assessed after challenge according to the schedule in Table 5. Physical 
observations will be performed. Venepuncture will be performed as per schedule of attendance 
(Tables 5-6). 
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. The severity 
of symptoms will be assessed using grading criteria summarised in APPENDIX C: Safety terms and 
administrative information. 
Antimalarial treatment will be instituted according to criteria described below (Table 2).  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 22 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
If the antimalarial treatment criteria have not yet been met, the treatment shall be initiated at day 21 
after challenge (D21). Subjects will be closely monitored until completion of antimalarial therapy, 
as described above. Summary of clinical assessment and laboratory including safety blood tests is 
shown in Table 5 and detail of blood volume and blood collection tube is shown in Table 6. 
B6.2.2.3 Antimalarial treatment 
A study physician will then immediately prescribe antimalarial treatment with chloroquine, in 
accordance with Table 3. Any side effect from the antimalarials will be rapidly dealt with in a 
controlled environment. They will continue to have blood films and qPCR once daily until clinically 
recovered AND two consecutive malaria blood films are negative (completion of the chloroquine 
treatment course). The volunteers will be then discharged from the hospital. Before discharge from 
the hospital, HIV-I and HIV-II antigen, HBV and HCV serology, COVID-19 testing (RT-PCR) and 
COVID-19 serology test will be carried out to confirm that subjects did not acquire COVID-19 during 
their inpatient stay.  
If any volunteer reaches day 21 post-challenge without a positive malaria blood film, they shall be 
started on a 3-day course of antimalarial (chloroquine). 
If any contraindications to chloroquine are identified, an alternative antimalarial dihydroartemisinin 
+ piperaquine or Malarone (atovaquone + proguanil) will be prescribed. If a volunteer is unable to 
tolerate an oral antimalarial, the appropriate parenteral antimalarial therapy will be prescribed.  
If a volunteer withdraws/is withdrawn from the study after challenge but before reaching the criteria 
for malaria treatment, then a complete, appropriate, curative course of antimalarial therapy 
must be completed. The importance of this will be emphasized to volunteers at screening. 
Table 2: Antimalarial treatment criteria. 
Malaria treatment Start 
Criteria  
 
Symptoms suggestive of malaria infection with the presence 
of blood parasitaemia by microscopy.  
 
Asymptomatic with ≥ 50,000 parasites/mL by microscopy           
 
          OR 
 
 
Asymptomatic and no parasitaemia until Day 21  
Note: In case the volunteers develop symptoms suggestive of malaria 
infection, but no parasitaemia, other causes of symptoms should be 
considered and treated as appropriate. 
Table 3: Chloroquine treatment dose* 
Weight 
Day 1 
Day 2 
Day 3 
CQ Tab 
CQ Tab 
CQ Tab 
50 kg or more 
4 
4 
2 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 23 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
* National Malaria Treatment Guideline 2019 (Thailand), CQ 150 mg base/tablet 
Supportive treatments and medications 
Any malaria symptoms (e.g., fever, nausea, vomiting) will be relieved accordingly. Paracetamol 
(500 mg orally up to four times a day) and dimenhydrinate (50 mg orally) will be prescribed 
according to clinical need. Intravenous fluid supplementation may be given following physician 
decision. 
All medications used in the trial will be recorded in the medical record.  
If the volunteers meet the criteria for severe malaria (by WHO definition) or fail to improve within 
48 hours of starting anti-malarial therapy, malaria experts in the Faculty of Tropical Medicine will 
be consulted. The volunteers will be managed according to National Malaria Treatment Guideline 
(Thailand) as well as the Hospital for Tropical Diseases’ standing order/ procedures/ guideline for 
severe malaria. 
The Investigators are able to treat any volunteer for malaria regardless of the blood film microscopy 
or qPCR result if they are clinically concerned or a volunteer wishes to withdraw from the study.  
B6.2.3. 
Follow up assessment phase (after completion of antimalarial treatment) 
On the antimalarial treatment end date, the volunteers will receive full counselling on the signs and 
symptoms that they have to be able to observe early and report in the diary card and seek medical 
advice. A diary card to self-document any abnormal symptoms will be given to them, and the contact 
channel with the study team will again be emphasized. After discharge from the hospital, the follow-
up assessment will be performed at FTMCTU as an out-patient on day 7 post treatment initiated     
(D Rx7), day 28 (D Rx28), day 60 (D Rx60), day 90 (D Rx90), day 180 (D Rx180), and day 1 year (D Rx 1yr.) 
post treatment initiation. On the follow-up days, diary cards will be reviewed and collected, a history 
taken using the clinical well-being checklist, a physical examination will be performed, and AE(s) 
assessed. Venipuncture will also be performed to detect malaria parasites by blood film and qPCR, 
and for malaria gametocyte qPCR, MFA, HIV-I and HIV-II antigen, HBV, HCV, CMV, and EBV 
serology, malaria immune response, CBC, and biochemistry according to Tables 5-6. Other study 
activities will be undertaken according to Table 5. 
During this follow up phase, the volunteers will be contacted fortnightly (biweekly) until D Rx90 by 
email / phone call / other social communications e.g., message (SMS), WhatsApp, Line, to ensure 
they remain well and asymptomatic. The volunteer will have to understand that they must stay in 
Bangkok until D Rx60 and always immediately contact the study team as soon as they observe any 
abnormal symptoms or would like to contact the team for any other reason. 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 24 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Table 4: Schedule of clinical and study procedures for volunteers pre-challenge  
Event 
Screening  
(within 14 days prior to Day 0) 
Screening 2/Admission 
(Day -1) 
Inclusion / Exclusion criteria 
X 
X 
Informed Consent Questionnaire 
X 
 
Informed consent 
X 
 
Medical History 
X 
X 
Physical Examination  
X 
X 
Body weight and heighta 
X 
X 
Urinalysis 
X 
 
Electrocardiogram 
X 
Vital signsb  
X 
X 
Blood typing ABO, Rhesus, Crossmatching 
X 
 
Duffy antigen for chemokine receptor (DARC) 
X 
CBC 
X 
X 
Haemoglobinopathies and thalassaemia traits*  
X 
 
G6PD*  
 
X 
Blood biochemistryc  
X 
X 
FBS 
X 
 
HIV, HBV, HCVd  
X  
Malaria diagnosis (Malaria antigen – blood film and 
qPCR)  
 
X 
X 
Malaria exposure* 
X 
 
Malaria immunology* 
X 
X 
β-HCG testing (women only) 
X 
(Serum pregnancy test) 
 
X 
(Serum pregnancy test) 
 
COVID-19 testing (RT-PCR) 
 
X 
COVID-19 serology test 
 
X 
CMV and EBV serology 
 
X 
Remark: 
* Tests will be tested for baseline information only and will not be used as exclusion criteria 
a Height will be measured only at screening visit. 
 
b Vital signs include blood pressure, pulse rate, respiratory rate and temperature.  
 
c Biochemistry including electrolytes (sodium, potassium, chloride, bicarbonate, calcium and magnesium), blood urea nitrogen 
(BUN), creatinine (Cr), eGFR, liver function tests (LFTs)  
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 25 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
d HIV-I and HIV-II antigen; anti-HCV antibody will be tested. HBV test includes HBsAg, anti-HBc, and anti-HBs.\ 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 26 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Table 5: Schedule of Clinic Attendances Post-challenge  
Event 
Admission Phase 
Follow-up Phase 
Timeline  
D0 
D1 –    
D qPCR+  
D after  qPCR+  –   
D reach treatment 
criteria 
D after start
CQ - D d/
DRx7 
DRx28 
DRx60 
DRx90 
DRx180 D Rx 1 yr. 
Window (days) 
0 
0 
0 
0 
±2 
±4 
±7 
±7 
±14 
±14 
Inclusion/exclusion 
criteria 
X 
 
 
 
 
 
 
 
 
 
Medical history            
(well-being check list)  
X 
X 
(once 
daily) 
X 
(Twice daily) 
X 
(once 
daily) 
X 
X 
X 
X 
X 
X 
Body weight  
X 
 
X 
(D reach treatment 
criteria) 
 
X 
X 
X 
X 
X 
X 
Physical examination   
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Urine pregnancy test 
(women only)  
 
 
X  
(before treatment)
 
 
 
 
 
 
 
Vital signs a 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Local & systemic 
events (AE/SAE) b 
X 
X 
X 
X 
X 
X 
X 
X  
X 
X 
Blood stage challenge 
X 
 
 
 
 
 
 
 
 
 
Treatment for Malaria c 
 
 
X (D reach 
treatment) 
X 
 
 
 
 
 
 
Diary card 
 
 
 
D d/c 
X 
X 
X 
X 
 
 
Malaria Blood Film  
 
X 
(daily) 
X 
(twice daily) 
X 
(daily) 
X 
X 
X 
X 
X 
X 
Malaria qPCR and 
Gametocyte qPCR 
 
X 
(daily) 
X 
(twice daily) 
X 
(daily) 
X 
X 
X 
X 
X 
X 
Haematology (CBC) 
 
D4,  
D qPCR+ 
X 
(D reach treatment 
criteria) 
  
X  
 
X 
X 
X 
X 
X 
Biochemistry d  
 
D qPCR+ 
X 
D reach treatment 
criteria) 
D d/c 
X 
X 
X 
X 
 
X 
Malaria Immunology  
 
 
D4,  
D qPCR+ 
X 
(D reach treatment 
criteria) 
 
X  
 
X 
X 
X 
X 
X 
MFA 
 
X 
(daily) 
X 
(twice daily) 
X 
(daily) 
X 
X 
X 
X 
X 
X 
COVID-19 testing (RT-
PCR) 
 
X e 
 
X 
(d/c) 
 
 
 
 
 
 
COVID-19 testing 
(serology test) 
 
 
X f 
 
 
 
 
 
 
 
EBV and CMV 
serology 
 
 
X g 
 
 
 
 
X 
HBV and HCV 
 
 
 
 
X h  
(D d/c) 
 
 
 
 
 
X h 
HIV-I, HIV-II 
 
 
 
 
X  
(D d/c) 
 
 
 
 
 
X 
Remark: 
a Vital signs include blood pressure, pulse rate, respiratory rate and body temperature. 
b AEs/SAEs will be collected until 1 year after antimalarial initiation. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 27 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
c This treatment period could occur on any day between D6 and D21. Blood draws will continue as per this Table for 
the relevant study day, up until the time-point of treatment. The treatment will be done on D21 if no criteria meet.  
d Biochemistry including electrolytes (sodium, potassium, chloride, bicarbonate), blood urea nitrogen (BUN), creatinine 
(Cr), liver function tests (LFTs) 
e In case volunteers have fever but malaria blood film and malaria qPCR are negative, COVID-19 will be tested. 
f COVID-19 serology test will be done on the day of discharge. 
g EBV and CMV serology will be done at 2-4 weeks after the inoculation. EBV and CMV serology including EBV 
VCA (IgM)/IgG antibody, anti EBNA and EA complex antibody, and CMV IgM/IgG antibody 
h Anti-HCV antibody will be tested. HBV test includes HBsAg, anti-HBc, and anti-HBs.\ 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH specimen collection 
Document No.:  FTM ECF-033-01 
Page 28 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
Table 6: Blood collection during study period 
 
Phase 
Clot blood 
EDTA blood 
NaF 
Total 
blood 
(mL) 
Accumulate 
blood volume 
(mL) 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
Screening 1 
6 
Blood typing, Cross 
matching  
24 
Malaria antigen (blood film*, qPCR) 
 
2 
Fasting 
blood sugar 
32 
32 
Blood biochemistry 
  
HBs profile, anti-HCV  
Duffy blood group 
Serum β-HCG 
CBC + Thalassemia 
Hemoglobinophathies 
  
 
 
  
Malaria exposure 
Malaria immunology 
HIV-I, HIV-II 
Screening 2 (D-1) 
10 
  
Serum β-HCG 
16 
CBC + G6PD screening test (FST) 
  
26 
58 
Blood biochemistry 
Malaria immunology 
CMV, EBV serology 
Malaria antigen (blood film*, qPCR) 
COVID-19 serology 
 
 
 
 
 
 
 
 
Admission D1 - 
 D qPCR+ (~D8) 
5 
Blood biochemistry (D 
qPCR+) 
60 
 
Malaria antigen (blood film*, qPCR, 
qGAM) (once daily)  
  
65 
123 
  
Membrane feeding (once daily)  
Malaria immunology (D4, D qPCR+),  
CBC (D4, D qPCR+)  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH specimen collection 
Document No.:  FTM ECF-033-01 
Page 29 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
Phase 
Clot blood 
EDTA blood 
NaF 
Total 
blood 
(mL) 
Accumulate 
blood volume 
(mL) 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
After D qPCR+ to  
D reach treatment 
criteria (D9~D16) 
9 
  
Blood biochemistry  
(D reach treatment criteria) 
78 
(48+16+
12+2) 
Malaria antigen (blood film*, qPCR, 
qGAM) (twice daily)  
  
87 
210 
CMV, EBV serology 
Membrane feeding (twice daily)  
Malaria immunology   
(D reach treatment criteria)  
CBC (D reach treatment criteria)  
D  After start CQ - D 
d/c 
6 
  
Blood biochemistry (D 
d/c) 
20 
Malaria antigen (blood film*, qPCR, 
qGAM) (once daily)  
 
26 
236 
COVID-19 serology 
(Dd/c) 
Membrane feeding (once daily)  
HBs profile, anti-HCV 
(Dd/c) 
HIV-I, HIV-II (Dd/c) 
DRx7 
  
5 
  
Blood biochemistry 
18 
  
Malaria antigen (blood film*, qPCR, 
qGAM)  
  
23 
259 
Membrane feeding  
Malaria immunology 
CBC  
DRx28 
5 
Blood biochemistry 
18 
Malaria antigen (blood film*, qPCR, 
qGAM)  
 
  
23 
282 
  
Membrane feeding  
Malaria immunology  
CBC  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH specimen collection 
Document No.:  FTM ECF-033-01 
Page 30 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
Phase 
Clot blood 
EDTA blood 
NaF 
Total 
blood 
(mL) 
Accumulate 
blood volume 
(mL) 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
Volume 
(mL) 
Test 
DRx60 
5 
Blood biochemistry 
18 
Malaria antigen (blood film*, qPCR, 
qGAM)  
 
  
23 
305 
  
Membrane feeding  
Malaria immunology 
CBC 
DRx90 
5 
Blood biochemistry 
18 
Malaria antigen (blood film*, qPCR, 
qGAM) 
 
  
23 
328 
  
Membrane feeding  
Malaria immunology 
CBC 
DRx180 
  
18 
Malaria antigen (blood film*, qPCR, 
qGAM)  
 
  
18 
346 
Membrane feeding   
Malaria immunology 
CBC 
D Rx 1 yr. 
9 
  
Blood biochemistry 
22 
Malaria antigen (blood film*, qPCR, 
qGAM) 
 
  
31  
377 
HBs profile, anti-HCV 
Membrane feeding  
EBV and CMV serology 
Malaria immunology 
 
CBC 
 
HIV-I, HIV-II 
 
 
 
* EDTA blood or Sodium Citrate blood 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 31 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
B6.2.4. Details of study clinical procedures/laboratory tests 
Procedures will be performed at the time points indicated in the schedule of procedures (Tables 4 - 
6). Additional procedures or laboratory tests may be performed, at the discretion of the investigators 
if clinically necessary. Study procedures will be monitored by the MORU Clinical Trials Support 
Group.  
B6.2.4.1. Clinical procedures 
B6.2.4.1.1. Body Weight and height 
Body Weight and/or height will be measured at the time points indicated in the schedule of 
procedures (Tables 4 & 5). 
B6.2.4.1.2. Vital signs 
Pulse rate (PR), blood pressure (BP), respiratory rate (RR) and body temperature (BT) will be 
measured at the time points indicated in the schedule of procedures (Tables 4 & 5). 
B6.2.4.1.3. Urinalysis 
Urine will be tested for the presence of clinically significant proteinuria, glucosuria or haematuria at 
the screening visit.  
B6.2.4.1.4. Electrocardiogram 
An electrocardiogram will be performed at the screening visit.  
B6.2.4.2. Laboratory procedures 
Tropical Medicine Diagnostic Reference Laboratory (TMDR) will be the central reception point for 
all blood samples collected from FTMCTU, and will follow a sample management system designed 
by the MORU Sample Management Centre. TMDR will aliquot diagnostic and research samples 
and be responsible for transfer to designated laboratories for testing and retrieve the result generated 
from the following laboratories. “In case any laboratories could not perform the tests specified below 
or stop providing the service, then the tests can be done at other validated laboratory.” 
 Diagnostic Laboratory Unit, Hospital for Tropical Diseases, Faculty of Tropical Medicine, 
Mahidol University 
1. Haematology: complete blood count (CBC) and blood grouping (ABO, Rhesus) 
2. G6PD fluorescence spot test 
3. Biochemistry: electrolytes, blood urea nitrogen (BUN), creatinine (Cr), liver function tests 
(LFTs), fasting blood sugar (FBS)   
4. Serum beta-human chorionic gonadotrophin (serum β-HCG)  
 
5. Diagnostic serology: HBs profile (HBsAg, anti-HBc, anti-HBs), anti-HCV  
6. Crossmatching 
7. Urinalysis 
8. Urine pregnancy test 
9. CMV, EBV serology 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 32 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 Tropical Medicine Diagnostic Reference Laboratory (TMDR)/ Microbiology and 
Immunology Department, Faculty of Tropical Medicine, Mahidol University 
1. COVID-19 serology (or other validated laboratories) 
2. COVID-19 PCR 
 Malaria Laboratory, Faculty of Tropical Medicine, Mahidol University 
1. Malaria microscopic examination 
 Molecular Tropical Medicine & Genetics Department, Faculty of Tropical Medicine, 
Mahidol University 
1. High volume qPCR for Malaria  
 ATGenes  
1. Duffy antigen  
2. Haematological diseases*: haemoglobinopathy analysis, CBC, haemoglobin 
electrophoresis and molecular studies for alpha and beta thalassaemia trait 
 Siriraj Hospital, Mahidol University 
1. qPCR for HIV-I, HIV-II 
 Mahidol Vivax Research Unit (MVRU), Faculty of Tropical Medicine, Mahidol University 
1. Background exposure to malaria*: Multiplex Bead Based Immunoassay will be performed 
to determine the magnitude of antibodies against recent exposure of 5-8 blood-stage P. vivax 
antigens 
2. Malaria and gametocyte qPCR 
3. Immunology: Immunological responses to P. vivax infection.  
4. Membrane feeding assay 
*This test will be done for data interpretation purpose. It will not be used as the inclusion/exclusion criteria. 
Samples may also be sent to collaborating laboratories within and outside Thailand for 
immunomonitoring and/or harmonisation of key immunological assays. There are two collaborating 
laboratories:  
(1) Ehime University, Matsuyama, Japan, and (2) Kenya Medical Research Institute (KEMRI), 
Kenya 
The AlphaScreen assay will be used to quantify malaria antigens-specific antibodies (up to 300 P. 
falciparum antigens and 300 P. vivax antigens). A seropositive cut-off will be set as half the lowest 
non-negative value of the assayed samples. Positive malaria history refers to a volunteer who 
previously had a malaria infection leading to antibodies in their serum which reactive to more than 
10% of the malaria antigens in the library. This may include human DNA and RNA analysis. 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 33 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
B6.2.5. 
Challenge infection performed in the trial participants  
B6.2.5.1. Description and administration of the study agents (which is the cryopreserved P. 
vivax infected erythrocytes)  
In this study, the study agent that we apply / administer to the volunteer is not a drug (since it is not 
a drug trial). It is “cryopreserved P. vivax infected erythrocytes”. The total volume of the diluted 
cryopreserved parasite that will be injected into each volunteer is 5 mL. 
B6.2.5.1.1. Source and preparation of the cryopreserved inoculum 
The cryopreserved P. vivax parasitized red blood cells that will be used were produced by 
investigator team in December 2020, as part of “A clinical study to assess the feasibility of a 
controlled human Plasmodium vivax malaria infection model through experimental sporozoite 
infection in Thai adults” (‘MIST 1 study’, certificate number 2020-038-01, approved by the FTM-
EC #TMEC19-067; OxTREC Ref 43-19. The information is as followed. 
Cryopreservation and storage of blood bank 
P. vivax infected blood processing was conducted within fumigated microbiological safety cabinets. 
First, the red cells were separated from plasma by centrifugation of the leukodepleted blood before 
mixing with GMP grade 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 1.5 mL cryovials and frozen at -80oC for one night 
before transferred to a dedicated liquid nitrogen tank. Confirmation of parasitic density within the 
blood collected for cryopreservation was performed via microscopy and quantitative PCR on the 
leukodepleted packed blood samples. Recently, there are 4 inoculum stocks. The information of the 
parasite from each donor is listed below. 
 
Donor no. 
Parasite density 
Blood film  
qPCR 
1 
6 asexual parasites/µL packed blood 
155,400 genome copies/mL  
2 
13 asexual parasites/µL packed blood 
988,600 genome copies/mL 
3 
51 asexual parasites/µL packed blood 
1,300,533 genome 
copies/mL 
4 
233 asexual parasites/µL packed blood 
3,300,133 genome 
copies/mL 
Sterility and screening of blood-borne and vector-borne infections of cryopreserved blood 
bank 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 34 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Sterility and screening for blood-borne and vector-borne infections of cryopreserved blood bank 
were performed to ensure the safety of cryopreserved blood bank. Real time PCR for HIV-1, HIV-
2, HBV, HCV, Dengue, chikungunya, zika, Japanese encephalitis, CMV and EBV, serology for 
HTLV-1, HTLV-2 and syphilis (TPHA), serology for filaria, and ELISA for mycoplasma were 
tested from plasma derived directly from blood bank. Blood collected for cryopreservation was sent 
to FTM hospital for testing the bacterial contamination using hemoculture technique. All tested 
results were negative for all four donors. 
B6.2.5.1.2. 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 local standard operating procedure. The processing 
of the inoculum will be carried out in MVRU’s dedicated clean room biosafety level II laboratory at 
the Hospital for Tropical Diseases, Faculty of Tropical medicine. Sample manipulations will be 
performed within a safety cabinet that has been fumigated, sterilised and dedicated for this purpose. 
Deglycerolizing will be performed by sequentially adding 0.1 volume of 12%, 10 volume of 1.6% 
and 10 volume of 0.9% saline solution. Either one whole vial, containing approximately 0.5 mL of 
red blood cells, one fifth of a vial, one tenth of the vial or one twentieth of a vial will be reconstituted 
in 0.9% saline, to a total volume of 5 mL. Five milliliters of each of the diluted inoculum will be 
drawn in 5 mL syringe and close with sterile cap. The inoculum will be kept in the warm transport 
box until inoculate to the volunteers. The inoculum must be injected within 4 hours of the inoculum 
being thawed. 
B6.2.5.1.3. Administering the study agent to the volunteers:  
The inoculation will take place at the Hospital for Tropical Diseases. The inoculum will be 
administered by injection into an indwelling intravenous cannula. 
The challenge steps:  
The volunteer(s) will be hospitalized a night before the inoculation day (D0) to ensure their safety / 
eligibility and to avoid the traffic burden. 
Inoculum will be transported directly from the MVRU clean room lab at 9th floor in a warm box and 
immediately transferred to a secure room within the Hospital for Tropical Diseases. 
The number of volunteers in each group depends on the availability of cryopreserved inoculum 
stocks. Two volunteers will be challenged with blood-stage inoculum per dose as shown in Table 7. 
All groups will receive an inoculum of parasitised red blood cells reconstituted in 0.9% normal 
saline, to a total volume of 5 mL, containing different dilution doses as specified below and as shown 
in the Table 7.  
Table 7 MIST 2 intervention details 
Dose finding for 2 cryopreserved inocula in Thai volunteers  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 35 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Participants 
per group 
Group 1 for 
inoculum#1 
Group 2 for 
inoculum#2 
Intervention 
8 volunteers 
  
2 
2 
Challenge with whole dose blood-stage inoculum 
(neat) 
2 
2 
Challenge with 1:5 dilution blood-stage inoculum  
2 
2 
Challenge with 1:10 dilution blood-stage inoculum 
2 
2 
Challenge with 1:20 dilution blood-stage inoculum 
Participant who withdraws or be withdrawn before inoculation is completed will be replaced. 
Whole dose: one whole vial, containing approximately 0.5 mL of red blood cells, will be 
reconstituted in 0.9% saline, to a total volume of 5 mL 
1:5 dilution:  one fifth of a vial will be reconstituted in 0.9% saline, to a total volume of 5 mL. 
1:10 dilution:  one tenth of a vial will be reconstituted in 0.9% saline, to a total volume of 5 mL. 
1:20 dilution:  one twentieth of a vial will be reconstituted in 0.9% saline, to a total volume of 5 mL. 
The reconstituted inoculum will be injected via an indwelling cannula, followed by a saline flush.  
One volunteer from each group (Phase A and B - Groups 1-4), receiving one of the four 
concentrations, will be administered the inoculum first. The inoculum will subsequently be 
administered to each of the second 4 volunteers from each group. This is in case the time between 
administration of the inoculum affects the viability of the parasites. 
Prior to challenge, all volunteers are briefed. The arms will be cleaned by using 70% alcohol 
according to the Inoculum Administration Work Instruction (WI). 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 36 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
B6.3 Schematic diagram of study design, procedures and stages, step-by-step 
 
    
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* COVID-19 (RT-PCR) will be tested on day -1 and during 
admission (during day 1 post challenge to D qPCR+), if 
volunteers present with a clinical feature(s), and malaria blood 
film and malaria qPCR are negative; and will be tested on 
discharge day. COVID-19 serology test will be performed on 
day -1 and day d/c. 
HBV and HCV serology and HIV-I, HIV-II antigen will be 
tested on day d/c. 
Eligible volunteers 
Obtaining approval from the Ethics Committees 
Informed consent be obtained 
Screening and eligibility assessment (day -14 to day -1) 
 
 History taking, vital signs and physical 
examination, body weight and height, ECG  
 Blood group ABO, RH Rhesus, Duffy blood group  
 CBC, blood diseases, FBS, blood biochemistry 
 HBV serological profile, HIV-I, HIV-II antigen, 
and anti-HCV  
 malaria diagnosis, malaria serology and 
immunological profiles 
 Serum pregnancy test (women only) 
 Urinalysis  
 Crossmatching 
Admission day (day -1) 
 
 History taking, vital signs and 
physical examination, body weight 
 G6PD screening, CBC, blood 
biochemistry 
 Malaria diagnosis, malaria serology 
and immunological profiles  
 Serum pregnancy test 
 COVID-19 testing* 
 EBV and CMV serology 
 
Blood stage challenge (at day 0) 
 
Well-being check list, vital signs and physical 
examination, body weight 
 
CBC and blood biochemistry  
 
Malaria blood film, malaria qPCR and gametocyte PCR, 
malaria immunology 
 
Membrane feeding assay 
 
Urine pregnancy test (before treatment for malaria) 
 
EBV and CMV serology (2-4 wks. after the inoculation) 
 
COVID-19 testing* 
 
If participants meet the criteria for treatment, the standard 
treatment for malaria will be initiated. 
(Chloroquine treatment) 
Discharge when clinically recovered AND two consecutive 
blood films are negative. 
 
(Diary card will be provided to subject for self-assessments.) 
Follow up phase (D Rx7, D Rx28, D Rx60, D Rx90, D Rx180, D Rx 1yr.) 
 
 
Well-being check list, 
vital signs and physical 
examination, body weight, diary card 
 
CBC and blood biochemistry 
 
Malaria blood film, malaria qPCR and gametocyte PCR, 
malaria immunology 
 
Membrane feeding assay, EBV and CMV serology  
 
HIV-I and HIV-II antigen, HBV and HCV serology 
Recruiting research participants 
Admission phase 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 37 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
B6.4 Specimen and data management 
 
Mahidol Vivax Research Unit (MVRU) will be the central reception point for all blood samples 
collected from FTMCTU, and will follow a sample management system designed by the MORU 
Sample Management Centre.  
 
The investigators will maintain and retain appropriate medical and research records and essential 
documents for this trial in compliance with ICH Good Clinical Practice (ICH GCP) E6 (R2) and 
regulatory and institutional requirements for safety and protection of confidentiality of volunteers. 
The Co-Principal Investigators are responsible for data management and for delegating the 
receiving, entering, cleaning, querying, analysing and storing all data that accrues from the study. 
The study staff will enter the data into the volunteers’ CRFs, which will be in a paper and/or 
electronic format. This includes safety data, laboratory data (both clinical and immunological) and 
outcome data. Data will be managed and stored in MACRO® database, a GCP-compliant electronic 
data capture system. A study data management plan will outline detailed procedures for data capture, 
storage, curation and preservation.  
The documentation, data and all other information generated will be held in strict confidence. Only 
authorized, trained study staff will have access to study records. The investigators will permit 
authorized representatives of the sponsor, ethical committee(s), regulatory authorities (if applicable), 
authorized representative of sponsor, 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. No information concerning the study or the data will be released to 
unauthorised third parties, without prior written approval of the Sponsor.  
B6.5 Data analysis 
The safety of the CHMI will be assessed by descriptive analysis of the frequency, incidence and 
nature of adverse events and serious adverse events arising during the study.  Since this is a feasibility 
study conducted in 2 volunteers per dosing group, formal statistical hypothesis testing will not be 
used for most analyses due to the limited sample size, and only a brief Statistical Analysis Plan 
(SAP) will be developed and finalized prior to database lock. 
B7. DATA AND SPECIMEN STORAGE AND/OR SHARING 
B7.1 Management of specimen/data archiving and of left-over samples 
Specimen storage  
With the volunteers’ informed consent, any leftover cells and serum will be stored for 10 years in 
the FTM/MORU Sample Management System for future immunological and genetic analysis of 
malaria-specific responses. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 38 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Data handling and record keeping 
Medical records and study documentation/data will be kept at study site for approximately 5 years 
after study completion. Data will be fully anonymized and stored indefinitely in a secure database. 
B7.2 Specimen/data sharing plan 
All data used for analysis will be de-identified; personal identifying information such as names and 
telephone numbers will not be used for analysis. Files containing identifiable information will be 
stored separate from other study data, in secure locations. Only the Sponsor’s representative, 
Investigators, the clinical monitor, the ethical committee(s) and the regulatory authorities will have 
access to these records. Photographs may be taken to be shown to other professional staff, used for 
educational purposes, or included in a scientific publication. Photographs will not include the 
volunteer’s face. And the volunteer’s written informed consent will be sought before photographs 
are taken. The photographs will be stored as confidential records, as above. 
De-identified data and results from blood analyses stored in our database may be shared with other 
researchers to use in the future.  
PART C: ETHICAL CONSIDERATION (describe only the responsibilities of the PI) 
C1. SIGNIFICANCE OF THE STUDY 
This study will be a proof of feasibility and safety of controlled blood-stage human P. vivax infection 
using cryopreserved inoculum. The optimum dose of inoculum determined in this study will be used 
in future CHMI and malaria vaccine studies. This study will also provide data on the immune 
response to malaria infection, gametocytemia and transmissibility of the gametocytes in volunteers. 
C2. BALANCE OF RISK AND BENEFIT  
C2.1 Risk of the study 
This section describes the foreseen risks to the volunteer including risk related to: 
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 venipuncture, which will not be documented as AEs if they occur. 
Inoculation 
This controlled blood-stage challenge will involve a transfusion of a blood inoculum. As with any 
transfusion, there are risks of blood-borne infections and allergic reaction to blood products. The 
risk of blood-borne infection was reduced during the collection of blood inoculum in MIST 1 by 
extensive screening for blood-borne infection in both the source patients and blood donation 
volunteers. Serious allergic reactions including anaphylaxis have not been seen in challenge studies 
to date, but for safety reason volunteers will be inoculated in an area where physicians and a 
defibrillator are immediately available.  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 39 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
As the inoculum is prepared from packed red cells, the volunteers have a risk of developing a 
transfusion reaction. This risk has been minimized by preparing the inoculum from donors with 
blood group O (universal donors), and the volume of blood to be used (0.5 mL packed red cells) is 
much smaller than that given in transfusion of one unit packed red cells (470 mL). Nevertheless, the 
volunteers will be monitored for this possibility during the first hour after the administration of the 
inoculum. 
Plasmodium vivax infection 
Base on previous studies, volunteers are likely to develop symptomatic malaria infection following 
challenge [18]. The 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 study schedule. We will admit 
the volunteers to the Hospital for Tropical Diseases, Faculty of Tropical Medicine, Mahidol 
University for close clinical monitoring, treatment (antimalarials) and symptomatic relief. A very 
small proportion of volunteers in previous P. vivax blood-stage and sporozoite-stage challenge 
studies have temporarily required intravenous fluid therapy for nausea and vomiting prior to 
treatment.  
C2.2 Preventive and alleviative measures for risk 
Phlebotomy 
Participants’ blood samples will be collected by well - trained medical personnel with aseptic 
technique to prevent infection or complications. 
Plasmodium vivax infection 
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.  
As described above, the volunteers will be hospitalized in the Hospital for Tropical Medicine, 
Faculty of Tropical Medicine, Mahidol University for close clinical monitoring, antimalarial 
treatment and symptomatic relief (e.g. fluids, analgesia and/or anti-emetics) until parasitaemia 
becomes negative. The treatment algorithm as well as the hospital guideline will be fully reviewed 
and strictly followed. The hospital will be officially informed prior to study initiation to ensure all 
the required medical facilities are in place. Clinical malaria expert at the Faculty of Tropical 
Medicine will be officially informed prior to study initiation. The medical monitors will be reachable 
throughout the whole study process.  
In addition, this blood-stage inoculation will cause P. vivax infection without liver-stage parasites, 
thus removing the risk of relapse.  
COVID-19 
Risk mitigation steps will be taken for all face to face research during the COVID-19 pandemic, 
following local guidance (Hospital for Tropical Diseases). 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 40 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
The subject going missing post-challenge  
The volunteers will be inpatients for entirety of the challenge phase, from the day before challenge 
until the completion of antimalarial treatment. It is unlikely that a volunteer will go absent during 
hospitalization (before antimalarial treatment is completed).  
C2.3 Benefits of the study 
The expected benefit is to provide information about the feasibility and safety of and immune 
response to P. vivax infection through experimental inoculation of cryopreserved infected 
erythrocytes, and on the optimal inoculation dose for future testing of pre-erythrocytic candidate 
vaccines. All of these will benefit patients with malaria infections in the future, and potentially lead 
to the development of a vaccine to prevent infections altogether. However, there are no direct 
benefits to the participants taking part in this study. 
C3. SAFETY  
C3.1 Safety experience from previous 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. This provides a 
comprehensive guide to the appropriate conduct of controlled human malaria infection studies [22]. 
Although there remain minor differences between centres in follow-up procedures in controlled 
human malaria infection trial conduct, there is consensus on 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. 
 Follow-up visits should be scheduled at least once daily, but may increase in frequency to 
twice daily, starting at day 5 post-sporozoite challenge. At all visits volunteers should be 
questioned about the occurrence of adverse events (AEs) and use of medication. 
 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 
control human challenge 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 antimalarial 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. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 41 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 After challenge, all volunteers should be followed until they have completely finished 
antimalarial 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 challenge trials 
which have an efficacy component and which have major potential safety concerns. 
Volunteer safety is of paramount importance, and we will follow these guidelines. The following 
measures are in place to safeguard volunteer safety: 
 Volunteers will only be enrolled in the study if investigator judges 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. 
 Before challenge, full contact details for each volunteer 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. Volunteers must also provide the Investigators with the name and 24 hour 
telephone number of a close friend(s), relative(s) or housemate(s) who live nearby and who 
will be kept informed of their whereabouts for the duration of the study. 
 The volunteers will be hospitalized throughout the challenge processes until clinically 
recovered AND the completion of the antimalarial course (chloroquine) AND until two 
consecutive negative blood films.  
 Volunteers will be able to contact a medically qualified member of the study team 24 hours 
a day throughout the study period. 
Expectations of volunteer events following blood stage challenge 
This part of the protocol aims to clearly review the events related to experience from the previous 
control human challenge studies in order to foresee the possible event(s) after the challenge that we 
may have to prepare to encounter within this study.  
 According to references No. 18-21 and personal communication with other investigators, all 
volunteers in P. vivax blood stage challenge studies developed infection within 3 weeks 
following the challenge. Most of AEs were mild to moderate grade and all were transient. 
No serious adverse events were reported. 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 42 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
C3.2 Safety Monitoring 
C3.2.1. Medical Monitor 
A Medical Monitor group, representing the Sponsor, will be appointed for oversight of safety in this 
clinical study. The Medical Monitor group will be responsible for safety assessments. The monitor 
will review the study prior to initiation and will be available to advise the Investigators on study-
related medical issues and to act as a representative for the welfare of the subjects. The medical 
monitor does not have direct involvement in the conduct of the study. All serious adverse events will 
be reported to the medical monitor within 24 hours of becoming aware of the event. The medical 
monitor group is responsible for the review of the safety data and communicate with the PI and/or 
the DSMB, as appropriate.  
The appointed medical monitor for the MIST 2 study is Dr. Lorenz Von Seidlein M.D.  
C3.2.2. Data and Safety Monitoring Board 
The Data Safety Monitoring Board (DSMB) will review the study prior to initiation; review the 
interim safety data reports; and review all serious adverse events (SAEs) according to DSMB charter. 
The Board may convene additional reviews if deemed necessary, on review of the safety data, as 
sent, periodically by the medical monitor. All SAEs will be reported by the site principal investigator 
to the DSMB at the same time as they are submitted to the ethics committees. The Co-Principal 
Investigator(s) will notify the Board and obtain a recommendation concerning continuation, 
modification, or termination of the study. The site principal investigator will submit written DSMB 
summary reports with recommendations to the ethics committees(s). The roles and responsibilities 
of the DSMB will be formalised in a charter agreed with the members of the DSMB. 
The DSMB will be independent of the Clinical Trials and Research Governance, Oxford University 
(Sponsor) and MIST programme team. The DSMB is charged with monitoring and evaluating the 
clinical data generated by this study, with a focus on safety, in and independent and objective 
manner. The main roles and responsibilities of the DSMB, DSMB procedures, and the DSMB 
meeting schedule are defined in the Data and Safety Monitoring Board Charter for the Malaria 
Infection Study in Thailand (MIST) Programme. 
C4. CONSIDERATION FOR VULNERABLE RESEARCH PARTICIPANTS 
Check whether your study involves any of the following vulnerable research participants.  
   Prisoners 
   Pregnant women 
   Mentally ill persons 
   Cancer or terminally ill patients 
   Neonates/infants/children (aged <18) 
   HIV/AIDS patients  
   Institutionalized persons e.g. military, students, etc. 
   Others (please specify)………………………………….…. 
   Not applicable 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 43 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
C5. INFORMED CONSENT ISSUES 
C5.1. Informed consent process 
The informed consent process: The investigator will explain the purpose of the study with the help 
of and consistent with the Participant Information Sheet (PIS) prior to any study related procedures 
being undertaken. The volunteer must personally sign and date the latest approved version of the 
informed consent form (ICF) before any study specific procedures are performed.  
The information sheet and informed consent form will be explained to the volunteers detailing no 
less than: the exact nature of the study; the implications and constraints of the protocol; the known 
side effects and any risks involved in taking part. The aims of the study and all procedures to be 
carried out will be explained. It will be emphasised that there is no direct benefit from 
participating the trial.  
 It will be clearly stated that participation is entirely voluntary and that refusing to participate 
will not involve any penalty or affect the volunteers’ right to receive standard medical care. 
 It will also be emphasized that if they do consent to participate and are enrolled, that they are 
free to withdraw from the study at any time, for any reason, without any penalty or prejudice 
to future care, and with no obligation to give the reason for withdrawal.  
The volunteer will have the opportunity to question the investigator, or other independent parties to 
decide whether or not they will participate in the study.  
If they do decide to participate, volunteers will be asked to complete a questionnaire testing their 
understanding of the trial before signing the consent. This helps to ensure that individuals 
understand the trial sufficiently to give informed consent. After the volunteer answers all questions 
in the questionnaire correctly, they will be asked to sign and date two copies of the consent form, 
one for them to take away and keep, and one to be stored in the investigator’s site file and retained 
at the study site. These forms 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 
following further discussion with the investigator. Only the subject who able to subsequently answer 
all questions in the questionnaire correctly will be asked to give the consent and will be screened for 
the trial. Only 2 attempts will be permitted in total. 
Hospital consent for HIV test: This will be obtained prior to counselling and HIV test using the 
standard HIV consent from of Hospital for Tropical Diseases. Pre and post counselling for HIV 
screening and reporting will be conducted with the support of physician from the Hospital for 
Tropical Diseases. In case a volunteer is found to be HIV positive, follow up measures including but 
not limited to providing counselling and treatment according to standard hospital procedure will be 
arranged through a physician from the Hospital for Tropical Diseases. 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 44 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
C5.2 Informed consent documentation 
Age 
Informed Consent 
Participant Information Sheet 
20 years and over (Adult) ICF for Adult 
Version7.0, dated 3 January 2024 
PIS for Adult 
Version 7.0, dated 3 January 2024 
ICF for Data Sharing/ Data and 
Leftover Specimen Storage from 
Current Study for Future Use 
Version 7.0 dated 3 January 2024  
ICF for Leftover Specimen 
Storage from Current Study for 
future genetic analysis 
Version 7.0 dated 3 January 2024 
20 years and over (Adult) HIV Pre-test counseling form dated 1 October 2011 
C5.3 Compensation for research participants 
  Yes, please provide details:  
 
 
Volunteers will be compensated for their time and for the inconvenience caused by 
procedures as below (Table 8). This amount of compensation is calculated based on the cost 
of living in Bangkok for time and traveling. According to the consensus recommended by 
the controlled human challenge study group meeting in 2015, compensation specifically for 
risk was also advised (Bambery B, Selgelid M, Weijer C, et al., Ethical criteria for human 
challenge studies in infectious disease. Public Health Ethics. 2016. 9; 92-103.) 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 45 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
Table 8: Estimated compensation amounts. 
Activity 
Compensation 
(THB) 
Number of visits 
Total 
(THB) 
1. Screening visit (D-14) 
2,000 
1 
2,000 
2. Admission per night 
2,000 
Around x 18 nights 
36,000 (estimated) 
3. DRx7 
1,500 
1 
1,500 
4. D Rx28 
1,500 
1 
1,500 
5. D Rx60  
1,500 
1 
1,500 
6. D Rx90 
1,500 
1 
1,500 
7. D Rx180 
1,500 
1 
1,500 
8. D Rx 1 yr. 
1,500 
1 
1,500 
Time in Trial 
(approx.) 
Maximum no. 
of visits 
Maximum volume of 
blood taken1 (mL) 
Total compensation amount (THB) 
1 year 
8 
 377 mL 
47,000 
Remark:  
1Maximum volume of blood taken is 377 mL in case volunteer received antimalarial treatment on day 16. 
In case a volunteer has to come for extra visit(s), they will be compensated for their time and for the 
inconvenience with 1,000 THB per day.  
  No, please provide reasons: ………………………………………………..……… 
 C5.4 Responsible and contact persons 
  Person(s) responsible for payment for treatment of complications and adverse effects 
  Person(s) including doctor(s) and/or contact address(es) and telephone number(s) for 
emergency use 
1. Dr. Borimas Hanboonkunupakarn, Department of Clinical Tropical Medicine, Faculty of 
Tropical Medicine, Mahidol University, Tel. (02) 354-9100 ext. 3160, Mobile (086) 970-5705  
2. Dr. Podjanee Jittamala, Department of Tropical Hygiene, Faculty of Tropical Medicine, 
Mahidol University, Tel. (02) 306-9157, Mobile (081) 956-3371. 
3. Dr. Kittiyod Poovorawan, Department of Clinical Tropical Medicine, Faculty of Tropical 
Medicine, Mahidol University, Tel. (02) 354-9100 ext. 1435, Mobile (083) 149-6864 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 46 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
PART D: APPENDIX A 
REFERENCES 
1. 
Baird JK. Evidence and implications of mortality associated with acute Plasmodium vivax 
malaria. Clin Microbiol Rev. 2013;26(1):36-57. 
2. 
WHO. World malaria report 2017. 2017. 
3. 
Rahimi BA TA, White NJ, Sirivichayakul C, Dondorp AM, Chokejindachai W Severe vivax 
malaria: a systematic review and meta-analysis of clinical studies since 1900. Malar J. 
2014;13:481. 
4. 
Price RN TE, Guerra CA, Yeung S, White NJ, Anstey NM. Vivax malaria: neglected and not 
benign. Am J Trop Med Hyg. 2007;77(6 Suppl):79-87. 
5. 
Watson J TW, Bancone G, Chu CS, Jittamala P, White NJ. Implications of current therapeutic 
restrictions for primaquine and tafenoquine in the radical cure of vivax malaria PLoS Negl 
Trop Dis. 2018;12(4):e0006440. 
6. 
Pybus BS, Marcsisin SR, Jin X, Deye G, Sousa JC, Li Q, et al. The metabolism of primaquine 
to its active metabolite is dependent on CYP 2D6. Malar J. 2013;12:212. 
7. 
Bennett JW, Pybus BS, Yadava A, Tosh D, Sousa JC, McCarthy WF, et al. Primaquine failure 
and cytochrome P-450 2D6 in Plasmodium vivax malaria. N Engl J Med. 2013;369(14):1381-
2. 
8. 
Grabowsky M. The billion-dollar malaria moment. Nature. 2008;451(7182):1051-2. 
9. 
Moorthy VS, Newman RD, Okwo-Bele JM. Malaria vaccine technology roadmap. Lancet. 
2013;382(9906):1700-1. 
10. Mueller I, Shakri AR, Chitnis CE. Development of vaccines for Plasmodium vivax malaria. 
Vaccine. 2015;33(52):7489-95. 
11. Payne RO, Griffin PM, McCarthy JS, Draper SJ. Plasmodium vivax Controlled Human 
Malaria Infection - Progress and Prospects. Trends Parasitol. 2017;33(2):141-50. 
12. Keith JA, Agostini Bigger L, Arthur PA, Maes E, Daems R. Delivering the promise of the 
Decade of Vaccines: opportunities and challenges in the development of high quality new 
vaccines. Vaccine. 2013;31 Suppl 2:B184-93. 
13. Pronker ES, Weenen TC, Commandeur H, Claassen EH, Osterhaus AD. Risk in vaccine 
research and development quantified. PLoS One. 2013;8(3):e57755. 
14. Knezevic I MV, Sheets RL. ,. WHO consultation on clinical evaluation of vaccines 17-18 July 
2014. Geneva, Switzerland. Vaccine. 2015;33(17):1999-2003. 
15. Stanisic DI, McCarthy JS, Good MF. Controlled Human Malaria Infection: Applications, 
Advances, and Challenges. Infect Immun. 2018;86(1). 
16. Roestenberg M, Hoogerwerf MA, Ferreira DM, Mordmüller B, Yazdanbakhsh M. 
Experimental infection of human volunteers. Lancet Infect Dis. 2018;18(10):e312-e22. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM ECF-033-01 
Page 47 of 60 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
17. Payne RO, Milne KH, Elias SC, Edwards NJ, Douglas AD, Brown RE, et al. Demonstration 
of the Blood-Stage Plasmodium falciparum Controlled Human Malaria Infection Model to 
Assess Efficacy of the P. falciparum Apical Membrane Antigen 1 Vaccine, FMP2.1/AS01. J 
Infect Dis. 2016;213(11):1743-51. 
18. McCarthy JS, Griffin PM, Sekuloski S, Bright AT, Rockett R, Looke D, et al. Experimentally 
induced blood-stage Plasmodium vivax infection in healthy volunteers. J Infect Dis. 
2013;208(10):1688-94. 
19. Griffin P, Pasay C, Elliott S, Sekuloski S, Sikulu M, Hugo L, et al. Safety and Reproducibility 
of a Clinical Trial System Using Induced Blood Stage Plasmodium vivax Infection and Its 
Potential as a Model to Evaluate Malaria Transmission. PLoS Negl Trop Dis. 
2016;10(12):e0005139. 
20. Collins KA, Wang CY, Adams M, Mitchell H, Robinson GJ, Rampton M, et al. A 
Plasmodium vivax experimental human infection model for evaluating efficacy of 
interventions. J Clin Invest. 2020;130(6):2920-7. 
21. Odedra A, Mudie K, Kennedy G, Watts RE, Rossignol E, Mitchell H, et al. Safety and 
feasibility of apheresis to harvest and concentrate parasites from subjects with induced blood 
stage Plasmodium vivax infection. Malaria Journal. 2021;20(1):43. 
22. Moorthy, V., Standardization of Design and Conduct of P. falciparum Sporozoite Challenge 
Trials. 2011. 
 
23. Robinson S, Harris A, Atkinson S, Atterbury C, Bolton-Maggs P, Elliott C, et al. The 
administration of blood components: a British Society for Haematology Guideline. Transfus 
Med. 2018;28(1):3-21. 
 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 48 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
APPENDIX B: SUBMISSION PACKAGE INCLUDES 
No. 
Submitted Documents 
1. 
Research Proposal Submission Form for a study involving human subject enrollment 
WITH specimen collection (FTM ECF-033-RR) 
2. 
Informed Consent Form and Participant Information Sheet 
3. 
Informed Consent Form for Data Sharing/ Data and Leftover Specimen Storage from 
Current Study for Future Use 
4. 
Informed Consent Form for Leftover Specimen Storage from Current Study for future 
genetic analysis 
5. 
HIV Pre-test Counseling Form 
6. 
Questionnaire to evaluate participant’s understanding of the study 
7. 
Recruitment material (flyer) 
8. 
Diary card 
9. 
Screening Form 
10. 
Case Record Form 
 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 49 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
APPENDIX C: SAFETY TERMS AND ADMINISTRATIVE INFORMATION 
 
1. SAFETY DETECTION, ASSESSMENT, DOCUMENTATION AND REPORTING 
The investigators and designated site staff is/are responsible for the detection, assessment, 
documentation and reporting of events meeting the criteria and definition of an adverse event (AE), 
or serious adverse event (SAE) as provided in this Research Proposal Submission Form. Volunteers 
will be instructed to contact the investigator immediately should the volunteer manifest any signs or 
symptoms they perceive.  
1.1 Definitions  
1.1.1 Adverse Event (AE): 
An AE is any untoward medical occurrences in a volunteer, which may occur during or after 
administration of a study intervention (in this case, vivax parasite challenge) and does not 
necessarily, have a causal relationship with the intervention. An AE can therefore be any unfavorable 
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. 
Any events occurring between screening and D-1 will be considered as baseline/, pre-existing 
conditions. This information will be recorded in the medical records.  
1.1.2 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 (i.e. results in death from any cause at any time). 
 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 serious 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). 
 Transfer of inpatient care to the intensive care unit, if the Investigators assess that a higher 
level of intervention and intense monitoring is required to manage symptoms following 
controlled human malaria infection, or drugs (over and above what can be provided by the 
Investigators in research bay). Hospitalization (including inpatient or outpatient 
hospitalization for an elective procedure) for a pre-existing condition that has not worsened 
unexpectedly does not constitute a SAE. 
 An important medical event (that may not cause death, be life threatening, or require 
hospitalisation) that may, based upon appropriate medical judgment, jeopardise the volunteer 
and/or require medical or surgical intervention to prevent one of the outcomes listed above.  
 Congenital anomaly or birth defect. 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 50 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
1.2 Clinical laboratory parameters and other abnormal assessments qualifying as adverse 
events or serious adverse events:  
In absence of a diagnosis, clinically abnormal laboratory findings (e.g. clinical chemistry, 
haematology, and urinalysis) or other abnormal assessments that are judged by the investigator to 
be clinically significant will be recorded as an AE or SAE if they meet the definition of an AE or 
SAE. Clinically significant abnormal laboratory findings or other abnormal assessments that are 
present at baseline and significantly worsen following the start of the study will also be reported as 
AEs or SAEs. 
The investigator will exercise his or her medical and scientific judgment in deciding whether an 
abnormal laboratory finding or other abnormal assessment is clinically significant. 
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 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. 
In addition, any COVID-19 infection in volunteers acquired during the in-patient stay following 
challenge will be considered an SAE and COVID-19 treatment will be provided to the volunteers 
free of charge. If volunteers have to stay in the hospital for SAE, admission compensation at 2,000 
THB per night will be provided to the volunteers. 
1.2.1. Causality assessment 
The investigator is obligated to assess the relationship between study procedure and/or antimalarial 
medications and the occurrence of each AE/SAE. The investigator will use clinical judgment to 
determine the relationship. Alternative plausible causes, such as natural history of the underlying 
diseases, concomitant therapy, other risk factors, and the temporal relationship of the event to the 
study procedure and antimalarial medications will be considered and investigated. The relationship 
of the adverse event with the study procedures will be categorized as unrelated, unlikely to be related, 
possibly related, probably related or definitely related (Table C1). An intervention-related AE refers 
to an AE for which there is a possible, probable or definite relationship to the study intervention. 
The delegated clinician will use clinical judgment to determine the relationship.  
 
Table C1: Guidelines for assessing the relationship of study intervention to an AE 
0 
Unrelated 
No temporal relationship to study intervention and 
Alternate aetiology (clinical state, environmental or other 
interventions); and 
Does not follow known pattern of response to CHMI, blood 
donation or drug. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 51 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
1 
Unlikely to be related 
Unlikely temporal relationship to study intervention and 
Alternate aetiology likely (clinical state, environmental or other 
interventions); and 
Does not follow known typical or plausible pattern of response 
to challenge, blood donation or drug. 
2 
Possibly related 
Reasonable temporal relationship to study intervention; or 
Event not readily produced by clinical state, environmental or 
other interventions; or 
Similar pattern of response to that seen with previous challenge, 
blood donation or similar drug. 
 
3 
Probably related 
Reasonable temporal relationship to study intervention; and 
Event not readily produced by clinical state, environment, or 
other interventions; or 
Known pattern of response seen with previous challenge, blood 
donation or drug. 
4 
Definitely related 
Reasonable temporal relationship to study intervention; and 
Event not readily produced by clinical state, environment, or 
other interventions; and 
Known pattern of response seen with previous challenge, blood 
donation or drug. 
1.2.2. Assessment of AE intensity 
Each adverse event will be graded by the investigator and designated study staff according to 
Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0.   
In the rare case that an adverse event is not graded in the CTCAE, then that event should be graded 
as follows: 
Grade 1: 
Mild AE 
Grade 2: 
Moderate AE 
Grade 3: 
Severe AE 
Grade 4: 
Life-threatening or disabling AE  
Grade 5:  
Death related to AE 
 
All AE/SAE will be collected throughout the first 3 month after challenge or until a satisfactory 
resolution occurs. All AEs that result in a volunteer’s withdrawal from the study will be followed up 
until a satisfactory resolution occurs, or until a non-study related causality is assigned (if the 
volunteer consents to this). In cases where the volunteer has to do AE self-detection and self- 
assessment, a simple grading system for the self-assessment on the diary card will be used (Table 
C2).  
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 52 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
Table C2: Severity grading criteria for the self-assessment for AEs. 
 
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 
1.2.3. Assessment of outcomes 
The investigator will assess the outcome of all AEs (including SAEs) recorded during the study as: 
 Recovered/resolved. 
 Recovering/resolving. 
 Not recovered/not resolved. 
 Recovered with sequelae/resolved with sequelae. 
 Fatal (SAEs only). 
During the inpatient phase, the adverse events (including all SAEs) will be assessed by the 
investigator and the delegated study staff.  
While the volunteer is followed as an outpatient, a diary card will be provided, and the volunteer 
will be instructed how to self-assess the cause and severity of AEs. This self-assessment symptom 
diary will be checked, discussed, collected and recorded at each clinic visit. 
1.3 Reporting procedures 
For SAEs 
In order to comply with current regulations on serious adverse event reporting to Ethics and 
regulatory authorities (if applicable), the event will be documented accurately and notification 
deadlines respected.  
SAEs will be reported to the medical monitor immediately (within 24 hours) of the Investigators’ 
being aware of their occurrence. 
SAEs will also be reported to ethics committees, the Data and Safety Monitoring Board (DSMB), 
and the regulatory authority (if applicable), in accordance with reporting requirements and according 
to required timelines.  
1.4 Events or outcomes not qualifying as adverse events or serious adverse events  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 53 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
Pregnancy 
Volunteers are informed that for the safety of volunteer and their child, pregnancy is prohibited until 
3 months after the challenge. Female study volunteers are asked to use appropriate contraceptive 
methods to prevent pregnancy while they participate in the study for 3 months after the injection of 
malaria infected erythrocytes. 
 
 
Contraceptive methods include: 
- Established use of oral, injected or implanted hormonal contraceptives 
- Intrauterine Device or Intrauterine System 
- Barrier methods (condoms or diaphragm with additional spermicide) 
- Male sterilisation and female sterilisation (with appropriate post-vasectomy documentation of 
absence of sperm in the ejaculate) 
- True abstinence, when this is in line with the preferred and usual lifestyle of the volunteer. 
Periodic abstinence (e.g., calendar, ovulation, symptothermal, post-ovulation methods) and 
withdrawal are not acceptable methods of contraception. 
 
Female volunteers will be tested for pregnancy immediately prior to challenge and during follow up 
as indicated in the schedule of procedures (Tables 4 and 5). 
 
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 any parasitaemia has been cleared by the 
antimalarial treatment. 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 consultants at 
Faculty of Tropical Medicine and the Mahidol Oxford Tropical Medicine Research Unit, 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 discontinued from the study as soon as we have confirmed that their 
parasitaemia has cleared. The volunteer will still be followed up until 3 months after challenge.  
During pregnancy the following should always be considered as an SAE: 
 Spontaneous abortion, (spontaneous pregnancy loss before/at 22 weeks of gestation) 
 Ectopic and molar pregnancy 
 Stillbirth (intrauterine death of fetus after 22 weeks of gestation). 
 Any early neonatal death (i.e. death of a live born infant occurring within the first 7 days of 
life).  

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 54 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
 Any congenital anomaly or birth defect identified in the offspring of a study subject (either 
during pregnancy, at birth or later) regardless of whether the fetus is delivered dead or alive. 
This includes anomalies identified by prenatal ultrasound, amniocentesis or examination of 
the products of conception after elective or spontaneous abortion. 
However, all pregnancies occurring in participants within 3 months of challenge will be considered 
a SAE.  
 
Furthermore, any SAE occurring as a result of a post-study pregnancy AND considered by the 
investigator to be reasonably related to the study procedures will be reported. While the investigator 
is not obligated to actively seek this information from former study volunteers, he/she may learn of 
a pregnancy through spontaneous reporting.  
 
All pregnancies occurring during the study will be reported to the DSMB. 
1.5 Safety monitoring 
Monitoring 
Monitoring will be performed using an established Monitoring Plan. Independent monitoring will 
be performed by Clinical Trials Supporting Group (CTSG), MORU. 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 Good Clinical Practice 
(GCP). The investigators 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. 
 
2. RANDOMIZATION 
This will be a double-blinded design. Participants will be recruited and randomised in blocks of size 
8 to one of the 4 inoculation dilutions using a computer program: whole dose blood-stage inoculum 
(neat) or 1:5 or 1:10 or 1:20 dilution blood-stage inoculum. Thus, by the end of each block, each of 
the inoculation dilution group will have 2 participants i.e. in the ratio 2:2:2:2.  
 
The randomization lists, will be prepared by MORU statistician and the randomization envelopes 
will be prepared by Clinical Trials Support Group (CTSG)/MORU. 
For each inoculation dilution group, 2 randomization numbers will be generated in blocks of size 8 
so, for the 4-dose inoculation arms in a ratio of 2:2:2:2 as follows:  
1. whole dose blood-stage inoculum (neat) 
2. 1:5 dilution blood-stage inoculum  
3. 1:10 dilution blood-stage inoculum 
4. 1:20 dilution blood-stage inoculum 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 55 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
Study volunteers will be assigned a randomization envelope, and thus will be randomly allocated to 
inoculation arm 1, 2, 3, or 4. This is a double-blinded study; volunteers, clinical investigators, and 
study staff who perform laboratory study endpoint will be blinded to inoculation arm allocation.  
Only MVRU staff who prepare the inoculum and CTSG statistical staff will know the concentration 
of each inoculum and these individuals will not be involved in providing volunteer care, 
investigation, and performing laboratory study endpoint. In addition, these individuals will not be 
involved in any other study assessment and evaluation. 
 
3. QUALITY CONTROL AND QUALITY ASSURANCE PROCEDURES  
Data will be evaluated for compliance with the protocol and accuracy in relation to source 
documents. Following written standard operating procedures and prespecified monitoring plan, the 
monitors will verify that the clinical study is conducted and data are generated, documented and 
reported in compliance with the current approved protocol, standard operating procedures, ICH GCP 
and relevant regulations. 
The Sponsor and authorised individuals may carry out audit to ensure compliance with the protocol, 
standard operating procedures, ICH GCP and relevant regulations. The study audits will be managed 
by an independent function according to standard operating procedures a prespecified audit plan.  
GCP inspections may also be undertaken by the regulatory authority or ethical committee(s) to 
ensure compliance with protocol and national regulations. The sponsor will assist in any inspections.  
4. ETHICS 
Ethical approval will be sought prior to commencing the study through the relevant Research Ethics 
Committees. Indemnity for the trial will be provided by the University of Oxford. SAEs will be 
reported to the medical monitor, DSMB, and the ethics committees. GCP certificate will be obtained 
by all staff/investigators prior to commencing the studies. 
 
The Investigators will ensure that this study is conducted in accordance with the principles of the 
Declaration of Helsinki 2013. The trial will adhere to the ICH GCP E6 (R2). 
 
The protocol, informed consent form, participant information sheet, and other written participant 
information/materials and Advertisement will be submitted to appropriate Research Ethics 
Committees (RECs), and regulatory authorities (if applicable) for written approval. The Principal 
Investigator (PI) will submit and, where necessary, obtain approval from the above parties for all 
amendments to the original approved documents. The Investigator will notify deviations from the 
protocol or SAEs occurring at the site to REC(s) in accordance with procedures.  
The principal investigator shall submit a report once a year throughout the study, or on request, to 
the ethic committees. In addition, an End of Study notification and final report will be submitted to 
the ethic committees. 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 56 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
 
5. SPONSOR: University of Oxford 
Contact information: University Offices, Wellington Square, Oxford, OX1 2JD, United Kingdom 
The study sponsor has a role in the design of the study; collection of data; management, analysis, 
and interpretation of data; writing of the report; and the decision to submit the report for publication. 
 
6. FUNDER 
The funder, the Wellcome Trust, had no role in the design of the study. The funder will not have any 
role in its execution, analyses, and interpretation of data or decision to submit results. 
 
7. TRIAL COMMITTEES 
Programme Steering Committee (PSC): The role of PSC is to provide overall supervision for the 
project on behalf of the Project Sponsor and Project Funder and to ensure that the project is 
conducted to the rigorous standards set out in the Research Governance Framework for Health and 
Social Care, and in the Guidelines for Good Clinical Practice. The composition and roles and 
responsibilities of the PSC, and the PSC meeting schedule, are defined in the PSC Terms of 
Reference. 
 
8. INSURANCE 
The University of Oxford has a specialist insurance policy in place - Newline Underwriting 
Management Ltd, at Lloyd’s of London – which would operate in the event of any volunteer 
suffering harm as a result of their involvement in the research. 
 
9. PUBLICATION POLICY 
All Investigators will be involved in reviewing drafts of the manuscripts, abstracts, press releases 
and any other publications arising from the study. Authorship will be determined in accordance 
with the International Committee of Medical Journal Editors (ICMJE) guidelines and other 
contributors will be acknowledged. 
The trial results for each volunteer will be communicated individually to that individual to inform 
his/her health history.  
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 57 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
APPENDIX D: LIST OF ABBREVIATIONS 
AE 
adverse event 
β-HCG 
beta–Human Chorionic Gonadotropin 
BP 
blood pressure 
BT 
body temperature 
BUN 
blood urea nitrogen 
CBC 
complete blood count 
CHIK 
chikungunya 
CHMI 
controlled human malaria infection 
CMV 
cytomegalovirus 
Cr 
creatinine 
CRF 
case record form  
CRO 
Contract Research Organization 
CSP 
circumsporozoite 
CTCAE 
Common Terminology Criteria for Adverse Events 
CTSG 
Clinical Trials Supporting Group 
CYP2D6 
enzyme predicting primaquine metabolism (p450 enzyme) 
DARC 
Duffy antigen receptor for chemokines 
DBP 
Duffy binding protein 
DEN 
Dengue 
DFA 
Direct Feeding Assay 
DSMB 
Data Safety Monitoring Board 
EBV 
Epstein-Barr virus 
ELISA 
Enzyme-linked Immunosorbent Assay 
FBS 
fasting blood sugar 
FTM 
Faculty of Tropical Medicine 
FTMCTU 
Clinical Therapeutics Unit (healthy volunteer ward), Faculty of Tropical 
Medicine 
FTMEC 
Ethics Committee, Faculty of Tropical Medicine 
G6PD 
glucose-6-phosphate dehydrogenase 
GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
HBsAg 
Hepatitis B surface antigen 
HBV 
Hepatitis B virus 
HCV 
Hepatitis C virus 
HIV 
Human Immunodeficiency virus 
HTLV 
Human T cell Lymphotrophic virus 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
JE 
Japanese Encephalitis 
KEMRI 
Kenya Medical Research Institute 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 58 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
 
 
LDL 
low density lipoprotein 
LFT 
liver function test 
MFA 
membrane feeding assay 
MORU 
Mahidol Oxford Tropical Medicine Research Unit 
MVRU 
Mahidol Vivax Research Unit 
OxTREC 
Oxford Tropical Research Ethics Committee 
PCR 
polymerase chain reaction 
P. falciparum 
Plasmodium falciparum 
PI 
Principal Investigator 
PIS 
Participant Information Sheet 
PMR 
parasite multiplication rate 
PQ 
primaquine 
PR 
pulse rate 
P. vivax 
Plasmodium vivax 
PvCSP 
P. vivax circumsporozoite 
qPCR 
quantitative polymerase chain reaction 
RBC 
red blood cell 
RR 
respiratory rate 
RUNMC 
Radboud University Nijmegen Medical Centre 
SAE 
serious adverse event 
SAP 
Statistical Analysis Plan 
SOP 
Standard Operating Procedure 
TMDR 
Tropical Medicine Diagnostic Reference Laboratory 
QIMR 
Queensland Institute of Medical Research 
ULN 
upper limit of normal 
USMMVP 
U.S. Military Malaria Vaccine Program 
WHO 
World Health Organization 
WI 
Work Instruction 
ZIK 
Zika virus 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 59 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
APPENDIX E: AMENDMENT HISTORY 
Amendmen
t No. 
Proposal 
Version 
No. 
Date issued 
Author(s) of 
changes 
Details of Changes made 
1 
v.4.0 
1 
November 
2021 
Salintip 
Chanchaivorawith, 
MPH 
1.The list of investigators is 
updated. 
 
2.PIS/ICF documentation is 
updated.  
2 
v.5.0 
7 February 
2022 
Salintip 
Chanchaivorawith, 
MPH 
1.For inclusion criteria,  
COVID-19 vaccination at 
least two doses of a 
vaccine(s) is included. 
 
2.For exclusion criteria, 
clarifying about COVID-19 
testing that must be 
diagnosed by PCR 
(nasopharyngeal swab). 
 
3.To include EBV and CMV 
serology testing in screening 
visit 3 (day -1) and during 2-
4 weeks after the inoculation. 
 
4.To include COVID-19 
serology Point-of-Care Test 
(POCT) in screening visit 3 
(day -1) and day -14 after the 
inoculation. 
 
5.Administrative procedures:  
one withdrawal 
criteria/discontinuation 
criteria about current 
COVID-19 infection during 
participant's admission in the 
hospital (after the 
inoculation) is added. 
 
 
6.Administrative procedure: 
to update criteria of COVID-
19 testing during participant's 
admission in the hospital. 
 

 
Research Proposal Submission Form  
for a study involving human subject enrollment WITH 
specimen collection 
Document No.:  FTM 
ECF-033-01 
Page 60 of 60 
 
 
 
MIST2_Research Proposal Submission Form, version 7.0, dated 3 January 2024 
 
 
3 
V6.0 
27 April 
2022 
Assist. Professor 
Borimas 
Hanboonkunupakar
n  
 
1) To remove Hemoglobin 
testing from screening1 
2) To change COVID-19 
serology POC test to 
COVID-19 serology  
3) To specify 2nd COVID-
19 serology test at Day 
discharge  
4) To revise blood 
collected volume 
according to the new 
laboratory requirement 
and new tests 
5) To revise laboratories 
and permission to 
perform lab test (s) at 
other validated  
6) laboratories   
7) Administrative changes 
and some typing errors 
 
4 
V7.0 
3 January 
2024 
Assist. Professor 
Borimas 
Hanboonkunupakar
n  
 
1) To update the number of 
inoculum stocks from MIST 
1 study (TMEC 19-067). 
2) To update the number of 
participants as per the 
updated stock of inoculum.  
 3) To perform additional 
laboratory tests as per DSMB 
suggestion.  
 - HIV-I and HIV-II antigen, 
HBV and  HCV serology 
testing before hospital 
discharge  
- HIV-I and HIV-II antigen, 
HBV, HCV, CMV, and EBV 
serology testing on 1 year 
follow-up phase 
4) To revise blood collection 
volume according to the 
additional laboratory test.  
5) Administrative changes