Assessment of Long Term Immunogenicity of Japanese Encephalitis Live Attenuated SA-14-14-2 Vaccine in Previously Vaccinated Bangladeshi Children and Antibody Response and Safety to a Booster Dose
Phase: PHASE4
Conditions: Encephalitis, Japanese
Sponsor: PATH

NCT02514746
Protocol:  Assessment of long term immunogenicity of Japanese encephalitis live attenuated 
SA-14-14-2 vaccine in previously vaccinated Bangladeshi children and antibody response and safety 
to a booster dose 
v2.1 dated 04 March 2018

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JEV07 Study Protocol, Version 2.1 
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TABLE OF CONTENTS 
RRC APPLICATION FORM ......................................................................................................................... 1 
Table of Contents ............................................................................................................................................ 11 
List of Abbreviations ...................................................................................................................................... 13 
A. Project Summary ...................................................................................................................................... 14 
B. Description of the research project ........................................................................................................... 15 
1.
Hypothesis to be tested ...................................................................................................................... 15 
2.
Specific Objectives ............................................................................................................................ 15 
2.1. Primary objective ....................................................................................................................... 15 
2.2. Secondary objectives ................................................................................................................. 15 
2.3. Exploratory objective ................................................................................................................ 15 
2.4. Outcome measures ..................................................................................................................... 15 
3.
Background of the Project including Preliminary Observations: ...................................................... 16 
3.1. Disease burden ........................................................................................................................... 16 
3.2. JE in Bangladesh ....................................................................................................................... 16 
3.3. Previous experience with CD-JEV ............................................................................................ 16 
3.4. Study rationale ........................................................................................................................... 17 
3.5. Potential risks and benefits ........................................................................................................ 18 
4.
Research Design and Methods ........................................................................................................... 18 
4.1. Study sites .................................................................................................................................. 20 
4.2. Study enrollment and withdrawal .............................................................................................. 20 
4.3. Study vaccine ............................................................................................................................. 22 
4.4. Study procedures ....................................................................................................................... 23 
4.5. Laboratory evaluations .............................................................................................................. 27 
5.
Sample Size Calculation and Outcome (Primary and Secondary) Variable(s) .................................. 28 
5.1. Serology variables ..................................................................................................................... 28 
5.2. Safety variables ......................................................................................................................... 28 
6.
Data Analysis ..................................................................................................................................... 28 
6.1. Immunogenicity ......................................................................................................................... 28 
6.2. Safety ......................................................................................................................................... 29 
6.3. Analysis sets .............................................................................................................................. 29 
7.
Data Safety Monitoring plan ............................................................................................................. 30 
7.1. Assessment of safety ................................................................................................................. 30 
7.2. Methods and timing for assessing, recording, and analyzing safety parameters ....................... 30 
7.3. Reporting procedures ................................................................................................................. 34 
7.4. Safety monitoring plan .............................................................................................................. 34 
7.5. Halting rules .............................................................................................................................. 34 
7.6. Data handling and record keeping ............................................................................................. 35 
7.7. Quality control and quality assurance........................................................................................ 36 
7.8. Clinical monitoring .................................................................................................................... 36 
7.9. Audits and inspections ............................................................................................................... 38 
8.
Ethical Assurance for Protection of Human rights ............................................................................ 38 
8.1. Ethical standard ......................................................................................................................... 38 
8.2. Institutional review board .......................................................................................................... 38 
8.3. Informed consent process .......................................................................................................... 38 
8.4. Inclusion of women and children .............................................................................................. 39 
8.5. Participant confidentiality.......................................................................................................... 39 
8.6. Study discontinuation ................................................................................................................ 40 
8.7. Future use of stored specimens .................................................................................................. 40 
9.
Use of Animals .................................................................................................................................. 40 

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10. Collaborative Arrangements .............................................................................................................. 40 
10.1. Publication policy ...................................................................................................................... 40 
11. Facilities Available ............................................................................................................................ 41 
11.1. Field site .................................................................................................................................... 41 
11.2. Laboratory facilities ................................................................................................................... 41 
12. Literature Cited .................................................................................................................................. 42 
 
 
 
 
 
 

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LIST OF ABBREVIATIONS 
AE 
adverse event 
CDIBP 
Chengdu Institute of Biological Products 
CD-JEV 
prequalified live, attenuated SA-14-4-2 Japanese encephalitis vaccine  
CI 
confidence interval 
CRF 
case report form 
CSR 
clinical study report 
DSMB 
data safety and monitoring board 
ELISA 
enzyme-linked immuno assay 
ERC 
ethical review committee 
FSC 
fixed site clinic 
GACVS 
WHO Global Advisory Committee on Vaccine Safety 
GCP 
good clinical practice 
GMC 
geometric  mean concentration 
GMP 
good manufacturing practice 
GMT 
geometric mean titer 
HDSS 
health and demographic surveillance system 
icddr,b 
International Center for Diarrheal Diseases Research, Bangladesh 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee on Medical Journal Editors 
IEC 
Independent or Institutional Ethics Committee 
IgM 
immunoglobulin M 
IRB 
Institutional Review Board 
ITT 
intent to treat analysis set 
JE 
Japanese encephalitis 
PHK 
primary hamster kidney 
PP 
per protocol analysis set 
PRNT 
plaque reduction neutralization test 
RRC 
research review committee 
SAE 
serious adverse event 
SAGE 
WHO Strategic Group of Experts 
SOP 
Standard Operating Procedure 
WIRB 
Western Institutional Review Board 
WHO 
World Health Organization 
 

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JEV07 Study Protocol, Version 2.1 
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A. PROJECT SUMMARY 
Principal Investigator: K. Zaman 
 
Research Protocol Title: Assessment of long term immunogenicity of Japanese encephalitis live attenuated SA-
14-14-2 vaccine in previously vaccinated Bangladeshi children and antibody response and safety to a booster 
dose 
Proposed start date: June 2015                                                                   Estimated end date: June 2017 
Japanese encephalitis (JE) virus, a mosquito-borne flavivirus, is the leading cause of viral neurological disease 
and disability across temperate and tropical zones of Asia. In 2012 PATH sponsored a double-blind, 
randomized controlled trial (icddr,b Protocol #11050, JEV05) among 818 Bangladeshi infants aged between 10 
and 12 months to compare the immunogenicity of three lots of live attenuated SA-14-14-2 Japanese 
encephalitis vaccine (CD-JEV) manufactured in the Chengdu Institute of Biological Products’ newly 
constructed Good Manufacturing Practice facility and one lot of CD-JEV manufactured in the pre-existing 
facility. Seroprotection rates (anti-JE neutralizing antibody titer >1:10) ranged from 80 to 86 percent for all four 
lots of vaccine, with an average geometric mean titer (GMT) of 56.1,2 These GMT and seroprotection rates were 
lower than those seen in other populations, including a study conducted in Philippines.3 
 
This study aims to understand the persistence of the antibody response to guide the Bangladesh Ministry of 
Health and Family Welfare, the World Health Organization, and other ministries of health in determining if and 
when a booster dose of CD-JEV may be advisable. The proposed study is a phase 4 open-label trial enrolling 
Bangladeshi children who had previously participated in JEV05 (icddr,b Protocol #11050). Eligible participants 
who still live in the area will be identified using the local health and demographic surveillance system and 
asked to participate.  
 
A 2 ml blood sample will be collected from all JEV05 participants in 2015 and 2016, which is three and four 
years after initial vaccination, respectively. The sera will be tested using a 50% plaque reduction neutralization 
test (PRNT-50); a JE neutralizing antibody titer > 1:10 will be considered seroprotective.1 In 2016, four years 
after receiving the initial dose of CD-JEV, eligible participants will be vaccinated with a second subcutaneous 
dose of CD-JEV. To evaluate the persistence of immunological memory, the response to this boosting dose will 
be measured in a serum sample collected 7 and 28 days after vaccination. This sample will be tested for anti-JE 
neutralizing antibodies. 
 
Following the booster dose, children will be monitored for immediate reactions during the first 30 minutes, 
solicited injection site and systemic adverse reactions for 7 days, unsolicited adverse events and serious adverse 
events for 28 days. 
 
 
 

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B. DESCRIPTION OF THE RESEARCH PROJECT 
 
1. HYPOTHESIS TO BE TESTED 
Does this research proposal involve testing of hypothesis: 
 No       
 Yes (describe below)  
 
The intention of this study is to describe the persistence of the antibody response three and four years post 
vaccination with live attenuated SA-14-14-2 Japanese encephalitis vaccine (CD-JEV) and to describe the kinetics 
of the antibody response after a booster dose. No hypotheses are defined. The results will be descriptive. 
 
2. SPECIFIC OBJECTIVES 
2.1. Primary objective 
To assess the long-term antibody response three and four years after CD-JEV vaccination among children who 
received a single dose of vaccine between 10-12 months of age. 
 
2.2. Secondary objectives 
To evaluate the antibody response following a booster dose of CD-JEV given at five years of age, among 
children who previously received a single dose of CD-JEV between 10 and 12 months of age. 
 
To determine the safety profile of a CD-JEV booster dose given to children four years after initial vaccination. 
 
2.3. Exploratory objective 
To further describe the antibody response to a booster dose by differentiating between a primary and secondary 
antibody response at 7 days post-booster dose. 
 
2.4. Outcome measures 
The immunologic endpoints will be: 
 
Seroprotection rate defined as the proportion of study participants with an anti-Japanese encephalitis (JE) 
neutralizing antibody titer ≥1:10 as measured by 50% plaque reduction neutralization test (PRNT-50). 
 
Geometric mean titer (GMT) of anti-JE neutralizing antibody as measured by PRNT-50.  
 
The safety endpoints will be: 
 
Frequency counts and percentage of participants reporting immediate unsolicited adverse events (AEs) 
occurring within 30 minutes of booster vaccination. 
 
Frequency counts and percentage of participants reporting solicited local and systemic AEs occurring within 
7 days of booster vaccination. 
 
Frequency counts and percentage of participants reporting unsolicited AEs and serious adverse events (SAEs) 
occurring within 28 days of booster vaccination. 
 
The exploratory immunologic endpoint will be: 
 
Proportion of participants with demonstrated seropositivity for anti-JE immunoglobulin M (IgM) prior to the 
booster dose and 7 days post booster dose as measured by enzyme-linked immuno assay (ELISA). 
Seropositivity will be defined per the ELISA manufacturer’s kit instructions.  
 

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3. BACKGROUND OF THE PROJECT INCLUDING PRELIMINARY OBSERVATIONS: 
3.1. Disease burden 
JE virus, a mosquito-borne flavivirus, is the leading cause of viral neurological disease and disability across 
temperate and tropical zones of Asia. The number of cases, the severity of disease, and the profound neurologic 
sequelae in survivors make JE the most important viral cause of encephalitis in Asia.4-6 In Asia, more than 750 
million children live in areas where they are at risk for JE virus infections.4,5 Although the incidence in children 
less than 15 years of age is higher than the rate in other age groups, adult cases may also be seen in JE-endemic 
areas.7 
 
3.2. JE in Bangladesh 
The first confirmed outbreak of JE in Bangladesh was recognized in 1977.8 However, JE was not identified again 
until 2003 when a new initiative was undertaken to assess the impact of JE disease.8,9 In collaboration with the 
U.S. Centers for Disease Control and Prevention, the International Center for Diarrheal Diseases Research in 
Bangladesh (icddr,b) conducted a hospital-based surveillance study in four tertiary hospitals in Dhaka, Rajshahi, 
Mymensigh, and Sylhet to identify the etiology of acute encephalitis cases. Between 2003 and 2005, 492 persons 
with acute encephalitis were serologically evaluated for JE virus infection and 4% had evidence of recent 
infection. In 2007, hospital-based surveillance was established at three tertiary hospitals in Rajshahi, Chitagong, 
and Khulna.9,10 Patients hospitalized with meningitis and/or encephalitis were tested for JE virus-specific 
Immunoglobulin M antibodies during an 11-month period. Of 632 patients with meningitis and/or encephalitis, 
6% had laboratory evidence of recent JE virus infection.10 These studies suggest that JE is an important cause of 
encephalitis in Bangladesh. 
3.3. Previous experience with CD-JEV 
CD-JEV has been used since 1988, with over 400 million doses administered internationally. The vaccine is 
currently licensed in twelve countries, and, in 2013, vaccine produced at a newly constructed Good 
Manufacturing Practice (GMP) compliant production facility was prequalified by the World Health Organization 
(WHO). Following prequalification, CD-JEV was also selected for use in the 2015 GAVI-supported catch-up 
campaigns in children less than 15-years-old. 
 
Like other parenterally administered vaccines, vaccination with CD-JEV vaccine may cause local and systematic 
side effects in some children. These include pain at the injection site for a short while after injection and swelling, 
redness, and tenderness at the injection site. Fever may occur and, in some cases, a rash may appear in the initial 
days following the vaccination. Because CD-JEV is a live attenuated vaccine, there is a theoretical risk that 
systemic disease including encephalitis could occur if the vaccine strain replicates unchecked. To date, this has 
not been reported with CD-JEV. No reports of severe allergic reactions or death have been associated with CD-
JEV. 
From 2005-2008, the WHO Global Advisory Committee on Vaccine Safety (GACVS) and the WHO Strategic 
Advisory Group of Experts (SAGE) on Immunization reviewed the safety of CD-JEV and other JE vaccines.11-14 
These reviews stated that CD-JEV is well tolerated across a wide age range and can safely be given to children as 
young as 9 months of age. The reviewers noted that no SAEs were attributed to CD-JEV in 12 clinical studies 
conducted from 1992 through 2008 in China, India, the Philippines, South Korea, Sri Lanka, and Thailand.3,15-26 
In 2013, the WHO GACVS reviewed the safety of CD-JEV again and concluded that it had an excellent safety 
profile.27 
 
Seroprotection rates following a single dose of CD-JEV (≥105.4 plaque-forming units) have been greater than 90% 
in most clinical trials. Several early Chinese studies have shown a dose-response gradient with 85% to 100% 
seroconversion after a single dose of CD-JEV vaccine was given to children 1 to 12 years of age.15,18,19,23-25 In one 
of the first immunogenicity studies performed outside of China, 95% of participants aged 9 to 15 months were 

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seroprotected within 90 days after CD-JEV vaccination. A Korean study of 68 children one to three years of age 
showed high seroconversion rates (96%) four weeks after one dose of JE live attenuated CD-JEV.21  
 
Although the number of clinical studies are limited, the neutralizing antibody response following immunization 
with vaccine remains high for up to four years after vaccination. A long-term study in Nepal demonstrated that 
90% of the study participants had a high level of neutralizing antibody four years after vaccination.28 When co-
administered with measles vaccine in healthy infants in the Philippines, the long-term immunogenicity of SA 14-
14-2 vaccine in infants as young as eight months was demonstrated. Specifically, seroprotective titers against JE 
were maintained in over 80% of study participants three years after vaccination (Table 1).3,16 
 
Table 1. Seropositivity rates and anti-measles IgG geometric mean concentrations and anti-JE geometric 
mean titers in Philippine infants vaccinated with JE and measles vaccines. 
 
In addition to the high seroprotection rates, CD-JEV is the only JE vaccine other than mouse brain-derived 
vaccine for which there is vaccine effectiveness data. A post-vaccination campaign case-control study in Nepal 
demonstrated that a single dose of CD-JEV was 99.3% effective (95% CI 94.9-100%) in preventing Japanese 
encephalitis when administered only days or weeks before exposure to infection.29 A follow-up case-control 
study in Nepal showed that vaccine effectiveness was 96.2% (95% CI 73.1-99.9) five years after the initial 
vaccination.30  
 
3.4. Study rationale 
In 2012 PATH sponsored a double-blind, randomized controlled trial among 818 Bangladeshi infants aged  
10 to 12 months to compare the immunogenicity of three lots of live attenuated CD-JEV manufactured in the 
Chengdu Institute of Biological Products’ (CDIBP) newly constructed GMP facility and one control lot of CD-
JEV manufactured in the pre-existing facility. The study was conducted in rural Matlab and in Mirpur, in urban 
Dhaka. All four lots of JE vaccine were well-tolerated by the Bangladeshi infants (“A clinical trial in healthy 
infants to assess lot-to-lot consistency of Japanese encephalitis live attenuated SA 14-14-2 vaccine manufactured 
in a new good manufacturing practice facility and non-inferiority with respect to an earlier product”, icddr,b 
Protocol # 11050).2 Seroprotection rates (anti-JE neutralizing antibody titer >1:10) ranged from 80% to 86% for 
all four lots of vaccine, with an average GMT of 56 (Table 2).2  
 
 
Group 1 (CD-JEV then MV) (N=88) 
Group 2 (concomitant) (N=222)a 
Group 3 (MV then CD-JEV) (N=180)b 
 
% seropositive 
(95% CI) 
GMC/GMT 
(95% CI) 
% seropositive 
(95% CI) 
GMC/GMT 
(95% CI) 
% seropositive 
(95% CI) 
GMC/GMT 
(95% CI) 
Measles vaccine 
response 
(anti-measles IgG) 
Day 0 
1.1  
(0.0-6.2) 
12.8  
(10.2-16.2) 
0.0  
(0.0-1.7)  
7.4  
(6.3-8.8) 
0.0 
(0.0-2.1)  
7.0  
(5.8-8.5) 
Day 28 
88.6  
(80.1-94.4)  
318.9 
(273.0-372.6) 
91.8  
(87.3-95.1)  
301.9  
(269.0-338.9) 
86.5  
(80.6-91.2)  
262.5  
(222.2-310.2) 
JE vaccine response 
(anti-JE neutralizing 
antibody) 
Day 0 
3.4  
(0.7-9.6) 
5.7 
(4.9-6.5) 
5.4 
(2.8-9.3) 
5.7 
(5.2-6.1) 
6.1 
(3.1-10.7) 
5.9 
(5.3-6.6) 
Day 28 
92.1 
(84.3-96.7) 
202.8 
(140.5-292.9) 
90.5 
(85.9-94.1) 
155.0 
(123.5-194.5) 
90.6 
(85.3-94.4) 
139.4 
(109.5-177.5) 
Year 1 
90.2  
(82.2-95.4) 
106.8 
(71.5-159.4) 
84.0 
(78.3-88.6) 
80.7 
(62.4-104.3) 
85.9 
(80.5-90.3) 
74.9 
(59.2-94.9) 
Year 2 
81.7  
(72.4-89.0) 
68.8  
(47.3-100.1) 
83.0 
(77.3-87.8) 
61.6 
(49.1-77.4) 
81.7 
(75.8-86.6) 
70.1 
(54.6-90.1) 
Year 3 
79.6  
(70.0-87.2) 
49.9 
(36.5-68.3) 
81.9 
(75.9-86.8) 
45.7 
(37.2-56.3) 
82.6 
(76.8-87.4) 
59.3 
(47.1-74.8) 
IgG: immunoglobulin G; CD-JEV: live, attenuated Japanese encephalitis SA-14-14-2 vaccine; MV: measles vaccine; N: number of infants in the per protocol 
analysis; CI: confidence interval; GMC: geometric mean concentration; mIU/mL: milli International Units per milliliter, seroprotection defined as ≥120 mIU/mL; 
GMT: geometric mean titer, seroprotection defined as JE neutralizing antibody titer ≥1:10 
a Among 222 participants in the per protocol population of Group 2, 219 had valid serology results for measles antibody during retesting. 
b Among 180 participants in the per protocol population of Group 3, 178 had valid serology results for measles antibody during retesting. 

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Table 2. Seroprotection rates and geometric mean antibody titers measured at 28 days post vaccination by 
treatment group (per protocol population). 
Study group (N) 
Group 1 (146) 
Group 2 (195) 
Group 3 (192) 
Group 4 (194) 
New Facility 
Group 2-4 (581) 
Number of seroprotected children (%) 
126 (86.3) 
160 (82.1) 
154 (80.2) 
164 (84.5) 
478 (82.3) 
95% Confidence intervals 
79.8–91.0 
76.1–86.8 
74.0–85.2 
78.8–89.0 
79.0–85.2 
Geometric mean antibody titer 
77.3 
52.8 
53.4 
62.8 
56.2 
95% Confidence intervals 
59.6–100.4 
42.9–65.1 
42.4–67.2 
50.3–78.4 
49.5–63.8 
Group 1 represents the existing facility lot; Groups 2, 3 and 4 the new facility lots. 
Seroprotection defined as JE neutralizing antibody titer ≥1:10 
 
This study was a requirement for WHO-prequalification; based on these data the vaccine was prequalified in 
October 2013. However, these GMT and seroprotection rates were lower than those seen in other study 
populations, including the study referenced above that was conducted in Philippines, which used the same 
reference laboratory to test the samples.3 While other study factors, such as the vaccine stability or lots used, may 
account for the results, different populations may respond differently to vaccines. For example, Bangladeshi 
children have been shown to have lower immune responses to other vaccines, including oral rotavirus vaccines, 
as compared to children in higher-resource settings.31 
 
This study aims to understand the persistence of immunity and the kinetics of the booster response. In addition, 
this study will determine the safety of a second dose of CD-JEV in this population. 
 
3.5. Potential risks and benefits 
3.5.1. Potential risks 
The child may experience discomfort and pain from venipuncture for collection of 2 ml of blood. Venipuncture is 
sometimes associated with discomfort during phlebotomy, and rarely, an infection at the site of phlebotomy if 
improperly conducted. The child may experience discomfort, pain, redness, swelling, and/or local hardness at the 
injection site. 
 
Like any vaccine, vaccination with CD-JEV may cause side effects in some children. The side effects of JE 
vaccination may include pain at the injection site for a short while after injection and swelling, redness, and 
tenderness at the injection site. Fever may occur and in some cases a rash may appear in the initial days following 
the vaccination. Because the vaccine is a live attenuated vaccine, there is a very small theoretical risk that 
encephalitis, or infection of the brain, could occur. However this has never been reported to occur with CD-JEV. 
Allergic reactions and death, while rare, are also possible with any vaccination. 
 
3.5.2. Potential benefits 
Children participating in this study will receive a booster dose of CD-JEV which should improve their protection 
against JE. Likewise, the participant will contribute information on long-term immunogenicity and response to a 
booster dose including needed safety information. This will facilitate potential policy decisions in Bangladesh 
and Southeast Asia regarding JE immunization programs. They will be significantly contributing to the 
advancement of this WHO-prequalified JE vaccine. By participating in this clinical trial they will help build the 
body of knowledge on CD-JEV and help ministries of health decide the best integrated vaccination schedule for 
CD-JEV to reduce and ultimately eliminate JE disease throughout Asia. 
 
 
4. RESEARCH DESIGN AND METHODS 
The proposed study is a phase 4 open-label trial enrolling up to 818 Bangladeshi children who had previously 
participated in JEV05, a PATH-supported clinical study conducted from May to December 2012 during which 
10- to 12-month-old infants were given a single dose of CD-JEV. Eligible JEV05 participants who still live in the 

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area will be identified using the local health and demographic surveillance system (HDSS) and asked to 
participate. Children will be divided into two groups based on whether they received vaccine from the old or new 
facility in JEV05: 
 
 
Group A (up to 655 eligible): children randomized to Groups 2-4 (new facility) in JEV05. 
 
Group B (up to 163 eligible): children randomized to Group 1 (old facility) in JEV05. 
 
An overview of the study activities is provided in Table 3. A 2 ml blood sample will be collected from all 
participants three and four years after initial vaccination. The sera will be tested using PRNT; JE neutralizing 
antibody titers >1:10 will be considered seroprotective.  
 
After the blood draw at Visit 02 occurring in May to December 2016, four years after their initial vaccination, 
both groups will receive a single subcutaneous booster dose of CD-JEV. Additional serum samples will be 
collected on Day 7 and Day 28 following the booster dose for measurement of serum anti-JE neutralizing 
antibody titers. 
 
Following the booster dose, children will be monitored for immediate reactions during the first 30 minutes, 
solicited injection site and systemic adverse reactions for 7 days, unsolicited adverse events and serious adverse 
events for 28 days. All safety events will be identified or observed by study staff during home visits, clinic visits, 
and/or reported by a parent or legal guardian. Solicited and systemic adverse reactions will be assessed through 
home visits 1 through 6 days following vaccination, and a fixed site clinic (FSC) visit on Day 7 following 
vaccination. Information regarding unsolicited AEs occurring in the 28 days following vaccination will be 
collected and recorded on a standard questionnaire during clinic visits. Events will be graded for severity and 
assessed for relatedness to vaccination by the investigator.  
 
The study will be open-label; no placebo control will be employed and safety assessments will be unblinded.  
 
Table 3. Schedule of events 
 
Y3 after JEV05 
±3 months after 
enrollment in 
JEV05 
Y4 after JEV05 
Visit 1  
(D0) 
Visit 02  
(D365±28) 
Visit 03 
through 
Visit 08 
(D366-371±0) 
Visit 09  
(D372+3) 
Visit 10  
(D393±7) 
Obtain informed consent 
X 
  
 
  
  
Collect baseline demographic information 
X 
  
 
  
  
Collect/review medical history 
X 
 X 
 
  
  
Perform physical exam 
X 
X 
 
X  
X 
Check inclusion/exclusion criteria 
X 
X 
 
  
  
Collect serum for long term response  
X 
X 
 
 
 
Collect serum for booster response 
 
 
 
X 
X 
Administer booster dose of CD-JEV 
  
X 
 
  
  
Observe for immediate reactions for 30 minutes 
  
X 
 
  
  
Inquire about child's well being 
  
  
X 
X 
X 
Record reported adverse events 
  
X 
X 
X 
X 
Record serious adverse events 
  
X 
X 
X 
X 
Exit participant from study 
  
  
 
  
X 
 

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4.1. Study sites 
The study will be conducted in the same low risk JE sites as the JEV05 study—rural Matlab and Mirpur (urban 
Dhaka). Matlab Upazila is a rural site located 55 kilometers southeast of Dhaka where icddr,b has been 
maintaining a HDSS since 1966. The HDSS area has two parts; an icddr,b intervention area where maternal, child 
health, and family planning services are offered and a comparison area where these services are provided by the 
government of Bangladesh (Figure 1). JEV05 participants who still live in the area will be identified through the 
HDSS and contacted for participation. The study team will consist of the principal investigator, central 
administrative team, medical officers, nurses, and laboratory personnel adequately trained to conduct the study. 
The central administrative team will be comprised of the study coordinator, field assistants, field officers, and a 
data manager.  
 
Figure 1: Matlab HDSS area (Intervention and comparison areas) 
 
 
4.2. Study enrollment and withdrawal 
4.2.1. Inclusion criteria at study entry (Visit 01) 
Each participant must satisfy all of the following inclusion criteria at study entry: 
 
Participant in JEV05 study and received one dose of CD-JEV. 
 
Resides in the Matlab HDSS or Mirpur area. 
 
At least one parent or guardian willing to provide written informed consent. 
 
4.2.2. Exclusion criteria at study entry (Visit 01) 
Potential study participants will be excluded if they have: 
 
Received a second dose of Japanese encephalitis vaccine within the past three years. 
 
Received immunoglobulins and/or any blood products within 90 days prior to enrollment. 
 
Been diagnosed with a primary or acquired immunodeficiency, including HIV infection within the past 
three years. 
 
 
 

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4.2.3. Exclusion criteria for Visit 02 at the time of blood draw 
Study participants will be excluded from further participation in the study if they have: 
 
Received a second dose of Japanese encephalitis vaccine within the past four years. 
 
Received immunoglobulins and/or any blood products within 90 days prior to Visit 02. 
 
Been diagnosed with a primary or acquired immunodeficiency, including HIV infection within the past 
year. 
 
4.2.4. Exclusion criteria for Visit 02 at the time of vaccination 
Study participants will be excluded from further participation in the study if they have: 
 
Received any other vaccine within 28 days prior to administration of the booster JE vaccine or planned 
vaccination during the 28 days after study vaccination. 
 
History of allergic disease or known hypersensitivity to any component of the study vaccine and/or 
following administration of vaccines included in the local program of immunization.  
 
Used any investigational or non-registered drug within 90 days prior to the administration of study 
vaccines or planned administration during the study period. 
 
Plan receipt of immunoglobulins and/or any blood products during the 28 days after the study 
vaccination. 
 
Received immunosuppressing or immune-modifying agents (including systemic corticosteroids 
equivalent to prednisone ≥0.5 mg/kg/day; topical and inhaled steroids are allowed) for 14 consecutive 
days or longer within 28 days prior to the administration of the booster JE vaccine. 
 
Acute or chronic, clinically significant pulmonary, cardiovascular, hepatic, or renal functional 
abnormality, as determined by medical history or physical examination, which in the opinion of the 
investigator, might interfere with the study objectives. 
 
Severe malnourishment as defined by World Health Organization weight-for-height tables (Z-score < -3). 
 
Acute illness at the time of vaccination (defined as the presence of a moderate or severe illness with fever 
[axillary temperature ≥38°C] or without fever [severity determined at the discretion of the investigator]. 
Acute illness is a temporary exclusion. Vaccination should be postponed until at least 7 days after 
recovery. 
 
Any condition or criterion that in the opinion of the investigator might compromise the well-being of the 
participant, compliance with study procedures, or interpretation of the outcomes of the study. 
 
4.2.5. Reasons for withdrawal 
After enrollment, a participant might be withdrawn from participation for several reasons: 
 
The investigator requests withdrawal because the participant: 
o Is not eligible or is no longer eligible; 
o Developed an AE or SAE requiring withdrawal; or  
o Is non-compliant. 
 
A participant voluntarily withdraws at his/her own request; 
 
A participant is lost to follow-up; or 
 
The trial is terminated prematurely by PATH. 
 
 
 

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4.2.6. Handling of withdrawals 
In case of premature withdrawal for any reason, the investigator should use his/her best effort to: 
 
Conduct an interview to determine if the participant has had any reaction or AE (serious or non-serious) 
that has led to the withdrawal during the three-week period after any study vaccination. Where possible, 
the investigator should visibly or physically assess any reported adverse reaction or AE. 
 
Document the reason for premature withdrawal on the case report form (CRF). 
 
Complete final visit CRF. 
 
The investigator must inform PATH within 7 days of becoming aware about of the premature termination of a 
volunteer’s participation in the trial. 
 
4.2.7. Termination of the trial 
The trial might be suspended at any time by PATH or by any ethical review committee overseeing this study for 
any safety concern. This includes, for example, and without limitation, an SAE resulting in death that is viewed 
to be related to receipt of study vaccine or an unusually high rate of SAEs. PATH may suspend the study in the 
event that study conduct is found to be below Good Clinical Practice (GCP) standards. 
 
In the event of the appearance of new data that indicate an increased level of risk to participants, the clinical trial 
will be suspended until PATH and all ethical review committees have reviewed relevant data and agreed that the 
trial may proceed. 
 
Withdrawal of a participant by the investigator may occur only if the participant is identified to have one of the 
listed exclusion criteria, or if continued participation of the participant threatens the participant’s well-being or 
the integrity of the study. Any participant with an adverse event related to involvement in the trial should 
continue to be followed for safety reasons until trial conclusion or resolution of the adverse event. 
 
4.3. Study vaccine 
4.3.1. Live, attenuated JE vaccine SA 14-14-2  
SA 14-14-2, the vaccine virus strain, was attenuated by an empirical process of serial passage of the wild-type SA 
14 virus in cell cultures of primary hamster kidney (PHK) cells and in animals (mice, hamsters) with successive 
plaque purifications (in primary chick embryo cells). SA 14-14-2 virus has 57 nucleotide changes compared to 
the parent SA 14 virus, resulting in 24 amino acid changes. Genetically-stable mutations resulting in 8 amino acid 
changes in the E protein gene are associated with reduced neurovirulence (attenuation).  
 
CD-JEV is a preparation of SA 14-14-2 grown in culture on a monolayer of PHK cells. After cultivation and 
harvest, an appropriate stabilizer is added to the virus suspension, which is then lyophilized. Live attenuated JE 
virus (strain SA 14-14-2), minimum essential medium, and human serum albumin are the major components of 
the final vaccine. 
 
The vaccine is prequalified by the WHO and produced in accordance with technical specifications in “Guidelines 
for the production and control of Japanese encephalitis vaccine (live) for human use” developed by the WHO 
Expert Committee on Biological Standardization, Chinese Pharmacopoeia and registered standard of Chinese 
State Food and Drug Administration (No.WS4-(ZB-072)-2010).32,33  
 
4.3.2. Acquisition 
The investigators or designee will be personally responsible for receiving the vaccine within Bangladesh and all 
subsequent management of study vaccines. Appropriate vaccine acquisition and handling standard operating 
procedures (SOPs) will be developed in collaboration with the investigator. Extra doses of vaccines will be 
provided and shall be used when the initially allocated dose is broken, damaged, or unusable. 

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PATH will coordinate direct shipment of CD-JEV from CDIBP to a central location in Bangladesh. The 
manufacturer, CDBIP, will donate these vaccines free of charge. The investigator or designee will receive the 
vaccine, provide customs clearance, and deliver the vaccine to a central facility under controlled cold-chain 
conditions. The investigators and their institution will receive no monetary compensation from the manufacturer 
for their participation in this study. 
 
4.3.3. Packaging and labeling  
The study vaccines will be provided in their commercial presentation. The commercial vial labels for these 
vaccines should remain intact. An additional label will be affixed by the manufacturer to each box and will bear 
the following information: 
 
Study number 
 
Legal information: “For Clinical Trial Use Only” 
 
4.3.4. Product storage and stability 
CD-JEV vaccine should be stored in a refrigerator or cold box at a temperature ranging from +2°C to +8°C and 
protected from light. Storage temperature should be monitored and documented on an appropriate form during 
the entire trial. 
 
The vaccines should never be frozen. In case of deep freezing or accidental disruption of the cold chain, the 
vaccine should not be administered and the investigator or the responsible person should contact the study 
monitor to receive further instructions. 
 
4.3.5. Dosage, preparation and administration of study vaccines 
CD-JEV will be supplied in 5-dose vials as a lyophilized powder that looks like a milky-white crisp cake. After 
reconstitution, it turns into a transparent orange-red liquid. Each 0.5 ml human dose for subcutaneous injection 
contains no less than 5.4 log PFU of live JE virus. A single dose of 0.5mL should be administered by 
subsubcutaneous injection with the rest of the vial discarded. 
 
The vaccines should be gently shaken prior to administration to assure homogenous suspension. CD-JEV will be 
administered subcutaneously into the anterolateral aspect of the right upper arm.  
 
Standard immunization practices should be observed and care should be taken to administer the injection. As 
with all injectable vaccines, appropriate medical treatment, staff, and supervision will be readily available in case 
of rare anaphylactic reactions following administration of the study vaccine and epinephrine 1:1000, 
diphenhydramine, and resuscitation equipment should be available in case of any anaphylactic reactions. 
 
4.3.6. Accountability procedures 
Study vaccine must be kept in a secure place. The investigator or the person in charge of vaccine management 
will maintain records of the vaccine's delivery to the trial site, the inventory at the site, the dose given to each 
participant, and the return or disposal of unused doses to PATH. Appropriate vaccine accountability SOPs will be 
developed in collaboration with the investigator. 
 
4.4. Study procedures 
4.4.1. Recruitment and screening information 
Participants of JEV05 will be identified from the computerized HDSS database and dataset from the previous 
study maintained at the field site in Matlab and Mirpur. Field workers will visit their homes and explain to the 
parent or legal guardian the study purpose and procedures. If the parent or legal guardian is interested in having 

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their child participate in the study, field staff will invite them to come to their nearby FSC on a scheduled 
enrollment day.  
 
All participants from JEV05 that continue to live in the study area are eligible to participate in this study. We 
propose to include children that were seronegative at 28 days because there is an indication from other studies 
that participants seronegative at D28 may generate a detectable antibody response at a later timepoint. 
Furthermore, we expect that some “seronegative” participants, can generate an anamnestic response. The serum 
collected at Day 7 following the booster dose will be used to determine whether previously vaccinated children 
who did not have any antibody at 3 and 4 years post CD-JEV vaccination can generate an anamnestic response 
(i.e., the children were immune to JE disease). Although serum neutralizing antibody is generally considered our 
best a correlate of protection, there are instances where vaccinated persons without antibody level are 
nevertheless truly immune and they are able to mount a complete and normal immune response should they 
become infected. The way to determine this is to look at the anamnestic immune response, i.e., short-term 
antibody response. This is an antibody response that is not due to a new primary response; it depends on having 
primed B and T lymphocytes ready to respond quickly. The magnitude of the response measured between Day 7 
and Day 28 will indicate whether the participant is mounting an anamnestic response to the booster or a primary 
response. 
 
4.4.2. Enrollment (Visit 01, 3 years after JEV05 vaccination ±3 months) 
When the parent or legal guardian brings their child to the FSCs on the scheduled day for enrollment, the study 
team will: 
  
 
Describe the study to the parents, including the study purpose, requirements, duration, and risks and 
benefits of the study. The parent or legal guardian will have the opportunity to ask any questions. 
 
Assess the family’s ability to comply with study requirements.  
 
Obtain written informed consent. Participation in the study is strictly voluntary, and those agreeing to 
participate must provide a written informed consent. The consent form will be in Bangla. The parent(s) or 
guardian(s) will be informed that they are free to refuse to have their child participate and also to 
withdraw their consent at any time. One parent’s or guardian’s signature is acceptable as informed 
consent for this study. If the parent or guardian is illiterate, a left thumb impression will be taken instead 
of signature in the presence of an impartial literate witness who is unrelated to the study. The impartial 
literate witness (hierarchically independent of the PI and not specified on the list of trial contributors) 
will also sign the informed consent form and attest that the information in the consent form and any other 
written information were explained to the parents or guardian and that the information was understood. 
Following the informed consent process, the child will be enrolled into the study. 
 
Collect demographic and address information. 
 
Collect pertinent medical history through parent interview, including data on child’s health and previous 
vaccinations and reaction to vaccinations and history of any chronic or recurrent medical conditions.  
 
Perform a physical exam (temperature, height and weight, a brief examination of the head, neck, heart, 
lungs, abdominal region, musculoskeletal system, and skin). 
 
Review study inclusion and exclusion criteria and determine eligibility. 
 
Assign the child a unique study participant number following confirmation of eligibility and consent.  
 
Collect 2 ml of blood with aseptic technique to detect anti-JE serum neutralizing antibody with the 
PRNT-50. 
 
Participants will be reminded that study staff will be contacting them in approximately one year for 
administration of a booster dose of CD-JEV. 

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4.4.3. Vaccine administration (Visit 02, 1 year after Visit 01 ±28 days) 
When the parent or legal guardian brings their child to the FSCs on the scheduled day for vaccination, the study 
team will: 
  
 
Describe the study to the parents, including the study purpose, requirements, duration, and risks and 
benefits of the study. The parent or legal guardian will have the opportunity to ask any questions. 
 
Reassess the family’s ability to comply with study requirements.  
 
Collect demographic and address information. 
 
Collect pertinent medical history through parent interview, including data on previous vaccinations and 
reaction to vaccinations and history of any chronic or recurrent medical and psychiatric conditions. 
 
Perform a physical exam (temperature, height and weight, a brief examination of the head, eyes, ears, 
nose, throat, neck, heart, lungs, abdominal region, lymph nodes, musculoskeletal system, and skin). 
 
Review exclusion criteria and determine eligibility for blood draw and booster vaccination. 
 
Collect 2 ml of blood from participants with aseptic technique to detect anti-JE serum neutralizing 
antibody and anti-JE IgM with PRNT-50 and ELISA, respectively. 
 
Administer CD-JEV vaccine subcutaneously in the anterolateral aspect of the right upper arm according 
to standard sterile techniques. 
 
Ask the parents to wait with the child in the clinic for 30 minutes after the vaccination(s) to assess 
immediate reactions. If the child experiences an adverse reaction or a SAE during this time period, he/she 
will be treated and the event will be recorded on the CRF. 
 
Inform the parents that they will be visited daily for the next 6 days in order to check on the welfare of 
the child and assess the occurrence of any local or systemic reactions. 
 
Remind the parents to seek medical care as appropriate and inform the investigator as soon as possible in 
the event of an adverse reaction or SAE. If the child visits a health care provider, the parents should be 
sure to inform him/her of the child’s participation in the study and present the provider with contact 
information for the investigator as indicated on the study identification card.  
 
Give the parents a study identification card that indicates that the child has been enrolled in a research 
study and given a study vaccine. The card will contain the following information: 
o Name, address, and contact phone numbers of local PI. 
o Vaccination clinic location. 
o Name of vaccine given to child. 
o Schedule of visits. 
o Indication that the child is in a vaccine trial and a request that medical personnel who treat the 
child contact the local PI within the study duration. 
 
The appropriate section(s) of the CRF will be completed. 
 
4.4.4. Home follow-up visits (Visit 03 through Visit 08, 1-6 days post vaccination ±0 day) 
A field worker will visit the particpant at home to: 
 
Inquire about the child’s health (any illnesses or medications taken or visits to a healthcare facility). 
 
Inquire about any reactions using a standard questionnaire to record the local and systemic reactogenicity 
after vaccination. 

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 
Remind the parents that they will be need to bring their child to the FSC on the 7th day (+ 3 days) following 
vaccination for a blood draw and to check on the welfare of the child. 
 
Remind the parents to seek medical care as appropriate and inform the investigator as soon as possible in 
the event of an adverse reaction or SAE. If the child visits a health care provider, the parents should be 
sure to inform him/her of the child’s participation in the study and present the provider with contact 
information for the investigator as indicated on the study identification card.  
 
Complete the appropriate section(s) of the CRF. 
 
4.4.5. Clinic follow-up visit (Visit 09, 7 days post vaccination +3 days) 
When the parent or legal guardian brings their child to the FSC/ Mirpur study clinic on the scheduled day for 
follow-up, the study team will: 
 
Inquire about the child’s health (any illnesses or medications taken or visits to a healthcare facility). 
 
Inquire about any reactions using a standard questionnaire to record the local and systemic reactogenicity 
after vaccination. 
 
Collect 2 ml of blood from participants with aseptic technique to detect anti-JE serum neutralizing 
antibody and anti-JE IgM with PRNT-50 and ELISA, respectively  
 
Inform parents that they will need to bring their child to the FSC at Matlab or Mirpir study clinic in 
approximately 28 days (±7 days) in order to check on the welfare of the child and assess the occurrence 
of any adverse events. 
 
Remind parents to seek medical care as appropriate and inform the investigator as soon as possible in the 
event of an adverse event or SAE. If the child visits a health care provider, the parents should be sure to 
inform him/her of the child’s participation in the study and present the provider with contact information 
for the investigator as indicated on the study identification card.  
 
Complete the appropriate section(s) of the CRF. 
 
4.4.6. Clinic follow-up visit (Visit 10, 28 days post vaccination ±7 days) 
A final study visit will be conducted 28 days after booster vaccination. The study team will: 
 
Inquire about the child’s health (any illnesses or medications taken or visits to a healthcare facility). 
 
Inquire about any adverse events. 
 
Collect 2 ml of blood from participants with aseptic technique to detect anti-JE serum neutralizing 
antibody with the PRNT-50.  
 
Complete the appropriate section(s) of the CRF. 
 
Exit the participant from the study. 
 
4.4.7. Booster Vaccine 
Although studies have shown that the JE live attenuated SA 14-14-2 vaccine (CD-JEV) produces long lasting 
immunity, China and India currently give a boosting dose at least one year after the primary vaccination. In a 
recent 2015 position paper regarding JE vaccines (Weekly Epidemiological record Feb 27, 2015; 
http://www.who.int.wer), WHO stated that "long-term immunogenicity studies would inform optimal dosing 
schedules for long-term protection, which may vary by location (based on natural boosting or other factors)". As 
an important research need for all of the newer JE vaccines, the proposed study will provide data for guiding 
policy decisions with regard to the booster dose for CD-JEV, as it would be impossible for routine immunization 
programs to select only seronegative children for boosting. Including participants seroprotected 3 years post 

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vaccination at the 4 year time point of booster dose is important to understand the kinetics of the antibody 
response in children with existing titers and provide information for optimal schedules. 
4.5. Laboratory evaluations 
4.5.1. Clinical laboratory evaluations 
There will be no clinical (safety) laboratory evaluations in this study. Laboratory assays specified in Section 4.5.2 
are for research purposes only and are not be used for clinical diagnosis. 
 
4.5.2. Serum assays 
4.5.2.1. Anti-JE neutralizing antibody titer 
Serum anti-JE neutralizing antibody titers will be measured using PRNT at the United States Army Medical 
Directorate, Armed Forces Research Institute of Medical Sciences in Bangkok, Thailand. The reported titer will 
be the PRNT-50 which is the highest serum dilution that is still capable of reducing the number of virus plaques 
in cell culture by 50% compared to virus growth in cell culture without any added serum. Seroprotection will be 
defined as a PRNT-50 titer of  ≥1:10.1  
 
4.5.2.2. Anti-JE IgM 
Anti-JE IgM will be measured on specimens collected prior to the booster dose and 7 days following the booster 
dose using ELISA at United States Army Medical Directorate, Armed Forces Research Institute of Medical 
Sciences in Bangkok, Thailand. Seropositivity will be defined per the manufacturer’s kit instructions. 
 
4.5.3. Specimen preparation, handling, and shipping 
4.5.3.1. Specimen collection 
Blood draws of 2 ml will be collected during study visits. The identity of the child will be verified immediately 
prior to the blood draw using the child’s study participant number. The study participant number and date of 
collection of the specimen will be recorded on all labels to be used in the study. One label will be affixed onto the 
vacutainer tube immediately prior to the blood sample drawing. 
 
After collection, blood samples must clot for 60 minutes to 2 hours at room temperature. Samples will then be 
kept in a cold box for transportation until processed in the site laboratory. All samples will be centrifuged on the 
same day of collection. 
 
4.5.3.2. Specimen preparation, handling, and storage 
After centrifugation, the serum will be aliquoted. To insure that samples are not mixed or inaccurately labeled, 
the child’s study participant number, date of blood collection, number of aliquots obtained, and the date and time 
of aliquoting will be specified on the sample identification log. A label will be affixed onto each of the aliquots. 
Comments may be made on the quality of samples (e.g., hemolyzed, contaminated).  
 
All serum aliquots will be kept in the -20ºC freezer at the local study site. Study staff will monitor and record 
temperature on the appropriate form during the entire trial. One serum aliquot will be shipped to United States 
Army Medical Directorate, Armed Forces Research Institute of Medical Sciences in Bangkok, Thailand. A 
second aliquot will be retained at the site as back up and shipped to a long term storage facility determined by the 
Sponsor at the completion of the study. All unused serum specimens will be stored at their assigned location for 5 
years following study completion. Samples will be destroyed after 5 years. Only authorized study staff should 
have access to the samples. Samples will not be used for unrelated, future studies without an institutional review 
board (IRB) approved amendment to the protocol.  
 
 
 

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4.5.3.3. Specimen shipment 
Specimens will be shipped according to the International Air Transport Association regulations after each study 
year.  
 
5. SAMPLE SIZE CALCULATION AND OUTCOME (PRIMARY AND SECONDARY) 
VARIABLE(S) 
No sample size will be calculated for this study. However, up to 818 participants (up to 655 in Group A and up to 
163 in Group B) who participated in JEV05 are expected to be enrolled in this study to evaluate the primary and 
secondary objectives.   
 
5.1. Serology variables 
Seroprotection for JE in this study is defined as percentage of participants with JE antibody titre ≥1:10 at Year 3, 
Year 4 prior to the booster dose, 7 days post-booster dose, and 28 days post-booster dose. GMTs of anti-JE 
neutralizing antibody will be calculated.  
 
Evaluation of long-term anti-JE antibody response three and four years after CD-JEV vaccination among children 
who received the single dose of vaccine will be made at Year 3 and Year 4 prior to the booster dose in terms of 
JE seroprotection rates. Additionally,the analysis will stratified by the seroprotection status of participants on Day 
28 post primary immunization in the previous JEV05 study. 
 
To further describe the antibody response to a booster dose, anti-JE IgM will be measured prior to the booster 
dose and 7 days post booster dose. Seropositivity will be defined per ELISA manufacturer’s instructions. 
 
5.2. Safety variables 
The safety profile will be evaluated in terms of frequency counts and percentage of participants experiencing the 
following: 
 
 
Immediate reactions occurring within 30 minutes after vaccination as observed by study staff. 
 
Solicited signs and symptoms occurring greater than 30 minutes after receipt of CD-JEV through 7 days post 
vaccination as observed by study staff or reported by the participants to study staff. 
 
All other adverse events occurring within one month of CD-JEV receipt as observed by study staff or 
reported by the participant’s parents to study staff. 
 
SAEs occurring within 28 days of CD-JEV receipt. 
 
 
6. DATA ANALYSIS 
A Statistical Analysis Plan will be specified prior to the first participant first visit and finalized before locking of 
the database. 
 
6.1. Immunogenicity 
Long-term assessment of the immune response to CD-JEV will be evaluated in terms of percentage of 
participants with JE neutralizing antibody titre ≥1:10 at Year 3 and Year 4, just prior to the booster dose. 
Additonally, the analysis will be stratified by the seroprotection status of participants on Day 28 post primary 
immunization in previous JEV05 study. 
 
Percentage of participants achieving seroprotection against JE along with 95% confidence interval (CI) will be 
calculated by group. The method for calculating the 95% CI will be based on the Wilson score method without 
continuity correction. In addition, the difference in JE seroprotection rates between Groups A and B will be 

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calculated along with the 95% CI of the difference using the Newcombe-Wilson method without continuity 
correction. 
 
Similarly, the immune response to the booster dose following a booster dose of CD-JEV given at Year 4 will be 
assessed in terms of percentage of participants with JE neutralizing antibody titre ≥1:10 at 7 days post booster 
dose and 28 days post booster dose. Percentage of participants achieving seroprotection against JE along with 
95% CI will be calculated using the Wilson score method without continuity correction. 
 
In addition, GMTs of the anti-JE neutralizing antibody responses and their associated 95% CI will be calculated 
at Year 3, at Year 4 prior to the booster dose, and at 7 and 28 days post booster dose using the t-test. The GMTs 
will be compared between Group A and Group B in terms of ratio of the GMTs using the t-test and 95% CI of 
ratio of GMTs will be calculated.  
 
The GMTs at 7 and 28 days post booster dose will be further compared to the GMTs at Year 4 prior to the 
booster dose using a paired t-test to evaluate the magnitude of increase in GMTs by group.   
 
Furthermore as part of exploratory analyses to differentiate between a primary or secondary immune response, 
the immune response to the booster dose will be assessed in terms of the percentage of participants with 
demonstrated seropositivity for anti-JE IgM prior to the booster dose and 7 days post booster dose. Percentage of 
participants with seropositivity against JE along with 95% CI will be calculated using the Wilson score method 
without continuity correction. 
 
6.2. Safety 
The safety profile will be evaluated in terms of frequency counts and percentage of patients experiencing specific 
events falling into one of the following four categories: 
 
 
Immediate reactions occurring within 30 minutes after vaccination will be listed at the participant-level and 
summarized by group. 
 
Solicited signs and symptoms occurring greater than 30 minutes after receipt of the booster dose through 7 
days post vaccination will be listed at the participant-level and summarized by severity for Groups A and B.  
Exact 95% CI will be provided. 
 
All other adverse events occurring within 28 days of the booster dose will be listed at the participant-level 
and summarized by severity and relationship to study vaccines for Groups A and B. 
 
SAEs occurring within 28 days of the booster dose will be listed at the participant-level and summarized by 
severity and relationship to study vaccines for Groups A and B. 
 
6.3. Analysis sets 
All immunogenicity analyses and summaries will be performed on the intention-to-treat (ITT) basis which will be 
considered as the primary approach to immunogenicity analyses. Per protocol (PP) analyses will also be 
conducted on all enrolled participants. Safety analyses will be conducted on the ITT basis. Analyses set 
definitions are provided below. 
 
6.3.1. Intention-to-Treat analysis set 
The ITT analysis will be conducted for analyses of serology using all enrolled participants in this study whether 
the participant took the booster dose or not. According to this analysis set, long-term assessment of the anti-JE 
antibody response to CD-JEV is evaluated using all participants enrolled in the study with at least one serology 
result in Year 3 or Year 4 prior to the booster dose. The ITT analysis set will be used to analyze the primary and 
secondary serology objectives. 
 

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6.3.2. Per-Protocol analysis set 
A participant is valid for the PP analysis set if s(he) satisfies the inclusion/exclusion criteria of the study at Visit 
01, has a valid serology result at Year 3 (Visit 01) and Year 4 (Visit 02) prior to vaccination, and had no 
significant deviation in blood collection at the two time points. The PP analysis set will be used to analyze the 
primary and secondary serology objectives. 
 
6.3.3. Safety analysis set 
The safety analysis set will include all enrolled participants who had the booster dose and will be used to analyze 
all safety measurements including solicited/ unsolicited AEs and SAEs collected in the study. 
 
 
7. DATA SAFETY MONITORING PLAN 
7.1. Assessment of safety 
7.1.1. Specification of safety parameters 
Study vaccine safety profiles will be parameterized as the proportion of participants experiencing specific adverse 
events categorized into one of the following four categories:  
 
Immediate reactions occurring within 30 minutes of booster dose, as observed by study staff or reported 
by the participant to study staff. 
 
Solicited signs and symptoms occurring greater than 30 minutes after the booster dose through 7 days 
post vaccination, as observed by study staff or reported by the participant to study staff. 
 
All other adverse events occurring within 28 days of receipt of the booster dose as observed by study 
staff or reported by the participant’s parents to study staff. 
 
All SAEs occurring within 28 days of receipt of the booster dose. 
 
7.2. Methods and timing for assessing, recording, and analyzing safety parameters 
7.2.1. Adverse events 
An adverse event is defined as any untoward medical occurrence in a patient or clinical investigation participant 
administered a pharmaceutical product which does not necessarily have a causal relationship with this treatment. 
An AE can therefore be any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medicinal product, whether or not considered related 
to the medicinal product. The occurrence of an AE may come to the attention of study personnel during study 
visits and interviews of a study participant presenting for medical care. 
 
Information to be collected on adverse events includes event description, time of onset, clinician’s assessment of 
severity, relationship to study product (assessed only by those with the training and authority to make a 
diagnosis), and time of resolution/stabilization of the event. 
 
Any medical condition that was present at the time that the patient was enrolled should not be reported as an AE. 
However, if it deteriorates during the study safety recording periods, it should be recorded as an AE.  
 
AEs should be graded for severity and relationship to vaccine. 
 
7.2.1.1. Severity of event 
Any AE which is not a local and systemic sign or symptom occurring within 7 days post booster dose will be 
assessed by the clinician to quantify intensity using the following guidelines34: 
 
 

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Grade 
Categorization 
Definition 
Grade 1 
Mild 
Symptoms causing no or minimal interference with usual social 
and functional activities. 
Grade 2 
Moderate 
Symptoms causing greater than minimal interference with usual 
social and functional activities. 
Grade 3 
Severe 
Symptoms causing inability to perform usual social and functional 
activities with intervention or hospitalization indicated. 
Grade 4 
Potentially life-threatening 
Symptoms causing inability to perform basic self-care functions 
OR medical or operative intervention indicated to prevent 
permanent impairment, persistent disability, or death. 
*Intervention defined as Medical, surgical, or other procedures recommended or provided by a healthcare professional for the treatment of an adverse 
event. 
 
Changes in the severity of an AE should be documented to allow an assessment of the duration of the event at 
each level of intensity to be performed. Adverse events characterized as intermittent require documentation of 
onset and duration of each episode. 
 
7.2.2. Relationship to study vaccines 
The clinician’s assessment of an AE's relationship to test vaccine is part of the documentation process, but it is 
not a factor in determining what is or is not recorded in the study. If there is any doubt as to whether a clinical 
observation is an AE, the event should be recorded. All AEs must have their relationship to study vaccines 
assessed using the following guidelines: 
 
Categorization 
Definition 
Definitely Related 
An adverse event or unanticipated problem clearly related to the research 
procedures. 
Possibly Related 
There is a reasonable possibility that the adverse event, incident, experience, or 
outcome may have been caused by the procedures involved in the research. 
Not Related 
Any adverse event or unanticipated problem clearly not related to study procedures. 
 
 
7.2.3. Reactogenicity 
Reactogenicity post vaccination will be assessed in the clinic for 30 minutes, during daily home visits 1 through 6 
days following the booster, and during a FSC visit 7 days following the booster. During the daily home visits and 
FSC visit, the site staff will use a structured reactogenicity reporting form for recording reactogenicity. 
 
Any reactogenicity will be managed in accordance with good medical practices by the clinical study site team, 
who will assess and treat or refer the child for medical care as appropriate. If needed to monitor or treat 
reactogenicity, additional study visits may be conducted. If any acute treatment or medical care is required as a 
result of harm caused by the study vaccine or study procedure, this will be provided by the site free of charge. All 
children reporting reactogenicity will be followed clinically until the reactogenicity resolves (returns to baseline) 
or stabilizes. Reactogenicity will be recorded as indicated in the CRF. Reactogenicity likely to be related to the 
product, whether serious or not, that persists at the end of the trial will be followed up by the PI until resolution or 
stabilization.  
 
The parents will be instructed to contact the study site staff to report any reactogenicity their child may 
experience. In the case of a life-threatening event, they will be instructed to seek immediate emergency care. 

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Where feasible and medically appropriate, parents of children will be encouraged to seek medical care for the 
child where the study physician is based, and to request that the physician be contacted upon their arrival. 
 
7.2.3.1. Immediate reactions 
All participants will be observed for immediate reactions for 30 minutes after vaccination, with appropriate 
medical treatment readily available in case of an anaphylactic reaction following the administration of study 
vaccine. Immediate reactions will be assessed by a study physician or appropriately trained medical staff. All 
reactions that occur during this time will be recorded on the CRF. Any immediate reaction which meets the 
criteria for a serious adverse event must also be documented on an SAE form. 
 
7.2.3.2. Local and systemic signs and symptoms 
All participants will be monitored for assessment of local and systemic signs and symptoms occurring from 30 
minutes through 7 days post vaccination. Study staff will inquire about specific (solicited) local and systemic 
signs and symptoms and other unsolicited adverse events. 
 
Using a standardized data collection instrument the following local and systemic signs and symptoms will be 
documented and graded on predefined scales based on functional assessment or magnitude of reaction.34 Any 
local and systemic reaction that meets the criteria for a serious adverse event must also be documented on an 
SAE form. 
 
Local reactions (at injection site) 
 
Ecchymosis (bruising) 
 
Erythema (redness) 
 
Edema (swelling) 
 
Induration (hardness) 
 
Pain/tenderness  
Local ecchymosis, erythema, edema, and induration, if present, will be graded as follows: 
Grade 1:  
≤2.5 cm in diameter. 
Grade 2:  
>2.5 cm in diameter with <50% of surface area of extremity segment involved (upper arm or 
thigh). 
Grade 3:  
≥50% of surface area of extremity segment involved OR ulceration OR secondary infection OR 
phlebitis OR sterile abscess OR drainage. 
Grade 4:  
potentially life-threatening consequences (e.g., abscess, exfoliative dermatitis, necrosis involving 
dermis or deeper tissue). 
 
Injection site pain/tenderness (pain without touching or tenderness as pain when the area is touched), if 
present, will be graded as follows:  
Grade 1:  
pain/tenderness causing no or minimal limitation of use of limb. 
Grade 2:  
pain or tenderness causing greater than minimal limitation of use of limb. 
Grade 3:  
pain/tenderness causing inability to perform usual social and functional activities. 
Grade 4:  
pain or tenderness causing inability to perform basic self-care function OR hospitalization 
indicated. 
 
 
 

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Systemic reactions: 
General Signs/Symptoms: 
 
Fever 
 
Change in eating habits 
 
Diarrhea 
 
Sleepiness 
 
Irritability 
 
Unusual crying 
 
Vomiting 
 
Fever if present will be graded as follows (axillary temperature):  
Grade 1:  
37.5°C to 37.9°C 
Grade 2:  
38.0°C to 38.4°C 
Grade 3:  
38.5°C to 40.0°C 
Grade 4:  
>40.0°C 
 
Change in eating habits, diarrhea, sleepiness, irritability, unusual crying, vomiting, and any other 
unsolicited reaction occurring through 7 days post booster vaccination will be graded as follows:  
Grade 1:  
symptoms causing no or minimal interference with usual social and functional activities. 
Grade 2:  
symptoms causing greater than minimal interference with usual social and functional activities. 
Grade 3:  
symptoms causing inability to perform usual social and functional activities with intervention or 
hospitalization indicated. 
Grade 4:  
symptoms causing inability to perform basic self-care functions OR medical or operative 
intervention indicated to prevent permanent impairment, persistent disability, or death. 
 
7.2.4. Serious adverse events 
An SAE is defined as an AE that meets one of the following conditions: 
 
Death. 
 
Life threatening (participant at immediate risk of death). (The term “life-threatening” in the definition of 
“serious” refers to an event in which the patient was at risk of death at the time of the event; it does not 
refer to an event which hypothetically might have caused death if it were more severe.) 
 
Requires inpatient hospitalization or prolongation of existing hospitalization. 
 
Results in a persistent or significant disability or incapacity. 
 
Important medical events that may not result in death, be life threatening, or require hospitalization may 
be considered SAEs when, based upon appropriate medical judgment, may jeopardize the participant and 
may require medical or surgical intervention to prevent one of the outcomes listed in this definition. 
Medical and scientific judgment should be exercised in deciding whether reporting is appropriate. 
 
All SAEs occurring within 28 days post vaccination must be reviewed and evaluated by a study physician (SAE 
relationship to study vaccine must be evaluated as outlined in Section 7.2.2) and recorded on an SAE form and 
reported (as specified in Section 7.3). SAEs, whether serious or not, that persist at the end of the trial will be 
followed up by the investigator until satisfactory resolution or until the investigator deems the event to be chronic 
or the participant to be stable. The investigator will document the date of final disappearance of the adverse event 

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on a data clarification form. All care for SAEs or care required as part of the study will be provided to study 
participants free of cost. 
 
7.3. Reporting procedures 
7.3.1.  Serious adverse events 
All SAEs occurring within 28 days post booster dose must be reported to both the local data safety and 
monitoring board (DSMB) and the Sponsor, even if the investigator considers that the SAE is not related to 
treatment. Reporting procedures to the local DSMB will follow established icddr,b policy, whereas reporting to 
the Sponsor will be described in detail in an SOP. To report to the Sponsor, the study clinician will complete a 
Serious Adverse Event Form within the following timelines of such events:  
 
 
All SAEs, regardless of relationship, will be recorded on the Serious Adverse Event Form and sent by 
email to the Sponsor and the contract research organization within 24 hours of site awareness, as outlined 
in a reporting SOP.  
 
SAEs reported to the local DSMB will be reviewed by the committee as per established icddr,b guidelines and 
will be reviewed during the scheduled meetings between the icddr,b, DSMB, and the PI. Details for how review 
of SAEs and other unanticipated problems will be conducted will be contained in an SOP. 
7.3.2. Regulatory reporting  
Collected SAEs will be reported to responsible ethical review committees according to requested timelines. 
Collected local and systemic reactions and adverse events not meeting the criteria of an SAE will be summarized 
at the end of the study, and a summary report will be sent to responsible ethical review committees and CDIBP 
manufacturers. 
 
7.4. Safety monitoring plan 
7.4.1.  Post vaccination follow-up 
Parents of children will be advised about possible short-term mild reactogenicity that may occur post vaccination. 
Parents will have the contact numbers of the study physician and community health workers. The parent will also 
be reminded to contact the study team if the child experiences any illness, has received any medical care, or 
requires any medical care. The Sponsor and the Ethical Review Committee (ERC) will be notified within 24 
hours of any SAEs.  
 
7.4.2. Data and safety monitoring board 
An independent, local DSMB will be constituted to monitor all SAEs. The DSMB chair will review all SAEs 
occurring 28 days post vaccination, including deaths. The DSMB will be instructed to use their clinical judgment 
in order to provide guidance or recommendations for stopping the study early due to a safety concern. The 
DSMB will evaluate the safety data from these children including SAEs. In addition, the DSMB will be asked to 
review in real time all SAEs suspected to be related to the product and provide recommendations to the icddr,b 
and PATH teams. 
 
7.5. Halting rules 
Given the experience with CD-JEV, no safety issues are anticipated which would result in the suspension of the 
study. However, the trial might be suspended at any time by PATH, the local DSMB, or any ethical review 
committee overseeing this study for any safety concern. This includes, for example, and without limitation, an 
SAE resulting in death, an unexpectedly high number of persons experiencing a similar event after receipt of any 
dose, or an unusually high rate of SAEs. PATH may suspend the study in the event that study conduct is found to 
be below GCP standards. 
 

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Should an SAE occur which is likely related to administration of study vaccine, the decision whether the study 
should continue per protocol, proceed with caution, be suspended pending further investigation, be discontinued, 
or be modified and then proceed will be made by PATH in consultation with the investigators, the DSMB, and/or 
the icddr,b Research Review Committee (RRC) and ERC. However, no rules will be prespecified to define these 
conditions.  
 
If the study is halted, participants will be contacted immediately explaining why the study has been halted and the 
implications for their medical well-being. 
 
In the event of the appearance of new data that indicate an increased level of risk to participants, the clinical trial 
will be suspended until PATH, the DSMB, and all ethical review committees have reviewed relevant data and 
agreed that the trial may proceed. 
 
7.6. Data handling and record keeping 
7.6.1.  Data management responsibilities 
The PI is responsible for ensuring the accuracy, completeness, legibility, and timeliness of the data reported. All 
source documents will be completed in a neat, legible manner in black ink to ensure accurate interpretation of 
data and clarity of reproduced copies. The study workers will be asked not to erase, overwrite or use correction 
fluid or tape on the original. An SOP will be available providing details of data management responsibilities. 
 
All source documents and laboratory reports will be reviewed by the clinical team and data entry staff, who will 
ensure that they are accurate and complete. Data collection will be the responsibility of the clinical trial staff at 
the site under the supervision of the PI. During the study, the PI will maintain complete and accurate 
documentation for the study and will advise and oversee local investigators who will conduct data management, 
entry, and quality review. Analysis and reporting of study data will be done by the trial statistician after data lock. 
 
7.6.2.  Data capture methods 
Clinical data and laboratory data will be recorded on CRFs. All children will be assigned a unique study 
participant number at study enrollment; this study participant number will be included on all forms and in the 
computer database and will serve to link study data to specific individuals. Data forms will be verified for 
accuracy, linked by study participant number, and managed using database management software (yet to be 
determined). 
 
7.6.3.  Study records 
In accordance with applicable regulatory requirements, following closure of the study, the investigator/institution 
will maintain a copy of study documents in a secure and designated location at the study site. Essential 
documents shall be retained for at least five years after the completion or discontinuation of the study. 
 
No records will be destroyed without the written consent of PATH. It is the responsibility of PATH to inform the 
PI when these documents no longer need to be retained. 
 
Study data collection forms (i.e., CRFs), informed consent documents, and adverse event-related medical records 
are the primary source documents for this study. Only authorized study staff and representatives of the 
monitoring team, the sponsor, independent ethical committee, and data monitoring committee and appropriate 
regulatory agencies may have direct access to source documents containing study data. 
 
 
 

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7.6.4. Protocol deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, GCP, or Manual of Procedures 
requirements. The noncompliance may be either on the part of the child or parent, the investigator(s), or the study 
site staff. As a result of deviations, corrective actions are to be developed by the site and implemented promptly.  
 
It is the responsibility of the site to use continuous vigilance to identify and report deviations within five working 
days of identification of the protocol deviation, or within five working days of the scheduled protocol-required 
activity. All deviations will be promptly reported to PATH.  
 
A completed copy of the Protocol Deviation Form will be maintained in the regulatory file. Protocol deviations 
will be sent to the local IRB/ERC per their guidelines. The PI and study staff are responsible for knowing and 
adhering to their IRB/ERC requirements. 
 
7.7. Quality control and quality assurance 
The study will be conducted in accordance with procedures identified in the protocol and SOPs. SOPs will be 
developed and revised as necessary. In addition, monitoring plans, data management plans, and quality 
management plans will be developed. 
 
Site monitoring will be conducted to ensure that human subject protection, study procedures, laboratory 
procedures, study intervention administration, and data collection processes are of high quality and meet sponsor, 
International Conference on Harmonisation (ICH) GCP, and regulatory standards, and that the study is conducted 
in accordance with the protocol and SOPs. 
 
All site staff will be required to have training in GCP, informed consent, and protocol implementation prior to 
work on the project. All study staff, including temporary staff such as nurses, phlebotomists, and interviewers 
will attend mandatory training prior to participant enrollment.  
 
7.8. Clinical monitoring 
7.8.1.  Site monitoring plan 
Qualified and appropriately trained individuals from PATH or designated by PATH will carefully monitor the 
study. The study monitor will periodically contact the site and perform on-site visits. The extent, nature and 
frequency of site visits will be based on such considerations as study objectives, study design and complexity, 
and enrollment rate; periodicity and nature of monitoring activities will be described in the Monitoring Plan. 
PATH may also include representatives of the Bill & Melinda Gates Foundation (The Foundation) in site contacts 
and visits, as appropriate, and will keep The Foundation program officer apprised of study progress. 
 
7.8.2.  Site initiation visit 
A site initiation visit will occur prior to approval to initiate the study. The study monitor and/or PATH 
representatives will verify and document that the study team has been properly informed about the trial, the 
materials to be used during the trial have been received and are appropriately stored, regulatory requirements are 
met and documented, and that approved study SOPs are in place. Prior to enrollment of participants at the study 
site, specific regulatory documents must be available, such as independent ethics committee (IEC) approvals, 
other IRB required approvals, and curriculum vitae for investigator and study staff.  
 
7.8.3.  Follow-up visits 
Monitoring will be conducted according to PATH’s requirements. The individuals responsible for monitoring the 
study will periodically review the progress of the study and should have access to all records necessary to ensure 
the ethical and safety conduct of the study and the integrity/validity of the recorded data. 
 

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During site visits and contacts, the monitor will: 
 
Check and assess the progress of the study. 
 
Review study data collected. 
 
Perform source data verification. 
 
Review trial master files for essential documents. 
 
Evaluate adherence to standard operating procedures. 
 
Identify any issues and address their resolution. 
 
This will be done in order to verify that: 
 
The data are authentic, accurate, and complete. 
 
The safety and rights of participants are protected. 
 
The study is conducted in accordance with the approved protocol (and any subsequent amendment) and 
all applicable regulatory requirements. 
 
As part of study conduct, the investigator agrees to allow the monitor or PATH representative direct access to all 
relevant documents and to allocate time to the monitor to discuss findings and any relevant issues. The 
investigator also agrees to allow representatives of the Foundation, authorized by PATH, to occasionally 
accompany the monitor or PATH representative during site visits. 
 
7.8.4.  Close-out visit 
Upon completion of the study, the study monitor or PATH representative and the investigator will conduct the 
following activities: 
 
Data clarification and/or resolution. 
 
Accounting, reconciliation, and destruction at sites of used and unused vaccines. 
 
Review of site study records for completeness. 
 
Return copy of study data to PATH. 
 
PATH and any regulatory body retain the right to temporarily suspend or prematurely discontinue this study at 
any time related to safety. If the study is stopped or suspended prematurely, PATH will inform the investigator as 
well as regulatory authorities about the decision and the reasons for termination or suspension. If such action is 
taken, all efforts must be made to ensure the safety of the participants enrolled in the study. The investigator will 
inform the responsible IEC and provide the reason for the suspension or termination. In case of premature study 
or study site closure, the monitor or PATH representative will conduct all activities as indicated above. 
 
7.8.5. Allowable schedule of study visits 
For clarity, per-protocol timing of key study visits is provided below in Table 4. No child should be dropped from 
the study solely on the basis of a visit falling outside of an allowable time-frame for that visit if there is good faith 
attempt by study staff and the child participant’s parent(s)/guardian(s) to adhere to the protocol. 
Table 4. Allowable Schedule 
Study Visit 
Should occur 
May occur 
Visit 01 
Three years after initial vaccination 
Three years ±3 months after initial vaccination 
Visit 02 
Day 365 
One year ±28 days after Visit 01 
Visit 03 
Day 366 
1 day after booster vaccination 
Visit 04 
Day 367 
2 days after booster vaccination 
Visit 05 
Day 368 
3 days after booster vaccination 
Visit 06 
Day 369 
4 days after booster vaccination 
Visit 07 
Day 370 
5 days after booster vaccination 

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Study Visit 
Should occur 
May occur 
Visit 08 
Day 371 
6 days after booster vaccination 
Visit 09 
Day 372 
7 to 10 days after booster vaccination 
Visit 10 
Day 393 
21 to 35 days after booster vaccination 
 
7.9. Audits and inspections 
For the purpose of compliance with applicable regulatory guidelines, it may be necessary for PATH or national or 
foreign regulatory authorities to conduct a site audit. This may occur at any time from study start to after the 
conclusion of the study. 
 
The investigator agrees to allow the auditor direct access to all relevant documents and to allocate time to the 
auditor to discuss findings and any relevant issues. 
 
National and foreign regulatory authorities may conduct a regulatory inspection of this study. If a regulatory 
authority requests an inspection, the investigator must inform PATH immediately about this request. The 
investigator agrees to allow the inspector(s) direct access to all relevant documents and to allocate time and the 
time of staff to the inspector(s) to discuss findings and any relevant issues. 
 
 
8. ETHICAL ASSURANCE FOR PROTECTION OF HUMAN RIGHTS 
8.1. Ethical standard 
The investigator will ensure that this study is conducted in full conformity with the current revision of the 
Declaration of Helsinki, ICH-GCP, and/or with local regulatory requirements, whichever affords the greater 
protection to the participant. 
 
8.2. Institutional review board 
The protocol will be reviewed and approved by all of the following bodies before recruitment begins: 
1. Western Institutional Review Board (WIRB). PATH subcontracts its ethical review for clinical trials 
to WIRB.  
2. The icddr,b RRC and ERC. 
 
All amendments will be approved in writing by WIRB and the icddr,b RRC and ERC before implementation and 
paperwork are shared between partners, as appropriate. No deviations from, or changes to, the protocol shall be 
initiated without prior written IRB approval by both WIRB and icddr,b of an appropriate amendment from all 
responsible IRBs, except when necessary to eliminate immediate hazards to the participants or when the 
change(s) involves only logistical or administrative aspects of the study (e.g., change of telephone number, etc.). 
The PI will sign the final approved protocol. 
8.3. Informed consent process 
Prior to receipt of study vaccine, written informed consent will be obtained from a parent/guardian for all child 
participants. Informed consent documents will embody the elements of consent as described in the Declaration of 
Helsinki and the ICH Harmonized Tripartite Guidelines for GCP. The consent form will be in Bangla. 
 
Original Informed Consent Forms must be kept on file by the investigator for possible inspection by regulatory 
authorities and PATH. The participant or the participant’s legally acceptable representative must receive a copy 
of the signed and dated Informed Consent Form(s), and any subsequent updates or amendments. The study 
monitor shall check the documentation of the individual Informed Consent Form(s) during each monitoring visit. 

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Parents will initially be briefed about the study by a field worker, since they are known to the household and 
routinely inform participants and parents/guardians about vaccine trials and surveillance.   
In the study clinic, the medical officer will ask if the child and parents/guardians have any questions about the 
study, read the informed consent form, and then conduct informed consent. One parent/guardian must sign the 
informed consent form. If the parent/guardian is illiterate, they may provide a thumb impression instead and an 
impartial literate witness (hierarchically independent of the PI and not specified on the list of trial contributors) 
will be present and attest to the parent/guardian consent. Consent will follow site-specific practices and be 
described in an SOP.  
 
8.4. Inclusion of women and children 
Children of any race/ethnicity and residing in the study area will be recruited for participation in the study. No 
special recruitment methods will be used to ensure certain levels of participation by any specific minorities 
residing in the source population. Investigators will emphasize during information sessions that girls and boys are 
at equal risk of JE and its complications. 
 
8.5. Participant confidentiality 
8.5.1.  Confidentiality of data 
By signing the protocol, the investigators agree that the study protocol, documentation, data, and all other 
information generated regarding the vaccines will be held in strict confidence. The investigators may divulge 
such information within regulatory restrictions and ethical considerations only to ethical review committees or 
similar expert boards or committees, and their affiliated institutions and employees, only under an appropriate 
understanding of confidentiality with such board or committee, and their affiliated institutions and employees. No 
information concerning the study or the data may be released to any unauthorized third party without prior 
written approval of PATH. Any regulatory agency deemed appropriate may consult study documents in order to 
verify case report form data. Investigators will ensure that all employees involved in the study respect the same 
rules.  
 
Documents and data pertaining to the study will be kept in a locked cabinet under the responsibility of the 
investigator. PATH will conduct periodic monitoring visits to ensure that the data is safe and stored in this secure 
place and that only those authorized study staff have access to the data. Study data will be kept for five years after 
completion of the study. 
 
Study participants will not be reported by name in any report or publication resulting from data collected in this 
study. 
 
8.5.2.  Confidentiality of patient records 
Participant confidentiality is strictly held in trust by the participating investigators, their staff, and the sponsor(s) 
and their agents. This confidentiality pertains to a participant’s personal and health information and extends to 
information on any biological specimen of the participant. Medical information about individual participants 
obtained during the course of this study is confidential and may not be disclosed to third parties, except 
authorized monitors, sponsors, auditors or inspectors. Confidentiality will be ensured by the use of participant 
numbers coding for the identification of each participant’s personal, health, and specimen information. Access to 
this linkage and other confidential data will be strictly controlled. 
 
The investigators will keep individual results confidential to the extent permitted by law. Information will not be 
released to anyone other than the participant, their parent or guardian, or their medical provider, unless required 
to do so by law. 
 
 
 

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8.5.3. Notification of primary care physician 
If agreed by the participant or participant’s legally acceptable representative, the investigator shall notify the 
participant’s primary care physician, if applicable, of participation in the study. Participant’s involvement in the 
study and study team contact numbers will be reported into the participant’s identification card as applicable. 
 
8.6. Study discontinuation 
There are no expected reasons that would result in suspension and/or termination of the study. Unexpected 
reasons include:  
 
 
New safety data about the investigational product resulting from this or any other trial becomes available.  
 
The sponsor or ERC overseeing this study suspends and/or terminates the study for any justified reason, 
including the event that early safety data reveal an unusually high rate of SAEs or in the event study 
conduct fails GCP standards. 
 
If the trial is prematurely terminated or suspended, the sponsor shall promptly inform the PI, the regulatory 
authorities, and the IRB/ERC of the reason for termination or suspension. Parents of children will be contacted 
immediately explaining why the study has been suspended or terminated and potential implications for children’s 
safety, and provided with the opportunity to ask questions. Contacts with parents will be documented. 
 
8.7. Future use of stored specimens 
Aliquots of participants’ serum specimens will be maintained for five years after the completion of the study. 
This is to allow the samples to be stored long enough to support any requests for additional information from 
WHO or other agencies required to register CD-JEV for use. Following completion of this period, all samples 
will be destroyed if additional studies are not requested. The specimens will be stored at a long term storage 
facility determined by the Sponsor . No personal identifying information associated with the study will be stored 
on site with the specimens. After the data analysis for the study is complete, the master linking document will be 
destroyed, thus removing the possibility of identifying the individual from whom the data and sample were 
collected. Samples will not be used for unrelated, future studies without appropriate ethics committee approvals 
and consent from participants (if deemed required by ethics committees and the master linking document with 
identifiers has not yet been destroyed). 
 
 
9. USE OF ANIMALS 
Not applicable. No animals will be used in this study.  
 
 
10. COLLABORATIVE ARRANGEMENTS 
10.1. Publication policy 
A Clinical Study Report (CSR) comprised of text and results tables reflecting all safety and immunogenicity data 
will be generated. The CSR will be reviewed, approved, and signed by the investigator. The CSR will be 
compliant with ICH guidelines. 
 
All data, documents, any recordings and information transferred by PATH to any contractor or obtained or 
prepared by any contractor, consultants, or persons associated by contractual relationships with any contractor 
during the trials, belong to icddr,b and PATH. 
 
All confidential information communicated to the investigators by PATH shall be kept strictly confidential by 
them or any other person connected with the study and shall not be disclosed, either orally or in written form, to 

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any third party without prior written consent of the organization of which the information is the exclusive 
property. 
 
Working with PATH representatives, the investigator is expected to publish the results of this research in a peer-
reviewed scientific journal. In no way may anyone or any organization prohibit the public dissemination of valid 
results of this trial. 
 
The International Committee of Medical Journal Editors (ICMJE) member journals have adopted a trials-
registration policy as a condition for publication. This policy requires that all clinical trials be registered in a 
public trials registry. It will be the responsibility of PATH representatives to register this trial in an acceptable 
registry (CT.GOV). ICMJE authorship criteria will be strictly followed for publication of any manuscript(s) 
arising from this trial. 
 
11. FACILITIES AVAILABLE 
Icddr,b has large multi-disciplinary international and national scientific research staff. Existing field, hospital, 
laboratory and office facilities will be used for this study. Icddr,b scientists have conducted a variety of vaccine 
studies (Polio, influenza, pneumococcal, cholera, Shigella, rotavirus, Japanese encephalitis, etc.). 
 
11.1. Field site 
The study will be conducted in rural Matlab and urban Mirpur, Dhaka. Icddr,b has been maintaining a field 
research centre at Matlab for about fifty years. Due to the presence of ongoing HDSS, clinic and laboratory 
facilities, effective referral systems and well-established infrastructure at Matlab, it offers excellent research 
facilities for this study. The HDSS is a regularly updated information system on the approximate population of 
220,000.  
 
11.2. Laboratory facilities 
Existing laboratory facilities in icddr,b will be used to process the samples. 
 
The specimens will be tested at the United States Army Medical Directorate, Armed Forces Research Institute of 
Medical Sciences, Bangkok, Thailand. 
 
 
 

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12. LITERATURE CITED 
 1.  Hombach J, Solomon T, Kurane I, Jacobson J, Wood D. Report on a WHO consultation on immunological 
endpoints for evaluation of new Japanese encephalitis vaccines, WHO, Geneva, 2-3 September, 2004. 
Vaccine. 2005;23(45):5205–5211.  
 2.  Zaman K, Naser AM, Power M, Yaich M, Zhang L, Ginsburg AS et al. Lot-to-lot consistency of live 
attenuated SA 14-14-2 Japanese encephalitis vaccine manufactured in a good manufacturing practice 
facility and non-inferiority with respect to an earlier product. Vaccine. 2014;32(46):6061–6066.  
 3.  Victor JC, Yao Y, Zhou B, Zhang L, Yoksan S, Kelly K et al. Corrigendum to "Comparison of the 
immunogenicity and safety of measles vaccine administered alone or with live, attenuated Japanese 
encephalitis SA 14-14-2 vaccine in Philippine infants". Vaccine. 2014;32(2):306–308.  
 4.  Fischer M, Lindsey N, Staples JE, Hills S. Japanese encephalitis vaccines: recommendations of the 
Advisory Committee on Immunization Practices (ACIP). Morbidity and Mortality Weekly Report. 
2010;59(RR-1):1–27.  
 5.  Halstead S, Jacobson J. Plotkin SA, Orenstein WA, Offit PA, editors. Japanese encephalitis vaccines. 
Vaccines. Fifth(17):311–352.  Saunders Elsevier; 2008.  
 6.  Igarashi A. Epidemiology and control of Japanese encephalitis. World Health Statistics Quarterly. 
1992;45(2-3):299–305.  
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