Flavivirus Cross-priming Potential of Live Attenuated Japanese Encephalitis (JE) Vaccine IMOJEV in Flavivirus naïve and Flavivirus Experienced Participants
Phase: NA
Conditions: Japanese Encephalitis
Sponsor: University of Liverpool

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Flavivirus cross-priming potential of live attenuated 
Japanese encephalitis (JE) vaccine IMOJEV® in 
flavivirus naïve and flavivirus experienced participants 
 
 
 
 
FlaviPrime Protocol  
V.6, 08/07/2021 
 
 
 
 
Study Sponsor(s): 
University of Liverpool 
2nd Floor Block D Waterhouse Building 
3 Brownlow Street 
Liverpool 
L69 3GL 
 
Clinicaltrials.gov: NCT03920111 
 
Research Ethics Ref: 19/NW/0330 
 
Sponsor Ref: UoL001462 
 
 
             

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Protocol Approval  
I, the undersigned, hereby approve this clinical study protocol:  
 
 
Authorised by Chief Investigator:  
 
 
 
 
 
Signature:  _______________________________________ 
Date: _____________ 
 
Dr Lance Turtle 
Senior Clinical Lecturer/Honorary Consultant in Infectious Diseases 
Institute of Infection and Global Health 
University of Liverpool 
 
 
 
Aug 9, 2021

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I, the undersigned, hereby approve this clinical study protocol:  
 
 
Authorised on behalf of Sponsor:  
 
 
 
 
 
 
 
 
Signature:  
_______________________________________ 
Date: _____________ 
 
Dr Neil French 
Head of Clinical Operations 
University of Liverpool 
Neil French (Aug 17, 2021 14:04 GMT+1)
Neil French
Aug 17, 2021

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I, the undersigned, hereby approve this clinical study protocol:  
 
 
 
Authorised on behalf of the Lead Statistician: 
 
 
 
 
 
Signature:  
______________________________________ 
Date: _____________ 
 
Richard Jackson 
Study Statistician 
University of Liverpool 
 
 
Richard Jackson (Aug 17, 2021 16:35 GMT+1)
Richard Jackson
Aug 17, 2021

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General Information 
This document describes the FlaviPrime study including detailed information about procedures and 
recruitment. The protocol should not be used as an aide-memoir or guide for the treatment of other patients. 
Every care was taken in its drafting, but corrections or amendments may be necessary. Any amendments 
will be circulated to the investigators participating in the study, but sites entering participants for the first time 
are advised to contact the coordinating centre (Liverpool Clinical Trials Centre) to confirm they have the most 
up to date version. Clinical problems relating to this study should be referred to the relevant Chief Investigator, 
Dr Lance Turtle, via the CTU. 
 
This protocol defines the participant characteristics required for study entry and the schedule of treatment 
and follow-up. Participant recruitment will be undertaken in compliance with this document and applicable 
regulatory and governance requirements. Waivers to authorise non-compliance are not permitted. 
Incidence of protocol non-compliance whether reported prospectively (e.g. where a treatment cannot be 
administered on a scheduled date as a result of public holidays) or retrospectively noted (e.g. as a result of 
central monitoring) are recorded as protocol deviations. These are monitored and reported to study oversight 
committees. 
 
The template content structure is consistent with the SPIRIT (Standard Protocol Item: Recommendations for 
Interventional Trials 2013) and has regard for the Health Research Authority guidance. Regulatory and ethical 
compliance information is located in section 14. 
 
The Liverpool Clinical Trials Centre has achieved full registration by the UK Clinical Research Collaboration 
(www.ukcrc.org) as their standards and systems were assessed by an international review panel as reaching 
the highest quality. The Liverpool Clinical Trials Centre has a diverse trial portfolio underpinned by 
methodological rigour, a GCP compliant data management system, and quality management system. 
 
 

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Contact Details: Institutions 
Sponsor : 
 
Study Management, Monitoring 
and Analysis: 
 
Clinical Laboratory: 
(central) 
 
Dr Neil French 
Head of Clinical Operations 
Clinical Directorate 
4th Floor Thompson Yates Building 
Faculty of Health and Life Sciences 
University of Liverpool 
Liverpool L69 3GB 
 
Telephone: 0151 794 8373 
E-mail: sponsor@liv.ac.uk 
Liverpool Clinical Trials Centre 
(LCTC) 
The University of Liverpool 
1st Floor, Block D 
Waterhouse Building 
3 Brownlow Street 
Liverpool 
L69 3GL 
 
Telephone: 0151 794 9766 
Email:  
flaviprime@liverpool.ac.uk 
Dr Lance Turtle, PhD, MRCP 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE  
 
Telephone: 0151 795 9668 
Email: lance.turtle@liverpool.ac.uk 
Statistics: 
Other Medical or Technical 
Departments and/or Institutions 
(1): 
Other Medical or Technical 
Departments and/or Institutions 
(2): 
Richard Jackson 
Liverpool Clinical Trials Centre 
The University of Liverpool 
1st Floor, Block D 
Waterhouse Building 
3 Brownlow Street 
Liverpool 
L69 3GL 
 
Telephone: 0151 795 5288 
Email: r.jackson@liv.ac.uk  
Dr Lance Turtle, PhD, MRCP 
University of Liverpool 
Institute of Infection, Veterinary and 
Ecological Sciences, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE  
 
Telephone: 0151 795 9642 
Email: lance.turtle@liverpool.ac.uk 
Prof Tom Solomon, PhD, FRCP 
University of Liverpool 
Institute of Infection, Veterinary and 
Ecological Sciences, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
 
Tel: 0151 795 9626 
Email: tsolomon@liverpool.ac.uk 
 
 
 

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Contact Details: Individuals 
Individual Authorised to Sign the 
Protocol 
and 
Protocol 
Amendments on behalf of the 
Sponsor: 
Chief Investigator (CI): 
 
Dr Lance Turtle, PhD, MRCP 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE  
 
Telephone: 0151 795 9668 
Email: lance.turtle@liverpool.ac.uk 
Dr Lance Turtle, PhD, MRCP 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE  
 
Telephone: 0151 795 9668 
Email: lance.turtle@liverpool.ac.uk 
 
 
In cases where the CI is unavailable to respond to urgent queries the following individual/s will act as cover: 
Medical Expert who will Advise 
on 
Protocol 
Related 
Clinical 
Queries: 
Medical Expert who will Evaluate 
SAE Reports: 
 
Prof Tom Solomon, PhD, FRCP 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
 
Tel: 0151 795 9626 
Email: tsolomon@liverpool.ac.uk 
 
Prof Neil French PhD, MRCP 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
 
Tel: 0151 795 9687 
Email: french@liverpool.ac.uk 
 
 
Additional Contacts:  
The contact details for the study oversight committee members and participating centres are detailed in 
documents supplementary to the protocol and stored in the Study Master File 
 
 
 

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 Table of Contents 
Protocol Approval ......................................................................................................................................... 2 
 
Table of Contents .............................................................................................................................. 8 
 
Protocol Overview ........................................................................................................................... 12 
 
Schematic of Study Design ......................................................................................................... 16 
 
Roles and Responsibilities .............................................................................................................. 17 
Sponsor .................................................................................................................................................. 17 
Funder 17 
 
Protocol Contributors .................................................................................................................. 18 
 
INTRODUCTION ............................................................................................................................ 20 
 
Background ................................................................................................................................. 20 
 
Rationale ..................................................................................................................................... 21 
 
Risk and Benefits ........................................................................................................................ 22 
 
Objectives ................................................................................................................................... 23 
Exploratory/Translational/Mechanistic Objective(s)....................................................... 23 
 
STUDY DESIGN ............................................................................................................................. 25 
 
Blinding ....................................................................................................................................... 25 
 
Study Setting ............................................................................................................................... 26 
 
ELIGIBILITY CRITERIA .................................................................................................................. 27 
 
Inclusion Criteria ......................................................................................................................... 27 
 
Exclusion Criteria ........................................................................................................................ 27 
 
Co-enrolment Guidelines ............................................................................................................. 28 
 
STUDY TREATMENT/INTERVENTIONS ....................................................................................... 29 
 
Introduction ................................................................................................................................. 29 
 
Treatment Name / Description ..................................................................................................... 29 
 
Manufacturing and Distribution .................................................................................................... 30 
 
Preparation, Dosage and Administration ..................................................................................... 30 
 
Accountability Procedures ........................................................................................................... 31 
 
Assessment of Compliance ......................................................................................................... 31 
 
Concomitant Medications/Treatments and Specific Restrictions .................................................. 31 
Medications Permitted .................................................................................................. 31 
Medications Not Permitted/ Precautions Required ........................................................ 31 
Data on Concomitant Medication .................................................................................. 32 
Data on Concomitant Medication .................................................................................. 32 
Specific Requirements/Restrictions .............................................................................. 32 
 
Overdose .................................................................................................................................... 33 
 
OUTCOMES ................................................................................................................................... 34 
 
PARTICIPANT TIMELINES AND ASSESSMENTS ......................................................................... 35 
 
Participant Identification and Screening ...................................................................................... 35 
 
Informed Consent ........................................................................................................................ 35 
Prospective Informed Consent Process ........................................................................ 35 
 
Screening, Eligibility Assessment and Confirmation .................................................................... 36 
 
Enrolment/Registration and Baseline Assessments .................................................................... 37 
 
Schedule for Assessments and Follow-up ................................................................................... 37 
Schedule of Assessments: ........................................................................................... 38 
Clinical Procedures....................................................................................................... 39 
Safety Assessments – Adverse Events (AE) ................................................................ 39 
Special Assays or Procedures ...................................................................................... 40 
 
Sampling ..................................................................................................................................... 41 
Sample Collection ........................................................................................................ 41 
Sample Shipment/Custodianship .................................................................................. 41 
Sample Storage and Handling ...................................................................................... 41 
 
Intervention Discontinuation and Participant Discontinuation/Withdrawal .................................... 41 
Premature Discontinuation of Study Intervention .......................................................... 42 
Participant Transfer ...................................................................................................... 43 
Loss to Follow-up ......................................................................................................... 43 

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Travel 43 
 
End of Study ............................................................................................................................... 43 
Study Discontinuation ................................................................................................... 44 
 
SAFETY REPORTING .................................................................................................................... 45 
 
Terms and Definitions ................................................................................................................. 45 
 
Assessment of Seriousness ........................................................................................................ 45 
 
Severity of Adverse Events ......................................................................................................... 46 
 
Assessment of “Causality” - Relationship to Study Treatment/Intervention .................................. 46 
 
Assessment of “Expectedness” ................................................................................................... 47 
Reference Safety Information / Information used to Assess Expectedness .................. 47 
 
Time Period for Active Monitoring of Safety Events ..................................................................... 47 
 
Notes on Safety Event Recording ............................................................................................... 48 
 
Reporting of Pregnancy .................................................................................................................. 49 
Notification of Deaths ................................................................................................... 50 
 
Reporting Procedures ................................................................................................................. 51 
Flowchart for Site Reporting Requirements of Adverse Events ..................................... 51 
Reporting Safety Events to the LCTC ........................................................................... 51 
Follow-up after Adverse Events .................................................................................... 51 
 
Investigator Reporting Responsibilities ........................................................................................ 52 
 
LCTC Responsibilities ................................................................................................................. 53 
Safety Reports .............................................................................................................. 54 
Urgent Safety Measures (USMs) .................................................................................. 54 
 
Contact Details and Out-of-hours Medical Cover......................................................................... 54 
 
STATISTICAL CONSIDERATIONS ................................................................................................ 55 
 
Introduction ................................................................................................................................. 55 
 
Sample Size ................................................................................................................................ 55 
 
Method of Registration ................................................................................................................ 55 
 
Outcome Measures ..................................................................................................................... 55 
 
Exploratory Objectives ................................................................................................................ 55 
 
Interim Analyses.......................................................................................................................... 56 
 
Analysis Plan .............................................................................................................................. 56 
 
DATA MANAGEMENT AND STUDY MONITORING....................................................................... 58 
 
Source Documents ...................................................................................................................... 58 
 
Data Collection Methods ............................................................................................................. 58 
 
Monitoring ................................................................................................................................... 58 
Central Monitoring ........................................................................................................ 58 
Clinical Site Monitoring ................................................................................................. 59 
 
Risk Assessment ......................................................................................................................... 59 
 
Confidentiality ............................................................................................................................. 59 
 
Quality Assurance and Control .................................................................................................... 60 
 
Records Retention ...................................................................................................................... 60 
 
REGULATORY AND ETHICAL CONSIDERATIONS ...................................................................... 62 
 
Statement of Compliance ............................................................................................................ 62 
 
Ethical Considerations................................................................................................................. 62 
 
Approvals .................................................................................................................................... 62 
 
Protocol Deviation and Serious Breaches ................................................................................... 62 
Non-Serious breaches .................................................................................................. 62 
Serious breaches ......................................................................................................... 62 
 
INDEMNITY .................................................................................................................................... 64 
 
PUBLICATION AND DISSEMINATION ........................................................................................... 65 
 
Publication Policy ........................................................................................................................ 65 
 
Dissemination to Key Stakeholders ............................................................................................. 65 
 
Data Sharing ............................................................................................................................... 65 
 
CHRONOLOGY OF PROTOCOL AMENDMENTS ......................................................................... 66 
 
Version 5.4 (21/06/2021) ............................................................................................................. 66 
 
Version 5.0 (08/03/2021) ............................................................................................................. 68 
 
Version 4.0 (18/12/2020) ............................................................................................................. 68 

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Version 3.0 (08/08/2020) ............................................................................................................. 69 
 
Version 1.2 (15/01/2020) ............................................................................................................. 72 
 
Version 2.0 (27/06/2019) ............................................................................................................. 72 
 
Version 1.1 (17/04/2019) ............................................................................................................. 72 
 
REFERENCES ............................................................................................................................... 73 
 
DOCUMENTS SUPPLEMENTARY TO THE PROTOCOL .............................................................. 75 
 
 

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Glossary 
 
AE 
Adverse Event 
CI 
Chief Investigator 
C 
Core protein 
CRF 
Case Report Form 
CTU 
Clinical Trials Unit 
DENV 
Dengue virus 
ELISpot 
Enzyme linked immunospot 
E 
Envelope Protein 
GCP 
Good Clinical Practice 
HBsAg 
Hepatitis B surface antigen 
HCV 
Hepatitis C virus 
HIV 
Human immunodeficiency virus 
HRA 
Health Research Authority 
IB 
Investigator’s Brochure 
IDSMC 
Independent Data and Safety Monitoring Committee 
IEC 
Independent Ethical Committee 
IFNγ 
Interferon-gamma 
IMP 
Investigational Medicinal Product 
IVIG 
Immunoglobulin therapy 
JE 
Japanese encephalitis 
JEV 
Japanese encephalitis virus 
LCTC 
Liverpool Clinical Trials Centre 
LREC 
Local Research Ethics Committee 
LUHFT 
Liverpool University Hospitals Foundation Trust 
MAIMP 
Manufacturing authorisation for investigational medical product 
MCRN CTU 
Medicines for Children Clinical Trials Unit 
MREC 
Multi-centre Research Ethics Committee 
NS 
Non-structural protein 
PFU                     Plaque forming unit 
PBMC  
Peripheral blood mononuclear cell 
PI 
Principal Investigator 
prM 
Pre-membrane protein 
PV 
Pharmacovigilance 
QP 
Qualified person 
R&D 
Research & Development 
RNA 
Ribonucleic acid 
RUAE 
Related Unexpected Adverse Event 
RUSAE 
Related Unexpected Serious Adverse Event 
SAE 
Serious Adverse Event 
UTR 
Untranslated region 
WHO 
World Health Organisation 
WNV 
West Nile Virus 
YFV 
Yellow Fever virus 
ZIKV 
Zika Virus 
 
 
 
 
 

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Protocol Overview 
 
Full Title:  
Flavivirus cross-priming potential of live attenuated Japanese
encephalitis (JE) vaccine IMOJEV® in flavivirus naïve and 
flavivirus experienced participants. Human experimental medicine 
study. 
Acronym: 
FlaviPrime 
Phase: 
IV 
Target Population: 
Healthy adults, aged 18 – 70 years 
Sample size: 
Up to 25 participants in total to be vaccinated: 
Group 1; up to 3-4 participants of any previous flavivirus exposure 
state 
Group 2; up to 6-10 flavivirus naïve participants 
Group 3; up to 8-12 flavivirus experienced participants 
Inclusion Criteria: 
 
1. A male or female adult between 18 and 70 years of age at 
consent. 
2. Written and informed consent obtained from participant and 
agreement of participant to comply with the requirements of 
the study 
3. Able to attend regularly to donate study blood samples for 
the duration of the study (8 weeks), no planned re-location 
or travel to a flavivirus endemic area during the study 
period. 
4. Satisfactory medical screen, as demonstrated by study 
screening document normal physical examination and 
normal screening blood tests 
5. Group 1: Any flavivirus exposure status; Group 2: No
previous flavivirus vaccination (JE, tick borne encephalitis or
yellow fever (YF)), no residence in a flavivirus endemic area
nor planned travel to a flavivirus endemic area during the
period of the study; Group 3: JE vaccine and/or yellow fever
vaccine or other proven flavivirus infection within the last 10
years or other proven flavivirus infection (lifetime). 
6. An efficacious method of contraception must be used 
during the study for 28 days after vaccination for women of 
childbearing potential (see section 11). 
Exclusion Criteria:  
 
1. Use of any investigational or non-registered drug within 5 
half-lives of the drug, or 30 days preceding administration 
of study JE vaccine, whichever is longer; or planned use 
during the study period. 

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2. Receipt of any investigational biologic agents with 
mechanisms of action that might affect the immune system, 
at the discretion of the CI and local PI. 
3. Administration of immunosuppressants or other immune-
modifying drugs within a period of six months before 
vaccination or at any time during the study period; 
participants who have received these agents may also be 
excluded at the discretion of the CI and local PI. 
4. Any confirmed or suspected immunosuppressive or 
immunodeficient condition. 
5. A family history of congenital or hereditary 
immunodeficiency. 
6. Any antiviral drug therapy within a period of 5 drug half-
lives or 30 days before vaccination, whichever is longer, or 
at any time during the study period. 
7. History of significant allergic reactions likely to be 
exacerbated by any component of the study vaccine, 
especially allergic disease or reactions to any previous 
dose of any vaccine. 
8. History of having received JE vaccine, yellow fever 
vaccine, tick-borne encephalitis vaccine or experimental 
flavivirus vaccine (group 2 only). 
9. Acute disease (for example acute infection) at the time of 
enrolment or vaccination, if symptoms are rated as 
anything more significant than a mild adverse event.  Entry 
into the study and/or vaccination may be deferred until the 
illness has resolved for at least one week.  
10. Acute or chronic, clinically significant in the opinion of the 
investigator, disease in any organ system, as determined 
by history, physical examination or laboratory testing. 
11. Presence of any inflammatory condition that might require 
immunomodulatory therapy. 
12. Recent blood donation (inclusion can be delayed under 
these circumstances; the participant should be enrolled 16 
weeks after their last blood donation. Each participant 
should give no more than 470 ml per 16 weeks, so regular 
blood donation should be suspended during the study and 
can re-commence 1 month after the last study sample). 

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13. Current or previous abattoir worker or sheep farmer in 
Scotland (risk of Louping ill virus exposure; group 2 only). 
14. Administration of immunoglobulins and/or any blood 
products within the three months preceding administration 
of vaccine, or planned administration during the study 
period. 
15. Seropositive for HIV. 
16. Pregnancy or Lactation.  
17. History of excessive alcohol consumption (>28 units per 
week), drug abuse or significant psychiatric illness. 
18. Any other condition or consideration that, in the opinion of 
the Investigator, would pose a health risk to the participant 
if they were enrolled in the study, or would otherwise 
interfere with the evaluation of the study aims (e.g. difficult 
venesection). 
Study Centres and 
Distribution: 
Single site – Liverpool University Hospitals NHS Foundation 
Trust (LUHFT, Royal Liverpool site) Clinical Research Unit 
(CRU) 
Participant Study 
Duration: 
Group 1: 28 days after vaccination; Groups 2 and 3: 8 weeks 
after vaccination 
Study Duration  
Participant duration: 
Group 1: 28 days  
Groups 2 and 3: 8 weeks  
IMP / Intervention: 
 
Intervention: Single dose live attenuated JE vaccine 
IMP: 
Form: 
Dose: 
Route: 
IMOJEV®  
Intra dermal – injection  
Single dose, 0.5mL (≥104.0 – 5.8 PFU) 
Subcutaneous 
No adjuvant or antimicrobial preservative has been added. 
Objectives: 
 
Exploratory/ 
Translational; 
Primary:  
Refer to section 8 for further 
details on endpoint/outcome 
measures 
1. To establish a human model system 
of JEV infection in healthy adult 
volunteers using live attenuated JE 
vaccine IMOJEV®. 
2. To sort and sequence individual 
responding B cells (plasmablasts) 
after vaccination with IMOJEV®, and 

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to generate human monoclonal 
antibodies to JEV.  
3. To generate JEV specific human 
monoclonal antibodies from the 
sequences derived in (2). 
4. To describe the development, 
specificity, cross-reactivity and 
function of the T cell response to 
IMOJEV®. 
5. To establish a sample bank for future 
work on cross-reactive and other 
responses to flaviviruses, flavivirus 
vaccines and other emergent 
viruses. 
Exploratory/ 
Translational; 
Secondary: 
Refer to section 8 for further 
details on endpoint/outcome 
measures 
1. To examine the specificity and cross-
reactivity of the antibody response 
after JE vaccination, using serum 
and human monoclonal antibodies. 
2. To determine whether there are 
epitopes which can serve as the 
target of broadly cross-neutralising 
antibody responses. 
 
 
 
 
 
 

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Schematic of Study Design 
Figure 1. Schematic of Study Design: 
 
Prior to enrolment — screening, flavivirus immune assays, clinical history, as part of the Acute emerging 
Virus Immunity Study (AVIS). 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 2. Schematic of Study Schedule: 
 
 
0
4
8
1
2
Vaccine
Study
week
FlaviPrime (IMOJEV) study schedule
Screening/
consent
= blood sample.
0
1
2
3
4
5
6
7
8
9
10
Study
day
= Single dose JE
vaccine IMOJEV
Vaccine
Screening/
consent
GROUP 1
GROUPS
2 & 3
Any Flavivirus status 
JE vaccine n = 3-4 
Screening 
Flavivirus naïve  
JE vaccine n = 6-8 
Cross-reactivity 
Approximately 100 monoclonal antibodies 
Group 1 
Group 2 
Group 3 
Flavivirus exposed 
JE vaccine n = 8-10 

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Roles and Responsibilities 
Sponsor 
The University of Liverpool is the Sponsoring organisation and is legally responsible for the study. They will 
formally delegate specific Sponsoring roles to the Chief Investigator and Clinical Trials Unit. 
 
Funder 
This study is funded as part of a Fellowship Award by the Wellcome Trust and managed by the Chief 
Investigator and Research Support Office at the University of Liverpool. 
 
 
Funder(s) 
 
Financial and Non-financial 
Support Given 
Role 
 
Wellcome Trust 
Up to £653,257 
This funding source had no role in the 
design of this study and will not have 
any 
role 
during 
its 
execution, 
analyses, interpretation of the data, or 
decision to submit results 
 
 
Chief Investigator: Dr Lance Turtle is the Chief Investigator for the study and is responsible for overall design 
and conduct of the study in collaboration with other members of the study team. 
 
Principal Investigators: In each participating centre a principal investigator will be identified to be 
responsible for identification, recruitment, data collection and completion of CRFs, along with follow up of 
study participants and adherence to study protocol at site. They will also be responsible for safety reporting 
and processing any applicable safety information. 
 
 
Clinical Trials Unit: The Liverpool Clinical Trials Centre (LCTC) at the University of Liverpool in collaboration 
with the Chief Investigator, will have overall management responsibility and will be responsible for study 
management activities including (but not limited to) study planning, budget administration, Study Master File 
management, safety reporting, data management, registration, statistical analysis, participating site 
coordination and IMP management.  
 
Oversight Committees 
 
FlaviPrime study is subject to oversight from the following committee: 
 
Study Management Group (SMG)  
A Study Management Group (SMG) will be formed comprising the Chief Investigator, other lead investigators 
(clinical and non-clinical) and members of the LCTC. The SMG are responsible for monitoring all aspects of 
the progress and conduct of the study and will be responsible for the day-to-day running and management 
of the study. The SMG will meet at least monthly at setup stage and then reduce to quarterly throughout the 
year unless more frequent meetings are required. 
 
Study Oversight Committee (SOC) 
The Study Oversight Committee will consist of an independent chairperson, 2/3 independent experts in the 
field of infectious diseases, biostatistician, including the CI and observers the role of the SOC is to provide 
overall supervision for the study and provide advice through its independent Chairman. The decision for the 
continuation of the study lies with the SOC and as such they will meet throughout the study (at least annually). 
 

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Protocol Contributors 
 
Name  
Affiliations 
Contribution to protocol 
Dr Lance Turtle 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
Development of protocol, clinical 
expertise 
Prof Tom Solomon 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
Development of protocol, clinical 
expertise 
Prof Neil French 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
Development of protocol, clinical 
expertise 
Richard Jackson 
The University of Liverpool 
Liverpool Clinical Trials Unit 
1st Floor, Block D 
Waterhouse Building 
3 Brownlow Street 
Liverpool 
L69 3GL 
Statistical expertise 
Kellie Platt 
The University of Liverpool 
Liverpool Clinical Trials Unit 
1st Floor, Block D 
Waterhouse Building 
3 Brownlow Street 
Liverpool 
L69 3GL 
Trial Management expertise 
Hayley Hardwick 
University of Liverpool 
Institute of Infection and Global 
Health, 
Ronald Ross Building, 
8 West Derby Street, 
Liverpool, L69 7BE 
Trial Management expertise 
Priya Francis 
Liverpool Clinical Trials Centre 
The University of Liverpool 
2nd Floor Institute in the Park 
Alder Hey Children’s NHS 
Foundation Trust 
Eaton Road 
Liverpool 
L12 2AP 
Trial Management expertise 
Mr Alex Astor 
The University of Liverpool 
Research Support Office 
2nd Floor, Block D 
Waterhouse building 
3 Brownlow Street 
Liverpool 
L69 3GL 
Sponsorship and protocol 
approval 

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Dr Mansour Yaïch 
 
Vaxyn Consulting, 
Paris 75001, 
Île-de-France, 
France  
Development of protocol 
 
 

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INTRODUCTION 
 
Background  
Japanese encephalitis and the genus Flavivirus 
Japanese encephalitis (JE) is the most important epidemic encephalitis in the world. JE is caused by JE virus 
(JEV), a member of the genus Flavivirus, which contains the most important mosquito borne viruses of man, 
including dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV) and Zika virus (ZIKV). 
Japanese encephalitis (JE) remains the most common epidemic encephalitis globally with 68,000 cases per 
year[1]. DENV causes 100 million illnesses annually[2], and keeps spreading. Yellow fever virus (YFV) 
outbreaks in 2015-16 almost exhausted the World Health Organisation (WHO) vaccine stockpile at one 
point[3]. Recent ZIKV related neurological syndromes have shown us how vulnerable we are to the emergence 
of new flaviviruses[4, 5]. For several important flavivirus diseases there are no good vaccines available. 
 
The flavivirus genome is 11 kb in size, positive sense single stranded RNA, and encodes a single 10.3 kb 
open reading frame flanked by untranslated regions. The open reading frame is translated as a single 
polypeptide which is then cleaved by host and viral proteases to produce three structural proteins (core, pre-
membrane and envelope) and seven non-structural (NS) proteins, named NS1, NS2A, NS2B, NS3, NS4A, 
NS4B and NS5 (figure 3)[6]. 
 
Figure 3. Flavivirus genomic organisation. The structural proteins core (C), pre-membrane (prM) and envelope (E) proteins are 
present in the viral particle, and hence are found in inactivated vaccines. The non-structural (NS) proteins are made during replication, 
and therefore can be produced upon infection by a live attenuated vaccine. UTR = untranslated region. 
Flaviviruses are closely related[6], leading to a significant degree of immunological cross-reactivity. For 
example, DENV is in fact four closely related viruses with immunologic cross-priming potential[7]. Although 
cross priming may be protective[8] it can also be detrimental — interfering with vaccine development, 
diagnosis and resulting in dengue illness that is more severe than the first infection. Beyond dengue there is 
also clear evidence that flavivirus cross priming can affect disease potential. Several studies show a degree 
of partial protection against JE in people who have already experienced DENV infection and mount a 
secondary type of immune response to JEV resulting in higher antibody levels[7, 8]. JE vaccination may also 
make DHF less severe[9]. However, one study suggests that the presence of anti-JEV neutralising antibody 
(but not JE vaccination) might predispose to worse dengue illness[10], so the interactions are clearly complex 
and not completely understood. 
 
Studies using human monoclonal antibody technologies (mAb)[11], have begun to give new insight into 
mechanisms of antibody cross-reactivity and protection against flaviviruses[11, 12]. Only a small fraction of 
antibodies against DENV are strongly neutralising[13], and these frequently recognise quaternary epitopes 
(where the antibody binds several E proteins)[11, 14, 15]. Many antibodies enhance infection[12], particularly 
highly cross-reactive antibodies against the prM protein if DENV[16] and the fusion loop of envelope (E) 
domain II (a conserved part of E protein important for cell entry)[11, 17].  
 
To date, there has been only one study of human mAb to JEV, which made a modest number of antibodies 
from people receiving inactivated vaccine[18]. Murine and primate monoclonals have been more extensively 
studied[19-21] but no quaternary epitopes arising from productive infection in humans have been determined. 
C
prM
E
NS1
NS2A
NS2B
NS3
NS4A
NS4B
NS5
5' UTR
3' UTR
Structural proteins
Non-structural proteins

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More efficiently neutralising murine mAb are often less cross-reactive[22], but, importantly, mice and humans 
mount quite different responses to flavivirus envelope proteins[23, 24]. 
 
Recent observations made during the Zika outbreak in South America have revealed at least one B cell 
epitope (called the envelope dimer epitope I, EDE I) that can serve as the target of human antibodies that 
can potently neutralise all four DENV serotypes and ZIKV[11, 25]. It is possible that by studying the response 
to a replication competent viral envelope protein in flavivirus experienced volunteers, we are able to uncover 
a similar epitope for the JE serogroup (JEV, St Louis encephalitis virus, West Nile virus, Murray Valley 
encephalitis virus, Usutu virus). Such an epitope could then be reverse engineered into a vaccine against 
multiple viruses, as is underway for DENV and ZIKV. 
 
Vaccines against JE 
Several different vaccines have been used to control JE since the 1950s[26] (table 1, reviewed in ref 17). 
Randomised controlled trials have shown the efficacy of JE vaccination, and showed that neutralising 
antibody correlated with protection[9]. Early JE vaccines were mostly derived from mouse brain, but this 
manufacturing method has now been replaced due to safety concerns and inferior immunogenicity[26].  
 
IMOJEV® (Sanofi Pasteur) is a Chimeric JE vaccine of JEV SA14-14-2 envelope and pre-membrane protein 
inserted into a yellow fever backbone (figure 4). IMOJEV® (formerly known as ChimeriVax-JE) is licensed in 
Australia and New Zealand for the prevention of JE in travellers, it is also marketed in some parts of Southeast 
Asia. 
 
Table 1. Vaccines against Japanese encephalitis.  
 
Description 
Type 
Virus strain 
Common name 
Developer/country of 
manufacture 
 
 
 
 
 
Older vaccines, no longer in use: 
 
 
 
 
Mouse brain 
Inactivated 
Nakayama 
BIKEN 
Japan, BIKEN 
Mouse brain 
Inactivated 
Nakayama 
Green Cross 
Korea, Green Cross 
Mouse brain 
Inactivated 
Beijing-1 
 
Japan 
Primary hamster kidney 
Inactivated 
P3 
 
China 
 
 
 
 
 
Vaccines currently available: 
 
 
 
 
Vero cell 
Inactivated 
P3 
 
China 
Primary hamster kidney 
Live 
attenuated 
SA14-14-2 
 
Chengdu Biological 
Products, China 
Vero cell 
Inactivated 
Beijing-1 
JEBIK®V 
Japan, BIKEN 
Vero cell 
Inactivated 
Beijing-1 
ENCEVAC® 
Japan, Kaketsuken 
Vero cell 
Inactivated 
SA14-14-2 
IC51, IXIARO 
Intercell, Valneva 
Vero cell 
Inactivated 
Kolar-821564XY 
JENVAC 
Bharat Biotech Int. ltd, 
India 
Yellow fever 17D recombinant 
vectored 
Live 
attenuated 
SA14-14-2 
(Envelope) 
Imojev, 
Chimerivax JE 
Acambis, Sanofi 
Pasteur 
 
 
Rationale 
There is a pressing need for a better experimental system to understand flavivirus antibody responses, 
beyond dengue, to make sure we are using current vaccines to greatest effect and to inform the development 
of next-generation vaccines. This study will use live chimeric JE vaccine IMOJEV® as a tool for flavivirus 
epitope discovery. This will allow experimental JEV infection using replication competent, live, attenuated 
virus as a model, in a setting where the flavivirus infection history of humans can be tightly controlled.  
 

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Figure 4. Schematic of IMOJEV® JE vaccine. The pre-membrane (pre-M) and envelope (E) proteins of JEV (orange) are inserted 
into the attenuated yellow fever 17D virus. 
 
Importantly, this study must be conducted using a replication competent vaccine, and not the inactivated 
vaccine available in Europe, IXIARO. This is because a replicating virus particle undergoes several different 
changes of state during the replication cycle[27], exposing epitopes that may not be present in an inactivated 
vaccine.  
The use of the live attenuated JE vaccine IMOJEV® in volunteers in the UK also permits the study of immune 
responses to JEV in completely flavivirus naïve subjects, something which is very difficult to achieve in 
flavivirus endemic areas where multiple viruses co-circulate and infection history is hard to determine 
accurately. By including a group of volunteers who have had previous flavivirus exposure (JE vaccine, yellow 
fever vaccine or dengue illness), it will also be possible to study the effect of previous flavivirus exposure on 
cross-reactive responses to JE vaccine in a highly controlled manner without the uncertainty of natural 
exposure arising from residence in an endemic area.  
 
The specificity and cross-reactivity of antibody responses will be studied by cloning antibodies from 
plasmablasts (developing antibody secreting cells that appear early in an immune response). Plasmablast 
responses precede the development of antibody responses in blood, and a large antibody response usually 
implies a large plasmablast response. Giving a booster dose of live JE vaccine to children who have been 
immunized with a primary course of inactivated JE vaccine produces a marked (approximately 18-fold) rise 
in neutralizing antibody titre[28], therefore we expect this vaccine also to generate a significant plasmablast 
response after vaccination. The plasmablast population is very enriched for cells specific to the antigen of 
interest, and therefore makes ideal starting material to generate human monoclonal antibodies against 
specific pathogens[11]. 
 
These experiments are the first step in determining the nature of the immune response to sequential viral 
infections in known order using replication competent vaccines as models of viral infection in humans. This 
will have important implications for the sequence of use of future JE and dengue vaccines, and represents 
the first step towards combining flavivirus vaccines beyond the tetravalent dengue vaccines into a universal 
flavivirus vaccine.  
 
 
Risk and Benefits 
 Potential Risks 
The below side effects are stated in the patient information leaflet that accompanies IMOJEV® 
 
Table 2 – side effects of IMOJEV® 
 
5' UTR
3' UTR
Structural proteins
Non-structural proteins
Capsid
pre-M
Envelope
NS1
NS2A
NS2B
NS3
NS4A
NS4B
NS5
JEV
Yellow fever
Yellow
fever

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We have received clarification from the MHRA that this study is not a CTIMP and therefore does not require 
a CTA application to be submitted.  
 
More detail regarding management of risks associated with this study are detailed in a separate Risk 
Assessment maintained in the Study Master File. 
 Potential Benefits 
Participants will develop immunity to Japanese encephalitis 2 weeks after vaccination. 
 
 
Objectives  
Scientific hypotheses to be tested and experimental approach 
This study aims to establish a unique human experimental medicine system which will be used to study 
flavivirus immune responses and for epitope discovery. As such, although safety will be monitored in the 
same way as a late phase clinical trial, there will be no primary or secondary endpoint in this study. 
 
This study will test the hypothesis that in previously flavivirus-exposed individuals, the antibody response is 
more broadly cross neutralising, and that this will lead to the identification of conserved virion surface epitopes 
that could be the target of second generation vaccines. All the scientific objectives of this study are 
exploratory. 
 
Exploratory/Translational/Mechanistic Objective(s) 
 
 Exploratory Primary Objectives  
Specific aims: 
 
1. To establish a human model system of JEV infection in healthy adult volunteers using live attenuated 
JE vaccine IMOJEV®. 
How likely
Tiredness
Feeling unwell
Injection site pain
Headache
Muscular pain
Feeling hot
Vomiting
Dizziness
Chills
Throat pain
Joint pain
Injection site redness Shortness of breath Diarrhoea
Injection site itching
Runny nose
Abdominal pain
Injection site swelling Cough
Nasal congestion
Injection site bruising Wheezing
Nausea
Rash
Uncommon 
(more than 1 in 
1000 persons 
and less than 1 in 
100 persons
Fever
Rare (more than 
1 in 10000 
persons and less 
than 1 in 1000 
persons) 
Viral infections
Very Common
(1 in 10 persons)
Common 
(more than 1 in 
100 persons and 
less that 1 in 10 
persons)
Side Effect

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2. To sort and sequence individual responding B cells (plasmablasts) after vaccination with IMOJEV®, 
and to generate human monoclonal antibodies to JEV.  
3. To generate JEV specific human monoclonal antibodies from the sequences derived in (2). 
4. To describe the development, specificity, cross-reactivity and function of the T cell response to 
IMOJEV®. 
5. To establish a sample bank for future work on cross-reactive and other responses to flaviviruses, 
flavivirus vaccines and other emergent viruses. 
 
 Exploratory Secondary Objectives 
1. To examine the specificity and cross-reactivity of the antibody response after JE vaccination, using 
serum and human monoclonal antibodies. 
2. To determine whether there are epitopes which can serve as the target of broadly cross-neutralising 
antibody responses. 
 
Experimentally, the fine specificity and cross-reactivity of the antibody response will be studied by cloning 
antibodies from plasmablasts (B cells responding to the vaccine) that have been single cell sorted by flow 
cytometry then sequenced at one week post vaccine. These human monoclonal antibodies will then be 
mapped on to the surface of the virus particle using established approaches[11], and tested to look for cross-
reactive antibodies. T cell responses to the vaccine will be studied using custom pools of synthetic peptides 
by ELISpot and flow cytometry. 

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STUDY DESIGN 
FlaviPrime is an open label, three arm, human experimental medicine study in healthy volunteers of any 
flavivirus background (group 1), flavivirus naïve (group 2) and flavivirus exposed (group 3) volunteers (figure 
1). Each group will receive a single dose of JE vaccine IMOJEV®. Group 1 will be sampled daily from days 5 
to 9 after vaccination, and groups 2 and 3 will follow an identical schedule (figure 2) with 4 sampling points 
over 8 weeks of follow-up. After the study, participants will be followed up in the observational Acute emerging 
Virus Immunity Study (AVIS), aiming for 5 years of follow up. 
 
Group 1:  up to 3-4 healthy adults. 
 
Group 2: up to 6-10 healthy adults who have never travelled to a flavivirus endemic area and are negative 
in screening tests for flavivirus immunity. 
 
Group 3: up to 8-12 healthy adults who have had JE vaccine and/or are previously flavivirus exposed, either 
through receiving yellow fever vaccine up to 5 years before the study, or from being diagnosed with a flavivirus 
illness (e.g. dengue or Zika). 
Group 1 will allow confirmation of the kinetics of the plasmablast response which has never been addressed 
for any flavivirus vaccine. The inclusion of groups 2 and 3 will allow comparison of the adaptive immune 
response to JEV in flavivirus naïve individuals, verses flavivirus experienced individuals. Because the 
hypothesis under test is that after multiple exposure the response is focused on more conserved epitopes 
compared with single exposure, the exact virus on the previous exposure is not critical. Group 3 more faithfully 
mimics the reality of residence in flavivirus endemic areas where there is exposure to multiple viruses and it 
is hard to determine with certainty which viruses have infected any given individual. 
 
Up to 25 participants will be studied (Group 1 = up to 3-4, Group 2 = up to 6-10, and Group 3 = up to 8-12). 
Groups can comprise male and/or female participants aged from 18 to 70 years at the time of consent. The 
group sizes do not need to be fixed because no sample size based comparisons are pre-specified. Any 
comparisons will be post-hoc and exploratory. We anticipate that antibodies made from group 3 will have 
wider virus specificity and therefore greater potential future application and interest; hence this group is 
allowed to be slightly larger. 
 
Each group will receive a single dose of live JE vaccine IMOJEV®. All doses will be administered at the 
LUHFT CRU facility in accordance with a bespoke vaccination administration working instruction. Group 1 
participants will be in the study for 28 days and will give 6 blood samples, one on the day vaccination and 
then daily on days 5 through to 9 post vaccination. Group 2 and 3 participants will be in the study for a total 
of 8 weeks and will be expected to attend study visits and provide six blood samples at baseline, and weeks 
1, 2, 4, and 8 (see Figure 2, Study Schedule). All participants will have Adverse Events (AEs) recorded while 
they are in the study. Sample collection in the Acute emerging Virus Immunity study (AVIS) will be temporarily 
suspended during FlaviPrime, participants will then revert back to AVIS once the FlaviPrime sampling 
collection period has finished. Participants will be followed up, aiming for 5 years of follow up, and have 
already been consented for storage of contact details for future studies. 
 
Blinding 
This is an open label study with no blinding requirements.  All researchers and participants know which 
treatment / intervention is being administered. 

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Study Setting 
Participants will be identified and recruited from an existing observational study (“Immune responses to Zika 
virus and other flaviviruses,” (Acute emerging Virus Immunity Study, AVIS) 16/NW/0170), by advertisement, 
and through Liverpool University Hospitals NHS Foundation Trust (LUHFT) Clinical Research Unit (CRU), for 
example using the consent4consent database. Follow up will occur at LUHFT. 
 Selection of Participating Sites 
FlaviPrime is a single centre study that will undertake intensive laboratory investigation of the immune 
response to IMOJEV®. Therefore, the study site must be close to the necessary facilities (single cell sorting, 
sequencing, etc.) and will only be open at one site, the LUHFT CRU, which has the necessary expertise for 
this study. 
 
The site will be opened to recruitment upon successful completion of all global (e.g. REC) and study-specific 
conditions (e.g. site personnel training requirements) and once all necessary documents have been returned 
to the CTU. Initiation of the site will be undertaken in compliance with LCTC internal processes. Conditions 
and documentation required will be detailed on a LCTC Green Light Checklist maintained in the TMF and 
must be fully completed prior to opening sites to recruitment.  
 Selection of Principal Investigators 
Principal Investigators will be required to demonstrate equipoise, relevant experience and commitment during 
early stage feasibility assessment. All investigators will have the particular medical expertise necessary to 
conduct the study in accordance to the protocol and all regulatory and ethical requirements. Written 
agreement to conduct research as such will be obtained prior to site initiation. 
 
A suitable co-investigator should be identified at each site to deputise in case of PI absence. 

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ELIGIBILITY CRITERIA 
The FlaviPrime study aims to recruit up to 25 participants based on sample size calculations described in 
Section 12.2. All patients must provide written, informed consent before any study procedures occur (see 
Section 9.2 for more information regarding informed consent processes) and must meet all eligibility criteria 
as described below. 
 
Inclusion Criteria 
Patients eligible for the study must comply with all of the following at registration: 
 
1. A male or female adult between 18 and 70 years of age at consent. 
2. Written and informed consent obtained from participant and agreement of participant to comply with 
the requirements of the study 
3. Able to attend regularly to donate study blood samples for the duration of the study (8 weeks), no 
planned re-location or travel to a flavivirus endemic area during the study period. 
4. Satisfactory medical screen, as demonstrated by study screening document normal physical 
examination and normal screening blood tests 
5. Group 1: Any flavivirus exposure status; Group 2: No previous flavivirus vaccination (JE, tick borne 
encephalitis or yellow fever (YF)), no residence in a flavivirus endemic area nor planned travel to a 
flavivirus endemic area during the period of the study; Group 3: JE vaccine and/or yellow fever vaccine 
or other proven flavivirus infection within the last 10 years or other proven flavivirus infection (lifetime). 
6. An efficacious method of contraception must be used during the study for 28 days after vaccination 
for women of childbearing potential (see section 11).  
 
Exclusion Criteria  
Any patient meeting any of the criteria listed below at baseline will be excluded from study registration: 
 
1. Use of any investigational or non-registered drug within 5 half-lives of the drug, or 30 days preceding 
administration of study JE vaccine, whichever is longer; or planned use during the study period. 
2. Receipt of any investigational biologic agents with mechanisms of action that might affect the immune 
system, at the discretion of the CI and local PI. 
3. Administration of immunosuppressants or other immune-modifying drugs within a period of six months 
before vaccination or at any time during the study period; participants who have received these agents 
may also be excluded at the discretion of the CI and local PI. 
4. Any confirmed or suspected immunosuppressive or immunodeficient condition. 
5. A family history of congenital or hereditary immunodeficiency. 
6. Any antiviral drug therapy within a period of 5 drug half-lives or 30 days before vaccination, whichever 
is longer, or at any time during the study period. 
7. History of significant allergic reactions likely to be exacerbated by any component of the study 
vaccine, especially allergic disease or reactions to any previous dose of any vaccine. 
8. History of having received JE vaccine, yellow fever vaccine, tick-borne encephalitis vaccine or 
experimental flavivirus vaccine (group 2 only). 

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9. Acute disease (for example acute infection) at the time of enrolment or vaccination, if symptoms are 
rated as anything more significant than a mild adverse event.  Entry into the study and/or vaccination 
may be deferred until the illness has resolved for at least one week.  
10. Acute or chronic, clinically significant in the opinion of the investigator, disease in any organ system, 
as determined by history, physical examination or laboratory testing. 
11. Presence of any inflammatory condition that might require immunomodulatory therapy. 
12. Recent blood donation (inclusion can be delayed under these circumstances; the participant should 
be enrolled 16 weeks after their last blood donation. Each participant should give no more than 470 
ml per 16 weeks, so regular blood donation should be suspended during the study and can re-
commence 1 month after the last study sample). 
13. Current or previous abattoir worker or sheep farmer in Scotland (risk of Louping ill virus exposure; 
group 2 only). 
14. Administration of immunoglobulins and/or any blood products within the three months preceding 
administration of vaccine, or planned administration during the study period. 
15. Seropositive for HIV. 
16. Pregnancy or Lactation.  
17. History of excessive alcohol consumption (>28 units per week), drug abuse or significant psychiatric 
illness. 
18. Any other condition or consideration that, in the opinion of the Investigator, would pose a health risk 
to the participant if they were enrolled in the study, or would otherwise interfere with the evaluation of 
the study aims (e.g. difficult venesection). 
 
 
Co-enrolment Guidelines 
Participants can enter FlaviPrime if they are participating in other studies, provided that: i) no experimental 
drug is taken five half lives before the screening and for the 8 week period after vaccination, ii) no 
experimental vaccine is given within 4 weeks before screening or after vaccination in this study, and iii) the 
total volume of blood drawn in a 16-week period does not exceed 470 ml (the same volume as would routinely 
be collected during blood donation). Participants will remain in the Acute and emerging Virus Immunity Study 
(AVIS). Enrolment into any new study should take place >8 weeks after vaccination in FlaviPrime, unless 
there is a pressing reason for enrolment, and the discretion of the chief investigator. In any event, the 
FlaviPrime study team should be informed of any new enrolment into a study during the 28 period (group 1) 
or 8 week period (groups 2 and 3) following vaccination. 
 

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STUDY TREATMENT/INTERVENTIONS 
 
Introduction 
Eligible patients in each group will receive a single dose of live, attenuated, JE vaccine IMOJEV®. 
IMP will be supplied by Sanofi Pasteur in accordance with all applicable guidelines. 
 
Treatment Name / Description 
 Live attenuated JE vaccine IMOJEV® 
 
Information on live attenuated JE vaccine IMOJEV® will be available to the investigators in the form of the 
Investigators Brochure (IB).  
 
The vaccine is a live attenuated virus. Following administration, the virus replicates locally and elicits 
neutralising antibodies and cell-mediated immune responses that are specific to the Japanese encephalitis 
(JE) virus. Available results indicate that protection is mainly mediated by neutralising antibodies. 
 
IMOJEV® is a monovalent, live attenuated viral vaccine. The virus was obtained via recombinant DNA 
technology. It is based on the 17D-204 yellow fever vaccine virus in which two genes have been replaced by 
the corresponding genes from Japanese encephalitis (JE) virus. These are the premembrane (prM) and 
envelope (E) coding sequences of the SA14-14-2 live attenuated JE vaccine virus. The immunising antigens 
are the prM and E proteins from the SA14-14-2 vaccine virus. 
 
Brand name / Active ingredient: 
IMOJEV® 
 
After reconstitution the active ingredients are: 
Live, attenuated, recombinant Japanese encephalitis virus: 4.0 
- 5.8 log10 Plaque Forming Units, Propagated in Vero cells. 
 
Excipients to the above active ingredients are: 
 
Mannitol  
 
Lactose  
 
Glutamic acid  
 
Potassium hydroxide  
 
Histidine  
 
Human Serum Albumin  
 
Sodium chloride  
 
Water for injections  
 
No adjuvant or antimicrobial preservative is added.  
 
Formulation: 
Powder for reconstitution / diluent 
The powder is a white to creamy white homogeneous cake 
which might be retracted from the sides of the vial. The diluent 
is a clear solution. After reconstitution, IMOJEV® is a colourless 
to amber suspension. 
 
Manufacturer: 
Sanofi Pasteur 
 

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Packaging, storage and stability: 
IMOJEV® is provided in separate packs, each pack contains: 
 
One vial containing a white to creamy white vaccine 
powder  
 
One vial (type I glass) each with a stopper (halo-butyl) 
and a flip off cap (aluminium/polypropylene) containing 
clear diluent 
 
One disposable syringe (polypropylene) with 2 separate 
needles (stainless steel) 
 
The vaccine boxes are to be stored in a refrigerator (2°C – 8°C) 
not be frozen.   
 
When out of the fridge the vials should be kept in the outer 
carton in order to protect from light. Do not use IMOJEV® after 
the expiry date which is stated on the carton after EXP. 
 
Supplier’s name: 
Sanofi Pasteur 
 
Regulatory Status: 
Licensed in Australia and New Zealand for the prevention of JE 
in travellers. 
It is also marketed in some parts of Southeast Asia. 
 
 
Manufacturing and Distribution 
IMOJEV® vaccine will be sourced from existing licensed stock in Australia where the vaccine is in routine 
use. The vaccine will be imported into the UK by Geryon Pharma, overseen by a Qualified Person (QP). The 
importation process will involve continual monitoring of temperature. After import, the vaccine will be stored 
by Geryon Pharma until the green light process is complete, at which point it will be transferred to the clinical 
trials pharmacy at LUHFT. The vaccine will be stored at 4°C in a monitored fridge in the clinical trials 
pharmacy. Upon prescription for use in the study, the vaccine will be dispensed by the pharmacy on a named 
participant basis to the CRU where it will be stored at 4°C in a monitored fridge until use. 
 
After use, any materials that have come into contact with the vaccine will be soaked in virkon overnight to 
ensure inactivation of the vaccine, in accordance with the controlled release of a GMO. 
 
 
Preparation, Dosage and Administration 
Using aseptic technique, IMOJEV® vaccine is reconstituted by injecting all the 0.4% sodium chloride solution 
(clear of colour) into the vial of freeze-dried vaccine, using the syringe and one of the needles provided in the 
carton. The vial is gently swirled.  
After complete dissolution, a 0.5 mL dose of the reconstituted suspension (colourless to amber) is withdrawn 
into this same syringe.  
 
For the administration injection, the syringe is fitted with the second needle provided in the package. The 
product should be used once reconstituted and must be discarded if it is not used within one hour of 
reconstitution. 
 
After use, any remaining vaccine and container must be disposed of safely, preferably by heat inactivation or 
incineration, according to locally agreed procedures. 

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Accountability Procedures 
Further detail on shipping, Qualified Person (QP) and vaccine management processes will be detailed in a 
bespoke Pharmacy Operating Manual.  In summary, shipping will be conducted under QP supervision. The 
vaccine will be shipped into the UK (destination specified in the bespoke Pharmacy Operating Manual) where 
it will undergo inspection including cold chain monitoring records, QP validation and release. It will then be 
transferred to the LUHFT CRU drug storage facilities and kept at monitored temperature according to 
standard protocols. 
 
All vaccine vials will be signed for on entry. In LUHFT, the study vaccine vials will be QP labelled with the 
protocol number and Clinicaltrials.gov number In addition, each vial will be given an individual ID to facilitate 
tracking and accountability. This process will be recorded in the vaccine accountability file. 
 
Upon use, the vaccine vial ID number, person using, time of removal from cold storage, time of reconstitution 
and fate of the vial will all be recorded in the accountability file. 
 
Assessment of Compliance 
The IMOJEV® vaccine dose will be administered by the PI or nominated members of the core clinical research 
team at the LUHFT CRU.  
 
The following measures will be employed to ensure treatment compliance:  
 
1. the vaccine will be administered under the supervision of suitably qualified LUHFT CRU staff 
or the study PI and delegated clinical staff 
 
Concomitant Medications/Treatments and Specific Restrictions 
This study is being conducted in healthy volunteers where the need for other medication is anticipated to be 
low. If an incidental diagnosis is made during the study (e.g. depression, hypertension) requiring drug 
treatment the CI/PI will review the individual situation and take a decision as to whether the participant should 
continue in a study. If the new medication required is not immunomodulatory, the presumption will be to 
continue in the study. Additional flavivirus vaccines should not be given during the study, but can be given 
once 8 weeks from the final dose of study vaccine has elapsed. 
 
Medications Permitted 
 
Permitted ‘rescue’ therapy (paracetamol, NSAIDs and anti-emetics) may be taken for either study related 
AEs or for any other reason, without the need to inform the investigators. All use of concomitant medication 
will be recorded throughout the study duration. The local PI and CI will need to be informed of any other 
medication needed during the study. The potential effects of concomitant medication will be reviewed by the 
investigators who will decide, along with the CI, whether the participant should remain in the study on a case 
by case basis.  
 
Medications Not Permitted/ Precautions Required 
 
Certain classes of medications, which would be expected to impair or alter immune responses, are not 
allowed to be taken during the study. These are: 
 
 
Corticosteroids, including inhaled steroids 
 
Colchicine 
 
Azathioprine, other cell cycle inhibitors, or chemotherapy agents 

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 
Methotrexate or other anti-metabolites 
 
Any biologic immunosuppressive or immunomodulatory drug 
 
Insulin/OHGD 
 
Any vaccine other than those being given as part of the study 
In the case of a participant needing such medication, the local PI and CI will review the case and decide 
whether to replace the participant or not. For example, if a participant has provided samples early in the 
study, replacement may not be necessary. 
Data on Concomitant Medication 
 
Data on Concomitant Medication 
 
Participants will be asked to keep a diary of any other medicine use while they are taking part in the study. 
This will be reviewed by a member of the core clinical research team at each of the study visits.     
 
Specific Requirements/Restrictions 
 
Participants will be asked to follow some restrictions: 
 
 
 
Further details 
Why 
Medication 
Not to take medication that will impair or alter 
their immune response, for example;  
- 
Corticosteroids, including inhaled steroids 
Colchicine 
- 
Azathioprine, other cell cycle inhibitors or    
chemotherapy agents 
- 
Methotrexate or other anti-metabolites  
Any biologic immunosuppressive or  
immunomodulatory drug,  
- 
Insulin or other oral anti-diabetic drug 
Any vaccine other than that being given as 
part of the study (COVID vaccines can be 
given) 
To ensure the response seen is due to 
the body, and not other medication.   
If it is necessary to take certain 
medications for participant health, 
participants will be asked to inform the 
study team as soon as they can.  
 
Further details 
Why 
Travel 
Not to travel to any country where Flaviviruses 
are found from screening till 8 weeks after the 
vaccination 
To limit exposure to a flavivirus which 
could create another response 
Alcohol 
Not to drink more than 28 units of alcohol a 
week e.g.   
- 
16 bottles of larger, beer or cider 
- 
2 and half bottles of wine 
Alcohol 
may 
affect 
they 
body’s 
response to the vaccine  
Other studies 
Not to take part in other studies until 8 weeks 
after receiving IMOJEV®  
To ensure the response we see if due 
to IMOJEV® and not another study  
Blood 
donation 
Not to donate blood for 3 months after 
vaccination 
There is a limit to the amount of blood 
a person can give in a 3 month period 
Contraception 
Female partners of childbearing potential of 
male participants in the study must use an 
acceptable method of contraception from the 
time of screening until 28 days after vaccination. 
 
Females of childbearing potential must use an 
acceptable method of contraception from the 
time of screening until 28 days after vaccination. 
 IMOJEV® is not suitable for use in 
pregnant women 

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Overdose 
Overdose is not possible due to the single dose nature of the study, and that the vaccine will be administered 
under supervision. 
 
Specific information on reporting adverse events can be found in Section 10. 

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OUTCOMES 
There will be no primary and secondary endpoint to the study, which is not aimed at licensure of a product. 
The scientific objectives of this study are to describe the priming and cross-reactivity of B and T cell responses 
to JE vaccine IMOJEV®. This will include mapping of specific B cell epitopes of the vaccine virus, which is 
performed by cloning antibodies from plasmablasts (responding B cells) at week 1 post vaccine. 
 
More details of the laboratory assays to be performed are given in section 9.5. In addition, the safety of study 
participants will be reviewed and monitored regularly by the study oversight committee.  
 
 

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PARTICIPANT TIMELINES AND ASSESSMENTS 
 
Participant Identification and Screening 
Participants will be identified from an existing observational study (“Acute emerging Virus Immunity Study,” 
(AVIS) 16/NW/0170), by advertisement, and through the LUHFT CRU, for example using the 
consent4consent database. AVIS is aimed at recruitment of people who have had flavivirus clinical illness, 
positive laboratory assays or previous vaccination, and controls, allowing identification of a pool of flavivirus 
exposed subjects for group 3 recruitment. 
 
Participants identified through AVIS will already have undergone the immunological profiling necessary for 
FlaviPrime. Therefore, the group allocation will already be known at the point of entry into FlaviPrime. 
Participants will be considered suitable for recruitment into FlaviPrime under the following conditions: 
 
Group 1 (any status): No special exposure criteria apply 
Group 2 (flavivirus naïve): No potential exposure to flaviviruses based on travel history 
Group 3 (flavivirus exposed): Confirmed flavivirus exposure, illness or vaccination 
 
Confirmed flavivirus exposure means travel history compatible with exposure (e.g. residence in a dengue 
endemic area) and positive neutralising antibody assay. Flavivirus illness means an illness compatible with 
a flavivirus infection (most commonly dengue), in combination with a positive PCR, or NS1 antigen test, or 
virus isolation, or IgM ELISA and neutralising antibody. 
 
Potential participants will be contacted (by telephone and/or email) by a member of the core clinical research 
team and invited to take part in FlaviPrime (consent for this contact is taken in AVIS). Participants identified 
through the consent4consent database or otherwise at the CRU can be consented at RLUH by a member of 
the study team or CRU staff, or elsewhere, and then donate blood samples and have the necessary flavivirus 
immunological assays performed in the University as per the Acute emerging Virus Immunity Study 
(16/NW/0170). Once identified, participants will be given written and verbal information on the FlaviPrime 
study, as well as an opportunity to ask questions. 
 
All potential participants will be given a unique screening ID which will subsequently be used to detail the 
reasons for the continuation or discontinuation at the screening stage.   
 
 Informed Consent  
 
Informed consent is a process initiated prior to an individual agreeing to participate in a study and continues 
throughout the individual’s participation. Written informed consent is required for all patients participating in 
CTU coordinated trials. The process should involve discussion between the potential participant and an 
individual knowledgeable about the research, the presentation of written material (e.g. information leaflet or 
consent document), and the opportunity for potential participants to ask questions and have these 
satisfactorily answered. In obtaining and documenting consent, the research team should comply with 
applicable regulatory requirements and should adhere to GCP and to the ethical principles that have their 
origin in the Declaration of Helsinki. Informed consent should be re-affirmed throughout the study and all 
discussions and consent should be documented appropriately. If a potential participant does not want to 
provide consent they do not have to give a reason. 
 
Prospective Informed Consent Process 
 

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Written informed consent will be sought from patients who will be approached by the study team and invited 
to consider participation.  
 
Patients will be invited to visit the LUHFT CRU, where a nominated member of the clinical research team will 
discuss participation in the study with them. A written information sheet that forms part of the ethically 
approved Information Sheet and Consent form will be provided. This includes a detailed explanation of the 
study and makes clear that the rights and welfare of the participants will be protected; it will be emphasised 
that consent may be declined or withdrawn at any time in the future without the quality of care being adversely 
affected. The research staff will facilitate verbal discussions about the research and the consent process, as 
well as providing answers to any questions that arise.  
 
After verbal and written information has been provided, the individual seeking consent will ensure that the 
patient has fully understood all the information and will ask if they are happy to consent to participation in the 
study.  
 
Where this is the case, written informed consent will be obtained by means of a dated patient’s signature on 
the consent form. This should be countersigned and dated by the person who obtained informed consent i.e. 
the PI or other appropriately qualified member of the research team who has been delegated this 
responsibility.   
 
The original signed document will be retained in the study site’s Investigator Site File (ISF) and copies will be 
made: 
 
One copy provided to the patients for their information,  
 
One copy transferred to the CTU via encrypted email / post 
 
One copy filed in the participant’s medical records paper/electronic. 
 
N.B. Details of the consent process (date, persons involved, version and type of information sheet and 
consent form used) must also be recorded directly into the participant’s medical records. 
 
Screening, Eligibility Assessment and Confirmation 
Once the participant has signed the ICF, the interviewing member of the clinical research team will carry out 
a more detailed medical review, drug history and travel history (to ensure no new flavivirus exposure). A 
physical examination and the collection of screening blood samples to ensure participants meet the full study 
eligibility criteria a HIV (and hepatitis B and C) test will be conducted. Hepatitis B and C tests are included to 
rule out non-specific immune effects of these viruses; hepatitis C is related to flaviviruses so may also give 
rise to specific cross-reactivity. Also if female a pregnancy blood test will be conducted. Both tests are 
completed for safety reasons as the safety of IMOJEV® has not been established in this patient population. 
 
Screening information recorded for all participants, regardless of their suitability for the study, will be retained 
and archived. 
 
Eligibility can only be confirmed by an appropriately qualified medical professional who is named on the 
delegation log and must not occur until fully informed consent is documented. Eligibility criteria are described 
in detail in Section 6. 
 
Eligibility confirmation must be documented in the participant’s medical notes and then on the trial’s Eligibility 
CRF. Details must include at a minimum who confirmed full eligibility, when this was confirmed, and when 
the participant was formally entered into the study (e.g. registration). 

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Enrolment/Registration and Baseline Assessments 
When satisfactory clinical screening results are received, the participant will be contacted (by telephone and 
/ or email) by a member of the core clinical research team and invited to attend the CRU where, prior to being 
registered on to the study, a final checklist of inclusion/exclusion criteria will be completed to ensure the 
potential participant’s eligibility. This will include confirmation that the participant has not travelled to a 
flavivirus endemic area since they were identified to be suitable in AVIS. Once eligibility has been confirmed 
by the PI at the CRU, the participant will be registered onto the study using an electronic registration platform. 
The minimum time between screening/explanation of the study and enrolment is 24 hours, but in practice this 
will usually be longer as the time needed to process routine HIV is >24 hours. The screening window is 8 
weeks as long as there is no change in the health of the potential participant. Documentation of reasons for 
non-inclusion is not necessary. 
 
Travel to a flavivirus endemic area or other potential flavivirus exposure between immunological testing in 
AVIS and enrolment would render a group 2 participant ineligible for the study, though theoretically if such 
an individual experienced a confirmed flavivirus infection during travel they would be eligible for Group 3.  
Travel to a flavivirus endemic area would mean the immunological assays would need to be repeated for a 
group 3 participant, though if this were done the participant would still be enrolled. 
 
Once a participant consents to taking part in the study, they will be registered on a bespoke electronic data 
capture system which will generate a unique screening ID for each participant.  This system will be used to 
capture the progress of screening for all participants. Each participant’s GP will be provided with a letter, 
notifying them of their patient’s involvement in the research study.  
 
At baseline, once screening is complete, participants will be invited to attend the CRU to confirm eligibility 
before vaccination. A baseline blood sample should be taken and peripheral blood mononuclear cell (PBMC) 
and serum cryopreserved. If a subject has not travelled to a flavivirus endemic area nor had any flavivirus 
vaccine, it is extremely unlikely that their immunological status will have changed. 
 
All participants at this stage will be registered to take part in the study and receive the vaccination.  
Participants will be registered by authorised members of the research team at the LUHFT CRU using a 
bespoke electronic data management system.  Registration using this system can take place 24 hours a day, 
7 days a week. 
 
Schedule for Assessments and Follow-up 
Participants recruited into Group 1 will remain on the study for 28 days post vaccination. Partipants recruited 
into Group 2 and Group 3 will remain on the study for 8 weeks post vaccination. Once consented to take 
part in the main study, participants will receive the first dose of the vaccine at baseline following the 
completion of final eligibility checks and the collection of a baseline blood sample.  Following this, participants 
will be asked to attend further clinic visits at the CRU on days 5 through 9 after vaccination for group 1, and 
at weeks 1, 2, 4 and 8 for groups 2 and 3, so that blood samples can be taken (55 – 75ml per visit) and any 
AEs can be assessed and recorded. T cell IFNγ-ELISpot intracellular cytokine staining (ICS) and flow 
cytometry assays will be performed at baseline and at weeks 1, 2, 4 and 8 after the vaccine in Groups 2 and 
3 participants, and at baseline and one week after the vaccine in group 1. Cells will be stained for plasmablast 
sorting for B cell ELISpot and single cell sequencing for assessment of the adaptive immune response at 
every timepoint for group 1, and at one week post vaccine for groups 2 and 3. Where possible, and where 
responses are identified, these will be mapped down to the peptide and later minimal epitope level and further 
characterised. Although the most crucial data regarding cross-reactivity are derived from the time point when 
responses are maximal (usually 1-2 weeks), the timing of responses can vary and these experiments have 
not been done in this population before, leading to uncertainty about when the maximal response is. For this 
reason, the assays will continue until week 8 post vaccine. 

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Details of the blood samples and volumes to be taken are shown in the Schedule of Assessments tables 
below. A slightly larger sample will be taken at week 1 post vaccine, to allow for single cell sorting of 
plasmablasts necessary to clone antibodies which will be performed at this time point. This time point can be 
revised based on the findings of group 1. 
Schedule of Assessments: 
 
Group 1 
  
 
 
 

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Groups 2 and 3 
 
 
SV = Study Visit (tolerance in +/- days is indicated for each visit). *At baseline, all procedures should be done before study 
intervention. **To be confirmed based on group 1. ***Only done in response to positive assays from JEV peptide pools 
 
 
Clinical Procedures 
 
The timings of all measurements to be performed during the study may be subject to change based on the 
ongoing review of safety and tolerability. All changes will be agreed with the Sponsor and documented in the 
study management folder.  Safety will be assessed through the reporting of Adverse Events (AE) and Serious 
Adverse Events (SAE) as detailed in Section 12. 
 
Safety Assessments – Adverse Events (AE) 
 
The condition of each participant will be monitored throughout the study, from the time of informed consent 
through to the final visit.  Participants will also be encouraged to spontaneously report adverse events 
occurring at any other time during the study. Participants will be issued an emergency contact card with the 
CRU 24 hour emergency telephone number on it which they can use to contact the CRU on call doctor at 
any time during the study if they feel unwell. Any adverse events and remedial action required will be recorded 
in the participant’s source data.  The nature, time of onset, duration and severity will be documented, together 
with the Investigators opinion of the relationship to the study drug. Any clinically significant events identified 
during the course of the study will be followed up until they are resolved or can be clinically explained.  

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Adverse event definitions, assignment, causality and procedures for reporting serious adverse events are 
detailed in Section 10 of this protocol.  
 
Vital Signs 
Supine blood pressure, pulse rate and oral body temperature will be measured at the time indicated in the 
study schedule.  Vital signs will also be performed at other times if judged clinically appropriate, if required 
by hospital policy or if the ongoing review of the data suggests a more detailed assessment of vital signs is 
required.  
 
Clinical Laboratory Evaluations 
Blood will be collected for clinical laboratory evaluations at the times indicated in the schedule.  Additional 
clinical laboratory evaluations will be performed at other times if judged to be clinically appropriate, required 
by hospital policy (e.g. MRSA) or if the ongoing review of the data suggests a more detailed assessment of 
the clinical laboratory safety evaluations is required. An investigator will perform a clinical assessment of all 
clinical laboratory data.   
 
Physical Examination 
A full physical examination or abbreviated physical examination will be performed at the times indicated in 
the study schedule.  The full physical examination will include the assessment of the following systems: 
general appearance, skin (site of vaccination), lymph nodes, HEENT (head, ears, eyes, nose and throat), 
neck, thorax / lungs, cardiovascular, abdomen, musculoskeletal and neurological.  The abbreviated physical 
examination will only involve the assessment of symptoms as directed by the participant. 
 
 
Special Assays or Procedures 
 
All information relating to laboratory assays and procedures employed within this study will be detailed in a 
separate bespoke Laboratory Manual.  This will include instructions for the preparation, handling and storage 
of specimens including required temperatures, aliquots of specimens, where they will be stored, and how 
they will be labelled.  All procedures will be detailed in separate protocols / working instructions which will be 
included in the laboratory manual together with plans in terms of the long-term access and consent for the 
future use of specimens.  
 
The aim of the experimental procedures will be to determine the epitope specificity, function and cross-
reactivity of the human antibody response to JE vaccine IMOJEV®. Additional assays will be conducted 
according to sample availability for characterising white cell populations by immunophenotyping /flow 
cytometry after vaccination. T cell responses to the vaccine will also be studied using IFNγ ELISpot assays 
and flow cytometry, because T cell responses help in the generation of potent antibody responses. 
 
PAXgene tubes will be stored for later RNA isolation. This will be used to track the evolution of specific B cell 
receptor sequences identified in the above experiments over time. If sufficient sample exists, transcriptomics 
and/or targeted RT-qPCR will be done to assess the relationship between early gene activation and the 
subsequent characteristics of the adaptive immune response[29]. PBMC samples will be collected for 8 weeks 
to enable deep analysis of the evolving B cell response over time. 
 
PBMC, sera and PAXgene/RNA samples will be stored for 25 years. The future intended use of these 
samples is address key questions on adaptive immunity to flaviviruses. For example, if a novel or previously 
obscure flavivirus were to emerge as a major human pathogen, such as Zika virus in 2015, we would be able 
to rapidly test the ability of our participants’ response to cross-react with such a novel threat. Therefore, the 
experiments that will be done using these samples cannot be pre-determined, as they will either be guided 
by previous results, or by new discoveries. However, they will be focussed on immunity to viruses/flaviviruses. 

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Samples will be stored in the liquid nitrogen facility (cells) and -80°C freezer (serum/plasma/PAXgene) in the 
Ronald Ross building, University of Liverpool, in compliance with the human tissue act. The ICF specifically 
addresses storage and future use of these samples, without the need for additional consent for experiments 
on immunity to viruses. 
 
Sampling 
 
Sample Collection 
 
The blood volumes that will be taken from each participant are detailed in the schedule of study assessments.   
The maximum blood volume to be taken per participant will be approximately 300ml, or 330ml for group 1, 
which is less than the volume of a single blood donation. 
 
People who are blood donors can participate in FlaviPrime, but they cannot give blood concomitantly to the 
study. The maximum amount of blood that can be given in blood donation is 470 ml per 16 weeks. Therefore, 
a participant who is a blood donor cannot join FlaviPrime until 16 weeks after their last blood donation. Four 
weeks after the last FlaviPrime study sample is taken, the participant becomes is able to donate blood again. 
 
Sample Shipment/Custodianship 
 
Laboratory safety samples will be analysed at the Royal Liverpool University Hospital Laboratories.  
Experimental blood samples relating to the outcomes of this study will be analysed within the Department of 
Clinical Infection, Microbiology and Immunology, University of Liverpool.  Samples will be transported from 
the CRU to the University of Liverpool in line with the study specific Material Transfer Agreement (MTA). 
Details of sample transport are in the bespoke laboratory manual for the study. Serological analyses will be 
performed in the laboratory of Dr Sutee Yoksan, Mahidol University, and Bangkok. Depending upon the initial 
results of the study, it may be beneficial to send samples to other laboratories, as is the case in the Acute 
emerging Virus Immunity Study.  
 
Sample Storage and Handling 
 
Samples given as part of the study, if they are not used in meeting the scientific objectives, will be stored for 
a period of 25 years. The samples can be used in future studies addressing the development of immunity to 
flaviviruses, and the cross-reactivity to flaviviruses, and to other emergent viruses, but not for other purposes 
(control antigens/mitogens will also be included). An example of such a use might be, testing for markers of 
immunity to a flavivirus or another virus that is undiscovered at the time of the study, but causes an outbreak 
in the future. 
 
These samples would be used for the purposes as outlined above in section 7.3.1, although these may apply 
to other viruses. Permission to send material to other laboratories, and the parameters of the purpose of this, 
will be included in the consent form at the start of the study. 
 
 
Intervention Discontinuation and Participant 
Discontinuation/Withdrawal 
In consenting to the study, participants are consented to study treatment, follow-up and data collection. If 
voluntary withdrawal occurs, the participant should be asked to allow continuation of scheduled evaluations, 

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complete an end-of-study evaluation, and be given appropriate care under medical supervision until the 
symptoms of any adverse event resolve or the subject’s condition becomes stable.   
 
The critical study data in FlaviPrime is derived from experimental blood samples. Although participants can 
refuse a blood sample at any time, refusal to give crucial samples (weeks 1, 2 and 4 weeks post JE vaccine 
IMOJEV®) would result in significant compromise to the study. Therefore, any participant who does not give 
these samples for any reason will be replaced with a new volunteer and removed from the study sampling 
schedule thereafter. The participant will remain in the study for safety. 
 
Premature Discontinuation of Study Intervention 
 
Once the single dose of JE vaccine IMOJEV® has been given, there is, in practice, no intervention to be 
withdrawn from, and participants can remain in the study. Foreseeable reasons why a participant may 
withdraw from the study generally apply to the taking of study blood samples, and include: 
 
i. 
Participant withdraws consent 
ii. Participant is unable to attend regularly for assessment and to donate week 1, 2 and 4 post 
vaccination samples for any reason 
iii. Loss of capacity during the study 
iv. Any other change in the participant’s condition that justifies the discontinuation of sample 
collection the study in the clinician’s opinion. 
 
A participant is free to withdraw from the study at any time.  In addition, the CI may decide, for reasons of 
medical prudence, to withdraw a participant.  In either event, the Sponsor will be notified and the date and 
reason(s) for the withdrawal will be documented in the participant’s source data. If a participant withdraws or 
is withdrawn, ideally they should remain in the study for collection of safety data and/or treatment of any AEs, 
until these have resolved.  
 
 
 
Participant Withdrawal from Follow Up  
 
If a participant wishes to withdraw from the study sampling schedule, they will be informed by the CRU staff 
of the importance of remaining on study follow-up for safety, or failing this, of allowing routine follow-up data 
to be used for study purposes. Generally, follow-up will continue unless the participant explicitly also 
withdraws consent for follow-up. The CI may also request that the participant return for an additional follow-
up visit, for the purpose of collecting safety data, or ensuring resolution/adequate treatment of an AE. 
 
Participants are free to withdraw from follow up at any time without providing a reason, though a reason 
should be recorded if one is given. Those who wish to withdraw from further follow-up will have the data 
collected up to the point of that withdrawal included in the analyses.  The participant will not contribute further 
data to the study and the CTU should be informed via email to the CTU and via completion of a Withdrawal 
CRF to be returned to the CTU within 7 days. 
 
If participants express a wish to withdraw from follow up, the research team at site should ascertain if this is 
for all elements of study follow-up, or if for example data from routine assessments can still be collected for 
the study. In the case of ongoing adverse events, participants should be given appropriate care under medical 
supervision until the symptoms of any adverse event resolve or the participant’s condition becomes stable. 

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Any SAEs will be notifiable to the CTU via processes detailed in Section 11 even if a participant has withdrawn 
from follow up. 
 
Participant Withdrawal from Study Completely  
 
Participants who withdraw from the study for other reasons have previously consented to follow-up in the 
study. Data up to this time can be included in the study if anonymised. They may need to reaffirm that they 
consent to follow-up through usual NHS mechanisms. If the participant explicitly states their wish not to 
contribute further data to the study, a withdrawal CRF should be completed.  
 
Participant Transfer 
 
For participants moving from the area, every effort should be made to encourage the participant to continue 
to attend follow-up if possible. If this is not possible, the participant will have to be withdrawn from the study. 
If the participant is withdrawn before the day 9 sample (group 1) / two-week sample (groups 2 and 3) has 
been donated, the participant should be replaced if possible (dependent on the availability of vaccine). This 
should be discussed with the Chief Investigator and LCTC.  
 
Loss to Follow-up 
 
If for any reason a participant does not complete the study they should attend a final visit / assessment with 
the study clinician as soon as possible.  If the study treatment is stopped before the end of the scheduled 
assessment period, the clinician may contact the participant to request additional information up to the point 
where the study treatment would be complete. 
 
Travel 
 
Participants in the study will be required not to travel to a JEV and DENV endemic area until the sampling 
schedule has reached study week (SW) 8.  
 
If any participant who does travel to a JEV and DENV endemic area before 8 weeks, consideration will be 
given to replacing them by the study oversight committee. They will remain in AVIS to gather additional data. 
For example, a participant who contracted a wild type flavivirus infection during the study would represent a 
natural experiment of great relevance to the principles under investigation in this study. 
 
 
End of Study 
The end of the study is defined to be the date on which data for all participants is frozen and data entry 
privileges are withdrawn from the study database (database is locked). Study enrolment may be stopped at 
a site when the total requested number of participants for the study have been recruited.  The Study Oversight 
Committee may recommend that the study be stopped prematurely.  In such circumstances of premature 
termination / suspension of the study, National Research Ethics Service (NRES) and HRA will be notified 
according to standard reporting guidelines 
 
Site and closure activities will be centrally coordinated and conducted in accordance with CTU processes 
regardless of whether the study closes as planned or prematurely. This includes activities such as: 
- 
End of Study notification to REC 
- 
Study-related materials reconciled and returned/disposed of as appropriate see section 7.5 for IMP 
- 
All site data entered onto the study database, discrepancies raised and satisfactory responses 
received 

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- 
Quality Control checks of the Investigator Site Files, Pharmacy Files and Study Master File as 
appropriate. 
 
Study Discontinuation 
 
If this study is prematurely discontinued (e.g., due to safety) all participants must be informed and the reason 
for the discontinuation should be written on the end of study form for each participant.  If a participant has 
been withdrawn completely from the study whilst is still ongoing, an end of study form should be completed.  
 
The need for stopping the study will be considered by the study Oversight Committee. The decision will be 
based upon participant safety. If any of the following scenarios occur with reasonable possibility of a causal 
relationship with the study vaccine, no further doses of study vaccine will be given. 
 
1. 
Clinically relevant signs or symptoms or intolerable adverse events of similar nature occurring 
in 2 or more participants that in the opinion of the CI warrant stopping, 
 
2. 
One or more participants report a serious adverse event considered by the CI, local PI, or 
delegated other physician who can assess causality and expectedness of AEs to be at least 
possibly related to the study drug. 
 

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  SAFETY REPORTING  
Safety reporting in clinical trials is a legal and ethical requirement and it is imperative that all applicable 
requirements detailed here are followed during the study. 
 
Terms and Definitions  
Adverse Event (AE) 
Any untoward medical occurrence, unintended disease or injury or any untoward clinical signs (including an 
abnormal laboratory finding) in subjects, users or other persons whether or not related to the study vaccine.  
 
Related Adverse Event (Related AE) 
An AE which resulted from administration of any of the research procedures – i.e. assessed as “probably”, 
“possibly” or “almost certainly” related to the study procedures or vaccine (see section 7) 
 
Related Unexpected Adverse Event (RUAE) 
A Related AE which is not expected, i.e. not consistent with the known effects of the study vaccine or 
research procedures (see section 7). 
 
Serious Adverse Event (SAE) 
An adverse event which meets the definition of “serious” (see section 10.2).  
 
Related Serious Adverse Event (Related SAE) 
A SAE which is assessed to be “probably”, “possibly” or “almost certainly” related to the study vaccine or 
study trial procedures (see section 7). 
 
Related Unexpected Serious Adverse Event (RUSAE) 
A Related SAE which is not expected, i.e. not consistent with the known effects of the study vaccine or 
study trial procedures (see section 7). 
 
Reference Safety Information (RSI) 
The information used for assessing whether an adverse reaction is expected (see section 10.5). This is 
contained in the Investigators Brochure (IB) for the vaccine. The RSI used to assess the expectedness of a 
SAE/SAR must be the current approved version at the time of onset of the SAR. The RSI for this trial is 
defined in section 10.5. 
 
Assessment of Seriousness 
The assessment of seriousness of safety events should be performed by an appropriately delegated, 
medically qualified member of the site research team. 
 
A #safety event (whether or not assessed as related to the study) is assessed as serious if it: 
 
 
Results in death;  
 
Is life-threatening (i.e. the investigator considers the event places the subject at immediate risk of 
death from the experience as it occurred (this does not include an adverse experience that, had it 
occurred in a more severe form, might have cause death); 
 
Requires hospitalisation or prolongation of existing hospitalisation (hospitalisation is defined as an 
inpatient admission, regardless of length of stay, even if the hospitalisation is a precautionary 
measure for continued observation);  
 
Results in persistent or significant disability or incapacity (substantial disruption of one’s ability to 
conduct normal life functions);  
 
Consists of a congenital anomaly or birth defect (in offspring of study participants, or their partners, 
regardless of time of diagnosis), or  

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 
Is otherwise considered medically significant by the investigator.  
 
#safety event / reaction applies apply to either AEs, or ARs, or Related AEs,  
 
 
Severity of Adverse Events 
All adverse events should be assessed for severity. The assignment of the severity/grading should be made 
by the investigator responsible for the care of the participant using the definitions in the table below: 
 
Table 1: Severity Grading 
Severity 
Description 
Mild 
Does not interfere with routine activities. 
Moderate 
Interferes with routine activities. 
Severe 
Impossible to perform routine activities. 
Life-Threatening  
Death 
 
 
N.B. A distinction is drawn between serious and severe AEs. Severity is a measure of intensity (see above) 
whereas seriousness is defined using the criteria in Section 10.2. Hence, a severe safety event need not 
necessarily be a “serious” safety event. 
 
Assessment of “Causality” - Relationship to Study 
Treatment/Intervention 
The assignment of the causality should be made by the investigator responsible for the care of the participant 
using the definitions in the table below: 
 
Table 2: Definitions of Causality 
Relationship 
Description 
Unrelated 
There is no evidence of any causal relationship.  
N.B. An alternative cause for the AE should be given 
Unlikely 
There is little evidence to suggest there is a causal relationship (e.g. the event 
did not occur within a reasonable time after administration of the study 
medication). There is another reasonable explanation for the event (e.g. the 
participant’s clinical condition, other concomitant treatment). 
Possibly 
There is some evidence to suggest a causal relationship (e.g. because the 
event occurs within a reasonable time after administration of the study 
medication). However, the influence of other factors may have contributed to 
the event (e.g. the participant’s clinical condition, other concomitant treatments). 
Probably 
There is evidence to suggest a causal relationship and the influence of other 
factors is unlikely. 
Almost certainly There is clear evidence to suggest a causal relationship and other possible 
contributing factors can be ruled out. 
 
Assessment of causality should be made based on known safety profile of IMOJEV®. In the case of discrepant 
views on causality between the treating investigator and others, the opinion of the treating investigator will 
never be downgraded and the REC will be informed of both points of view. Events that are assessed as being 
possibly, probably or almost certainly related will be reported as having a reasonable possibility of being 

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related, and events assessed as unrelated or unlikely will be reported as having no reasonable possibility of 
being related.  
 
Assessment of causality should be made based on known safety profiles of IMP, or IB and known risk profiles 
of other drugs in the same class. If any doubt about the causality exists the local investigator should inform 
the LCTC who will notify the Chief Investigator. In the case of discrepant views on causality between the 
treating investigator and others, the opinion of the treating investigator will never be downgraded and the 
REC will be informed of both points of view. 
 
 
Assessment of “Expectedness” 
The Chief Investigator for the FlaviPrime study is responsible for determining whether a safety event is 
expected or unexpected, however a Chief Investigator will not assess their own participants, these 
participants will be assessed by the Medical Reviewer/Clinical Coordinator. There is no requirement for a 
reporting investigator to make an assessment of expectedness. 
 
An event will be considered unexpected if it is not listed within the current and approved RSI for the study at 
the time of the event’s onset. The nature, severity, or frequency of the event should be considered – if this is 
not consistent with that described for the type of event in the RSI the event should be assessed as 
unexpected.  
 
Reference Safety Information / Information used to Assess Expectedness  
 
Document name: Australian product information  
 IMOJEV® Japanese encephalitis vaccine (live, attenuated) 
 
IMOJEV IB section “Adverse Effects”  “Data in adult populations” 
 
The current version of the RSI documents applicable to this study will be made available to all clinicians 
involved in assessing the causality of adverse events, as well as the ethics committee and sponsor.  
 
Time Period for Active Monitoring of Safety Events 
IMPORTANT: Any safety events occurring after the end of the below described “active monitoring” period 
which meet the definition of serious and are recorded for this study must continue to be reported by sites to 
the LCTC in accordance with the timeframes and procedures described in section 11.3. The same processes 
established for SAEs within the active monitoring period should be followed for these events. 
 
All new serious adverse events should be reported from the point of consent until 28 days after administration 
of the study vaccine.  Depending on the nature of the event, the reporting procedures below should be 
followed.  Any questions concerning adverse event reporting should be directed to LCTC in the first instance. 
Group 1 participants, who have their last study visit 9 days after vaccination, will be encouraged to report any 
adverse events that occur up to 28 days to the LUHFT CRU by phone.  
 
AEs that occur during this timeframe should be reported to LCTC via the AE form. When reporting an increase 
in severity for an existing AE, the AE form should be changed to the new grade and the reason why (increase 
in severity) included. For a decrease in severity, no action is to be taken unless the symptoms have fully 
resolved (in which case the AE/SAE should be updated until resolved. 
 

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Although the period of adverse event monitoring in this study is 28 days, if the site team become aware of an 
adverse event that occurs outside this time and there are reasonable grounds to suspect the event is related 
to the vaccine or to participation in the study, it would be good practice to report this event to LCTC.  
 
 
Notes on Safety Event Recording  
The following events must be recorded for the purposes of the study: 
 
An exacerbation of a pre-existing illness 
 
An increase in frequency or intensity of a pre-existing episodic event/condition 
 
A condition (even though it may have been present prior to the start of the study) detected after study 
drug administration 
 
Continuous persistent disease or symptoms present at baseline that worsens following the 
administration of the study/study treatment 
 
Laboratory abnormalities that require clinical intervention or further investigation (unless they are 
associated with an already reported clinical event). 
 
Abnormalities in physiological testing or physical examination that require further investigation or 
clinical intervention 
 
Injury or accidents 
 
Pregnancy (See section 11 for more details) 
 
Do not record: 
 
Medical or surgical procedures - the condition which leads to the procedure is the adverse event  
 
Pre-existing disease or conditions present before treatment that do not worsen 
 
Situations where an untoward medical occurrence has occurred e.g. cosmetic elective surgery 
 
Overdose of medication without signs or symptoms* 
 
The disease being treated or associated symptoms/signs unless more severe than expected for the 
participant’s condition 
*N.B. If overdose occurred with resulting signs and symptoms that meet the protocol criteria for 
AE/SAE/SAR/RUSAE then they should be reported accordingly and the overdose highlighted to the LCTC 
team.  
 
The events above do not need recording as the study population is healthy, and no disease is being treated 
by administration of this vaccine. The study is considered low risk. Any health event not related to the vaccine 
and not occurring during the study period (i.e. not an AE) is definitely unrelated to use of the vaccine, which 
is already in routine clinical use with a well understood safety profile. 
 

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 Reporting of Pregnancy 
If a participant or their partner becomes pregnant during study treatment or within 28 days following treatment, 
a Pregnancy Report Form must be completed and forwarded to the LCTC within 24 hours of learning of its 
occurrence and the investigational product will be permanently discontinued in an appropriate manner. 
(Should you need a paper copy of the Pregnancy Report Form please contact the Study Coordinator). 
Protocol-required procedures for study discontinuation and follow-up must be performed on the subject 
unless contraindicated by pregnancy.   
 
IMOJEV® has not been studied in pregnancy, so if a participant were to fall pregnant, there are no data upon 
which to base an opinion as to the likely effects. However, in order to have an effect, the participant would 
have to actually be pregnant at the time of vaccine administration, or when vaccine related viraemia is 
present. This lasts up to 6 days after vaccination[30], so a participant would have to fall pregnant very quickly 
after study enrolment for there to be any possibility of an effect on the pregnancy. The immune effects of the 
pregnancy could have an effect on the study data, but the risk of harm to the participant and the pregnancy 
would be extremely small. It should be noted that yellow fever vaccine (which is more toxic than IMOJEV) 
has been used in pregnancy during vaccination campaigns to contain outbreaks and there have not been 
reports of adverse effects on the fetus[31]. 
 
On pregnancy outcome, the final Pregnancy Report Form should be completed and forwarded to LCTC 28 
days after the outcome. The final Pregnancy Report Form is used to determine outcome, including 
spontaneous or voluntary termination, details of the birth, and the presence or absence of any birth defects, 
congenital abnormalities, or maternal and/or new-born complications. Pregnancy follow-up information on 
this form also includes an assessment of the possible relationship to the study intervention of any pregnancy 
outcome. Pregnancy outcomes should also be collected for the female partners of male participants 
participating in the study. Infants should be followed for a minimum of 8 weeks. Consent to report information 
regarding these pregnancy outcomes should be obtained from the mother prior to completion and faxing of 
the final Pregnancy Report Form. Any SAE experienced during pregnancy must be reported on the SAE 
form.  
 
The LCTC will report all pregnancies to the study Sponsor and National Research Ethics Service. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Please Report 
All pregnancies using the Pregnancy Report Form. 
This should be reported within 24 hours of becoming aware. 
ALSO 
Pregnancy Outcomes must also be reported by completing the Final 
Pregnancy Report Form. 
This should be completed 28 days following the outcome. 
Completed forms should be sent via SECURE email to 
lctcsafe@liverpool.ac.uk 

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Sexually active women of child bearing potential (WOCBP) must use an effective method of birth control 
during the course of the study, in a manner such that risk of failure is minimised.  
 
Before study enrolment, WOCBP must be advised of the importance of avoiding pregnancy during study 
participation and the potential risk factors for an unintentional pregnancy in accordance with the information 
contained in the Participant Information Sheet.  
 
All WOCBP MUST have a negative pregnancy test within 72 hours before receiving treatment. The 
minimum sensitivity of the pregnancy test must be 25 IU/L or equivalent units of HCG. If the pregnancy test 
is positive, the subject must not be enrolled in the study. In addition, all WOCBP should be instructed to 
contact the investigator immediately if they suspect they might be pregnant (e.g. missed or late menstrual 
period) at any time during study participation. 
 
Female participants who are of non-child bearing potential (i.e. due to being post-menopausal for at least 1 
year (confirmed by FSH assessment) or permanently sterile following hysterectomy, bilateral salpingectomy, 
bilateral oophorectomy) will not be required to use contraception. 
 
Female participants must be willing to use a highly effective method of birth control (i.e. contraceptive 
measure with a failure rate of <1% per year), from the time of Screening until 28 days after vaccination. Highly 
effective methods of contraception include: 
 
Placement of intrauterine device or intrauterine system 
 
Established use of oral, injected or implanted hormonal methods of contraception associated with 
inhibition of ovulation 
 
Additional barrier method (i.e. diaphragm or cervical vault cap used in conjunction with spermicide 
gel, foam, cream, film or suppository). 
 
Male sterilisation (with the appropriate post-vasectomy confirmation of surgical success). For female 
participants on the study, the vasectomised male partner should be the sole partner for that 
participant. 
 
Bilateral tubal ligation 
 
Female partners of childbearing potential of male participants in the study must use an acceptable method 
of contraception from the time of screening until 28 days after vaccination. Other acceptable methods of 
contraception include: 
 
Any highly effective method of contraception listed above for female participants 
 
Progesterone-only oral contraception, where inhibition of ovulation is not the primary mode of action 
 
Cap, diaphragm, or sponge with spermicide 
  
For participants who practice true abstinence, when this is in line with the preferred and usual lifestyle of the 
participant, contraceptive requirements do not apply. For participants who are exclusively in same-sex 
relationships, contraceptive requirements do not apply. Male participants with pregnant partners should use 
condoms from the time of vaccination and for two weeks afterwards. 
 
 
Notification of Deaths 
 
If the research team become aware of the death of a participant (whether related to the study or not) this 
should be notified to the LCTC using the appropriate CRF within 24 hours of becoming aware. 
 

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Reporting Procedures 
All safety events which are recorded for the study should be reported following the procedures detailed below. 
The occurrence of a safety event may come to the attention of research staff during routine study visits, from 
the participant’s notes, directly from the participant or by other means. Note that reporting procedures vary 
dependent on the nature of the incident (i.e. “serious” events are to be reported to LCTC in an expedited 
manner). Any questions concerning adverse event reporting should be directed to the LCTC in the first 
instance. A flowchart is given below to aid in determining reporting procedures for different types of adverse 
events. 
 
Flowchart for Site Reporting Requirements of Adverse Events 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Reporting Safety Events to the LCTC 
All safety events (whether or not assessed as serious / related / expected) should be recorded on an Adverse 
Event Form; multiple events can be recorded on one form.  
 
Safety events which are assessed as “serious” must also be recorded in more detail on Serious Adverse 
Event Forms; a single form is used for each individual event (i.e. a single diagnosis), though multiple 
symptoms can be recorded. Each SAE should have a corresponding record on the participant’s AE form. 
Where additional information is received by site after initial submission to LCTC, this should be provided on 
a follow-up form within 5 days. Serious Adverse Event Forms collect data regarding the nature of event, date 
of onset, severity, corrective therapies given, outcome and causality; all serious events reported to LCTC will 
be reviewed by the Chief Investigator or Medical Reviewer/Clinical Coordinator, and assessed for causality 
and expectedness. 
 
Follow-up after Adverse Events 
All reportable adverse events should be followed until satisfactory resolution or until the investigator 
responsible for the care of the participant deems the event to be chronic or the participant to be stable. 
Adverse Event (AE) 
(Occurring from consent until 28 days after discontinuation of study vaccine) 
Report to CTU within 24 hours  
using the SAE form 
Complete AE CRF  
Submit CRF as per routine schedule 
Expected / 
Unexpected  
Related AE / RUAE 
Expected /  
Unexpected  
SAE 
Expected /  
Unexpected  
AE 
Yes 
Yes 
Yes 
No 
No 
No 
Expected /  
Unexpected  
Related SAE / RUSAE 
 
Is the  
serious event 
related? 
Is the  
event   
serious? 
 
Is the  
event 
related? 

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When reporting “serious” safety events the investigator responsible for the care of the participant should 
apply the following criteria to provide information relating to event outcomes:  
 
resolved 
 
resolved with sequelae (specifying with additional narrative) 
 
not resolved/ongoing 
 
ongoing at final follow-up 
 
fatal or unknown. 
 
Investigator Reporting Responsibilities 
The PI is responsible for ensuring that all safety events requiring recording on this study which the local 
research team becomes aware of are reported to LCTC. It is the responsibility of the PI/Co-PI as recorded 
on the Delegation Log (medically qualified person) to assess the seriousness and causality of events.  
 
All safety events must be recorded on an AE form and transferred to LCTC within seven days of the site 
team becoming aware of the event. 
 
Safety events which meet the definition of “serious” must be reported in more detail to the LCTC on an SAE 
form and reported immediately and in no circumstances later than 24 hours from becoming aware 
unless the SAE is specified in the protocol as not requiring immediate reporting where they will be 
appropriately processed. 
 
The SAE form should be completed by an appropriately delegated member of the research team; the 
assessments of seriousness and causality must be performed by an appropriately medically qualified person. 
Minimum reporting information must be provided in initial reports for all studies.  
 
Minimum information required for reporting: 
 
Study identifier 
 
Study centre 
 
Patient number 
 
A description of the event 
 
Date of onset 
 
Current status 
 
Whether study treatment was 
discontinued 
 
The reason why the event is 
classified as serious 
 
Investigator assessment of the 
association between the event 
and study treatment 
 
 
N.B. In the absence of a delegated medically qualified person the form should be completed and signed by 
an alternative member of the research site study team and submitted to the LCTC. As soon as possible 
thereafter the responsible investigator should check the SAE form, make amendments as appropriate, sign 
and re-send to the LCTC. The initial report shall be followed by detailed follow-up reports as appropriate.  
 
Safety events should be reported to the site R&D team in accordance with local policy. 
 
 Reporting an Initial or Follow-up SAE 
The investigator should ensure the actions below are completed for all reportable SAEs: 
 
1) Research sites should telephone the appropriate study co-ordinator / data manager on telephone 
number 0151 794 9766 to advise that an SAE report has been submitted as soon as possible.  

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2) The SAE form should be transferred securely to lctcsafe@liverpool.ac.uk (within 24 hours) to 
the CTU 
 
3) The responsible investigator must notify their R&D department of the event (as per standard local 
governance procedures). 
 
4) The participant must be identified by study number, age or month and year of birth and initials only. 
The participant’s name should not be used on any correspondence. 
 
5) SAEs must be subsequently followed up in line with the processes below: 
 
Follow up must continue until clinical recovery is complete and laboratory results have 
returned to normal, or until the event has stabilised (see Section 11.7.4). N.B. Follow-up may 
continue after completion of protocol treatment if necessary. 
 
Follow-up information is noted on a new SAE form to be transferred securely to the CTU as 
soon as more information becomes available 
 
Tick the appropriate box on the new SAE form to identify the type of report; this is dependent 
on resolution status of the SAE e.g. follow-up / final. 
 
6) Extra, annotated information and/or copies of anonymised test results may be provided separately. 
 
Participant safety incidents that take place in the course of research should be reported to the National 
Reporting and Learning System (NRLS) by each participating NHS Trust in accordance with local reporting 
procedures.  This also includes reporting any NIMPs. 
 
 
LCTC Responsibilities 
The study Sponsor, University of Liverpool, have delegated to LCTC the duty of onward reporting of safety 
events to REC, regulatory authorities. SOPs will be followed to ensure appropriate reporting as detailed 
below. 
 
All “serious” safety events will be forwarded to the Chief Investigator or Medical Reviewer/Clinical Coordinator 
by LCTC within 24 hours of receiving the minimum information from site. The CI or Medical Reviewer/Clinical 
Coordinator will review information provided by site and for all events assessed as “related” will provide an 
assessment of “expectedness”. 
 
Safety events which are assessed as “serious”, “related” and “unexpected” (i.e. RUSAEs) will be onward 
reported by LCTC to the ethics committee within 15 days of the LCTC first becoming aware of the event.   
 
Additionally, RUSAEs will be reported to the study Sponsor(s) and Principal Investigators of participating 
sites. 
 
A list of all safety events recorded for the study will also be reported annually by LCTC to the ethics committee 
and Independent Data Safety & Monitoring Committee. 
 
Any concerns raised by the TSC/IDSMC or inconsistencies regarding safety reporting noted at a given site 
may prompt additional training at sites, with the potential for the LCTC to carry out site visits if there is 
suspicion of unreported AEs and SAEs in participant case notes. Additional training will also be provided if 
there are unacceptable delays in safety reporting timelines. 
 

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Safety Reports 
Safety reports will be generated during the course of the study which allows for monitoring of safety event 
including reporting rates and safety events by participant group. The LCTC will send Annual Progress Reports 
(APRs) containing a list of all SAEs to the IDSMC, regulatory authorities and main REC. If any safety reports 
identify issues that have implications for the safety of study participants, the PIs at all institutions participating 
in the study will be notified. 
 
Urgent Safety Measures (USMs) 
An urgent safety measure (USM) is a procedure to protect clinical study participants from any immediate 
hazard to their health and safety but has not previously been defined by the protocol. It can be put in place 
prior to authorisation by the REC.  
 
The LCTC will notify the REC immediately and, in any event, within 3 days that such a measure has been 
taken and the reasons why it has been taken. The initial notification to the REC will be by telephone (ideally 
within 24 hours) and a notice in writing will be sent within 3 days, setting out the reasons for the USM and 
the plan for further action. After discussion with the REC, further action will be agreed, which may include 
submission of a substantial amendment, a temporary halt, or permanent termination of the study. 
 
Following notification, if a substantial amendment is required this must be submitted as soon as possible to 
the REC. If the study is temporarily halted it may not recommence until authorised to do so by the REC. If 
the study is permanently terminated before the date specified for its conclusion (in the original applications 
to REC), the Sponsor should notify the REC within 15 days of the date of termination by submitting the formal 
End of Study Notification. 
 
Contact Details and Out-of-hours Medical Cover 
As IMOJEV® has a well-established safety profile, emergency and out-of-hours medical care will be in line 
with usual LUHFT CRU arrangements and local standard practice; no special provision is required for 
FlaviPrime participants. All participants will be provided with a CRU contact card with a 24-hour emergency 
telephone number for the CRU on call doctor, and a copy of the information sheet which includes information 
about their participation and contact details for the local research team who may be contacted if necessary. 
During office hours, the CI, LUHFT CRU, or delegate are able to provide medical advice in relation to 
participation using the contact details listed at the beginning of this document. 
 

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 STATISTICAL CONSIDERATIONS 
 
Introduction 
In the following section a brief overview of the statistical considerations relating to the design and conduct of 
the study. No formally pre-specified statistical analyses will be conducted as part of the study. 
 
Sample Size   
As this is a study with an exploratory endpoint, there is no formal hypothesis to be tested or a formal sample 
size calculation based on powering a hypothesis to some degree of accuracy. In order to allow sufficient 
material for antibody cloning, the study aims to collect single B cell receptor repertoire sequencing data on > 
1000 sorted plasmablasts from 20-25 participants. This sample size is broken down as: 
o Group 1; up to 3-4 participants of any previous flavivirus exposure state 
o Group 2; up to 6-10 flavivirus naïve participants 
o Group 3; up to 8-12 flavivirus experienced participants 
 
A total of 20 participants is expected to be vaccinated, though this figure can be increased up to a maximum 
of 25 if the critical B cell data are not obtained (e.g. the number of plasmablasts sorted for single B cell 
receptor repertoire sequencing is > 1000, or of there is technical failure of any kind). 20 participants was 
chosen as a pragmatic figure which would be feasible for generating sufficient antibody sequence data and 
monoclonal antibodies, which are extremely labour intensive in the laboratory, it being possible to make many 
hundreds of antibodies from a single individual.  
 
Method of Registration 
This study does not include participant randomisation.  All participants who have given written informed 
consent and have been found to comply with the inclusion and exclusion criteria will be registered onto the 
electronic registration database (in accordance with the LCTC registration operating procedures) by 
authorised research staff.  In the unlikely event that the back-up registration process is activated, participant 
registration will be performed by site staff with a second-line check being performed by authorised LCTC 
staff.   
 
Outcome Measures 
This study aims to establish a unique human experimental medicine system which will be used to study 
flavivirus immune responses and for epitope discovery and there will be no primary or secondary endpoint in 
this study. 
 
Exploratory Objectives 
The objective of this study is to describe the priming of B cells to JE vaccine IMOJEV®  including mapping of 
specific B cell epitopes of the vaccine virus, by generating a panel of monoclonal antibodies from single cell 
sorted plasmablasts at one week post vaccine. 
 
To determine the ability of B cell responses primed by JE vaccine IMOJEV® to cross-react with other 
flaviviruses, by neutralisation assay (B cells). 

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Interim Analyses 
As there are no formal hypotheses being tested, there are no formal stopping rules (other than safety) or 
mechanisms defined here to stop the study prior to the planned end of the study.  The study does have a 
formal Oversight Committee who will be able to review at regular intervals all accumulating data.  The main 
responsibility of this committee will be to review the recruitment of participants, the collection of all essential 
data, to assess participant safety and to provide recommendations regarding the administration of a second 
dose of the JE IMOJEV® vaccine, based on the magnitude of the immune response to the vaccine. 
 
 
Analysis Plan 
Analyses for the study are under the remit of the CI and will not be performed by the LCTC.  The analysis 
being carried out is exploratory in nature, the details in terms of the methodology for this analysis may be 
altered during the course of the study. The analysis will be largely descriptive including (but not limited to) 
summaries of 
 
% plasmablasts (defined as CD38hi, CD27hi) in the recently activated B cell population (CD19+, 
CD20lo). 
 
Number of plasmablasts sorted 
 
% of JEV specific cells, measured by B cell ELISpot 
 
Number of antibodies made that bind JEV  
 
Proportion of virion binding antibodies that neutralise 
 
Proportion that are specific for the fusion loop epitope 
 
JEV neutralisation/neutralisation of other viruses before and after vaccination (DENV1-4, ZIKV, WNV) 
Reporting guidelines for analyses are detailed in section 12.7.1 to 12.7.6. 
 Participant Groups for Analysis 
As this is not a study comparing an intervention on participant outcome, final analysis will take place on a 
per-protocol basis, removing any participants who have any major protocol deviations. 
 Significance Levels 
As this is an exploratory study, no formal levels of significance are set.  All statistics presented will be 
presented alongside 95% confidence intervals so as to give an indication of the level of precision only. 
 Missing Data 
The likelihood of missing data are small given the small number of participants in this study.  Final analysis 
will take place on a complete-case basis with no adjustments made (e.g. multiple imputation) in the case of 
missing data. 
 Exposure to Vaccine 
Exposure to vaccine will be assessed by the assessment of compliance. Because the study involves a single 
dose of vaccine, only participants who have been vaccinated will be included in the subsequent sampling 
schedule and analyses. 
 Trigger for Final Analysis 
Analysis of study data will take place once all participants have received the planned follow-up and all data 
are available for analysis. 

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 Data descriptions 
Continuous data shall be summarised as median, inter-quartile ranges and ranges.  Categorical data shall 
be summarised as frequencies of counts and associated percentages. 
 

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 DATA MANAGEMENT AND STUDY MONITORING 
For the FlaviPrime study the responsibilities for Data Management and monitoring are delegated to the LCTC  
Separate Data Management and Study Monitoring Plans have been developed which provide detail 
regarding the internal processes that will be conducted at the LCTC throughout the study. Justification for the 
level of monitoring is provided within those documents and the study-specific risk assessment. All data will 
be managed as per local LCTC processes and in line with all relevant regulatory, ethical and legal obligations. 
 
 
Source Documents 
The case report form (CRF) will be considered the source document for data where no prior record exists 
and which is recorded directly in the bespoke CRF. A FlaviPrime source document list will be produced for 
each site to be kept in the ISF and provide detail of what constitutes FlaviPrime-specific source data. 
 
Date(s) of informed consent processes (including date of provision of patient information, randomisation 
number and the fact that the patient is participating in a clinical study (including possible treatment arms) 
should be added to the patient’s medical record chronologically.  
 
Data Collection Methods 
Participant eCRFs/ CRFs will be provided to sites for local completion by members of the research team 
trained and delegated the duty. Study staff named at each site will enter data from source documents 
corresponding to a participant’s visit onto the relevant CRF in the participant’s folder The CRF is the primary 
data collection instrument for the study so all data requested on the CRF must be recorded and all missing 
data must be explained. A copy of all CRFs should be retained at site. Any corrections should be made in 
accordance with GCP. 
 
For participant registration: Data are to be entered into a bespoken LCTC electronic database by members 
of the research team at site. Training will be provided prior to any data entry. 
 
Monitoring 
Monitoring is conducted to ensure protection of patients participating in the study and all aspects of the study 
(procedures, laboratory, study intervention administration and data collection) are of high quality and 
conducted in accordance with sponsor and regulatory requirements. 
 
A detailed Study Monitoring Plan will be developed and agreed by the SMG and CI to describe who will 
conduct the monitoring, at what frequency monitoring will be done, and what level of detail monitoring will be 
conducted. This will be dependent on the documented risk assessment of the study which determines the 
level and type of monitoring required for specific hazards. All processes may be subject to monitoring, e.g. 
enrolment, consent, adherence to study interventions, accuracy and timeliness of data collection etc.  
 
Study Oversight Committees related to the monitoring of the study are detailed in Roles and Responsibilities 
see section 3. 
 
Central Monitoring 
There are a number of monitoring features in place at the LCTC to ensure reliability and validity of the study 
data, to be detailed in the study monitoring plan. Data will be entered into a validated database and during 
data processing there will be checks for missing or unusual values (range checks) and for consistency within 

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participants over time. Other data checks relevant to patient rights and safety will also be regularly performed 
as per LCTC processes. Any suspect data will be returned to the site in the form of data queries. Data query 
forms will be produced at the LCTC from the study database and sent either electronically or through the post 
to a named individual (as listed on the site delegation log). Sites will respond the queries providing an 
explanation/resolution to the discrepancies and return the data query forms to the LCTC. The forms will then 
be filed along with the appropriate CRFs and the appropriate corrections made on the database.  
 
Site monitoring visits may be ‘triggered’ in response to concerns regarding study conduct, participant 
recruitment, outlier data or other factors as appropriate.  
 
Clinical Site Monitoring 
 
In order to perform their role effectively, monitors and persons involved in Quality Assurance and Inspection 
will need direct access to primary subject data, e.g. participant records, laboratory reports, appointment 
books, etc. Each PI therefore permits study related monitoring, audits, ethics committee review and regulatory 
inspections by providing direct access to source data/documents. As this affects the participant’s 
confidentiality, this fact is included on the PIS and ICF. 
 Risk Assessment 
In accordance with the LCTC Standard Operating Procedures and the requirements of the sponsor, a risk 
assessment has been completed in partnership with:  
 
 
Representatives of the Study Sponsors (University of Liverpool)  
 
Chief Investigator  
 
Members of the  Study Management Group 
 
Study Coordinator  
 
Statistician  
 
LCTC Operational Director  
 
In conducting this risk assessment, the contributors considered potential patient, organisational and study 
hazards, the likelihood of their occurrence and resulting impact should they occur.  
 
The outcome of the risk assessment is categorised based upon the potential risk associated with the study 
intervention in accordance with MRC/DH/MHRA Project on Risk-adapted Approaches to the Management of 
Clinical Trials of Investigational Medicinal Products based on the following categories:  
 
 
Type A: No higher than that of standard medical care  
 
Type B: Somewhat higher than that of standard medical care  
 
Type C: Markedly higher than that of standard medical care 
 
Non-CTIMP 
 
The FlaviPrime study is using a vaccine that is not licensed in an EU member State.  However is licensed for 
use in Australia since August 2010, Thailand since October 2010 and Malaysia 2013. The MHRA have 
confirmed that this study is classified as a Non-CTIMP. 
 
Confidentiality 
This study will collect personal data (e.g. participant names), including special category personal data (i.e. 
participant medical information) and this will be handled in accordance with all applicable data protection 
legislation. Data (including special category) will only be collected, used and stored if necessary for the study 

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(e.g. evidencing provision of consent, for data management and central monitoring, statistical analysis, 
regulatory reporting, etc.). At all times, this data will be handled confidentially and securely. 
 
CRFs will be labelled with a unique study screening number. Verification that appropriate informed consent 
is obtained will be enabled by the provision of copies of participant’s signed informed consent forms being 
supplied to the CTU by recruiting sites. This transfer of identifiable data is disclosed in the PISC. 
N.B. Consent forms must be transferred separately to any other study documentation to ensure the 
pseudonymisation of special category data is maintained. 
 
Site-specific study-related information will be stored securely and confidentially at sites and all local relevant 
data protection policies will be adhered to.  
 
The LCTC as part of The University of Liverpool will preserve the confidentiality of participants taking part in 
the study. The University of Liverpool is registered as a Data Controller with the Information Commissioners 
Office.  
 
Breaches of data protection principles or regulations identified by LCTC will be notified promptly to the study 
Sponsor and The University of Liverpool’s Data Protection Officer and appropriate processes followed. 
 
 
Quality Assurance and Control 
To assure protocol compliance, ethical standards, regulatory compliance and data quality, as a minimum, the 
following will occur:  
 
The PI and other key staff from each centre will attend initiation training, which will incorporate 
elements of study-specific training necessary to fulfil the requirements of the protocol. 
 
The SOC will determine the minimum key staff required to be recorded on the delegation log in order 
for the centre to be eligible to be initiated. 
 
The TC at the LCTC will verify appropriate approvals are in place prior to initiation of a centre and the 
relevant personnel have attended the study specific training. A greenlight checklist will verify all 
approvals are in place prior to study initiation at LCTC and the individual centre.  
 
The study will be conducted in accordance with procedures identified in the protocol. 
 
The Study Oversight Committee (SOC) will provide independent oversight of the study. 
 
SOC will monitor screening, consent rate and compliance with the protocol. 
 
Data quality checks and monitoring procedures will be undertaken in line with the study Data 
Management Plan. 
 
Records Retention 
The retention period for the FlaviPrime data and information is a minimum of 5 years after issue of the final 
report, or longer at Sponsor’s request. 
 
The PI at each investigational site must make arrangements to store the essential study documents (as 
defined by ICH GCP guidelines) including the Investigator Site File, the applicable participant medical records 
and Pharmacy Site File, for the full length of the study’s retention period and will arrange for confidential 
destruction at the end of this period as instructed by the Sponsor / CTU. 
 
The PI is also responsible for archiving all relevant source documents so that the study data can be compared 
against source data after completion of the study (e.g. in case of inspection from authorities). They must 
ensure the continued storage of the documents, even if they, for example, leave the clinic/practice or retire 
before the end of required storage period. Delegation of responsibility for this must be documented in writing. 
 

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All other persons and organisations involved in the study will be responsible for storing and archiving the 
parts of the TMF relevant to their delegated duties (e.g. laboratories, IMP manufacturers and distributors, 
third-party vendors providing randomisation and IMP allocation systems, etc.). 
 
The LCTC undertakes to archive as per their contractual requirements; documents will be archived in 
compliance with the principles of GCP. All electronic CRFs and study data will be archived onto an 
appropriate media for long term accessible storage. Hard copies of data will be boxed and transferred to 
secure premises where unique reference numbers are applied to enable confidentiality, tracking and retrieval. 

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 REGULATORY AND ETHICAL CONSIDERATIONS 
 
Statement of Compliance 
This study is designed to comply with the guideline developed by the International Conference on 
Harmonisation (ICH) for Good Clinical Practice (GCP) and will be conducted in compliance with the protocol, 
LCTC Standard Operating Procedures and EU Directive 2001/20/EC, transposed into UK law as the UK 
Statutory Instrument 2004 No 1031: Medicines for Human Use (Clinical Trials) Regulations 2004. 
 
Ethical Considerations 
The study will be conducted to conform to the principles of the Declaration of Helsinki as adopted by the 18th 
World Medical Assembly, 1964, and subsequent amendments (Tokyo (1975), Venice (1983), Hong Kong 
(1989) and South Africa (1996).  
 
The study will be conducted in accordance with the EU Directive 2001/20/EC, the Medicines for Human Use 
(Clinical Trials) Regulations 2004 and the principles of Good Clinical Practice.  
 
Participants will be asked to consent that data are recorded, collected, stored and processed and may be 
transferred to other countries, in accordance with any national legislation implementing the EU Data 
Protection Directive (95/46/EC).  
 
This study may be terminated at the request of the CI, Study Oversight Committee, or National Research 
Ethics Committee (REC) if, during the course of the study, concerns about the safety of further dosing 
emerge.  
 
The CI will update the ethics committee of any new information related to the study vaccine when appropriate. 
 
Approvals 
The study protocol has received the favourable opinion of the North West – Greater Manchester Central 
Multi-centre Research Ethics Committee (MREC) but all participating sites must undergo site specific 
assessment of capacity and capability.  A copy of all site approval documents and a copy of the PISC on 
local headed paper should be forwarded to LCTC before patients are entered. The LCTC should receive a 
confirmation of capacity and capability for each new centre via the site’s R&D department. 
 
 
Protocol Deviation and Serious Breaches 
Deviations from, breaches or violations of, or non-compliance to either the protocol, the conditions or 
principles of GCP, and relevant regulatory and ethical e.g.  REC requirements are handled based on their 
nature and severity. 
 
Non-Serious breaches 
Protocol deviations and other non-serious breaches of GCP etc. will be managed according to local site and 
LCTC procedures as appropriate. They will be reported to study oversight committee. 
 
Serious breaches 
A breach of the protocol or GCP is ‘serious’ if it meets the definition of being “likely to affect to a significant 
degree the safety or physical or mental integrity of the study participants, or the scientific value of the study”. 
This assessment can only be determined by the Sponsor. 
 

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If any persons involved in the conduct of the study become aware of a potential serious breach, they must 
immediately report this to the LCTC who will in turn notify the Sponsor. The Sponsor will assess the breach 
and determine if it meets the criteria of a ‘serious’ breach.  
 
The Sponsor may seek advice from medical expert members of the oversight committee in determining 
whether or not the breach is likely to affect to a significant degree the safety, physical or mental integrity of 
participants. The Sponsor may seek advice from the study statistician and CI in determining whether or not 
the breach is likely to significantly affect the scientific value of the study. However, the Sponsor retains 
responsibility for the assessment of whether or not a breach meets the definition of ‘serious’ and is subject 
to expedited reporting to REC. 
 
Breaches confirmed as ‘serious’ will be reported to REC within 7 days by the CTU on behalf of the Sponsor 
and notified to the SOC at their next meeting. Any requests for additional information from the Sponsor, SOC, 
or REC, will be promptly actioned by the relevant member(s) of the research team and open communication 
will be maintained to ensure appropriate corrective actions are taken and documented. Incidents of protocol 
non-compliance will be recorded as protocol deviations, the incidence of which are monitored and reported 
to the study oversight committee.  
 

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 INDEMNITY 
FlaviPrime - IMOJEV® is sponsored by the University of Liverpool and co-ordinated by the LCTC in the 
University of Liverpool. The University of Liverpool does not hold insurance against claims for compensation 
for injury caused by participation in a clinical study and they cannot offer any indemnity.  
 
As this is an investigator-initiated study, The Association of the British Pharmaceutical Industry (ABPI) 
guidelines for patient compensation by the pharmaceutical industry do not apply. However, in terms of liability, 
NHS Trust and Non-Trust Hospitals have a duty of care to patients treated, whether or not the patient is 
taking part in a clinical study and they are legally liable for the negligent acts and omission of their employees. 
Compensation is therefore available in the event of clinical negligence being proven. 
 
Clinical negligence is defined as: 
 
“A breach of duty of care by members of the health care professions employed by NHS bodies or by others 
consequent on decisions or judgments made by members of those professions acting in their professional 
capacity in the course of their employment, and which are admitted as negligent by the employer or are 
determined as such through the legal process”. 
 
 
 
 
 

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 PUBLICATION AND DISSEMINATION 
 
Publication Policy 
The results from this study will be analysed together and published as soon as possible. Individual 
researchers must undertake not to submit any part of their individual data for publication without the prior 
consent of the Study Management Group. 
 
The Study Management Group will form the basis of the Writing Committee and advise on the nature of 
publications. 
The 
Uniform 
Requirements 
for 
Manuscripts 
Submitted 
to 
Biomedical 
Journals 
(http://www.icmje.org/) will be respected. All publications shall include a list of participants, and if there are 
named authors, these should include the study’s Chief Investigator(s), Statistician(s) and Study Manager(s) 
involved at least. If there are no named authors (i.e. group authorship) then a writing committee will be 
identified that would usually include these people, at least.  
 
The members of the Study Oversight Committee should be listed with their affiliations in the 
Acknowledgements / Appendix of the main publication. All publications arising from the FlaviPrime Study will 
be open access, according to the conditions of the funder of the Study, the Wellcome Trust 
(https://wellcome.ac.uk/funding/managing-grant/open-access-policy). 
 
 
Dissemination to Key Stakeholders 
On completion of the research, a Final Study Report will be prepared and submitted to the REC. The results 
of FlaviPrime will be published regardless of the magnitude or direction of effect. 
 
Data Sharing 
At the end of the study, after the primary results have been published, the anonymised individual participant 
data (IPD) and associated documentation (e.g. protocol, statistical analysis plan, annotated blank CRF) will 
be prepared in order to be shared with external researchers. All requests for access to the IPD will be 
reviewed by an internal committee at the CTU and discussed with the Chief Investigator in accordance with 
the CTU policy on data sharing. Data will be shared with the AVIS study, which already collects many of the 
same data, to facilitate the investigation of how the response to IMOJEV is modulated by pre-existing immune 
responses to flaviviruses, which have been determined in the flavimmune substudy of AVIS.   
 

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 CHRONOLOGY OF PROTOCOL AMENDMENTS 
 
Version 6 (08/07/2021) 
Summary of Amendments from Protocol V5.0 to Protocol V6.0 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
Throughout 
Throughout 
Change to version and date of document. 
Throughout 
Throughout 
Sample size amended from ‘up to 20’ to ‘up to 25’ 
Number of participants in group 2 amended from 6-8 to 6-10 
Number of participants in group 3 amended from 8-10 to 8-12 
Throughout 
Throughout 
‘Patient’ amended to ‘participant’ as necessary. 
Throughout 
Throughout 
Study duration for group 1 clarified as 28 days. 
Throughout 
Throughout 
Flavimmune study updated to AVIS study, as a reflection of 
study name change. 
Throughout 
Throughout 
References to MedDRA/CTCAE removed – these were 
included in error. 
N/A 
Table of 
Contents and 
Glossary 
Updated to reflect changes throughout protocol. 
2 
Protocol 
overview – 
Inclusion 
Criteria 
Inclusion criterion 6 updated to clarify contraception must be 
used for 28 days after vaccination. 
2 
Protocol 
overview – 
Exclusion 
Criteria 
Exclusion criterion 9 (‘detectable anti-Flavirus neutralizing 
antibodies in screening tests, group 2 only’) removed; all 
subsequent criteria re-numbered. 
5 
Study design 
Text amended to clarify that sample collection in AVIS study will 
be temporarily suspended during participation in FlaviPrime. 
6.1 
Inclusion 
Criteria 
Inclusion criterion 6 updated to clarify contraception must be 
used for 28 days after vaccination. 
6.2 
Exclusion 
Criteria 
Exclusion criterion 9 (‘detectable anti-Flavirus neutralizing 
antibodies in screening tests, group 2 only’) removed; all 
subsequent criteria re-numbered. 
6.3 
Co-enrolment 
guidelines 
Co-enrolment guidelines amended to confirm participants can 
join FlaviPrime provided certain certain criteria are met. 

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7.7 
Concomitant 
medications/ 
treatments and 
specific 
restrictions 
Clarification added that COVID vaccines can be given to 
participants, and participants should inform study team if they 
must take certain medications. 
9.1 
Participant 
identification 
and screening 
‘Negative neutralising antibody’ removed as a condition for 
participants to enter group 2. 
9.4 
Enrolment/ 
registration and 
baseline 
assessments 
Addition of GP notification added – participant GP will receive 
a letter to notify them of patient involvement in FlaviPrime. 
9.5 
Schedule for 
assessments 
and follow-up 
Clarification of laboratory assessments and re-insertion of table 
showing correct schedule of assessments (note, these tables 
were used in a separate Schedule of Assessments document, 
as part of FlaviPrime Amendment 4; they have now been re-
added to the protocol). 
9.7 
Intervention  
Discontinuation 
and Participant 
Discontinuation/
Withdrawal 
Participant transfer section updated to clarify that replacing 
transferred participants is dependent on vaccine availability . 
10.6 
Time period for 
active 
monitoring of 
safety events 
Clarficiation added that group 1 participants should report AEs 
that occur 28 after vaccination to site, by phone (group 1 last 
study visit to site is 9 days after vaccination). 
 
Clarification added that, if site become aware of AEs occurring 
outside of the 28 day AE monitoring window, these events can 
be reported to LCTC. 
11 
Reporting of 
pregnancy 
Contact email address for reporting events updated to LCTC 
Central Safety Team (lctcsafe@liverpool.ac.uk). 
 
Error in birth control window corrected to 28 days after 
vaccination. 
11.2.1 
Reporting an 
initial or follow-
up SAE 
Contact email address for reporting events updated to LCTC 
Central Safety Team (lctcsafe@liverpool.ac.uk). 
12 
Statistical 
considerations 
Section updated throughout to reflect: there are no formally pre-
specificied statistical analyses conducted; analyses are under 
the remit of the Chief Investigator and not performed by LCTC; 
and statistical analyses plan will not be place due to the 
exploratory nature of the analyses.  
12.2 
Sample size 
Sample size amended from ‘up to 20’ to ‘up to 25’ 
Number of participants in group 2 amended from 6-8 to 6-10 
Number of participants in group 3 amended from 8-10 to 8-12 
 
Information added to explain sample size figure chosen. 
13.2 
Data collection 
methods 
Reference to screening log removed – this was not omitted 
previously, in error.  
16.3 
Data sharing 
Statement added to clarify data sharing with AVIS study. 

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Version 5.0 (08/03/2021) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Version 4.0 (18/12/2020) 
17 
Chronology of 
protocol 
amendments 
Chronology updated to reflect document history; history of 
previous protocol changes also added. 
Summary of Amendments from Protocol V4.0 to Protocol V5.0 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
Throughout 
Throughout 
Change to version and date of document. 
1 
Table of 
Contents 
Updated to reflect current document. 
9.5 
Schedule of 
assessments 
(table) 
Table removed from protocol due to typographical errors. This 
Schedule of Assessments table is now provided as a 
supplementary document to the study protocol. 
17 
Chronology of 
protocol 
amendments 
Chronology updated to reflect document history.  
Summary of Amendments from Protocol V3.0 to Protocol V4.0 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
Throughout 
Throughout 
Change to version and date of document 
N/A 
Contact details 
Sponsor Representative details updated. Institute of Infection 
and Global Health update to Institute of Infection, Veterinary 
and Ecological Sciences; contact number for Dr Turtle updated 
Section 2 
Overview 
Study phase corrected. Study duration for group 1 specified 
(previously missing) 
Section 2.1 
Schematic of 
study design 
Figure 2 updated to include the baseline blood sample for group 
1 which was previously missing 
Section 3 
Roles and 
responsibilities – 
clinical trials unit 
LCTC defined (Liverpool Clinical Trials Centre) 
Section 3 
Oversight 
committees 
“Trial” corrected to “study” throughout 

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Version 3.0 (08/08/2020) 
This version made major changes, merging the changes from v2.0 and v1.2, resolving version control issues 
and transferring to the current LCTC template. 
 
Section 4.3 and 
4.4 
Introduction 
Minor typographical errors corrected 
9.5 
Schedule for 
assessments 
and follow up 
Study duration for group 1 specified (previously missing) 
9.5 
Schedule of 
assessments 
(table) 
Table updated to correct typographical errors - removal of X 
indicating 2nd HIV/HBV/HCV test (was never planned) and 
removal of travel history at followup visits for group 1 (never 
planned). 
17 
Chronology of 
protocol 
amendments 
Chronology updated to reflect document history.  
Summary of Amendments from Protocol V1.2 and V2.0 to Protocol V3.0 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
Throughout 
Throughout 
Change to version and date of document 
Complete overhaul of document due to the introduction of a 
new LCTC protocol SOP/template 
Correction of typographical errors 
Conflicts between versions 1.2 and 2.0 resolved (previously 
approved) 
‘Trial’ replaced with ‘study’ 

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Site name updated to Liverpool University Hospitals NHS 
Foundation Trust (LUHFT, Royal Liverpool site) (this is as a 
result of merger with Aintree) 
T cell and associated IFNγ-ELISpot and intracellular cytokine 
staining terms have been updated for clarification of the 
techniques that will be used to study the T cell response to the 
vaccine 
N/A 
Contact Details: 
Institutions 
Update to contact details 
N/A 
Table of 
Contents and 
Glossary 
Updated to reflect changes throughout protocol 
2 
Protocol 
Overview 
Overview updated to reflect changes within protocol 
2.1 
Schematic of 
Study Design 
New schedule for group 1 (newly added to protocol v3.0), no 
changes for groups 2 and 3, aside from group name 
4.4 
Objectives 
Clarification that all objectives are exploratory, change to 
wording of primary objectives 3 and 4 (though scientific-
meaning has not changed), addition of primary objective 5  (“To 
establish a sample bank for future work on cross-reactive and 
other responses to flaviviruses, flavivirus vaccines and other 
emergent viruses”). 
Addition of 2 secondary objectives (“To examine the specificity 
and cross-reactivity of the antibody response after JE 
vaccination, using serum and human monoclonal antibodies” 
and “To determine whether there are epitopes which can serve 
as 
the 
target 
of 
broadly 
cross-neutralising 
antibody 
responses.”) 
Clarification of which objectives are primary and which are 
secondary 
5 
Study Design 
Addition of another patient group (now 3 groups in total; group 
1 = participants of any previous flavivirus exposure, group 2 = 
flavivirus naïve participants, group 3 = flavivirus experiences 
participants) 
 Increase in the sample size from 6 to ‘up to 20.’ 
Change to timelines as a result of new group. 
6 
Eligibility Criteria 
Inclusion criterion 3 – typographical error corrected regarding 
study duration 
Inclusion criterion 5 –  due to the introduction of another patient 
group, group-specific criteria amended to reflect new group 
numbers 
Exclusion criteria 8, 9 and 14 –  due to the introduction of 
another patient group, group-specific criteria amended to 
reflect new group numbers 
7.3 
Manufacturing 
and Distribution 
Information added that omitted from the previous protocol 
7.7 
Specific 
Requirements/ 
Restrictions 
Table of study-specific restrictions added (this was omitted from 
previous versions of the protocol in error, but was present in the 
PISC) 

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9.1 
Participant 
Identification 
and Screening 
Clarification text added confirming that participants identified 
through consent4consent database at LUHFT CRU can be 
consented at site, donate blood and have flavivirus 
immunological assays performed as per the Flavimmune study 
9.3 
Screening, 
Eligibility 
Assessment and 
Confirmation 
Hepatitis B and C tests added at screening to reduce non-
specific effects from these viruses, and text added to explain 
the rationale behind this 
9.5 
Schedule of 
Assessments 
Schedule of Assessments added to reflect the 3 study groups 
9.5 
Adverse Events 
Added in the use of an emergency contact card that participants 
will received so they can contract the CRU doctor on call 
9.6 
Sampling 
Slight increase in blood volumes collected 
Change in  name of University of Liverpool department who will 
analyse blood 
Added in use of blood samples for other emergent viruses 
(specifically proposed up front consent for experiments relating 
to newly emerging viruses, based on experience with SARS-
CoV-2) 
9.7 
Participant 
Transfer 
Correction of error to process for participant transfers – now 
clarified that participants will have to be withdrawn if they are 
unable to attend follow-up visits, and if withdrawn before the 
day 9 sample (group 1) two-week sample (groups 2 and 3) has 
been taken, participant should be replaced. 
10 
Safety 
Reporting 
Overhaul of Safety Reporting section to refer to non-CTIMP 
processes, and new LCTC reporting processes 
11 
Reporting of 
Pregnancy 
 Information added regarding pregnancy, the possibility of 
vaccine viraemia, and the low likelihood of these coinciding has 
been resolved between protocol v2.0 and v1.2 (previously 
approved) 
11.4 
Contact  Details 
and Out-of-
hours Medical 
Cover 
Information added regarding out-of-hours medical cover (this 
was omitted from previous versions in error) 
12.7 
Statistical 
Analysis Plan 
Removal of ‘proportion binding virion versus NS1’ from list of 
descriptive statistics that will be calculated 
13 
Data 
Management 
and Study 
Monitoring 
Change from central monitoring to on-site monitoring (LCTC 
will only carry out monitoring if triggered) 
17 
Chronology of 
Protocol 
Amendments 
Chronology of protocol amendments updated for clarification of 
document history 
18 
References 
Additional references added 

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Page 72 of 75 
 
Version 1.2 (15/01/2020) 
This version was created from v1.1 in error. V1.2 was submitted as part of Amendment 1. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Version 2.0 (27/06/2019) 
This was the original approved version of the protocol (REC favourable opinion received 25/07/19). 
 
 
 
 
 
 
 
 
 
 
 
Version 1.1 (17/04/2019) 
This was the original version of the protocol submitted to the REC and considered on 21st May 2019. 
 
 
Summary of Amendments from Protocol V1.1 to Protocol V1.2 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
Section 4.2 
(v1.2) 
Exclusion 
criteria 
Lymphocyte count < 1.0 x 109/L left in the protocol, in error 
Section 5.3 
(v1.2) 
Screening 
Minor wording changes relating to pregnancy inserted at the 
request of the REC upon review of PISC 
Section 9.2.3 
(v1.2) 
Reporting of 
pregnancy 
Clarification of the likely effects on pregnancy inserted at the 
request of the REC upon review of PISC 
Summary of Amendments from Protocol V1.1 to Protocol V2.0 
Protocol 
Section 
Number 
Protocol 
Section Title 
Summary of Changes 
4.2 (v1.1, now 
section 6.2) 
Exclusion 
criteria 
Lymphocyte count < 1.0 x 109/L removed (had been left in 
accidentally from a previous version of the protocol that 
used a different vaccine) 

FlaviPrime Protocol V.6, 08/07/2021 
(Based on TM001TEM01: LCTC Protocol Template, v1.0, 20/02/2020) 
 
 
 
IRAS ID: 264195 
 
Page 73 of 75 
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FlaviPrime Protocol V.6, 08/07/2021 
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IRAS ID: 264195 
 
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 DOCUMENTS SUPPLEMENTARY TO THE PROTOCOL 
 
Documents referenced within the protocol are separately maintained and version controlled. Any of the 
supplementary documents subject to CA and / or Ethical review are submitted as separate version controlled 
documents.