An Open-Label, Phase 2 Trial to Investigate the Humoral and Cell-Mediated Immune Responses and Safety of a Tetravalent Dengue Vaccine Candidate (TDV) Administered Subcutaneously in Flavivirus-Naïve and Dengue-Immune Healthy Adults
Phase: PHASE2
Conditions: Dengue Fever
Sponsor: Takeda

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Takeda’s Tetravalent Dengue Vaccine Candidate 
Trial No. DEN-210 
Clinical Study Report 
Page 1 of 427
CONFIDENTIAL 
16.1.1 Protocol and Protocol Amendments 
DEN-210 Protocol Amendment 4, Version 5.0, 18 December 2020...............................................2 
DEN-210 Protocol Amendment 3, Version 4.0, 11 March 2020...................................................88 
DEN-210 Protocol Amendment 2, Version 3.0, 06 December 2019...........................................172 
DEN-210 Protocol Amendment 1, Version 2.0, 23 January 2019...............................................257 
DEN-210 Original Protocol, Version 1.0, 02 May 2018 .............................................................346 
0001
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Trial No. DEN-210 
Page 4 of 88 
Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
1.3 
Protocol Amendment Summary of Changes 
This document describes the changes in reference to the Protocol incorporating 
Amendment No. 1. 
1.3.1 Amendment History 
Date 
Amendment 
Number 
Amendment 
Type 
Region 
02 May 2018 
Initial 
Protocol 
Not applicable 
Local (United States of America) 
23 January 2019 
1 
Non-substantial 
Local (United States of America) 
1.3.2 Summary of Changes 
Amendment to Protocol Version 1.0 dated 02 May 2018 
Rationale for the Amendment: 
This protocol has been amended to implement modifications with the aim of accelerating 
the recruitment process and to anticipate on subject drop-outs. In particular, participation of 
more than one trial center has been made possible and the overall sample size is increased 
by 10% to 22 subjects in each trial group, with the aim of 20 subjects being evaluable in 
each trial group. 
Other modifications 
Trial days 93, 95, 98, 101, and 104 have been corrected into trial days 94, 96, 99, 102 and 
105, respectively. 
Update of literature list. 
Administrative changes. 
Minor grammatical and editorial changes are included for clarification and consistency 
purposes only. 
New text is shown in bold italics and any deleted text is marked using strikethrough. 
Section 
Description of change 
Title 
page 
Date: 02 May 2018 23 January 2019 
Version: Version 1.0 2.0 (supersedes Version 1.0) 
Section 
1.2 
SIGNATURES 
, PhD, MSc PV 
 
Takeda Pharmaceuticals International AG 
, HN BC 
, MPH 
 
Vaccines Business Unit 
Takeda Vaccines, Inc. 
, MD 
, Dengue Clinical Development 
Vaccine Business Unit 
Takeda Pharmaceuticals International AG USA, Inc. 
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Trial No. DEN-210 
Page 5 of 88 
Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
Section 
Description of change 
Sections 
2.0 and 
4.2 
This trial will be conducted at a single trial centers in the United States of 
America (USA) using a pre existing pool of involving healthy adults who are 
either flavivirus-naïve or seropositive for DENV-1 or DENV-3 (ie, serology 
consistent with primary infection with either DENV-1 or DENV-3). The 
serological status of subjects will either has have been determined by the trial 
center prior to and outside the scope of this trial or will be determined through 
screening within the scope of this trial. 
Sections 
2.0, 6.1 
This is an open-label, phase 2 trial in 40 44 healthy adult subjects aged 18 to 60 
years (inclusive) to investigate the immunogenicity and safety of SC 
administration of a 2-dose regimen of TDV. 
Subjects will be enrolled in 2 trial groups based on results from serological 
testing performed either by the trial centers prior to and outside the scope of 
this trial or through screening within the scope of this trial (up to 35 days 
[5 weeks] prior to Day 1[M0]): 
-
Group 1: approximately 20 22 flavivirus-naïve subjects.
-
Group 2: approximately 20 22 dengue-immune subjects (ie,
approximately 10 11 subjects with serology consistent with primary
infection by DENV-1 and approximately 10 11 subjects with serology
consistent with primary infection by DENV-3).
All subjects will be followed-up for 9 months post second vaccination so the 
trial duration will be approximately 360 days (12 months) or 13 months for 
each subject depending on whether serological testing with regard to the 
inclusion criteria for the trial is performed outside or within the scope of this 
trial, respectively. 
Section 
2.0 
Subject Population: 
Planned Number of Subjects: 40 44 
Planned Number of Trial Arms: 2 trial groups (20 22 subjects per trial group); 
2-dose regimen (1 trial vaccine dose on Day 1 [M0] and 1 trial vaccine dose on
Day 90 [M3]), SC route.
Estimated Total: 40 44 enrolled subjects.
Note: SAEs and MAAEs are collected continuously throughout the trial.
M=month, MAAE=medically attended adverse event, SAE=serious adverse event, V=visit
(a) A screening visit is only applicable if serological testing with regard to the inclusion criteria for the trial is performed within the scope of this trial.
V1
Day 1 (M0)
Enrollment
(N=44)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V2
Day 4
Blood 
sample
V3
Day 6
Blood 
sample
V4
Day 9
Blood 
sample
V5
Day 12
Blood 
sample
V6
Day 15
Blood 
sample
V7
Day 30 
(M1)
Blood 
sample
V8
Day 60 
(M2)
Blood 
sample
V9
Day 90 (M3)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V19
Day 360 (M12)
Blood sample
Safety follow-up 
(SAEs and MAAEs)
V16
Day 150
(M5)
Blood 
sample
V15
Day 120
(M4)
Blood 
sample
V14
Day 105
Blood 
sample
V13
Day 102
Blood 
sample
V12
Day 99
Blood 
sample
V11
Day 96
Blood 
sample
V10
Day 94
Blood 
sample
V17
Day 180
(M6)
Blood 
sample
V18
Day 270
(M9)
Blood 
sample
Screening Visit (a)
Blood sample
Up to 35 days (5 weeks) prior to 
Day 1 (M0)
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Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
Section 
Description of change 
Sections 
2.0 and 
7.1 
4. Group 1 only: immunologically naïve to dengue, Zika, Yellow Fever (YF),
Japanese Encephalitis (JE), West Nile (WN) (based on negative results for
detection of anti-DENV, anti-Zika, anti-YF, anti-JE, anti-WN antibodies) as
documented by serological testing performed either by the trial center prior to
and outside the scope of this trial or through screening within the scope of this
trial (up to 35 days [5 weeks] prior to Day 1 [M0]).
5. Group 2 only: serology consistent with primary infection with either DENV-1
or DENV-3 (defined as detectable neutralizing antibodies against DENV-1 or
DENV-3 only, or titers for DENV-1 or DENV-3 ≥4-times higher than titers for
the 2 other dengue serotypes) as documented by serological testing performed
either by the trial center prior to and outside the scope of this trial or through
screening within the scope of this trial (up to 35 days [5 weeks] prior to Day 1
[M0]).
Section 
2.1 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to 
Day 90 (M3)] and Visits 10 to 19 [Day 94 to Day 360 [M12]) 
Screening visit 
(a) 
V1 
Up to 35 days 
(5 weeks) prior to 
D1 (M0) 
D1 
M0 
Informed consent(a) 
X 
X (b) 
Assessment of eligibility criteria(bc) 
X 
X 
Trial group assignment (flavivirus-naïve or dengue-immune)(cd) 
X 
Demographics, medical history, and prior medication/ vaccination 
X 
X(e) 
Medical history 
X 
X 
Prior medication/vaccination 
X 
X 
Concomitant medications/vaccinations(df) 
X 
X 
Review of systems 
X 
Complete physical examination(eg) 
X 
X 
Targeted physical examination(fh) 
X 
Vital signs(gi) 
Pregnancy test(hj) 
X 
X 
Pregnancy avoidance guidance(ik) 
X 
Blood sampling for serological testing (2.5 mL) 
X 
X 
Blood sample for humoral immune response (20 mL)(jl, km) 
Blood sample for T cell-mediated immune response (60 mL)(jl) 
X 
Blood sample for B cell-mediated immune response (30 mL)(jl) 
X 
Blood sample for innate immune response (5 mL)(jl) 
X 
Blood sample for vaccine viremia (5 mL)(jl) 
X 
Check criteria for delay of trial vaccine administration 
X 
Check contraindications for trial vaccine administration 
X 
Trial vaccine administration(ln) 
X 
Injection site evaluation(mo) 
X 
Diary 
card (np) 
Distribution 
X 
Review/collection of solicited and unsolicited AEs 
X 
AEs leading to trial vaccine withdrawal or trial discontinuation, 
SAEs, MAAEs(oq) 
X 
AEs=Adverse Events, D=Day, M=Month, MAAEs=Medically Attended Adverse Events, NA=Not Applicable, SAEs= 
Serious Adverse Events, V=Visit 
(a)
A screening visit is only applicable if serological testing with regard to the inclusion criteria for the trial is 
performed within the scope of this trial. 
(b)
Up to 28 days prior to Day 1 (Month 0 [M0]).
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Trial No. DEN-210 
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Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
Section 
Description of change 
Section 
2.1 
(c)
(b) After informed consent has been obtained, eligibility of the subject will be assessed by review of 
inclusion/exclusion criteria at Screening or prior to TDV administration on Day 1 (M0), as applicable. 
(d)
(c) Subjects will be enrolled in Group 1 (flavivirus-naïve subjects) or Group 2 (dengue-immune subjects with
serology consistent with primary infection with either wild type dengue virus [DENV]-1 or DENV-3) based on 
serological testing performed either by the trial center prior to and outside the scope of this trial or within the
scope of this trial (up to 35 days [5 weeks] prior to Day 1 [M0]). 
(e)
Not applicable if a Screening visit has been performed.
(f)
(d) All medications and vaccine history from 1 month (minimum 28 days) prior to administration of each trial 
vaccine dose up to 1 month (minimum 28 days) thereafter, steroids and immunostimulants within 60 days prior to 
Day 1 (M0), immunoglobulins and blood products within 3 months prior to Day 1 (M0), and immunosuppressive 
therapy within 6 months prior to Day 1 (M0). Concomitant medication/vaccination will be collected throughout 
the trial. 
(g)
(e) Physical examination including measurement of weight and height; Body Mass Index (BMI) will be calculated.
Measurement of height is only required at Screening or at Day 1 (M0), as applicable. 
(h)
(f) Subjects may undergo a targeted symptom-directed physical examination. Clinically significant changes from
the Baseline examination should be recorded in the subject’s source documents and electronic Case Report Form
(eCRF).
(i)
(g) Vital signs including (but not limited to) the measurement of systolic blood pressure/diastolic blood pressure,
heart rate, and body temperature.
(j)
(h) Pregnancy testing (serum or urine) for females of childbearing potential. Results must be confirmed and 
documented as negative prior to each trial dose administration. Additional pregnancy tests may be performed
during the trial if deemed necessary by the investigator.
(k)
(i) Females of childbearing potential who are sexually active will be reminded during trial visits to adhere to 
acceptable contraceptive methods up to 6 weeks after the last dose of TDV (Day 90 [Month 3] (M3]) + 6 weeks).
Subjects will be provided with information on acceptable methods of contraception as part of the subject informed
consent process and will be asked to sign a consent form stating that they understand the requirements for 
avoidance of pregnancy and donation of ova. During the course of the trial, subjects of childbearing potential will
receive continued guidance with respect to the avoidance of pregnancy.
(l)
(j) All blood samples on days of vaccination (Day 1 [M0] and Day 90 [M3]) should be taken prior to 
administration of TDV.
(m) (k) Dengue neutralizing antibodies and anti-dengue Non-Structural protein 1 (NS1) antibodies for all subjects, and
T cell epitope mapping in a subset of subjects with Interferon-gamma Enzyme-Linked Immunospot
(IFN- γELISpot) responses >500 Spot Forming Cells/106 cells and availability of sufficient cells).
(n)
(l) Subjects will receive TDV by subcutaneous injection.
(o)
(m) Injection site pain, erythema, and swelling assessed by trial staff for 30 minutes post-vaccination. 
(p)
(n) Diary cards (paper or electronic) will be distributed for the recording of 1) solicited AEs including solicited
local (injection site) reactions for 7 days (day of administration + 6 days) and solicited systemic events for 14 days
(day of administration + 13 days) following administration of each trial vaccine dose, and 2) unsolicited AE for 28
days (day of administration + 27 days) following administration of each trial vaccine dose. The investigator will 
categorize events by severity (mild, moderate or severe) and will assess causality to vaccine administration
(“related” or “not related”). For persistent/prolonged solicited local (injection site) reactions or systemic events
observed as continuing on Day 8 or 15, respectively, following each trial vaccination, the end date will be captured
on the “Adverse Event” eCRF to permit a separate analysis from the unsolicited AEs. Any solicited local (injection
site) reaction or systemic event that resolves before 8 or 15 days, respectively, following each trial vaccination, but
recurs at a later time (ie, if discontinues), should be recorded as an unsolicited AE on the “Adverse Event” eCRF.
(q)
(o) MAAEs and SAEs will be collected for the trial duration.
(r)
(p) The Final Visit will be performed on Day 360 (Month 12 [M12]). If a subject terminates trial participation 
earlier, Day 360 (M12) procedures should be performed at their last trial visit, if possible.
V19 
D360 (p r) 
M12 
Sections 
2.0 and 
6.3 
The trial duration for each subject will be at least approximately 360 days 
(12 months) including vaccination (Day 1 [M0] and Day 90 [M3]) and follow-
up through Day 360 (M12). This duration will be approximately 13 months if 
serological testing with regard to the inclusion criteria for the trial is 
performed within the scope of this trial (ie, up to 35 days [5 weeks] prior to 
Day 1 [M0]). 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
Trial No. DEN-210 
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Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
Section 
Description of change 
Sections 
1.2, 4.2, 
15.0, 
and 15.2 
International Council for Harmonization of Technical Requirements for 
Pharmaceuticals for Human Use (ICH) E6 Good Clinical Practice (GCP): 
Consolidated Guideline [1 2]. 
Sections 
2.0, 2.1, 
5.2, 6.1, 
and 
9.3.5 
Days 93 94, 95 96, 98 99, 101 102, and 104 105. 
Section 
9.1.7 
Days 9394, 9596, and 104 105. 
Section 
9.1.10 
Days 9596, 98 99, 101 102, and 104 105. 
Section 
9.1.4 
Physical examinations must be performed by a qualified health professional in 
accordance with local regulations and as listed within the Site Responsibility 
Delegation Log. A complete physical examination will be performed at 
Screening (if applicable) and prior to vaccination on Day 1 (M0) and 
Day 90 (M3). 
Measurement of height is only required at Screening or Day 1 (M0), as 
applicable. 
Section 
9.1.6 
9.1.6 Screening 
A blood sample (2.5 mL) will be collected at the screening visit (up to 35 days 
[5 weeks] prior to Day 1 [M0)]). A screening visit is only applicable if 
serological testing with regard to the inclusion criteria (see Section 7.1) is 
performed within the scope of this trial. All blood samples will be processed, 
labeled and stored according to the Laboratory Manual or other appropriate 
guideline provided to the trial site. 
Note: The renumbering of subsequent heading 3 sections following addition of 
this section is not documented. 
Section 
9.1.11 
For female subjects of childbearing potential, serum or urine pregnancy testing 
will be performed at Screening (if applicable) and prior to each trial dose 
administration (Day 1 [M0] and Day 90 [M3]). 
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Protocol Version 2.0 
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CONFIDENTIAL 
Section 
Description of change 
Section 
9.3.1 
Section 9.3.1 Procedures at the Screening Visit (up to 35 days [5 weeks] prior 
to Day 1 [M0]). 
Note: A screening visit is only applicable if serological testing with regard to 
the inclusion criteria (see Section 7.1) is performed within the scope of this 
trial. 
1. Before performing any trial procedure, the signed informed consent form
needs to be obtained. Refer to Section 9.1.1.
2. Check inclusion and exclusion criteria. Refer to Section 7.1 and
Section 7.2, respectively.
3. Collect demographic data, medical history, and prior
medication/vaccination. Refer to Section 9.1.2.
4. Collect concomitant medications/vaccinations. Refer to Section 9.1.2.
5. Perform a complete physical examination. Refer to Section 9.1.4.
6. Perform pregnancy testing (serum or urine) for females of childbearing
potential. Refer to Section 9.1.11
7. Collect blood sample. Refer to Section 9.1.6.
Blood should be taken from the subject using an aseptic venipuncture
technique. Refer to the detailed collection and handling procedures
outlined in the Procedures Manual.
The site should schedule the next trial visit with the subject. 
The subject will receive a written reminder of the next trial visit. 
Note: The renumbering of subsequent heading 3 sections following addition of 
this section is not documented. 
Section 
9.3.2 
1. Before performing any trial procedure, the signed informed consent form
needs to be obtained (Day 1 [M0] unless obtained at Screening). Refer to
Section 9.1.1.
Section 
9.4 
In this trial, specimens will be collected for serological testing as described in 
Section 9.1.6, immune response testing as described in Section 9.1.6 9.1.7. and 
Section 9.1.7 9.1.8, and for clinical safety as described in Section 9.1.9 9.1.10. 
Section 
15.4.2 
Primary Completion of Trial completion corresponds to the date on which the 
final subject was examined or received an intervention for the purposes of final 
collection of data (Last Subject Last Visit) for the primary outcome, whether 
the clinical trial concluded according to the pre specified protocol or was 
terminated. 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
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Page 10 of 88 
Protocol Version 2.0 
23 January 2019 
CONFIDENTIAL 
Section 
Description of change 
Section 
16.0 
10. Sanofi Pasteur, 2011. http://www.dengue.info [Accessed 22 October 2017]
Wichmann O, Vannice K, Asturias EJ, de Albuquerque Luna EJ, Longini I,
Lopez AL, et al. Live-attenuated tetravalent dengue vaccines: The needs and
challenges of post-licensure evaluation of vaccine safety and effectiveness.
Vaccine. 2017. 35(42):5535-42.
12. Sanofi Pasteur, 2017. http://mediaroom.sanofi.com/sanofi-updates-
information on dengue vaccine [Accessed 12 March 2018] Sridhar S, Luedtke
A, Langevin E, Zhu M, Bonaparte M, Machabert T, et al. Effect of dengue
serostatus on dengue vaccine safety and efficacy. N Engl J Med. 2018.
379(4):327-40.
Section 
16.0 
13. World Health Organization. Weekly Epidemiological Record. 2018.
93:329-44. Available at http://www.who.int/wer [Accessed 22 October 2018].
16. Huang CY, Butrapet S, Pierro DJ, Chang GJ, Hunt AR, Bhamarapravati
N, et al. Chimeric dengue type 2 (vaccine strain PDK-53)/dengue type 1 virus
as a potential candidate dengue type 1 virus vaccine. J Virol.
2000;74(7):3020-8.
18 20 WHO Drug Dictionary. http://www.wow.com/wiki/WHO_Drug_ 
Dictionary [Accessed 22 November 2017] Available at 
http://www.who.int/medicines/ publications/druginformation/en/ [Accessed 
23 January 2019] 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
Trial No. DEN-210 
Page 4 of 84 
Protocol Version 3.0 
06 December 2019 
CONFIDENTIAL 
1.3 
Protocol Amendment Summary of Changes 
This document describes the changes in reference to the Protocol incorporating 
Amendment No. 2. 
1.3.1 Amendment History 
Date 
Amendment 
Number 
Amendment 
Type 
Region 
02 May 2018 
Initial 
Protocol 
Not applicable 
Local (United States of America) 
23 January 2019 
1 
Non-substantial 
Local (United States of America) 
06 December 2019 
2 
Substantial 
Local (United States of America) 
1.3.2 Summary of Changes 
Amendment to Protocol Version 2.0 dated 23 January 2019 
Rationale for the Amendment: 
This protocol has been amended because incidence estimates of dengue among travelers 
indicates that although the incidence rate is variable (due to the multifactorial nature of 
dengue transmission in endemic countries), overall, the risk of dengue transmission among 
travelers is considered to be low (1-4). As a result, the criterion #22 that excludes subjects 
from participating in the trial based on planned travel to dengue endemic countries during 
the conduction of the trial has been removed. Removal of this exclusion criteria is expected 
to address, at least to some extent, the slow enrollment rate experienced in the trial. 
In addition, the screening window for subjects screened within the scope of this trial has 
been increased to 70 days (10 weeks) to allow enrollment of subjects falling outside the 
previous defined screening window of 35 days (5 weeks). 
1.
Baaten GG, Sonder GJ, Zaaijer HL, van Gool T, Kint JA, van den Hoek A. Travel-related dengue virus
infection, The Netherlands, 2006-2007. Emerg Infect Dis 2011;17(5):821-8.
2.
Cobelens FG, Groen J, Osterhaus AD, Leentvaar-Kuipers A, Wertheim-van Dillen PM, Kager PA. Trop
Med Int Health 2002;7(4):331-8.
3.
Potasman I, Srugo I, Schwartz E. Dengue seroconversion among Israeli travelers to tropical countries.
Emerg Infect Dis 1999;5(6):824-7.
4.
Ratnam I, Black J, Leder K, Biggs BA, Matchett E, Padiglione A, et al. Incidence and seroprevalence of 
dengue virus infections in Australian travellers to Asia. Eur J Clin Microbiol Infect Dis 2012;31(6):1203-
10. 
Other modifications: 
–
Administrative change
–
Update of literature list
–
Minor editorial changes are included for consistency purposes only.
New text is shown in bold italics and any deleted text is marked using strikethrough. 
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Protocol Version 3.0 
06 December 2019 
CONFIDENTIAL 
Section 
Description of change 
Title page Date: 23 January 2019 06 December 2019 
Version: Version 2.0 3.0 (supersedes Version 1.0 2.0) 
1.2 
, PhD 
, MSc 
 
Vaccines Statistics 
Global Statistics 
Takeda Pharmaceuticals International AG 
Sections 
2.0, 6.1 
Subjects will be enrolled in 2 trial groups based on results from serological 
testing performed either by the trial center prior to and outside the scope of 
this trial or through screening within the scope of this trial (up to 35 70 days [5 
10 weeks] prior to Day 1[M0]): 
All subjects will be followed-up for 9 months post second vaccination so the 
trial duration will be approximately 360 days (12 months) or 13 14.5 months 
for each subject depending on whether serological testing with regard to the 
inclusion criteria for the trial is performed outside or within the scope of this 
trial, respectively. 
Sections 
2.0, 6.1, 
Figure 
2.a,
Figure 6.a
-
Group 2: approximately 22 dengue-immune subjects (ie, approximately 11
subjects with serology consistent with primary infection by DENV-1 and
approximately 11 subjects with serology consistent with primary infection
by DENV-3).
Note: SAEs and MAAEs are collected continuously throughout the trial.
M=month, MAAE=medically attended adverse event, SAE=serious adverse event, V=visit
(a) A screening visit is only applicable if serological testing with regard to the inclusion criteria for the trial is performed within the scope of this trial.
V1
Day 1 (M0)
Enrollment
(N=44)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V2
Day 4
Blood 
sample
V3
Day 6
Blood 
sample
V4
Day 9
Blood 
sample
V5
Day 12
Blood 
sample
V6
Day 15
Blood 
sample
V7
Day 30 
(M1)
Blood 
sample
V8
Day 60 
(M2)
Blood 
sample
V9
Day 90 (M3)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V19
Day 360 (M12)
Blood sample
Safety follow-up 
(SAEs and MAAEs)
V16
Day 150
(M5)
Blood 
sample
V15
Day 120
(M4)
Blood 
sample
V14
Day 105
Blood 
sample
V13
Day 102
Blood 
sample
V12
Day 99
Blood 
sample
V11
Day 96
Blood 
sample
V10
Day 94
Blood 
sample
V17
Day 180
(M6)
Blood 
sample
V18
Day 270
(M9)
Blood 
sample
Screening Visit (a)
Blood sample
Up to 35 70 days (5 10 weeks) 
prior to Day 1 (M0)
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Protocol Version 3.0 
06 December 2019 
CONFIDENTIAL 
Section 
Description of change 
Sections 
2.0 and 
7.1 
4. Group 1 only: immunologically naïve to dengue, Zika, Yellow Fever (YF),
Japanese Encephalitis (JE), West Nile (WN) (based on negative results for
detection of anti-DENV, anti-Zika, anti-YF, anti-JE, anti-WN antibodies) as
documented by serological testing performed either by the trial center prior to
and outside the scope of this trial or through screening within the scope of this
trial (up to 35 70 days [5 10 weeks] prior to Day 1 [M0]).
5. Group 2 only: serology consistent with primary infection with either
DENV-1 or DENV-3 (defined as detectable neutralizing antibodies against
DENV-1 or DENV-3 only, or titers for DENV-1 or DENV-3 ≥4-times higher
than titers for the 2 other dengue serotypes) as documented by serological
testing performed either by the trial center prior to and outside the scope of
this trial or through screening within the scope of this trial (up to 35 70 days [5
10 weeks] prior to Day 1 [M0]).
Sections 
2.0 and 
7.2 
22. Planned travel (during the trial conduct) to any area endemic for dengue.
Section 
2.1, 
Table 2.a 
Table 2.a Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to 
Day 90 (M3)] and Visits 10 to 19 [Day 94 to Day 360 [M12])
Screening 
visit (a) 
V
1 
V
2 
V
3 
V
4 
V
5 
V
6 
V
7 
V
8 
V
9 
V 
10 
V
11 
V
12 
V
13 
V
14 
V
15 
V
16 
V
17 
V
18 
V
19 
Visit window 
(days) 
Up to 35 
70 days  
(5  10 
weeks)  
prior to D1 
(M0) 
Record travel 
history and 
planned travel to 
any country 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
x  
x 
(d) Subjects will be enrolled in Group 1 (flavivirus-naïve subjects) or Group 2 (dengue-immune subjects with
serology consistent with primary infection with either wild type dengue virus [DENV]-1 or DENV-3) based on 
serological testing performed either by the trial center prior to and outside the scope of this trial or within the scope of 
this trial (up to 35 70 days [5 10 weeks] prior to Day 1 [M0]). 
Diary card(o p)
AEs leading to trial vaccine withdrawal or trial discontinuation, SAEs, 
MAAEs(p q) 
Sections 
2.0 and 
6.3 
The trial duration for each subject will be at least approximately 360 days 
(12 months) including vaccination (Day 1 [M0] and Day 90 [M3]) and follow-
up through Day 360 (M12). This duration will be approximately 13 
14.5 months if serological testing with regard to the inclusion criteria for the 
trial is performed within the scope of this trial (ie, up to 35 70 days [5 
10 weeks] prior to Day 1 [M0]). 
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Protocol Version 3.0 
06 December 2019 
CONFIDENTIAL 
Section 
Description of change 
Section 
9.1.6 
A blood sample (2.5 mL) will be collected at the screening visit (up to 
35 70 days [5 10 weeks] prior to Day 1 [M0)]). A screening visit is only 
applicable if serological testing with regard to the inclusion criteria (see 
Section 7.1) is performed within the scope of this trial. All blood samples will 
be processed, labeled and stored according to the Laboratory Manual or other 
appropriate guideline provided to the trial site.
Section 
9.3.1 
9.3.1 Procedures at the Screening Visit (up to 35 70 days [5 10 weeks] prior to 
Day 1 [M0]). 
Record travel history and planned travel to any country. 
Sections 
9.3.2 and 
9.3.5 
Record travel history and planned travel to any country. 
Section 
16.0 
2. International Council for Harmonisation of Technical Requirements for
Pharmaceuticals for Human Use (ICH). ICH harmonised guideline. Integrated
Addendum to ICH E6 (R1): Guideline for Good Clinical Practice E6 (R2).
Available at http://www.ich.org/fileadmin/ Public_Web_Site/ICH_Products/
Guidelines/Efficacy/E6/E6_R2_Step_4_2016_109.pdf.
https://database.ich.org/sites/default/files/E6_R2_Addendum.pdf [Accessed
23 January 06 December 2019].
6. World Health Organization, 2018. Dengue and severe dengue. Fact Sheet.
Available at http://www.who.int/mediacentre/factsheets/fs117/en/
https://www.who.int/en/news-room/fact-sheets/detail/dengue-and-severe-
dengue [Accessed 23 January 06 December 2019].
7. World Health Organization, 1997. Dengue hemorrhagic fever: diagnosis,
treatment, prevention and control, 2nd Edition. Geneva. Available at
https://www.who.int/csr/resources/publications/dengue/Denguepublication/en/
[Accessed 23 January 06 December 2019].
8. World Health Organization, 2009. Dengue guidelines for diagnosis,
treatment, prevention and control. Available at
http://www.who.int/tdr/publications/documents/dengue-diagnosis.pdf.
[Accessed 23 January 06 December 2019].
13. World Health Organization. Weekly Epidemiological Record. 2018.
93:329-44. Available at http://www.who.int/wer [Accessed 23 January 06
December 2019].
17. Policy and communication bulletin – The clinical center. Guidelines for
limits of blood drawn for research purposes in the clinical center. Manual
transmittal sheet, no. M95-9 (rev.), 2009. Available at
https://irb.research.chop.edu/.../files/documents/g_nih_blooddraws [Accessed
23 January 06 December 2019].
20. WHO Drug Dictionary. Available at
http://www.who.int/medicines/publications/druginformation/en/ [Accessed 23
January 06 December 2019].
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Page 4 of 83 
Protocol Version 4.0 
11 March 2020 
CONFIDENTIAL 
1.3 
Protocol Amendment Summary of Changes 
This document describes the changes in reference to the Protocol incorporating 
Amendment No. 3. 
1.3.1 Amendment History 
Date 
Amendment 
Number 
Amendment 
Type 
Region 
02 May 2018 
Initial Protocol 
Not applicable 
Local (United States of America) 
23 January 2019 
1 
Non-substantial 
Local (United States of America) 
06 December 2019 
2 
Substantial 
Local (United States of America) 
11 March 2020 
3 
Substantial 
Local (United States of America) 
1.3.2 Summary of Changes 
1. Amendment to Protocol Version 3.0 dated 06 December 2019
2. Rationale for the Amendment:
3. This protocol amendment is due to the slow enrollment rate experienced in the trial for
Group 2 (DENV-1 or DENV-3 positive subjects) while complete enrollment of the
planned number of subjects in Group 1 (flavivirus-negative subjects) has been achieved
(n=22). Group 2 would create a subject population potentially eligible for inclusion in a
future DENV-1/DENV-3 human challenge trial. Since Group 1 is considered a key
group for the characterization of immune responses to Takeda’s Tetravalent Dengue
Vaccine Candidate (TDV) (primary and secondary trial objectives), it was decided to
stop the recruitment of subjects in Group 2. As a result, the criterion that excludes
subjects from participating in the trial based on planned travel to dengue endemic
countries during the conduct of the trial which was removed from Protocol Version 3.0
dated 06 December 2019 is reinstated since 1) removal of this criterion was previously
done to facilitate enrollment of subjects in Group 2, and 2) travel to dengue endemic
regions is undesirable for subjects in Group 1 and Group 2. As the criterion of
authorized travel to dengue endemic countries is no longer included, recording of the
travel history and planned trips to any country is no longer necessary.
4. Protocol Version 3.0 dated 06 December 2019 has not been submitted to any
Institutional Review Boards.
5. Other modifications:
–
Administrative change
–
Correction in the unit for the non-structural protein 1 (NS1) antibody test result.
–
Removal of reference to the Procedures Manual as it is not applicable for the trial.
–
A minor editorial change is included for consistency purposes only.
–
Update of literature list
6. New text is shown in bold italics and any deleted text is marked using strikethrough.
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Protocol Version 4.0 
11 March 2020 
CONFIDENTIAL 
Section 
Description of change 
Title page Date: 06 December 2019 11 March 2020 
Version: Version 3.0 4.0 (supersedes Version 2.0 3.0) 
Section 
1.2 
, MPH 
 
 
Vaccine Business Unit 
Takeda Vaccines, Inc 
Sections 
2.0, 6.1, 
Figure 
2.a,
Figure 6.a
Note: The actual number of subjects in Group 2 will be lower than the 
planned 22 subjects as it was decided to stop the recruitment of subjects in 
Group 2. 
Sections 
2.0 and 
7.2 
7. 22. Planned travel (during the trial conduct) to any area endemic for
dengue.
Sections 
2.1, 9.3.1, 
9.3.2 and 
9.3.5, 
Table 2.a 
Record travel history and planned travel to any country 
Sections 
2.0 and 
5.2.3 

Average EC50 concentration (relative units/mL) of anti-dengue NS1
antibodies for each of the 4 dengue serotypes measured by ELISA using
blood samples collected from all subjects post first vaccination on Days
15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on
Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12).
Section 
9.1.5 
Body temperature measurement will be described in the Procedures Manual. 
Sections 
9.3.1 
9.3.2, and 
9.3.5 
Refer to the detailed collection and handling procedures outlined in the 
Procedures Manual. 
Note: SAEs and MAAEs are collected continuously throughout the trial.
Note: The actual number of subjects in Group 2 will be lower than the planned 22 subjects as it was decided to stop the recruitment of subjects in Group 2.
M=month, MAAE=medically attended adverse event, SAE=serious adverse event, V=visit
(a) A screening visit is only applicable if serological testing with regard to the inclusion criteria for the trial is performed within the scope of this trial.
V1
Day 1 (M0)
Enrollment
(N=44)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V2
Day 4
Blood 
sample
V3
Day 6
Blood 
sample
V4
Day 9
Blood 
sample
V5
Day 12
Blood 
sample
V6
Day 15
Blood 
sample
V7
Day 30 
(M1)
Blood 
sample
V8
Day 60 
(M2)
Blood 
sample
V9
Day 90 (M3)
Group 1 (flavivirus-naïve): 
(N=22)
Group 2 (dengue-immune): 
(N=22)
Pre-vaccination blood sample
TDV vaccination
Safety assessments
V19
Day 360 (M12)
Blood sample
Safety follow-up 
(SAEs and MAAEs)
V16
Day 150
(M5)
Blood 
sample
V15
Day 120
(M4)
Blood 
sample
V14
Day 105
Blood 
sample
V13
Day 102
Blood 
sample
V12
Day 99
Blood 
sample
V11
Day 96
Blood 
sample
V10
Day 94
Blood 
sample
V17
Day 180
(M6)
Blood 
sample
V18
Day 270
(M9)
Blood 
sample
Screening Visit (a)
Blood sample
Up to 70 days (10 weeks) prior
to Day 1 (M0)
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Protocol Version 4.0 
11 March 2020 
CONFIDENTIAL 
Section 
13.1 
Data reviews will be conducted prior to the planned interim analysis and final 
analysis. This These reviews will assess the accuracy and completeness of the 
trial database, subject evaluability, and appropriateness of the planned 
statistical methods. 
Section 
16.0 
2. International Council for Harmonisation of Technical Requirements for
Pharmaceuticals for Human Use (ICH). ICH harmonised guideline. Integrated
Addendum to ICH E6 (R1): Guideline for Good Clinical Practice E6 (R2).
Available at https://database.ich.org/sites/default/files/E6_R2_Addendum.pdf
[Accessed 06 December 2019 11 March 2020].
8. 6. World Health Organization, 2018. Dengue and severe dengue. Fact
Sheet. Available at https://www.who.int/en/news-room/fact-
sheets/detail/dengue-and-severe-dengue [Accessed 06 December 2019 11
March 2020].
7. World Health Organization, 1997. Dengue hemorrhagic fever: diagnosis,
treatment, prevention and control, 2nd Edition. Geneva. Available at
https://www.who.int/csr/resources/publications/dengue/Denguepublication/en/
[Accessed 06 December 2019 11 March 2020].
8. World Health Organization, 2009. Dengue guidelines for diagnosis,
treatment, prevention and control. Available at
http://www.who.int/tdr/publications/documents/dengue-diagnosis.pdf.
[Accessed 06 December 2019 11 March 2020].
13. World Health Organization. Weekly Epidemiological Record. 2018.
93:329-44. Available at http://www.who.int/wer [Accessed 06 December 2019
11 March 2020].
9. 17. Policy and communication bulletin – The clinical center. Guidelines
for limits of blood drawn for research purposes in the clinical center.
Manual transmittal sheet, no. M95-9 (rev.), 2009. Available at
https://irb.research.chop.edu/.../files/documents/g_nih_blooddraws
[Accessed 06 December 2019 11 March 2020].
10. 20. WHO Drug Dictionary. Available at
http://www.who.int/medicines/publications/druginformation/en/ [Accessed
06 December 2019 11 March 2020].
0093
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An Open-Label, Phase 2 Trial to Investigate the Humoral and Cell-Mediated Immune 
Responses and Safety of a Tetravalent Dengue Vaccine Candidate (TDV) Administered 
Subcutaneously in Flavivirus-Naïve and Dengue-Immune Healthy Adults 
Immunogenicity and Safety of TDV in Flavivirus-Naïve and Dengue-Immune Adults 
Sponsor: 
Takeda Vaccines, Inc. 
40 Landsdowne Street 
Cambridge, MA 02139 
USA 
Trial Identifier: 
DEN-210 
IND Number: 
014292 
EudraCT Number: 
Not Applicable 
Trial Vaccine 
Name: 
Takeda’s tetravalent dengue vaccine candidate (TDV) comprised of 
a molecularly characterized, attenuated dengue serotype 2 strain 
(TDV-2), a dengue serotypes 2/1 chimeric strain (TDV-1), a 
dengue serotypes 2/3 chimeric strain (TDV-3), and a dengue 
serotypes 2/4 chimeric strain (TDV-4). 
Takeda Approval 
Date: 
18 December 2020 
Version: 
Version 5.0 (supersedes Version 4.0) 
CONFIDENTIAL PROPERTY OF TAKEDA 
This document is a confidential communication of Takeda. Acceptance of this document constitutes the agreement 
by the recipient that no information contained herein will be published or disclosed without written authorization 
from Takeda except to the extent necessary to obtain informed consent from those persons to whom the 
investigational product may be administered. Furthermore, the information is only meant for review and 
compliance by the recipient, his or her staff, and applicable institutional review committees, and regulatory 
agencies to enable conduct of the trial. 
0002
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Trial No. DEN-210 
Page 2 of 85 
Protocol Version 5.0 
18 December 2020 
CONFIDENTIAL 
1.0 
ADMINISTRATIVE INFORMATION 
1.1 
Contacts 
Issue 
Contact 
Serious adverse event and 
pregnancy reporting 
IQVIA Integrated Safety Management Lifecycle 
Safety 
Fax: +1 919 800 0122 
Hotline: +91 80 71311011 
E-mail: TakedaDensafety@Quintiles.com
1.2 
Approval 
REPRESENTATIVES OF TAKEDA 
This trial will be conducted with the highest respect for the individual subjects in accordance 
with the requirements of this clinical trial protocol and in accordance with the following: 

The ethical principles that have their origin in the Declaration of Helsinki [1].

International Council for Harmonisation of Technical Requirements for Pharmaceuticals
for Human Use (ICH) E6 Good Clinical Practice (GCP): Consolidated Guideline [2].

All applicable laws and regulations, including, but not limited to those related to data
privacy and clinical trial disclosure.
SIGNATURES 
{See appended signature page} 
{See appended signature page} 
, PharmD, DPhil 
, Clinical Development 
Vaccine Business Unit 
Takeda Pharmaceuticals International AG 
Date 
, PhD, MSc PV 
Takeda Pharmaceuticals International AG 
Date 
{See appended signature page} 
{See appended signature page} 
, MSc 
Vaccines Statistics 
Global Statistics 
Takeda Pharmaceuticals International AG 
Date 
Vaccine Business Unit 
Takeda Vaccines, Inc. 
Date 
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Protocol Version 5.0 
18 December 2020 
CONFIDENTIAL 
INVESTIGATOR AGREEMENT 
I confirm that I have read and that I understand this protocol, the Investigator’s Brochure 
(IB), and any other product information provided by the sponsor. I agree to conduct this trial 
in accordance with the requirements of this protocol and protect the rights, safety, privacy, 
and well-being of trial subjects in accordance with the following: 

The ethical principles that have their origin in the Declaration of Helsinki [1].

ICH E6 GCP: Consolidated Guideline [2].

All applicable laws and regulations, including, but not limited to those related to data
privacy and clinical trial disclosure.

Regulatory requirements for reporting Serious Adverse Events (SAEs) defined in
Section 10.4.4 of this protocol.

Terms outlined in the Clinical Trial Site Agreement.

Appendix A – Responsibilities of the Investigator.
I further authorize that my personal information may be processed and transferred in 
accordance with the uses contemplated in Appendix B of this protocol. 
Signature of Investigator 
Date 
Investigator Name (print or type) 
Investigator’s Title 
Location of Facility (City, State) 
Location of Facility (Country) 
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Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
1.3 
Protocol Amendment Summary of Changes 
This document describes the changes in reference to the Protocol incorporating 
Amendment No. 4. 
1.3.1 Amendment History 
Date 
Amendment 
Number 
Amendment 
Type 
Region 
02 May 2018 
Initial Protocol 
Not applicable 
Local (United States of America) 
23 January 2019 
1 
Non-substantial 
Local (United States of America) 
06 December 2019 
2 
Substantial 
Local (United States of America) 
11 March 2020 
3 
Substantial 
Local (United States of America) 
18 December 2020 
4 
Substantial 
Local (United States of America) 
1.3.2 Summary of Changes 
Amendment to Protocol Version 4.0 dated 11 March 2020 
Rationale for the Amendment: 
This protocol amendment is to remove the planned interim analysis due to the short time 
period between the projected dates for the database lock for the interim analysis (March 
2021) and final analysis (August 2021). The interim analysis would have provided safety 
and immunogenicity data when all subjects have completed the Day 120 (M4) visit. Given 
the anticipated timing of the database lock for both analyses, it is considered that the 
interim analysis is no longer needed. 
Other modifications: 
 Addition of a clarification that, due to the COVID-19 pandemic, alternative monitoring 
approaches may be used. 
 Addition of a clarification that some of the test results may not be available prior to 
database lock because trial laboratory procedures are not carried out during the 
COVID-19 pandemic. 
 Following modifications were made for alignment with protocol template Version 4.0: 
- Update in responsibilities of the Signatory Investigator. 
- Addition of trial risk management. 
 Administrative change 
 Few minor editorial changes. 
New text is shown in bold italics and any deleted text is marked using strikethrough. 
 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-210 
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Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
Section 
Description of change 
Title page Takeda Approval Date: 11 March 18 December 2020 
Version: Version 4.0 5.0 (supersedes Version 3.0 4.0) 
1.2 
, MD 
, Clinical 
Development 
Vaccine Business Unit 
Takeda Pharmaceuticals USA, Inc. 
, PharmD, DPhil 
, Clinical Development 
Vaccine Business Unit 
Takeda Pharmaceuticals International 
AG 
1.3 
This document describes the changes in reference to the Protocol incorporating 
Amendment No. 3 4. 
2.0 
Interim Analysis: 
An interim analysis of immunogenicity and safety data is planned when all 
subjects have completed the Day 120 (M4) visit. 
No interim analysis is planned. 
2.1 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 
[Day 1 (M0) to Day 90 (M3)] and Visits 10 to 19 [Day 94 to 
Day 360 [M12]) 
Note: When a site visit cannot be carried out due to the COVID-19 
pandemic, telephone contacts will be made for subjects who are still under 
monitoring for safety reporting. 
3.2 
The sponsor will select a Signatory Principal Investigator from the 
investigators who participate in the trial. Selection criteria for this the 
Principal iInvestigator will include significant knowledge of the trial protocol, 
the investigational vaccine, their expertise in the therapeutic area and the 
conduct of clinical research as well as trial participation. Takeda will select a 
Signatory Investigator from the investigators who participate in the trial. 
The Signatory Principal Investigator will be required to review and sign the 
clinical protocol. The Signatory Investigator will also be required to review 
and sign the cClinical sStudy rReport (CSR) and by doing so agrees that it 
accurately describes the results of the trial. 
3.3 
CSR 
Clinical Study Report 
MHRA 
Medicines and Healthcare Products Regulatory Agency of the 
United Kingdom 
QTL 
Quality Tolerance Limits 
6.1 
Refer to Section 13.2 for the planned interim analysis. 
9.3.7 
Not applicable. 
When a site visit cannot be carried out due to the COVID-19 pandemic, 
telephone contacts will be made for subjects who are still under monitoring 
for safety reporting. Refer also to Section 14.1. 
12.1 
When a site visit cannot be carried out due to the COVID-19 pandemic, 
telephone contacts will be made for subjects for whom monitoring for safety 
reporting is still ongoing. Refer also to Section 14.1. 
0006
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Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
Section 
Description of change 
13.1 
A Statistical Analysis Plan (SAP) will be prepared and finalized prior to the 
planned interim analysis database lock. This document will provide further 
details regarding the definition of analysis variables and analysis methodology 
to address all trial objectives. 
Data reviews will be conducted prior to the planned interim analysis and final 
analysis database lock. These reviews will assess the accuracy and 
completeness of the trial database, and subject evaluability, and appropriateness 
of the planned statistical methods. 
As trial laboratory procedures are not carried out during the COVID-19 
pandemic there is a possibility that not all pre-defined test results will be 
available prior to database lock. Thus, there is a risk for database 
unlock/relock and additional data may be added in CSR amendments or 
addenda to the final CSR. 
13.2 
An interim analysis of immunogenicity and safety data is planned when all 
subjects have completed the Day 120 (M4) visit. 
No interim analysis is planned. 
14.0 
14.1 Trial-Site Monitoring Visits 
In the event a monitor cannot visit the site in a timely manner due to the 
COVID-19 pandemic, alternative monitoring approaches such as remote 
source data verification (SDV) or telephone contact may be used to ensure 
data quality and integrity and maintain subject safety. Alternative monitoring 
approaches should be used only where allowed by the local Health Authority 
and when approved by the IRB/IEC. During remote monitoring, the monitor 
should focus on trial activities that are essential to the safety of trial subjects 
and/or data reliability. 
14.4 Trial Risk Management 
The ICH E6 addendum (R2) guidance encourages a risk-based approach to 
the management of clinical trials and includes requirements for risk control 
and risk reporting. Takeda or designee established Quality Tolerance Limits 
(QTL) taking into consideration the medical and statistical characteristics of 
the variables and the statistical design of the trial. This process was 
performed according to Takeda internal procedures. 
At the end of the trial, the quality management approach implemented will be 
described in the CSR. If applicable, the CSR will summarize important 
deviations from the predefined QTL and the remedial actions taken. 
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Trial No. DEN-210 
Page 7 of 85 
Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
Section 
Description of change 
16.0 
2. International Council for Harmonisation of Technical Requirements for 
Pharmaceuticals for Human Use (ICH). ICH harmonised guideline. Integrated 
Addendum to ICH E6 (R1): Guideline for Good Clinical Practice E6 (R2). 
Available at https://database.ich.org/sites/default/files/E6_R2_Addendum.pdf. 
[Accessed 11 March 18 December 2020]. 
6. World Health Organization, 2018. Dengue and severe dengue. Fact Sheet. 
Available at https://www.who.int/en/news-room/fact-sheets/detail/dengue-and-
severe-dengue [Accessed 11 March 18 December 2020]. 
7. World Health Organization, 1997. Dengue hemorrhagic fever: diagnosis, 
treatment, prevention and control, 2nd Edition. Geneva. Available at 
https://www.who.int/csr/resources/publications/dengue/Denguepublication/en/ 
[Accessed 11 March 18 December 2020]. 
8. World Health Organization, 2009. Dengue guidelines for diagnosis, 
treatment, prevention and control. Available at 
http://www.who.int/tdr/publications/documents/dengue-diagnosis.pdf. 
[Accessed 11 March 18 December 2020]. 
13. World Health Organization. Weekly Epidemiological Record. 2018. 
93:329-44. Available at http://www.who.int/wer [Accessed 11 March 18 
December 2020]. 
17. Policy and communication bulletin – The clinical center. Guidelines for 
limits of blood drawn for research purposes in the clinical center. Manual 
transmittal sheet, no. M95-9 (rev.), 2009. Available at 
https://irb.research.chop.edu/.../files/documents/g_nih_blooddraws [Accessed 
11 March 18 December 2020]. 
20. WHO Drug Dictionary. Available at 
http://www.who.int/medicines/publications/druginformation/en/ [Accessed 11 
March 18 December 2020]. 
Appendix
 A 
13. Review and provide a signature as approval of the content of the clinical 
study report CSR 
0008
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Trial No. DEN-210 
Page 8 of 85 
Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
TABLE OF CONTENTS 
1.0 
ADMINISTRATIVE INFORMATION .................................................................. 2 
1.1 
Contacts.............................................................................................................. 2 
1.2 
Approval ............................................................................................................ 2 
1.3 
Protocol Amendment Summary of Changes...................................................... 4 
1.3.1 Amendment History ..................................................................................... 4 
1.3.2 Summary of Changes ................................................................................... 4 
2.0 
TRIAL SUMMARY .............................................................................................. 12 
2.1 
Schedule of Trial Procedures ........................................................................... 22 
3.0 
TRIAL REFERENCE INFORMATION ............................................................... 26 
3.1 
Trial-Related Responsibilities .......................................................................... 26 
3.2 
Principal Investigator ....................................................................................... 26 
3.3 
List of Abbreviations ....................................................................................... 27 
3.4 
Corporate Identification ................................................................................... 29 
4.0 
INTRODUCTION ................................................................................................. 30 
4.1 
Background ...................................................................................................... 30 
4.2 
Rationale for the Proposed Trial ...................................................................... 31 
5.0 
TRIAL OBJECTIVES AND ENDPOINTS .......................................................... 33 
5.1 
Objectives ........................................................................................................ 33 
5.1.1 Primary Objective ...................................................................................... 33 
5.1.2 Secondary Objectives................................................................................. 33 
5.1.3 Exploratory Objectives .............................................................................. 33 
5.2 
Endpoints ......................................................................................................... 34 
5.2.1 Primary Endpoint ....................................................................................... 34 
5.2.2 Secondary Endpoints ................................................................................. 34 
5.2.3 Exploratory Endpoints ............................................................................... 35 
6.0 
TRIAL DESIGN AND DESCRIPTION ............................................................... 36 
6.1 
Trial Design ..................................................................................................... 36 
6.2 
Justification for Trial Design, Dose, and Endpoints ........................................ 38 
6.3 
Planned Duration of Subject’s Expected Participation in the Entire Trial ...... 39 
6.4 
Premature Termination or Suspension of Trial or Investigational Site ........... 39 
6.4.1 Criteria for Premature Termination or Suspension of the Trial ................. 39 
6.4.2 Criteria for Premature Termination or Suspension of Investigational Sites
.................................................................................................................... 40 
6.4.3 Procedures for Premature Termination or Suspension of the Trial or the 
Participation of Investigational Site(s)....................................................... 40 
7.0 
SELECTION AND DISCONTINUATION/WITHDRAWAL OF SUBJECTS ... 41 
7.1 
Inclusion Criteria ............................................................................................. 41 
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7.2 
Exclusion Criteria ............................................................................................ 41 
7.3 
Criteria for Delay of Trial Vaccine Administration ......................................... 44 
7.4 
Early Termination of a Subject’s Trial Participation ....................................... 44 
7.5 
Criteria for Premature Discontinuation of Trial Vaccine Administration ....... 45 
8.0 
CLINICAL TRIAL MATERIAL MANAGEMENT ............................................. 47 
8.1 
Trial Vaccine and Materials ............................................................................. 47 
8.1.1 Dosage Form, Manufacturing, Packaging, and Labeling .......................... 47 
8.1.2 Storage ....................................................................................................... 47 
8.1.3 Dose and Regimen ..................................................................................... 48 
8.2 
Trial Vaccine Assignment and Dispensing Procedures ................................... 48 
8.2.1 Precautions to be Observed when Administering the Trial Vaccine ......... 48 
8.3 
Randomization Code Creation and Storage ..................................................... 49 
8.4 
Trial Vaccine Blind Maintenance .................................................................... 49 
8.5 
Unblinding Procedure ...................................................................................... 49 
8.6 
Accountability and Destruction of Sponsor-Supplied Trial Vaccine and Other 
Clinical Trial Materials .................................................................................... 49 
9.0 
TRIAL PLAN ........................................................................................................ 51 
9.1 
Trial Procedures ............................................................................................... 51 
9.1.1 Informed Consent....................................................................................... 51 
9.1.2 Demographics, Medical History and Prior Medications ............................ 51 
9.1.3 Documentation of Trial Entrance/Randomization ..................................... 52 
9.1.4 Physical Examination................................................................................. 52 
9.1.5 Vital Signs .................................................................................................. 53 
9.1.6 Screening.................................................................................................... 53 
9.1.7 Immunogenicity Assessments .................................................................... 53 
9.1.8 Processing, Labeling and Storage of Biological Samples ......................... 54 
9.1.9 Safety Assessments .................................................................................... 54 
9.1.10 Clinical Safety Laboratory Variables ......................................................... 54 
9.1.11 Contraception and Pregnancy Avoidance Procedure ................................. 54 
9.1.12 Pregnancy ................................................................................................... 55 
9.1.13 Documentation of Subjects Who are Not Randomized ............................. 55 
9.2 
Monitoring Subject Compliance ...................................................................... 55 
9.3 
Schedule of Observations and Procedures ....................................................... 55 
9.3.1 Procedures at the Screening Visit (up to 70 days [10 weeks] prior to Day 1 
[M0]) .......................................................................................................... 56 
9.3.2 Pre-Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) .................... 56 
9.3.3 Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) .......................... 57 
9.3.4 Post Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) .................. 57 
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9.3.5 Site Visits after Vaccination (Post First Vaccination: Days 4, 6, 9, 12, 15, 
30 [M1], and 60 [M2]; Post Second Vaccination: Days 94, 96, 99, 102, 
105, 120 [M4], 150 [M5], 180 [M6], 270 [M9], and 360 [M12]) ............. 59 
9.3.6 Phone Contacts - Reminder Calls .............................................................. 60 
9.3.7 Phone Contacts – Safety Call ..................................................................... 60 
9.3.8 Final (End of Trial) Visit ........................................................................... 60 
9.3.9 Post-Trial Care ........................................................................................... 60 
9.4 
Biological Sample Retention and Destruction ................................................. 60 
10.0 
ADVERSE EVENTS ............................................................................................. 61 
10.1 
Definitions........................................................................................................ 61 
10.1.1 Adverse Events .......................................................................................... 61 
10.1.2 Solicited Adverse Events ........................................................................... 61 
10.1.3 Adverse Events of Special Interest ............................................................ 63 
10.1.4 Medically Attended Adverse Events ......................................................... 63 
10.1.5 Serious Adverse Events ............................................................................. 63 
10.2 
Causality of Adverse Events ............................................................................ 63 
10.2.1 Relationship to Trial Procedures ................................................................ 64 
10.2.2 Outcome of Adverse Events ...................................................................... 64 
10.3 
Additional Points to Consider for Adverse Events .......................................... 64 
10.4 
Procedures ........................................................................................................ 65 
10.4.1 Collection and Reporting of Adverse Events............................................. 65 
10.4.2 Collection and Reporting of Solicited Adverse Events ............................. 66 
10.4.3 Collection and Reporting of Adverse Events of Special Interest/Medically 
Attended Adverse Events ........................................................................... 67 
10.4.4 Collection and Reporting of Serious Adverse Events................................ 67 
10.5 
Follow-up Procedures ...................................................................................... 67 
10.5.1 Adverse Events .......................................................................................... 67 
10.5.2 Serious Adverse Events ............................................................................. 67 
10.5.3 Safety Reporting to Investigators, Investigational Review Boards or 
Independent Ethics Committees, and Regulatory Authorities ................... 68 
10.5.4 Post-Trial Events ........................................................................................ 68 
11.0 
TRIAL-SPECIFIC REQUIREMENT(S) ............................................................... 69 
11.1 
Trial-Specific Committees ............................................................................... 69 
11.1.1 Data Monitoring Committee ...................................................................... 69 
12.0 
DATA HANDLING AND RECORD KEEPING ................................................. 70 
12.1 
CRFs (Electronic) ............................................................................................ 70 
12.2 
Record Retention ............................................................................................. 70 
13.0 
STATISTICAL METHODS .................................................................................. 72 
13.1 
Statistical and Analytical Plans ........................................................................ 72 
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13.1.1 Analysis Sets .............................................................................................. 72 
13.1.2 Analysis of Demographics and Other Baseline Characteristics ................ 72 
13.1.3 Immunogenicity Analysis .......................................................................... 73 
13.1.4 Safety Analysis .......................................................................................... 73 
13.2 
Interim Analysis and Criteria for Early Termination ....................................... 74 
13.3 
Determination of Sample Size ......................................................................... 74 
14.0 
QUALITY CONTROL AND QUALITY ASSURANCE ..................................... 75 
14.1 
Trial-Site Monitoring Visits ............................................................................. 75 
14.2 
Protocol Deviations .......................................................................................... 75 
14.3 
Quality Assurance Audits and Regulatory Agency Inspections ...................... 75 
14.4 
Trial Risk Management.................................................................................... 76 
15.0 
ETHICAL ASPECTS OF THE TRIAL ................................................................. 77 
15.1 
Institutional Review Board and/or Independent Ethics Committee Approval 77 
15.2 
Subject Information, Informed Consent, and Subject Authorization .............. 78 
15.3 
Subject Confidentiality .................................................................................... 79 
15.4 
Clinical Trial Registration, Publication and Disclosure Policy ....................... 79 
15.4.1 Clinical Trial Registration.......................................................................... 79 
15.4.2 Clinical Trial Results Disclosure ............................................................... 79 
15.4.3 Publication of Trial Results ....................................................................... 79 
15.5 
Insurance and Compensation for Injury ........................................................... 80 
16.0 
REFERENCES ...................................................................................................... 81 
 
LIST OF IN-TEXT TABLES 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to 
Day 90 (M3)] and Visits 10 to 19 [Day 94 to Day 360 [M12]) ................. 22 
Table 10.a 
Solicited Local (Injection Site) Reactions and Systemic Events ............... 62 
Table 10.b 
Severity Scales for Solicited Safety Parameters ........................................ 62 
 
LIST OF IN-TEXT FIGURES 
Figure 2.a 
Schematic of DEN-210 Trial Design ......................................................... 14 
Figure 6.a 
Schematic of Trial Design ......................................................................... 37 
 
LIST OF APPENDICES 
Appendix A 
Responsibilities of the Investigator ............................................................ 83 
Appendix B 
Investigator Consent to Use of Personal Information ................................ 85 
 
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2.0 
TRIAL SUMMARY 
Name of Sponsor: 
Takeda Vaccines, Inc. 
40 Landsdowne Street 
Cambridge, MA 02139 
USA 
Product Name: 
Takeda’s Tetravalent Dengue Vaccine Candidate (TDV) 
Trial Title: An Open-Label, Phase 2 Trial to Investigate the Humoral and Cell-Mediated Immune Responses 
and Safety of a Tetravalent Dengue Vaccine Candidate (TDV) Administered Subcutaneously in Flavivirus-
Naïve and Dengue-Immune Healthy Adults 
IND No.: 014292 
EudraCT No.: Not applicable 
Trial Identifier: DEN-210 
Phase: 2 
Blinding Schema: Open Label 
Indication: Prevention of dengue fever of any severity due to any serotype. 
Background and Rationale: 
Dengue fever is caused by infection with the wild type dengue virus (DENV), a ribonucleic acid virus that 
occurs as 4 recognized serotypes, DENV-1, DENV-2, DENV-3, or DENV-4. These 4 DENVs are 
transmitted to humans by mosquitoes (primarily Aedes aegypti), and are endemic in Asia, Central and South 
America, the Caribbean, the Pacific Islands, and parts of Africa. There are an estimated 390 million dengue 
infections per year worldwide, which is close to 4 times the previous World Health Organization (WHO) 
estimate of 50 to 100 million cases. Every year, around 500,000 cases of Dengue Hemorrhagic Fever (DHF) 
require hospitalization with an estimated death rate of 2.5%, primarily in children. It is estimated that 
3.9 billion people are at risk of dengue infection. 
Dengue fever is clinically defined as an acute febrile illness with 2 or more of the following manifestations: 
headache, retro-orbital pain, myalgia, arthralgia, rash, hemorrhagic manifestations, or leukopenia, and 
occurrence at the same location and time as other confirmed cases of dengue fever. The most severe forms of 
dengue infection – DHF and Dengue Shock Syndrome (DSS) – are life threatening. Primary infection with 
any one of the 4 dengue serotypes is thought to result in life-long protection from re-infection by the same 
serotype but does not protect against a secondary infection by one of the other 3 dengue serotypes which 
may lead to an increased risk of severe disease (DHF/DSS). 
Treatment of dengue fever is based solely on medical management of signs and symptoms, with fluid 
replacement required for hemorrhagic or shock cases. An antiviral therapy for DENV infection is not 
available at this time. Preventive measures that rely on mosquito control and individual protection are of 
limited efficacy, complex to implement and questionable in terms of cost-effectiveness. There is a great 
unmet global public health need for a safe and effective vaccine to reduce the morbidity and mortality 
associated with dengue disease. Vaccine development has focused on tetravalent vaccines that provide 
protection against all 4 dengue serotypes simultaneously since all 4 dengue serotypes commonly co-circulate 
in endemic areas. A first dengue vaccine (Chimeric Yellow fever virus Dengue virus-Tetravalent Dengue 
Vaccine [CYD-TDV]) has been approved (year 2015) in some countries in Asia and Latin America. Initial 
findings showed that vaccine efficacy was different between serotypes and depended on dengue pre-
exposure status. Additionally, recent analyses found that people who had not been infected by dengue virus 
before vaccination had a higher risk of getting severe disease when they were infected with dengue virus 
after vaccination with CYD-TDV. Hence, there is a continued unmet public health need for safer and more 
efficacious dengue vaccines. 
Takeda’s Tetravalent Dengue Vaccine Candidate (TDV) - Background: 
Takeda’s TDV consists of 1 molecularly characterized, attenuated dengue serotype 2 virus strain and 
3 chimeric recombinant dengue virus strains expressing surface antigens corresponding to DENV serotypes 
1– 4. The dengue serotype 2 strain (TDV-2) is based upon the attenuated laboratory-derived DENV-2 virus 
strain, originally isolated at Mahidol University, Bangkok, Thailand and generated by 53 serial passages in 
Primary Dog Kidney (PDK) cells (DENV-2 PDK-53). The chimeric, attenuated vaccine strains for dengue 
serotypes 1, 3 and 4 were engineered by substituting the structural genes, pre-Membrane (prM) and 
Envelope (E), of TDV-2 with the prM and E genes of the DENV virus strains, DENV-1 16007, 
DENV- 3 16562 or DENV-4 1036 virus, respectively. Thus, TDV is comprised of 4 dengue virus strains: 
a molecularly characterized, attenuated dengue serotype 2 strain (TDV-2), a dengue serotypes 2/1 chimeric 
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strain (TDV-1), a dengue serotypes 2/3 chimeric strain (TDV-3), and a dengue serotypes 2/4 chimeric strain 
(TDV-4). 
Nonclinical studies carried out in mice and nonhuman primates have demonstrated an acceptable safety, 
immunogenicity, and efficacy profile of Takeda’s TDV. Additionally, data from completed phase 1 and 
phase 2 clinical trials in humans have shown satisfactory reactogenicity, safety and immunogenicity profiles 
of Takeda’s TDV in healthy adults in nonendemic areas as well as in healthy adults and children in endemic 
areas in Asia and Latin America. Ongoing and completed phase 2 clinical trials have enabled the selection of 
a final TDV dose (lyophilized formulation), and a 2-dose vaccination regimen (2 single doses) administered 
3 months (ie, 90 days) apart by subcutaneous (SC) injection for use in the ongoing pivotal program. 
The current Investigator’s Brochure contains additional product information and a more detailed review of 
nonclinical studies and clinical trials. 
Rationale for the Proposed Trial: 
The purpose of this trial is to characterize the magnitude, quality, and evolution over time of humoral and 
cell-mediated immune responses to TDV when administered as a 2-dose vaccination regimen (2 single doses 
3 months [ie, 90 days] apart). Furthermore, this trial aims to characterize the relationships between 
TDV- specific innate, humoral and cellular immune responses. This characterization will aid in the 
understanding of vaccine efficacy and in the identification of potential determinants for protection against all 
4 dengue serotypes in the ongoing phase 3 TDV clinical program. This trial will be conducted at trial centers 
in the United States of America involving healthy adults who are either flavivirus-naïve or seropositive for 
DENV-1 or DENV-3 (ie, serology consistent with primary infection with either DENV-1 or DENV-3). The 
serological status of subjects will either have been determined by the trial center prior to and outside the 
scope of this trial or will be determined through screening within the scope of this trial. 
Although certain parameters of TDV-specific humoral and cell-mediated immune responses have been 
evaluated to some extent in phase 1 and other phase 2 clinical trials with TDV, the present trial is the only one 
that will allow a comprehensive analysis of TDV-specific humoral, T cell-mediated, B cell-mediated, innate, 
and early immune responses. In particular, these responses following 2 doses of TDV in both flavivirus-naïve 
and dengue-immune adults have not been evaluated yet in any other single trial in the TDV clinical program. 
Furthermore, conduct of the present trial would create a subject population, potentially eligible for inclusion 
in a future DENV-1/DENV-3 human challenge trial. Such a trial may be considered at a later date by the 
sponsor if deemed appropriate to further guide TDV development. 
The present trial will be conducted in accordance with the protocol, International Council for Harmonization 
of Technical Requirements for Pharmaceuticals for Human Use (ICH) and Good Clinical Practice (GCP) 
Guidelines, and applicable regulatory requirements. 
Trial Design: 
This is an open-label, phase 2 trial in 44 healthy adult subjects aged 18 to 60 years (inclusive) to investigate 
the immunogenicity and safety of SC administration of a 2-dose regimen of TDV. 
Subjects will be enrolled in 2 trial groups based on results from serological testing performed either by the 
trial center prior to and outside the scope of this trial or through screening within the scope of this trial (up to 
70 days [10 weeks] prior to Day 1 [Month 0 (M0)]): 
- 
Group 1: approximately 22 flavivirus-naïve subjects. 
- 
Group 2: approximately 22 dengue-immune subjects (ie, subjects with serology consistent with 
primary infection by DENV-1 and subjects with serology consistent with primary infection by 
DENV-3). 
Note: The actual number of subjects in Group 2 will be lower than the planned 22 subjects as it was decided 
to stop the recruitment of subjects in Group 2. 
TDV will be administered on Day 1 (M0) and on Day 90 (Month 3 [M3]) in both Groups 1 and 2. 
The blood sampling schedule is summarized in Figure 2.a. In compliance with the National Institutes of 
Health guidance, the total amount of blood that will be drawn from trial participants over an 8-week period 
will not exceed 10.5 mL/kg or 550 mL (whichever is smaller). 
All subjects will be followed-up for 9 months post second vaccination so the trial duration will be 
approximately 360 days (12 months) or 14.5 months for each subject depending on whether serological 
testing with regard to the inclusion criteria for the trial is performed outside or within the scope of this trial, 
respectively. The trial schedule is presented in Figure 2.a. 
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Figure 2.a 
Schematic of DEN-210 Trial Design 
 
Immunogenicity evaluations: 
 
Dengue neutralizing antibodies will be measured using blood samples collected from all subjects 
pre first vaccination (Day 1 [M0]) and on Days 15, 30 (Month 1 [M1]), and 60 (Month 2 [M2]); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (Month 4 [M4]), 150 (Month 5 [M5]), 
180 (Month 6 [M6]), 270 (Month 9 [M9]), and 360 (Month 12 [M12]). 
 
T cell-mediated immune response will be measured using blood samples collected from all subjects 
pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 
B cell-mediated immune response will be measured using blood samples collected from all subjects 
pre first vaccination (Day 1 [M0]) and on Day 30 (M1); pre second vaccination (Day 90 [M3]) and on 
Days 105, 120 (M4), 180 (M6), 270 (M9), and 360 (M12). 
 
T cell epitopes will be mapped using blood samples collected from all subjects pre first vaccination 
(Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on 
Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 
Anti-dengue non-structural protein 1 (NS1) antibodies will be measured using blood samples collected 
from all subjects pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); pre second 
vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 
360 (M12). 
 
The innate immune response will be measured using blood samples collected from all subjects pre first 
vaccination (Day 1 [M0]) and on Days 4 and 6; pre second vaccination (Day 90 [M3]), and on Days 94 
and 96. 
Safety evaluations: 
 
Diary cards (paper or electronic) will be distributed for the recording of: 
- 
Solicited Adverse Events (AEs): 
- 
Solicited local (injection site) reactions for 7 days following administration of each trial 
vaccine dose on Day 1 (M0) and Day 90 (M3) (day of administration + 6 days). These will 
include: injection site pain, injection site erythema, and injection site swelling. 
- 
Solicited systemic events for 14 days following administration of each trial vaccine dose on 
Day 1 (M0) and Day 90 (M3) (day of administration + 13 days). These will include: fever, 
headache, asthenia, malaise, and myalgia. 
- 
Unsolicited AEs for 28 days following administration of each trial vaccine dose on Day 1 (M0) and 
Day 90 (M3) (day of administration + 27 days). 
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 
Serious Adverse Events (SAEs), Medically Attended Adverse Events (MAAEs), and AEs leading to 
trial vaccine withdrawal or trial discontinuation will be collected for the trial duration. MAAEs are 
defined as AEs leading to an unscheduled medical visit to or by a healthcare professional including 
visits to an emergency department, but not fulfilling seriousness criteria. 
Vaccine viremia evaluation: 
 
Vaccine viremia will be measured using blood samples collected from all subjects pre first vaccination 
(Day 1 [M0]) and on Days 6, 9, 12, 15, and 30 (M1); pre second vaccination (Day 90 [M3]) and on 
Days 96, 99, 102, 105, and 120 (M4). 
Data collection will be by electronic Case Report Form (eCRF). 
Primary Objective: 
 
To assess the neutralizing antibody response (Geometric Mean Titers [GMT]) against each dengue 
serotype post-vaccination (Microneutralization Test 50% [MNT50]). 
Secondary Objectives: 
Immunogenicity 
 
To assess the magnitude (Interferon-gamma Enzyme-Linked Immunospot [IFN-γ ELISpot]) and 
polyfunctionality (Intracellular Cytokine Staining [ICS]) of the T cell-mediated immune response 
post-vaccination. 
 
To assess vaccine viremia post-vaccination and the integrity of main attenuation mutations. 
Safety 
 
To describe the safety of 2 doses of TDV in healthy subjects aged 18 to 60 years (inclusive). 
Exploratory Objectives: 
 
To assess the neutralizing antibody response against each dengue serotype post-vaccination (dengue 
Reporter Virus Particle [RVP] test). 
 
To assess the B cell-mediated immune response post-vaccination (Quad-color FluoroSpot). 
 
To assess the anti-dengue NS1 antibody response post-vaccination (Enzyme-Linked ImmunoSorbent 
Assay [ELISA]). 
 
To map the epitopes of the dengue antibodies post-vaccination (IFN-γ ELISpot response). 
 
To assess the innate immune response post-vaccination. 
 
To characterize the relationships between TDV-specific innate, humoral and cellular immune responses 
on an individual level post-vaccination (MNT50, dengue RVP, Quad color FluoroSpot, NS1 antibody 
ELISA, IFN- γ ELISpot, ICS, and exploratory assays). 
 
To characterize the relationships between vaccine viremia (quantitative Reverse Transcription-
Polymerase Chain Reaction [qRT-PCR]) and TDV-specific innate, humoral and cellular immune 
responses on an individual level post-vaccination (MNT50, dengue RVP, Quad color FluoroSpot, 
NS1 antibody ELISA, IFN-γ ELISpot, ICS, and exploratory assays). 
Subject Population: 
Healthy Subjects: yes 
Age Range: ≥18 to ≤60 years 
Planned Number of Subjects: 44 
Planned Number of Trial Arms: 2 trial groups (22 subjects per trial group); 2-dose regimen (1 trial vaccine 
dose on Day 1 [M0] and 1 trial vaccine dose on Day 90 [M3]), SC route. 
Estimated Total: 44 enrolled subjects. 
Note: The actual number of subjects in Group 2 will be lower than the planned 22 subjects as it was decided 
to stop the recruitment of subjects in Group 2. 
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Key Inclusion Criteria: 
1. The subject is aged ≥18 to ≤60 years. 
2. Male or female. 
3. Subjects who are in good health at the time of entry into the trial as determined by medical history, 
physical examination (including vital signs) and clinical judgment of the investigator. 
4. Group 1 only: immunologically naïve to dengue, Zika, Yellow Fever (YF), Japanese Encephalitis (JE), 
West Nile (WN) (based on negative results for detection of anti-DENV, anti-Zika, anti-YF, anti-JE, 
anti- WN antibodies) as documented by serological testing performed either by the trial center prior to 
and outside the scope of this trial or through screening within the scope of this trial (up to 70 days 
[10 weeks] prior to Day 1 [M0]). 
5. Group 2 only: serology consistent with primary infection with either DENV-1 or DENV-3 (defined as 
detectable neutralizing antibodies against DENV-1 or DENV-3 only, or titers for DENV-1 or 
DENV- 3 ≥4-times higher than titers for the 2 other dengue serotypes) as documented by serological 
testing performed either by the trial center prior to and outside the scope of this trial or through 
screening within the scope of this trial (up to 70 days [10 weeks] prior to Day 1 [M0]). 
6. The subject signs and dates a written, informed consent and any required privacy authorization prior to 
the initiation of any trial procedures, after the nature of the trial has been explained according to local 
regulatory requirements. 
7. Subjects who can comply with trial procedures and are available for the duration of follow-up. 
Key Exclusion Criteria: 
1. Subjects with a clinically active significant infection (as assessed by the investigator) or body 
temperature ≥38°C (100.4°F) within 3 days of the intended date of vaccination (consider whether 
applicable as an exclusion criterion or criterion for delay of trial vaccine administration, see below). 
2. Subjects with a known hypersensitivity or allergy to any of the trial vaccine components (including 
excipients). 
3. Subjects with behavioral or cognitive impairment or psychiatric disease that, in the opinion of the 
investigator, may interfere with the subject’s ability to participate in the trial. 
4. Subjects with any history of progressive or severe neurologic disorder, seizure disorder or neuro-
inflammatory disease (eg, Guillain-Barré syndrome). 
5. Subjects with any illness, or history of any illness that, in the opinion of the investigator, might interfere 
with the results of the trial or pose additional risk to the subjects due to participation in the trial. 
6. Known or suspected impairment/alteration of immune function (consider whether applicable as an 
exclusion criterion or criterion for delay of trial vaccine administration, see below), including: 
a) Chronic use of oral steroids (equivalent to 20 mg/day prednisone ≥12 weeks and/or ≥2 mg/kg body 
weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0) (use of inhaled, intranasal, or 
topical corticosteroids is allowed). 
b) Receipt of parenteral steroids (equivalent to 20 mg/day prednisone ≥12 weeks and/or ≥2 mg/kg 
body weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0). 
c) Administration of immunoglobulins and/or any blood products within 3 months prior to Day 1 (M0) 
or planned administration during the trial. 
d) Receipt of immunostimulants within 60 days prior to Day 1 (M0). 
e) Immunosuppressive therapy such as anti-cancer chemotherapy or radiation therapy within 6 months 
prior to Day 1 (M0). 
f) 
Known Human Immunodeficiency Virus (HIV) infection or HIV-related disease. 
g) Hepatitis C virus infection. 
h) Genetic immunodeficiency. 
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7. Abnormalities of splenic or thymic function. 
8. Subjects with a known bleeding diathesis, or any condition that may be associated with a prolonged 
bleeding time. 
9. Subjects with any serious chronic or progressive disease according to the judgment of the investigator 
(eg, neoplasm, hematologic malignancies, insulin dependent diabetes; cardiac, renal, or hepatic disease). 
10. Subjects with Body Mass Index (BMI) greater than or equal to 35 kg/m2 (=weight in kg/ height in 
meters2). 
11. Subjects participating in any clinical trial with another investigational product 30 days prior to 
Day 1 (M0) or intending to participate in another clinical trial at any time during the conduct of this 
trial. 
12. Subjects who have received any other vaccines within 14 days (for inactivated vaccines) or 28 days (for 
live vaccines) prior to enrollment in this trial or who are planning to receive any vaccine within 28 days 
of trial vaccine administration (consider whether applicable as an exclusion criterion or criterion for 
delay of trial vaccine administration, see below). 
13. Subjects who have used antipyretics and/or analgesic medications within 24 hours prior to vaccination. 
The reason for their use (prophylaxis versus treatment) must be documented. Trial entry should be 
delayed to allow for a full 24 hours to have passed since last use of antipyretics and/or analgesic 
medications (consider whether applicable as an exclusion criterion or criterion for delay of trial vaccine 
administration, see below). 
14. Subjects involved in the trial conduct or their first-degree relatives. 
15. Subjects with history of substance or alcohol abuse within the past 2 years. 
16. Female subjects who are pregnant or breastfeeding. 
17. Female subjects of childbearing potential1 who are sexually active with men and have not used any of 
the “acceptable contraceptive methods” 2 for at least 2 months prior to Day 1 (M0). 
18. Female subjects of childbearing potential who are sexually active with men and refuse to use an 
acceptable contraceptive method up to 6 weeks after the last dose of trial vaccine 
(Day 90 [M3] + 6 weeks). In addition, female subjects must be advised not to donate ova during this 
period. 
19. Any positive or indeterminate pregnancy test. 
20. Previous participation in any clinical trial of a dengue candidate vaccine, except for subjects who 
received placebo in those trials, or previous and planned vaccination (during the trial conduct) against 
dengue. 
21. Planned vaccination (during the trial conduct) against any non-dengue flavivirus (eg, Zika, YF, JE, WN, 
tick-borne encephalitis, or Murray-Valley encephalitis). 
22. Planned travel (during the trial conduct) to any area endemic for dengue. 
There may be instances when individuals meet all entry criteria except one that relates to transient clinical 
circumstances (eg, body temperature elevation or recent use of excluded medication[s] or vaccine[s]). Under 
these circumstances, eligibility for trial enrollment may be considered if the appropriate window for delay 
has passed, inclusion/exclusion criteria have been rechecked, and if the subject is confirmed to be eligible. 
                                                 
1 Defined as status post onset of menarche and not meeting any of the following conditions: menopausal for at 
least 2 years, status after bilateral tubal ligation for at least 1 year, status after bilateral oophorectomy for at least 
1 year or status after hysterectomy. 
2 One or more of the following: hormonal contraceptives (such as oral, injection, transdermal patch, implant, 
cervical ring), barrier method (condom with spermicide or diaphragm with spermicide) every time during 
intercourse, intrauterine device, monogamous relationship with vasectomized partner (partner must have been 
vasectomized for at least 6 months prior to the subject’s enrollment [Day 1 (M0)]). Other contraceptive methods 
may be considered in agreement with the sponsor and will be approved by the appropriate ethics committee. 
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CONFIDENTIAL 
Criteria for delay of vaccination: 
After enrollment, subjects may encounter clinical circumstances that warrant a delay in the administration of 
trial vaccine. These situations are listed below. In the event that a subject meets a criterion for delay of trial 
vaccine administration, the subject may receive the trial vaccine once the window for delay has passed as 
long as the subject is otherwise eligible for trial participation. 
If any of the conditions below occur at the time scheduled for the second TDV administration 
(Day 90 [M3]), the second dose may be administered at a later date as long as the subject is otherwise 
eligible to continue trial participation. In certain situations, the period of delay may lead to deviation from 
the time window for the second dose at Day 90 (M3). The decision to vaccinate in those situations will be 
made by the investigator. 
The following clinical circumstances warrant a delay for administration of the vaccination (consider whether 
applicable as criterion for delay of trial vaccine administration or an exclusion criterion, see above): 
 
Subjects with a clinically active significant infection (as assessed by the investigator) or body 
temperature ≥38°C (100.4°F) within 3 days of the intended date of vaccination. 
 
Subjects who have received any other vaccines within 14 days (for inactivated vaccines) or 28 days (for 
live vaccines) prior to enrollment in this trial or who are planning to receive any vaccine within 28 days 
of trial vaccine administration. 
 
Known or suspected altered or impaired immune function as specified under the exclusion criteria. 
 
Subjects who have used antipyretics and/or analgesic medications within 24 hours prior to vaccination. 
The reason for their use (prophylaxis versus treatment) must be documented. Trial vaccine 
administration should be delayed to allow for a full 24 hours to have passed between having used 
antipyretics and/or analgesic medications and trial vaccine administration. 
Criteria for contraindication to vaccination at Day 90 (M3): 
There are also circumstances under which receipt of the second trial vaccine dose at Day 90 (M3) is 
contraindicated in this trial. These circumstances include but are not limited to anaphylaxis or severe 
hypersensitivity reactions following the first TDV vaccination at Day 1 (M0). If these reactions occur, the 
subject must not receive the TDV vaccination at Day 90 (M3) but will be encouraged to continue trial 
participation for safety follow-up. 
Trial Vaccine: 
The investigational vaccine is Takeda’s TDV, a tetravalent dengue vaccine comprised of 1 molecularly 
characterized, attenuated dengue virus strain, and 3 chimeric dengue virus strains with potencies of not less 
than 3.3, 2.7, 4.0, and 4.5 log10 plaque forming units per dose of TDV-1, TDV-2, TDV-3, and TDV-4, 
respectively. 
Route of administration: SC route 
Duration of the Trial and Subject Participation: 
The trial duration for each subject will be at least approximately 360 days (12 months) including vaccination 
(Day 1 [M0] and Day 90 [M3]) and follow-up through Day 360 (M12). This duration will be approximately 
14.5 months if serological testing with regard to the inclusion criteria for the trial is performed within the 
scope of this trial (ie, up to 70 days [10 weeks] prior to Day 1 [M0]). 
Criteria for Evaluation and Analyses: 
Primary Endpoint: 
 
GMT of neutralizing antibodies (by MNT50) for each of the 4 dengue serotypes using blood samples 
collected from all subjects post first vaccination on Days 15, 30 (M1), and 60 (M2); pre second 
vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
Secondary Endpoints: 
 
Frequency (percentage of subjects) and magnitude (number of Spot Forming Cells [SFC]/106 Peripheral 
Blood Mononuclear Cells [PBMC]) of IFN-γ ELISpot responses to TDV using blood samples collected 
from all subjects post first vaccination on Days 15, 30 (M1) and 60 (M2); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). Cellular 
immune response is defined as an IFN-γ ELISpot response that is >3 times higher compared with 
background (no peptide) and ≥50 spots per 106 PBMC. 
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CONFIDENTIAL 
 
Phenotype characteristics of cellular immune responses to TDV by ICS using blood samples collected 
post first vaccination on Days 15, 30 (M1) and 60 (M2); pre second vaccination (Day 90 [M3]) and on 
Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). Markers will include Cluster of 
Differentiation (CD)4, CD8, IFN-γ, tumor necrosis factor-alpha and interleukin-2. This endpoint will be 
evaluated in a subset of subjects with IFN- γ ELISpot responses >500 SFC/106 cells and availability of 
sufficient cells. 
 
Incidence, duration, and level of vaccine viremia for each of the 4 dengue serotypes measured by 
qRT- PCR using blood samples collected from all subjects post first vaccination on Days 6, 9, 12, 15, 
and 30 (M1); pre second vaccination (Day 90 [M3]) and on Days 96, 99, 102, 105, and 120 (M4). 
 
Frequency and percentage of subjects with solicited local (injection site) reactions for 7 days (day of 
administration + 6 days) and solicited systemic events for 14 days (day of administration + 13 days) 
following administration of each trial vaccine dose (Day 1 [M0] and Day 90 [M3]). 
 
Frequency and percentage of subjects with any unsolicited AEs for 28 days (day of administration + 
27 days) following administration of each trial vaccine dose (Day 1 [M0] and Day 90 [M3]). 
 
Frequency and percentage of subjects with SAEs throughout the trial. 
 
Frequency and percentage of subjects with MAAEs throughout the trial. 
Exploratory Endpoints: 
 
Average Effective Concentration 50 ([EC50], dengue RVP) of neutralizing antibodies for each of the 
4 dengue serotypes using blood samples collected from all subjects post first vaccination on Days 15, 
30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 
180 (M6), 270 (M9), and 360 (M12). 
 
Seropositivity rates (percentage of subjects) from dengue RVP for each of the 4 dengue serotypes using 
blood samples collected from all subjects post first vaccination on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 
360 (M12). 
 
Seropositivity rates (percentage of subjects) from dengue RVP for multiple (2, 3 or 4) dengue serotypes 
using blood samples collected from all subjects post first vaccination on Days 15, 30 (M1), and 
60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). 
 
Average number of memory B cells expressing type-specific and cross-reactive dengue-specific 
antibodies/106 PBMC measured by Quad-color FluoroSpot using blood samples collected from all 
subjects post first vaccination on Day 30 (M1); pre second vaccination (Day 90 [M3]) and on Days 105, 
120 (M4), 180 (M6), 270 (M9), and 360 (M12). 
 
Average concentration (relative units/mL) of anti-dengue NS1 antibodies for each of the 4 dengue 
serotypes measured by ELISA using blood samples collected from all subjects post first vaccination on 
Days 15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 
150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 
Epitope mapping of the IFN-γ ELISpot responses to TDV on an individual level using blood samples 
collected from all subjects post first vaccination on Days 15, 30 (M1), and 60 (M2); pre second 
vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
This endpoint will be evaluated in a subset of subjects with IFN- γ ELISpot responses >500 SFC/106 
cells and availability of sufficient cells. 
 
Gene expression profiles on an individual level using blood samples collected from all subjects post first 
vaccination on Days 4 and 6; pre second vaccination (Day 90 [M3]) and on Days 94 and 96. 
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Protocol Version 5.0 
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CONFIDENTIAL 
Statistical Considerations: 
Summaries will be provided by trial group (Group 1 and Group 2) and further for Group 2 by subgroups 
(DENV- 1 or DENV-3 seropositive). Unless specified otherwise, number of subjects with non-missing 
observations, mean, SD, median, minimum and maximum will be presented for continuous data. Frequency 
and percent will be presented for categorical data. 
Analysis Sets 
Safety Set: The Safety Set will consist of all subjects who received at least 1 dose of trial vaccine. 
Full Analysis Set (FAS): The FAS will include all subjects who received at least 1 dose of trial vaccine and 
for whom a valid pre-dose and at least one valid post-dose blood sample is taken. 
Per–Protocol Set (PPS): The PPS will include all subjects in the FAS who have no major protocol 
violations. The major protocol violation criteria will be defined as part of the data review prior to database 
lock. The categories of major protocol violations include: (1) not meeting selected entry criteria, 
(2) receiving prohibited therapies, (3) not receiving 2 doses of TDV, or receiving the second vaccination 
inadmissibly outside of the visit window, and (4) other major protocol violations that may be identified 
during data review prior to database lock. 
Analysis of Demographics and Other Baseline Characteristics 
Age, gender, race, and other Baseline characteristics will be summarized descriptively. 
Immunogenicity Analysis 
For the primary immunogenicity endpoint, descriptive statistics and 95% CIs will be provided for each 
applicable visit. Summaries will be provided by trial group (Group 1 and Group 2) and further for Group 2 
by subgroups (DENV-1 or DENV-3 seropositive). The primary immunogenicity analyses will be based on 
the PPS; supportive analyses may be provided based on the FAS. 
Similar descriptive analyses as for the primary immunogenicity endpoint will be provided for the secondary 
and exploratory endpoints for each applicable assay at all relevant time points, based on the PPS. Supportive 
analyses based on the FAS may also be provided for selected endpoints. 
Antibody titers will be analyzed under log-normal distribution assumption. GMT will be calculated for each 
relevant time point as anti-logarithm of arithmetic mean of natural log transformed titers. The 95% CI for 
GMT will be calculated as the anti-log transformation of upper and lower limits for a 2-sided CI of the mean 
of the log-transformed titers (based on Student’s t-distribution). 
Other quantitative data will be summarized descriptively on the original scale. 
Seropositivity rates will be accompanied with 95% CIs calculated by exact (Clopper-Pearson) method. 
Relationship between immunogenicity measures will be assessed using non-parametric correlation 
coefficient, as appropriate. Graphical methods will also be explored. Further details will be provided in the 
Statistical Analysis Plan (SAP). 
Handling of missing data will be described in the SAP. 
Safety Analysis 
All safety data will be summarized descriptively using the Safety Set, by trial group (Group 1 and Group 2) 
and further for Group 2 by subgroups (DENV-1 or DENV-3 seropositive). 
Solicited AEs 
For each solicited AE, the number and percentage of subjects with local (injection site) reactions and 
systemic events will be summarized by event severity for each day after each vaccination (ie, Day 1 [M0] 
through Day 7 for local [injection site] reactions and Day 1 [M0] through Day 14 for systemic events), and 
overall. Summaries of first onset of each event and the number of days subjects reported experiencing each 
event will also be provided. For subjects with more than 1 episode of the same event, the maximum severity 
will be used for tabulations. 
Persistent/prolonged solicited local (injection site) reactions or systemic events continuing on Day 8 or 
Day 15, respectively, following each trial vaccination will be assessed separately. Unless otherwise 
specified, these events will not be included in the analyses/tabulations of unsolicited AEs and will have 
separate listings. 
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Protocol Version 5.0 
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CONFIDENTIAL 
Unsolicited AEs 
Unsolicited AEs will be coded using the Medical Dictionary for Regulatory Activities (MedDRA), and 
summarized by System Organ Class (SOC) and Preferred Term (PT). Unsolicited AEs will be summarized 
for 28 days following each vaccination (day of vaccination + 27 days) as follows: by PT including events 
with frequency greater than a pre-defined threshold (the percentage will be specified in the SAP); by SOC 
and PT; by SOC, PT, and severity; and by SOC, PT, and relationship to TDV. Subjects reporting more than 
1 occurrence for the term (level) being summarized will be counted only once. 
AEs leading to trial vaccine withdrawal or trial discontinuation will be collected and summarized for the 
entire trial period. 
SAEs and MAAEs 
SAEs and MAAEs will be coded using MedDRA, and summarized by SOC and PT for the entire trial 
period. 
Other Safety Observations 
Vaccine viremia (incidence, duration, and level) will be evaluated descriptively at all applicable time points. 
Relationship between vaccine viremia and immunogenicity measures will be assessed using non-parametric 
correlation coefficient, as appropriate. Graphical methods will also be explored. Further details will be 
provided in the SAP. 
Sample Size Justification: 
This trial is designed for a descriptive evaluation of the endpoints that does not require hypotheses testing. 
Therefore, the sample size was not determined based on formal statistical power calculations. 
Interim Analysis: 
No interim analysis is planned. 
Data Monitoring Committee (DMC): 
A DMC will have oversight of this trial. The DMC functions at a program level and further information is 
available in the DMC Charter. 
DEN-210 Version 5.0 (18 December 2020) 
 
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Protocol Version 5.0 
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CONFIDENTIAL 
2.1 
Schedule of Trial Procedures 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to Day 90 (M3)] and Visits 10 to 19 [Day 94 to 
Day 360 [M12]) 
 
Screening visit (a) 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
V8 
V9 
 
 
D1 
M0 
D4 
D6 
D9 
D12 
D15 
D30 
M1 
D60 
M2 
D90 
M3 
Visit window (days) 
Up to 70 days 
(10 weeks) prior to 
D1 (M0) 
1 day 
(±NA) 
4 days 
(±1) 
after V1 
6 days 
(±1) 
after V1 
9 days 
(±1) 
after V1 
12 days 
(±1) 
after V1 
15 days 
(-1/+2) 
after V1 
30 days 
(-1/+7) 
after V1 
60 days 
(±5) 
after V1 
90 days 
(-4/+7) 
after V1 
Informed consent 
X 
X (b) 
 
 
 
 
 
 
 
 
Assessment of eligibility criteria (c) 
X 
X 
 
 
 
 
 
 
 
 
Trial group assignment (flavivirus-naïve or dengue-
immune) (d) 
 
X 
 
 
 
 
 
 
 
 
Demographics 
X 
X (e) 
 
 
 
 
 
 
 
 
Medical history 
X 
X 
 
 
 
 
 
 
 
 
Prior medication/vaccination 
X 
X 
 
 
 
 
 
 
 
 
Concomitant medications/vaccinations (f) 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Review of systems 
 
X 
 
 
 
 
 
 
 
X 
Complete physical examination (g) 
X 
X 
 
 
 
 
 
 
 
X 
Targeted physical examination (h) 
 
 
 
 
 
 
 
X 
 
 
Vital signs (i) 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Pregnancy test (j) 
X 
X 
 
 
 
 
 
 
 
X 
Pregnancy avoidance guidance (k) 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Blood sampling for serological testing (2.5 mL) 
X 
 
 
 
 
 
 
 
 
 
Blood sample for humoral immune response 
(20 mL) (l, m) 
 
X 
 
 
 
 
X 
X 
X 
X 
Footnotes are on last table page. 
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Protocol Version 5.0 
18 December 2020 
 
CONFIDENTIAL 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to Day 90 (M3)] and Visits 10 to 19 [Day 94 to 
Day 360 [M12]) (continued) 
 
Screening visit (a) 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
V8 
V9 
 
 
D1 
M0 
D4 
D6 
D9 
D12 
D15 
D30 
M1 
D60 
M2 
D90 
M3 
Visit window (days) 
Up to 70 days 
(10 weeks) prior to 
D1 (M0) 
1 day 
(±NA) 
4 days 
(±1) 
after V1 
6 days 
(±1) 
after V1 
9 days 
(±1) 
after V1 
12 days 
(±1) 
after V1 
15 days 
(-1/+2) 
after V1 
30 days 
(-1/+7) 
after V1 
60 days 
(±5) 
after V1 
90 days 
(-4/+7) 
after V1 
Blood sample for T cell-mediated immune response 
(60 mL) (l) 
 
X 
 
 
 
 
X 
X 
X 
X 
Blood sample for B cell-mediated immune response 
(30 mL) (l) 
 
X 
 
 
 
 
 
X 
 
X 
Blood sample for innate immune response (5 mL) (l) 
 
X 
X 
X 
 
 
 
 
 
X 
Blood sample for vaccine viremia (5 mL) (l) 
 
X 
 
X 
X 
X 
X 
X 
 
X 
Check criteria for delay of trial vaccine 
administration 
 
X 
 
 
 
 
 
 
 
X 
Check contraindications for trial vaccine 
administration 
 
X 
 
 
 
 
 
 
 
X 
Trial vaccine administration (n) 
 
X 
 
 
 
 
 
 
 
X 
Injection site evaluation (o) 
 
X 
 
 
 
 
 
 
 
X 
Diary card (p) 
Distribution 
 
X 
 
 
 
 
 
 
 
X 
Review/collection of solicited and 
unsolicited AEs 
 
 
X 
X 
X 
X 
X 
X 
 
 
AEs leading to trial vaccine withdrawal or trial 
discontinuation, SAEs, MAAEs (q) 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Footnotes are on last table page. 
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Protocol Version 5.0 
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CONFIDENTIAL 
Table 2.a 
Schedule of Trial Procedures (Screening Visit, Visits 1 to 9 [Day 1 (M0) to Day 90 (M3)] and Visits 10 to 19 [Day 94 to 
Day 360 [M12]) (continued) 
 
V10 
V11 
V12 
V13 
V14 
V15 
V16 
V17 
V18 
V19 
 
D94 
D96 
D99 
D102 
D105 
D120 
M4 
D150 
M5 
D180 
M6 
D270 
M9 
D360 (r) 
M12 
Visit window (days) 
4 days 
(±1) 
after V9 
6 days  
(±1) 
after V9 
9 days  
(±1) 
after V9 
12 days 
(±1) 
after V9 
15 days 
(-1/+2) 
after V9 
30 days  
(-1/+7) 
after V9 
60 days 
(±5) 
after V9 
90 days 
(±7) 
after V9 
180 days  
(-7/+14) 
after V9 
270 days 
(±14) 
after V9 
Concomitant medications/vaccinations (e) 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Targeted physical examination (g) 
 
 
 
 
 
X 
 
 
 
X 
Vital signs (h) 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Pregnancy avoidance guidance (j) 
X 
X 
X 
X 
X 
X 
 
 
 
 
Blood sample for humoral immune response 
(20 mL) (l) 
 
 
 
 
X 
X 
X 
X 
X 
X 
Blood sample for T cell-mediated immune response 
(60 mL) 
 
 
 
 
X 
X 
X 
X 
X 
X 
Blood sample for B cell-mediated immune response 
(30 mL) 
 
 
 
 
X 
X 
 
X 
X 
X 
Blood sample for innate immune response (5 mL) 
X 
X 
 
 
 
 
 
 
 
 
Blood sample for vaccine viremia (5 mL) 
 
X 
X 
X 
X 
X 
 
 
 
 
Diary card (p) 
Review/collection of solicited and 
unsolicited AEs 
X 
X 
X 
X 
X 
X 
 
 
 
 
AEs leading to trial vaccine withdrawal or trial 
discontinuation, SAEs, MAAEs (q) 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
AEs=Adverse Events, D=Day, M=Month, MAAEs=Medically Attended Adverse Events, NA=Not Applicable, SAEs= Serious Adverse Events, V=Visit 
Note: When a site visit cannot be carried out due to the COVID-19 pandemic, telephone contacts will be made for subjects who are still under monitoring for safety reporting. 
(a) A screening visit is only applicable if serological testing with regard to the inclusion criteria for the trial is performed within the scope of this trial. 
(b) Up to 28 days prior to Day 1 (Month 0 [M0]). 
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CONFIDENTIAL 
(c) After informed consent has been obtained, eligibility of the subject will be assessed by review of inclusion/exclusion criteria at Screening or prior to TDV administration on 
Day 1 (M0), as applicable. 
(d) Subjects will be enrolled in Group 1 (flavivirus-naïve subjects) or Group 2 (dengue-immune subjects with serology consistent with primary infection with either wild type dengue 
virus [DENV]-1 or DENV-3) based on serological testing performed either by the trial center prior to and outside the scope of this trial or within the scope of this trial (up to 
70 days [10 weeks] prior to Day 1 [M0]). 
(e) Not applicable if a Screening visit has been performed. 
(f) All medications and vaccine history from 1 month (minimum 28 days) prior to administration of each trial vaccine dose up to 1 month (minimum 28 days) thereafter, steroids and 
immunostimulants within 60 days prior to Day 1 (M0), immunoglobulins and blood products within 3 months prior to Day 1 (M0), and immunosuppressive therapy within 
6 months prior to Day 1 (M0). Concomitant medication/vaccination will be collected throughout the trial. 
(g) Physical examination including measurement of weight and height; Body Mass Index (BMI) will be calculated. Measurement of height is only required at Screening or at 
Day 1 (M0), as applicable. 
(h) Subjects may undergo a targeted symptom-directed physical examination. Clinically significant changes from the Baseline examination should be recorded in the subject’s source 
documents and electronic Case Report Form (eCRF). 
(i) Vital signs including (but not limited to) the measurement of systolic blood pressure/diastolic blood pressure, heart rate, and body temperature. 
(j) Pregnancy testing (serum or urine) for females of childbearing potential. Results must be confirmed and documented as negative prior to each trial dose administration. Additional 
pregnancy tests may be performed during the trial if deemed necessary by the investigator. 
(k) Females of childbearing potential who are sexually active will be reminded during trial visits to adhere to acceptable contraceptive methods up to 6 weeks after the last dose of 
TDV (Day 90 [Month 3] (M3]) + 6 weeks). Subjects will be provided with information on acceptable methods of contraception as part of the subject informed consent process 
and will be asked to sign a consent form stating that they understand the requirements for avoidance of pregnancy and donation of ova. During the course of the trial, subjects of 
childbearing potential will receive continued guidance with respect to the avoidance of pregnancy. 
(l) All blood samples on days of vaccination (Day 1 [M0] and Day 90 [M3]) should be taken prior to administration of TDV. 
(m) Dengue neutralizing antibodies and anti-dengue non-structural protein 1 (NS1) antibodies for all subjects, and T cell epitope mapping in a subset of subjects with Interferon-
gamma Enzyme-Linked Immunospot (IFN-γ ELISpot) responses >500 Spot Forming Cells/106 cells and availability of sufficient cells). 
(n) Subjects will receive TDV by subcutaneous injection. 
(o) Injection site pain, erythema, and swelling assessed by trial staff for 30 minutes post-vaccination. 
(p) Diary cards (paper or electronic) will be distributed for the recording of 1) solicited AEs including solicited local (injection site) reactions for 7 days (day of administration + 
6 days) and solicited systemic events for 14 days (day of administration + 13 days) following administration of each trial vaccine dose, and 2) unsolicited AE for 28 days (day of 
administration + 27 days) following administration of each trial vaccine dose. The investigator will categorize events by severity (mild, moderate or severe) and will assess 
causality to vaccine administration (“related” or “not related”). For persistent/prolonged solicited local (injection site) reactions or systemic events observed as continuing on 
Day 8 or 15, respectively, following each trial vaccination, the end date will be captured on the “Adverse Event” eCRF to permit a separate analysis from the unsolicited AEs. 
Any solicited local (injection site) reaction or systemic event that resolves before 8 or 15 days, respectively, following each trial vaccination, but recurs at a later time (ie, if 
discontinues), should be recorded as an unsolicited AE on the “Adverse Event” eCRF. 
(q) MAAEs and SAEs will be collected for the trial duration. 
(r) The Final Visit will be performed on Day 360 (Month 12 [M12]). If a subject terminates trial participation earlier, Day 360 (M12) procedures should be performed at their last 
trial visit, if possible. 
 
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CONFIDENTIAL 
3.0 
TRIAL REFERENCE INFORMATION 
3.1 
Trial-Related Responsibilities 
The sponsor will perform all trial-related activities with the exception of those identified in the 
Trial-Related Responsibilities template. The vendors identified in the template for specific 
trial-related activities will perform these activities in full or in partnership with the sponsor. 
3.2 
Principal Investigator 
Selection criteria for the Principal Investigator will include significant knowledge of the trial 
protocol, the investigational vaccine, their expertise in the therapeutic area and the conduct of 
clinical research as well as trial participation. Takeda will select a Signatory Investigator from 
the investigators who participate in the trial. The Signatory Investigator will be required to 
review and sign the clinical protocol. The Signatory Investigator will also be required to review 
and sign the Clinical Study Report (CSR) and by doing so agrees that it accurately describes the 
results of the trial. 
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3.3 
List of Abbreviations 
AE(s) 
Adverse Event(s) 
BMI 
Body Mass Index 
CD 
Cluster of Differentiation 
CFR 
Code of Federal Regulations 
CSR 
Clinical Study Report 
CYD-TDV 
Chimeric Yellow fever virus Dengue virus-Tetravalent Dengue 
Vaccine 
DENV 
Wild type dengue virus 
DENV-1, -2, -3, -4 
Wild type dengue virus serotypes 1, 2, 3, and 4 
DHF 
Dengue Hemorrhagic Fever 
DMC 
Data Monitoring Committee 
DSS 
Dengue Shock Syndrome 
E 
Envelope 
EC50 
Effective Concentration 50 
eCRF 
electronic Case Report Form 
ELISA 
Enzyme-Linked ImmunoSorbent Assay 
FAS 
Full Analysis Set 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
GMT 
Geometric Mean Titers 
HIV 
Human Immunodeficiency Virus 
IB 
Investigator’s Brochure 
ICH 
International Council for Harmonization of Technical Requirements 
for Pharmaceuticals for Human Use 
ICS 
Intracellular Cytokine Staining 
IEC 
Independent Ethics Committee 
IFN-γ 
Interferon-gamma 
IFN-γ ELISpot 
Interferon-gamma Enzyme-Linked Immunospot 
Inc 
Incorporated 
IND 
Investigational New Drug 
IRB 
Institutional Review Board 
JE 
Japanese Encephalitis 
M0, 1, 2, 3, 4, 5, 6, 9, 12 
Month 0, 1, 2, 3, 4, 5, 6, 9, 12 
MAAEs 
Medically Attended Adverse Events 
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MedDRA 
Medical Dictionary for Regulatory Activities 
MHRA 
Medicines and Healthcare Products Regulatory Agency of the United 
Kingdom 
MNT50 
Microneutralization Test 50% 
NS1 
Non-structural protein 1 
PBMC 
Peripheral Blood Mononuclear Cells 
PDK 
Primary Dog Kidney 
PMDA 
Pharmaceuticals and Medical Devices Agency of Japan 
PPS 
Per-Protocol Set 
prM 
pre-Membrane 
PT 
Preferred Term 
qRT-PCR 
quantitative Reverse Transcription-Polymerase Chain Reaction 
QTL 
Quality Tolerance Limits 
RVP 
Reporter Virus Particle 
SAE(s) 
Serious Adverse Event(s) 
SAP 
Statistical Analysis Plan 
SC 
Subcutaneous 
SFC 
Spot Forming Cells 
SOC 
System Organ Class 
SOP 
Standard Operating Procedures 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TDV 
Takeda’s Tetravalent Dengue Vaccine Candidate 
Tetravalent Dengue Vaccine Candidate 
TDV-1 
Dengue serotypes 2/1 chimeric strain 
TDV-2 
Molecularly characterized, attenuated dengue serotype 2 strain 
TDV-3 
Dengue serotypes 2/3 chimeric strain 
TDV-4 
Dengue serotypes 2/4 chimeric strain 
US(A) 
United States (of America) 
WHO 
World Health Organization 
WN 
West Nile 
YF 
Yellow Fever 
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3.4 
Corporate Identification 
TV 
Takeda Vaccines, Inc. 
 
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4.0 
INTRODUCTION 
4.1 
Background 
Dengue fever is caused by infection with the wild type dengue virus (DENV), a ribonucleic acid 
virus that occurs as 4 recognized serotypes, DENV-1, DENV-2, DENV-3, or DENV-4. These 
4 DENVs are transmitted to humans by mosquitoes (primarily Aedes aegypti), and are endemic 
in Asia, Central and South America, the Caribbean, the Pacific Islands, and parts of Africa. 
There are an estimated 390 million dengue infections per year worldwide, which is close to 
4 times the previous World Health Organization (WHO) estimate of 50 to 100 million cases. 
Every year, around 500,000 cases of Dengue Hemorrhagic Fever (DHF) require hospitalization 
with an estimated death rate of 2.5%, primarily in children. It is estimated that 3.9 billion people 
are at risk of dengue infection [3-6]. 
Dengue fever is clinically defined as an acute febrile illness with 2 or more of the following 
manifestations: headache, retro-orbital pain, myalgia, arthralgia, rash, hemorrhagic 
manifestations, or leukopenia, and occurrence at the same location and time as other confirmed 
cases of dengue fever. The most severe forms of dengue infection – DHF and Dengue Shock 
Syndrome (DSS) – are life threatening. Primary infection with any one of the 4 dengue serotypes 
is thought to result in life-long protection from re-infection by the same serotype but does not 
protect against a secondary infection by one of the other 3 dengue serotypes which may lead to 
an increased risk of severe disease (DHF/DSS) [5-8]. 
Treatment of dengue fever is based solely on medical management of signs and symptoms, with 
fluid replacement required for hemorrhagic or shock cases. An antiviral therapy for DENV 
infection is not available at this time. Preventive measures that rely on mosquito control and 
individual protection are of limited efficacy, complex to implement and questionable in terms of 
cost-effectiveness. There is a great unmet global public health need for a safe and effective 
vaccine to reduce the morbidity and mortality associated with dengue disease. Vaccine 
development has focused on tetravalent vaccines that provide protection against all 4 dengue 
serotypes simultaneously since all 4 dengue serotypes commonly co-circulate in endemic 
areas [3-9]. A first dengue vaccine (Chimeric Yellow fever virus Dengue virus-Tetravalent 
Dengue Vaccine [CYD-TDV]) has been approved (year 2015) in some countries in Asia and 
Latin America [10]. Initial findings showed that vaccine efficacy was different between 
serotypes and depended on dengue pre-exposure status [11]. Additionally, recent analyses found 
that people who had not been infected by dengue virus before vaccination had a higher risk of 
getting severe disease when they were infected with dengue virus after vaccination with 
CYD- TDV [12]. Hence, there is a continued unmet public health need for safer and more 
efficacious dengue vaccines [13]. 
Takeda’s Tetravalent Dengue Vaccine Candidate (TDV) - Background: 
Takeda’s TDV consists of 1 molecularly characterized, attenuated dengue serotype 2 virus strain 
and 3 chimeric recombinant dengue virus strains expressing surface antigens corresponding to 
DENV serotypes 1–4. The dengue serotype 2 strain (TDV-2) is based upon the attenuated 
laboratory-derived DENV-2 virus strain, originally isolated at Mahidol University, Bangkok, 
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Thailand and generated by 53 serial passages in Primary Dog Kidney (PDK) cells 
(DENV- 2 PDK- 53) [14]. The chimeric, attenuated vaccine strains for dengue serotypes 1, 3 
and 4 were engineered by substituting the structural genes, pre-Membrane (prM) and 
Envelope (E), of TDV-2 with the prM and E genes of the DENV virus strains, DENV-1 16007, 
DENV- 3 16562 or DENV-4 1036 virus, respectively [15, 16]. Thus, TDV is comprised of 
4 dengue virus strains: a molecularly characterized, attenuated dengue serotype 2 strain (TDV-2), 
a dengue serotypes 2/1 chimeric strain (TDV-1), a dengue serotypes 2/3 chimeric strain 
(TDV-3), and a dengue serotypes 2/4 chimeric strain (TDV-4). 
Nonclinical studies carried out in mice and nonhuman primates have demonstrated an acceptable 
safety, immunogenicity, and efficacy profile of Takeda’s TDV. Additionally, data from 
completed phase 1 and phase 2 clinical trials in humans have shown satisfactory reactogenicity, 
safety and immunogenicity profiles of Takeda’s TDV in healthy adults in nonendemic areas as 
well as in healthy adults and children in endemic areas in Asia and Latin America. Ongoing and 
completed phase 2 clinical trials have enabled the selection of a final TDV dose (lyophilized 
formulation), and a 2-dose vaccination regimen (2 single doses) administered 3 months 
(ie, 90 days) apart by subcutaneous (SC) injection for use in the ongoing pivotal program. 
The current IB contains additional product information and a more detailed review of nonclinical 
studies and clinical trials. 
4.2 
Rationale for the Proposed Trial 
The purpose of this trial is to characterize the magnitude, quality, and evolution over time of 
humoral and cell-mediated immune responses to TDV when administered as a 2-dose 
vaccination regimen (2 single doses 3 months [ie, 90 days] apart). Furthermore, this trial aims to 
characterize the relationships between TDV-specific innate, humoral and cellular immune 
responses. This characterization will aid in the understanding of vaccine efficacy and in the 
identification of potential determinants for protection against all 4 dengue serotypes in the 
ongoing phase 3 TDV clinical program. This trial will be conducted at trial centers in the United 
States of America involving healthy adults who are either flavivirus-naïve or seropositive for 
DENV-1 or DENV-3 (ie, serology consistent with primary infection with either DENV-1 or 
DENV-3). The serological status of subjects will either have been determined by the trial center 
prior to and outside the scope of this trial or will be determined through screening within the 
scope of this trial. 
Although certain parameters of TDV-specific humoral and cell-mediated immune responses have 
been evaluated to some extent in phase 1 and other phase 2 clinical trials with TDV, the present 
trial is the only one that will allow a comprehensive analysis of TDV-specific humoral, T cell-
mediated, B cell-mediated, innate, and early immune responses. In particular, these responses 
following 2 doses of TDV in both flavivirus-naïve and dengue-immune adults have not been 
evaluated yet in any other single trial in the TDV clinical program. Furthermore, conduct of the 
present trial would create a subject population, potentially eligible for inclusion in a future 
DENV-1/DENV-3 human challenge trial. Such a trial may be considered at a later date by the 
sponsor if deemed appropriate to further guide TDV development. 
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The present trial will be conducted in accordance with the protocol, ICH and GCP Guidelines, 
and applicable regulatory requirements [2]. 
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5.0 
TRIAL OBJECTIVES AND ENDPOINTS 
5.1 
Objectives 
5.1.1 Primary Objective 
 To assess the neutralizing antibody response (Geometric Mean Titers [GMT]) against each 
dengue serotype post-vaccination (Microneutralization Test 50% [MNT50]). 
5.1.2 Secondary Objectives 
Immunogenicity 
 To assess the magnitude (Interferon-gamma Enzyme-Linked Immunospot [IFN-γ ELISpot]) 
and polyfunctionality (Intracellular Cytokine Staining [ICS]) of the T cell-mediated immune 
response post-vaccination. 
 To assess vaccine viremia post-vaccination and the integrity of main attenuation mutations. 
Safety 
 To describe the safety of 2 doses of TDV in healthy subjects aged 18 to 60 years (inclusive). 
5.1.3 Exploratory Objectives 
 To assess the neutralizing antibody response against each dengue serotype post-vaccination 
(Reporter Virus Particle [RVP] test). 
 To assess the B cell-mediated immune response post-vaccination (Quad-color FluoroSpot). 
 To assess the anti-dengue non-structural protein 1 (NS1) antibody response post-vaccination 
(Enzyme-Linked ImmunoSorbent Assay [ELISA]). 
 To map the epitopes of the dengue antibodies post-vaccination (IFN-γ ELISpot response). 
 To assess the innate immune response post-vaccination. 
 To characterize the relationships between TDV-specific innate, humoral and cellular immune 
responses on an individual level post-vaccination (MNT50, dengue RVP, Quad color 
FluoroSpot, NS1 antibody ELISA, IFN- γ ELISpot, ICS, and exploratory assays). 
 To characterize the relationships between vaccine viremia (quantitative Reverse 
Transcription-Polymerase Chain Reaction [qRT-PCR]) and TDV-specific innate, humoral 
and cellular immune responses on an individual level post-vaccination (MNT50, dengue RVP, 
Quad color FluoroSpot, NS1 antibody ELISA, IFN-γ ELISpot, ICS, and exploratory assays). 
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5.2 
Endpoints 
5.2.1 Primary Endpoint 
 GMT of neutralizing antibodies (by MNT50) for each of the 4 dengue serotypes using blood 
samples collected from all subjects post first vaccination on Days 15, 30 (Month 1 [M1]), 
and 60 (Month 2 [M2]); pre second vaccination (Day 90 [Month 3, (M3)]) and on Days 105, 
120 (Month 4 [M4]), 150 (Month 5 [M5]), 180 (Month 6 [M6]), 270 (Month 9 [M9]), and 
360 (Month 12 [M12]). 
5.2.2 Secondary Endpoints 
 Frequency (percentage of subjects) and magnitude (number of Spot Forming Cells [SFC]/106 
Peripheral Blood Mononuclear Cells [PBMC]) of IFN-γ ELISpot responses to TDV using 
blood samples collected from all subjects post first vaccination on Days 15, 30 (M1) and 
60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 
180 (M6), 270 (M9), and 360 (M12). Cellular immune response is defined as an 
IFN- γ ELISpot response that is >3 times higher compared with background (no peptide) and 
≥50 spots per 106 PBMC. 
 Phenotype characteristics of cellular immune responses to TDV by ICS using blood samples 
collected post first vaccination on Days 15, 30 (M1) and 60 (M2); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
Markers will include Cluster of Differentiation (CD)4, CD8, IFN-γ, tumor necrosis 
factor-alpha and interleukin-2. This endpoint will be evaluated in a subset of subjects with 
IFN- γ ELISpot responses >500 SFC/106 cells and availability of sufficient cells. 
 Incidence, duration, and level of vaccine viremia for each of the 4 dengue serotypes 
measured by qRT-PCR using blood samples collected from all subjects post first vaccination 
on Days 6, 9, 12, 15, and 30 (M1); pre second vaccination (Day 90 [M3]) and on Days 96, 
99, 102, 105, and 120 (M4). 
 Frequency and percentage of subjects with solicited local (injection site) reactions for 7 days 
(day of administration + 6 days) and solicited systemic events for 14 days (day of 
administration + 13 days) following administration of each trial vaccine dose (Day 1 [M0] 
and Day 90 [M3]). 
 Frequency and percentage of subjects with any unsolicited Adverse Events (AEs) for 28 days 
(day of administration + 27 days) following administration of each trial vaccine dose 
(Day 1 [M0] and Day 90 [M3]). 
 Frequency and percentage of subjects with SAEs throughout the trial. 
 Frequency and percentage of subjects with Medically Attended Adverse Events (MAAEs) 
throughout the trial. 
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5.2.3 Exploratory Endpoints 
 Average effective concentration 50 ([EC50], dengue RVP) of neutralizing antibodies for each 
of the 4 dengue serotypes using blood samples collected from all subjects post first 
vaccination on Days 15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on 
Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 Seropositivity rates (percentage of subjects) from dengue RVP for each of the 4 dengue 
serotypes using blood samples collected from all subjects post first vaccination on Days 15, 
30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 
150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 Seropositivity rates (percentage of subjects) from dengue RVP for multiple (2, 3 or 4) dengue 
serotypes using blood samples collected from all subjects post first vaccination on Days 15, 
30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 
150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 Average number of memory B cells expressing type-specific and cross-reactive dengue-
specific antibodies/106 PBMC measured by Quad-color FluoroSpot using blood samples 
collected from all subjects post first vaccination on Day 30 (M1); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 180 (M6), 270 (M9), and 360 (M12). 
 Average concentration (relative units/mL) of anti-dengue NS1 antibodies for each of the 
4 dengue serotypes measured by ELISA using blood samples collected from all subjects post 
first vaccination on Days 15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) 
and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 Epitope mapping of the IFN-γ ELISpot responses to TDV on an individual level using blood 
samples collected from all subjects post first vaccination on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). This endpoint will be evaluated in a subset of subjects with 
IFN- γ ELISpot responses >500 SFC/106 cells and availability of sufficient cells. 
 Gene expression profiles on an individual level using blood samples collected from all 
subjects post first vaccination on Days 4 and 6; pre second vaccination (Day 90 [M3]) and on 
Days 94 and 96. 
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6.0 
TRIAL DESIGN AND DESCRIPTION 
6.1 
Trial Design 
This is an open-label, phase 2 trial in 44 healthy adult subjects aged 18 to 60 years (inclusive) to 
investigate the immunogenicity and safety of SC administration of a 2-dose regimen of TDV. 
Subjects will be enrolled in 2 trial groups based on results from serological testing performed 
either by the trial center prior to and outside the scope of this trial or through screening within 
the scope of this trial (up to 70 days [10 weeks] prior to Day 1[M0]): 
- Group 1: approximately 22 flavivirus-naïve subjects. 
- Group 2: approximately 22 dengue-immune subjects (ie, subjects with serology 
consistent with primary infection by DENV-1 and subjects with serology consistent with 
primary infection by DENV-3). 
Note: The actual number of subjects in Group 2 will be lower than the planned 22 subjects as it 
was decided to stop the recruitment of subjects in Group 2. 
TDV will be administered on Day 1 (M0) and on Day 90 (M3) in both Groups 1 and 2. 
The blood sampling schedule is summarized in Figure 6.a and Table 2.a. In compliance with the 
National Institutes of Health guidance, the total amount of blood that will be drawn from trial 
participants over an 8-week period will not exceed 10.5 mL/kg or 550 mL (whichever is smaller) 
[17]. 
All subjects will be followed-up for 9 months post second vaccination so the trial duration will 
be approximately 360 days (12 months) or 14.5 months for each subject depending on whether 
serological testing with regard to the inclusion criteria for the trial is performed outside or within 
the scope of this trial, respectively. 
A schematic of the trial design is included as Figure 6.a. A schedule of trial procedures is 
provided in Section 2.1. 
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Figure 6.a 
Schematic of Trial Design 
 
Immunogenicity evaluations (see also Section 9.1.7): 
 Dengue neutralizing antibodies will be measured using blood samples collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). 
 T cell-mediated immune response will be measured using blood samples collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). 
 B cell-mediated immune response will be measured using blood samples collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Day 30 (M1); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 180 (M6), 270 (M9), and 360 (M12). 
 T cell epitopes will be mapped using blood samples collected from all subjects pre first 
vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); pre second vaccination 
(Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 Anti-dengue NS1 antibodies will be measured using blood samples collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). 
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 The innate immune response will be measured using blood samples collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Days 4 and 6; pre second vaccination 
(Day 90 [M3]) and on Days 94 and 96. 
Safety evaluations: 
 Diary cards (paper or electronic) will be distributed for the recording of: 
- Solicited AEs: 
- Solicited local (injection site) reactions for 7 days following administration of each 
trial vaccine dose on Day 1 (M0) and Day 90 (M3) (day of administration + 6 days). 
These will include: injection site pain, injection site erythema, and injection site 
swelling. 
- Solicited systemic events for 14 days following administration of each trial vaccine 
dose on Day 1 (M0) and Day 90 (M3) (day of administration + 13 days). These will 
include: fever, headache, asthenia, malaise, and myalgia. 
- Unsolicited AEs for 28 days following administration of each trial vaccine dose on 
Day 1 (M0) and Day 90 (M3) (day of administration + 27 days). 
 SAEs, MAAEs, and AEs leading to trial vaccine withdrawal or trial discontinuation will be 
collected for the trial duration. MAAEs are defined as AEs leading to an unscheduled 
medical visit to or by a healthcare professional including visits to an emergency department, 
but not fulfilling seriousness criteria. 
Vaccine viremia evaluation: 
 Vaccine viremia will be measured using blood samples collected from all subjects pre first 
vaccination (Day 1 [M0]) and on Days 6, 9, 12, 15, and 30 (M1); pre second vaccination 
(Day 90 [M3]) and on Days 96, 99, 102, 105, and 120 (M4). 
Data collection will be by electronic Case Report Form (eCRF). 
6.2 
Justification for Trial Design, Dose, and Endpoints 
The trial design and the collection of solicited AEs (local [injection site] reactions and systemic 
events), unsolicited AEs (non-serious AEs and SAEs), and MAAEs following trial dose 
administration are consistent with vaccine evaluation trials. 
Ongoing and completed phase 2 trials have enabled the selection of a final TDV dose 
(lyophilized formulation) and a 2-dose vaccination regimen 3 months apart by SC injection for 
use in Takeda’s dengue pivotal study program. 
The timing of the primary and secondary endpoints after vaccination is consistent with previous 
trials with TDV. Dengue neutralizing antibodies have been generally accepted as the immune 
response endpoint for dengue vaccine trials. 
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The blood sampling schedule and exploratory analyses will allow a comprehensive analysis of 
the kinetics (onset, longevity) of the various immune responses. 
Considering that the measurements of the immune responses are unlikely to be influenced by the 
knowledge of the trial group assignment, a placebo group for comparison has not been included 
in this trial. 
Involvement of healthy adults who are either flavivirus-naïve or seropositive for DENV-1 or 
DENV-3 in the present trial could create a subject population eligible for participation in a 
potential future DENV-1/DENV-3 human challenge trial if the sponsor deems this necessary in 
the further development of the trial vaccine. 
Justification of the sample size is included in Section 13.3. The rationale for the proposed trial is 
given in Section 4.2. 
The current IB contains additional product information and a more detailed review of nonclinical 
studies and clinical trials. 
6.3 
Planned Duration of Subject’s Expected Participation in the Entire Trial 
The trial duration for each subject will be at least approximately 360 days (12 months) including 
vaccination (Day 1 [M0] and Day 90 [M3]) and follow-up through Day 360 (M12). This duration 
will be approximately 14.5 months if serological testing with regard to the inclusion criteria for 
the trial is performed within the scope of this trial (ie, up to 70 days [10 weeks] prior to 
Day 1 [M0]). 
6.4 
Premature Termination or Suspension of Trial or Investigational Site 
6.4.1 Criteria for Premature Termination or Suspension of the Trial 
The trial will be completed as planned unless one or more of the following criteria that require 
temporary suspension or early termination of the trial are satisfied. 
 New information or other evaluation regarding the safety or efficacy of the investigational 
vaccine that indicates a change in the known risk/benefit profile, such that the risk/benefit is 
no longer acceptable for subjects participating in the trial. 
 The Data Monitoring Committee (DMC) recommends that the trial should be suspended or 
terminated. 
 Significant deviation from GCP that compromises the ability to achieve the primary trial 
objectives or compromises subject safety. 
 The sponsor decides to terminate or suspend the trial. 
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6.4.2 Criteria for Premature Termination or Suspension of Investigational Sites 
A trial site may be terminated prematurely or suspended if the site (including the investigator) is 
found in significant deviation from GCP, protocol, or contractual agreement, is unable to ensure 
adequate performance of the trial, or as otherwise permitted by the contractual agreement. 
6.4.3 Procedures for Premature Termination or Suspension of the Trial or the 
Participation of Investigational Site(s) 
In the event that the sponsor, an Institutional Review Board (IRB)/Independent Ethics 
Committee (IEC) or regulatory authority elects to terminate or suspend the trial or the 
participation of the investigational site, a trial-specific procedure for early termination or 
suspension will be provided by the sponsor; the procedure will be followed by the investigational 
site during the course of termination or trial suspension. 
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7.0 
SELECTION AND DISCONTINUATION/WITHDRAWAL OF SUBJECTS 
All entry criteria, including test results, need to be confirmed prior to first vaccination. 
7.1 
Inclusion Criteria 
Subject eligibility is determined according to the following criteria: 
1. The subject is aged ≥18 to ≤60 years. 
2. Male or female. 
3. Subjects who are in good health at the time of entry into the trial as determined by medical 
history, physical examination (including vital signs) and clinical judgment of the 
investigator. 
4. Group 1 only: immunologically naïve to dengue, Zika, Yellow Fever (YF), Japanese 
Encephalitis (JE), West Nile (WN) (based on negative results for detection of anti-DENV, 
anti-Zika, anti-YF, anti-JE, anti-WN antibodies) as documented by serological testing 
performed either by the trial center prior to and outside the scope of this trial or through 
screening within the scope of this trial (up to 70 days [10 weeks] prior to Day 1 [M0]). 
5. Group 2 only: serology consistent with primary infection with either DENV-1 or DENV-3 
(defined as detectable neutralizing antibodies against DENV-1 or DENV-3 only, or titers for 
DENV-1 or DENV-3 ≥4-times higher than titers for the 2 other dengue serotypes) as 
documented by serological testing performed either by the trial center prior to and outside the 
scope of this trial or through screening within the scope of this trial (up to 70 days [10 weeks] 
prior to Day 1 [M0]). 
6. The subject signs and dates a written, informed consent and any required privacy 
authorization prior to the initiation of any trial procedures, after the nature of the trial has 
been explained according to local regulatory requirements. 
7. Subjects who can comply with trial procedures and are available for the duration of 
follow-up. 
7.2 
Exclusion Criteria 
Any subject who meets any of the following criteria will not qualify for entry into the trial: 
1. Subjects with a clinically active significant infection (as assessed by the investigator) or body 
temperature ≥38°C (100.4°F) within 3 days of the intended date of vaccination (consider 
whether applicable as an exclusion criterion or criterion for delay of trial vaccine 
administration, see Section 7.3). 
2. Subjects with a known hypersensitivity or allergy to any of the trial vaccine components 
(including excipients). 
3. Subjects with behavioral or cognitive impairment or psychiatric disease that, in the opinion 
of the investigator, may interfere with the subject’s ability to participate in the trial. 
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4. Subjects with any history of progressive or severe neurologic disorder, seizure disorder or 
neuro-inflammatory disease (eg, Guillain-Barré syndrome). 
5. Subjects with any illness, or history of any illness that, in the opinion of the investigator, 
might interfere with the results of the trial or pose additional risk to the subjects due to 
participation in the trial. 
6. Known or suspected impairment/alteration of immune function (consider whether applicable 
as an exclusion criterion or criterion for delay of trial vaccine administration, see 
Section 7.3), including: 
a) Chronic use of oral steroids (equivalent to 20 mg/day prednisone ≥12 weeks and/or 
≥2 mg/kg body weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0) 
(use of inhaled, intranasal, or topical corticosteroids is allowed). 
b) Receipt of parenteral steroids (equivalent to 20 mg/day prednisone ≥12 weeks and/or 
≥2 mg/kg body weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0). 
c) Administration of immunoglobulins and/or any blood products within 3 months prior to 
Day 1 (M0) or planned administration during the trial. 
d) Receipt of immunostimulants within 60 days prior to Day 1 (M0). 
e) Immunosuppressive therapy such as anti-cancer chemotherapy or radiation therapy 
within 6 months prior to Day 1 (M0). 
f) Known Human Immunodeficiency Virus (HIV) infection or HIV-related disease. 
g) Hepatitis C virus infection. 
h) Genetic immunodeficiency. 
7. Abnormalities of splenic or thymic function. 
8. Subjects with a known bleeding diathesis, or any condition that may be associated with a 
prolonged bleeding time. 
9. Subjects with any serious chronic or progressive disease according to the judgment of the 
investigator (eg, neoplasm, hematologic malignancies, insulin dependent diabetes; cardiac, 
renal, or hepatic disease). 
10. Subjects with Body Mass Index (BMI) greater than or equal to 35 kg/m2 (=weight in 
kg/ height in meters2). 
11. Subjects participating in any clinical trial with another investigational product 30 days prior 
to Day 1 (M0) or intending to participate in another clinical trial at any time during the 
conduct of this trial. 
12. Subjects who have received any other vaccines within 14 days (for inactivated vaccines) or 
28 days (for live vaccines) prior to enrollment in this trial or who are planning to receive any 
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vaccine within 28 days of trial vaccine administration (consider whether applicable as an 
exclusion criterion or criterion for delay of trial vaccine administration, see Section 7.3). 
13. Subjects who have used antipyretics and/or analgesic medications within 24 hours prior to 
vaccination. The reason for their use (prophylaxis versus treatment) must be documented. 
Trial entry should be delayed to allow for a full 24 hours to have passed since last use of 
antipyretics and/or analgesic medications (consider whether applicable as an exclusion 
criterion or criterion for delay of trial vaccine administration, see Section 7.3). 
14. Subjects involved in the trial conduct or their first-degree relatives. 
15. Subjects with history of substance or alcohol abuse within the past 2 years. 
16. Female subjects who are pregnant or breastfeeding. 
17. Female subjects of childbearing potential who are sexually active with men and have not 
used any of the “acceptable contraceptive methods” for at least 2 months prior to Day 1 (M0). 
a) Of “childbearing potential” is defined as status post onset of menarche and not meeting 
any of the following conditions: menopausal for at least 2 years, status after bilateral 
tubal ligation for at least 1 year, status after bilateral oophorectomy for at least 1 year or 
status after hysterectomy. 
b) “Acceptable birth control methods” are defined as one or more of the following: 
I. 
Hormonal contraceptives (such as oral, injection, transdermal patch, implant, 
cervical ring). 
II. 
Barrier method (condom with spermicide or diaphragm with spermicide) every 
time during intercourse. 
III. 
Intrauterine device. 
IV. 
Monogamous relationship with vasectomized partner (partner must have been 
vasectomized for at least 6 months prior to the subject’s enrollment [Day 1 (M0)]). 
Other contraceptive methods may be considered in agreement with the sponsor and 
will be approved by the appropriate ethics committee. 
18. Female subjects of childbearing potential who are sexually active with men and refuse to use 
an acceptable contraceptive method up to 6 weeks after the last dose of trial vaccine 
(Day 90 [M3] + 6 weeks). In addition, female subjects must be advised not to donate ova 
during this period (see Section 9.1.11). 
19. Any positive or indeterminate pregnancy test (see Section 9.1.12). 
20. Previous participation in any clinical trial of a dengue candidate vaccine, except for subjects 
who received placebo in those trials, or previous and planned vaccination (during the trial 
conduct) against dengue. 
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21. Planned vaccination (during the trial conduct) against any non-dengue flavivirus (eg, Zika, 
YF, JE, WN, tick-borne encephalitis, or Murray-Valley encephalitis). 
22. Planned travel (during the trial conduct) to any area endemic for dengue. 
There may be instances when individuals meet all entry criteria except one that relates to 
transient clinical circumstances (eg, body temperature elevation or recent use of excluded 
medication[s] or vaccine[s]). Under these circumstances, eligibility for trial enrollment may be 
considered if the appropriate window for delay has passed, inclusion/exclusion criteria have been 
rechecked, and if the subject is confirmed to be eligible. 
7.3 
Criteria for Delay of Trial Vaccine Administration 
After enrollment, subjects may encounter clinical circumstances that warrant a delay in the 
administration of trial vaccine. These situations are listed below. In the event that a subject meets 
a criterion for delay of trial vaccine administration, the subject may receive the trial vaccine once 
the window for delay has passed as long as the subject is otherwise eligible for trial participation. 
If any of the conditions below occur at the time scheduled for the second TDV administration 
(Day 90 [M3]), the second dose may be administered at a later date as long as the subject is 
otherwise eligible to continue trial participation. In certain situations, the period of delay may 
lead to deviation from the time window for the second dose at Day 90 (M3). The decision to 
vaccinate in those situations will be made by the investigator. 
The following clinical circumstances warrant a delay for administration of the vaccination 
(consider whether applicable as criterion for delay of trial vaccine administration or an exclusion 
criterion, see Section 7.2): 
 Subjects with a clinically active significant infection (as assessed by the investigator) or body 
temperature ≥38°C (100.4°F) within 3 days of the intended date of vaccination. 
 Subjects who have received any other vaccines within 14 days (for inactivated vaccines) or 
28 days (for live vaccines) prior to enrollment in this trial or who are planning to receive any 
vaccine within 28 days of trial vaccine administration. 
 Known or suspected altered or impaired immune function as specified under the exclusion 
criteria (see Section 7.2, exclusion criterion no. 6). 
 Subjects who have used antipyretics and/or analgesic medications within 24 hours prior to 
vaccination. The reason for their use (prophylaxis versus treatment) must be documented. 
Trial vaccine administration should be delayed to allow for a full 24 hours to have passed 
between having used antipyretics and/or analgesic medications and trial vaccine 
administration. 
7.4 
Early Termination of a Subject’s Trial Participation 
Under some circumstances, a subject’s trial participation may be terminated early. This means 
that no further trial procedures (including data collection) will be performed on that subject 
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beyond the specific date of early termination of trial participation. The primary reason for early 
termination of the subject’s trial participation should be documented using the following 
categories. While the subject has no obligation to provide a reason for withdrawing consent, 
attempts should be made to determine the underlying reason for the withdrawal and, where 
possible, the primary underlying reason should be documented. 
For screen failure subjects, refer to Section 9.1.13. 
1. Adverse Event: The subject has experienced an AE (irrespective of being related/unrelated to 
the trial vaccine[s], or trial-related procedures) that requires early termination because 
continued participation imposes an unacceptable risk to the subject’s health and/or the 
subject is unwilling to continue participation because of the AE. If the subject is unwilling to 
continue because of the AE, the primary reason for early termination of trial participation in 
this case will be ‘withdrawal due to AE’ and not ‘withdrawal of consent’, see below. Any 
ongoing AEs leading to early termination of trial participation should be followed up by the 
investigator until resolution or stabilization. 
2. Lost to follow-up: The subject did not return to the clinic and at least 3 attempts to contact 
the subject were unsuccessful. 
3. Withdrawal of consent: The subject wishes to withdraw from the trial. The primary reason 
for early termination will be “withdrawal of consent” if the subject withdraws from 
participation due to a non-medical reason (ie, reason other than AE). The reason for 
withdrawal, if provided, should be recorded in the eCRF. 
4. Premature trial termination by the sponsor, a regulatory agency, the IEC/IRB, or any other 
authority. 
If the clinical trial is prematurely terminated by the sponsor, the investigator is to promptly 
inform the trial subjects and local IEC/IRB and should assure appropriate follow-up for the 
subjects. The primary reason for early termination in this case will be “trial termination”. 
5. Subject’s death during trial participation. 
6. Other. 
7.5 
Criteria for Premature Discontinuation of Trial Vaccine Administration 
Criteria for contraindication to vaccination at Day 90 (M3): 
There are also circumstances under which receipt of the second trial vaccine dose at 
Day 90 (M3) is contraindicated in this trial. These circumstances include but are not limited to 
anaphylaxis or severe hypersensitivity reactions following the first TDV vaccination at 
Day 1 (M0). If these reactions occur, the subject must not receive the TDV vaccination at 
Day 90 (M3) but will be encouraged to continue trial participation for safety follow-up. 
Early termination of a subject’s trial participation will by default prevent the subject from 
receiving further doses of trial vaccine, as the subject will no longer be participating in the trial. 
In addition to criteria for early termination of a subject’s participation (see Section 7.4), other 
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situations may apply in which subjects may continue participating in the trial (eg, contributing 
safety data according to protocol) but trial vaccine administration is discontinued. Even if the 
subject is deemed ineligible to receive further doses of trial vaccine, all efforts should be made to 
continue the collection of safety data according to protocol. 
In addition, the primary reason for premature discontinuation of trial vaccine administration 
should be recorded in the “End of Trial Vaccine Administration” eCRF using the following 
categories: 
1. Adverse Event: The subject has experienced an AE (irrespective of being related/unrelated to 
the trial vaccine or trial-related procedures) for which subsequent trial vaccine 
administration(s) impose an unacceptable risk to the subject’s health, but the subject will 
continue trial participation for safety, or a subset of other trial procedures. 
2. Lost to follow-up: The subject did not return to the clinic and at least 3 attempts to contact 
the subject were unsuccessful. 
3. Withdrawal of consent: The subject wishes to withdraw from the trial. The primary reason 
for early termination will be “withdrawal of consent” if the subject withdraws from 
participation due to a non-medical reason (ie, reason other than AE). The reason for 
withdrawal, if provided, should be recorded in the eCRF. 
4. Premature trial termination by sponsor, a regulatory agency, the IEC/IRB, or any other 
authority. 
If the clinical trial is prematurely terminated by the sponsor, the investigator is to promptly 
inform the trial subjects and local IEC/IRB and should assure appropriate follow-up for the 
subjects. The primary reason for early termination in this case will be “trial termination”. 
5. Subject´s death during trial participation. 
6. Protocol deviation: A protocol deviation is any change, divergence, or departure from the 
trial design or procedures of a trial protocol. The subject may remain in the trial unless 
continuation in the trial jeopardizes the subject’s health, safety or rights (see Section 7.4). 
7. Pregnancy: Any subject who, despite the requirement for adequate contraception, becomes 
pregnant during the trial will not receive further trial vaccine administrations. Pregnant 
subjects should, however, be asked to continue participating in the trial contributing data to 
the safety follow-up according to protocol. In addition, the site should maintain contact with 
the pregnant subject and complete a “Clinical Trial Pregnancy Form” as soon as possible. 
The subject should be followed-up until the birth of the child, or spontaneous or voluntary 
termination; when pregnancy outcome information becomes available, the information 
should be captured using the same form. Data obtained from the “Clinical Trial Pregnancy 
Form” will be captured in the safety database. 
8. Other. 
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8.0 
CLINICAL TRIAL MATERIAL MANAGEMENT 
This section contains information regarding all trial vaccines and materials provided directly by 
the sponsor, and/or sourced by other means, that are required by the trial protocol, including 
important sections describing the management of clinical trial material. 
8.1 
Trial Vaccine and Materials 
The investigational vaccine is Takeda’s TDV, a tetravalent dengue vaccine comprised of 
1 molecularly characterized, attenuated dengue virus strain, and 3 chimeric dengue virus strains 
with potencies of not less than 3.3, 2.7, 4.0, and 4.5 log10 plaque forming units per dose of 
TDV-1, TDV-2, TDV-3, and TDV-4, respectively. 
8.1.1 Dosage Form, Manufacturing, Packaging, and Labeling 
Manufacturing of monovalent bulk vaccine substances of TDV, mixing of the 4 TDV vaccine 
substances, filling into vials, and lyophilization of TDV is done at IDT Biologika GmbH, 
Germany. 
Lyophilized TDV is presented in a single-dose 2 mL glass vial with a grey bromo butyl rubber 
stopper and flip-top aluminum over seal. 
TDV diluent (37 mM sodium chloride solution) is a clear, colorless solution provided in a 
single-use 2 mL glass vial or a single-use 1 mL glass syringe and is used to reconstitute the 
lyophilized TDV to deliver a 0.5 mL dose. 
TDV and TDV diluent vials are packaged together into single dose dispensing cartons. The units 
will be labeled to contain pertinent trial information in local languages. 
The sponsor will supply the trial site with TDV and TDV diluent packaged into single dose 
dispensing cartons. The units and cartons will be labeled with pertinent trial information in local 
languages. Further details can be found in the Pharmacy Manual. 
8.1.2 Storage 
The trial vaccine (TDV) and TDV diluent will be shipped in refrigerated containers at 2°C to 
8°C. From receipt and prior to use, TDV and TDV diluent must be protected from light and 
stored at 2°C to 8°C in a refrigerator. Do not freeze. 
All clinical trial material must be kept in an appropriate, limited-access, secure place until it is 
used or returned to the sponsor or designee for destruction. All sponsor-supplied vaccines must 
be stored under the conditions specified on the label and remain in the original container until 
dispensed. A daily temperature log of the vaccine storage area must be maintained every 
working day. Temperature excursions must be reported to the sponsor as soon as possible and 
use of these vaccines requires sponsor approval. 
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8.1.3 Dose and Regimen 
All subjects in Groups 1 and 2 are to receive 1 trial vaccine dose on Day 1 (M0) and 1 trial 
vaccine dose on Day 90 [M3]). 
The 0.5 mL trial vaccine doses will be prepared and administered by the pharmacist or vaccine 
administrator according to the instructions in the Pharmacy Manual or per sponsor instructions. 
TDV will be administered by the SC route. 
8.2 
Trial Vaccine Assignment and Dispensing Procedures 
Refer to Section 8.6 for accountability of sponsor-supplied vaccines. 
If sponsor-supplied vaccine is lost or damaged, the site can request a replacement. Expired 
vaccines must not be administered. 
Prior to vaccination, a subject must be determined to be eligible for trial vaccination and it must 
be clinically appropriate in the judgment of the investigator to vaccinate. Eligibility for 
vaccination prior to first trial vaccine administration is determined by evaluating the entry 
criteria outlined in this protocol (see Section 7.1 and Section 7.2). 
Eligibility for subsequent trial vaccination is determined by following the criteria outlined in 
Section 7.3. 
Trial vaccine should not be administered to individuals with known hypersensitivity to any 
component of the vaccine. 
8.2.1 Precautions to be Observed when Administering the Trial Vaccine 
Prior to trial vaccine administration, a subject must be determined to be eligible to receive trial 
vaccine and it must be clinically appropriate in the judgment of the investigator to administer the 
trial vaccine. 
First, trial eligibility is evaluated according to the entry criteria outlined in this protocol 
(Sections 7.1 and 7.2). Once eligibility is confirmed, the subject is able to receive the first trial 
vaccination. 
Prior to subsequent trial vaccine administration, site staff must determine if the subject is eligible 
to receive vaccination by evaluating the criteria outlined in Sections 7.3, 7.4, and 7.5. 
Standard immunization practices are to be observed and care should be taken to administer the 
injection by the SC route. In addition, WHO recommendations to reduce anxiety and pain at the 
time of vaccination should be followed [18]. Before administering the trial vaccine, the 
vaccination site is to be disinfected with a skin disinfectant (eg, 70% alcohol). Allow the skin to 
dry. DO NOT inject intravascularly. 
As with all injectable vaccines, trained medical personnel and appropriate medical treatment 
should be readily available in case of anaphylactic reactions following vaccination. For example, 
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epinephrine 1:1000, diphenhydramine, and/or other medications for treating anaphylaxis should 
be available. These rescue medications will not be supplied by the sponsor. 
8.3 
Randomization Code Creation and Storage 
Not applicable. 
8.4 
Trial Vaccine Blind Maintenance 
Not applicable. 
8.5 
Unblinding Procedure 
Not applicable. 
8.6 
Accountability and Destruction of Sponsor-Supplied Trial Vaccine and Other 
Clinical Trial Materials 
Vaccine supplies will be counted and reconciled at the site before being locally destroyed or 
returned to the sponsor or designee as noted below. The site will maintain source documents. 
The investigator or designee must ensure that the sponsor-supplied trial vaccine is used in 
accordance with the approved protocol and is/are administered only to subjects enrolled in the 
trial. To document appropriate use of sponsor-supplied trial vaccine (TDV including TDV 
diluent), the investigator must maintain records of all sponsor-supplied trial vaccine delivery to 
the site, site inventory, administration and use by each subject, and return to the sponsor or 
designee. 
Upon receipt of sponsor-supplied trial vaccine, the investigator or designee must verify the 
contents of the shipments against the packing list. The verifier should ensure that the quantity is 
correct, the trial vaccine is received within the labeled storage conditions (ie, no cold chain break 
has occurred during transit), and is in good condition. If quantity and conditions are acceptable, 
investigator or designee will acknowledge receipt of the shipment by signing bottom half of the 
packing list and forward the list per instructions provided on the form. 
If there are any discrepancies between the packing list versus the actual product received, the 
sponsor or designee must be contacted to resolve the issue. The packing list should be filed in the 
Pharmacy Investigator Site File by a qualified investigator designee. 
The pharmacist (or designated individual) at the site must maintain 100% accountability for 
sponsor-supplied trial vaccine (TDV including TDV diluent) and other clinical trial material 
received and administered during their entire participation in the trial. Accountability includes, 
but is not limited to: 
 Verifying that actual inventory matches documented inventory. 
 Verifying that the log is completed for the vaccine lot used to prepare each dose. 
 Verifying that all trial vaccine kits used are documented accurately on the log. 
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 Verifying that required fields are completed accurately and legibly. 
If any dispensing errors or discrepancies are discovered, the sponsor must be notified 
immediately. 
The pharmacist (or designated individual) at the site must record the current inventory of all 
sponsor-supplied vaccine (TDV including TDV diluent) on a sponsor-approved vaccine 
accountability log. The following information will be recorded at a minimum: protocol number 
and title, name of investigator, site identifier and number, description of sponsor-supplied trial 
vaccine, expiry and/or retest date, and amount. The log should include all required information 
as a separate entry for each subject to whom sponsor-supplied trial vaccine is administered. 
The investigator will be notified of any expiry date or retest date extension of trial vaccine or 
clinical trial material during the trial conduct. On expiry date notification from the sponsor or 
designee, the site must complete all instructions outlined in the notification, including 
segregation of expired clinical trial material for return to the sponsor or its designee for 
destruction. 
Prior to site closure or at appropriate intervals throughout the trial, before any trial vaccine or 
clinical trial materials are returned to the sponsor or designee for destruction, a representative 
from the sponsor will perform clinical trial material accountability and reconciliation. The 
investigator will retain a copy of the documentation regarding clinical trial material 
accountability, return and/or destruction, and originals will be sent to the sponsor or designee. 
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9.0 
TRIAL PLAN 
9.1 
Trial Procedures 
The following sections describe the trial procedures and data to be collected. For each procedure, 
subjects are to be assessed by the same investigator or site personnel whenever possible. The 
Schedule of Trial Procedures is located in Section 2.1. All procedures must be performed by 
qualified and trained staff. 
9.1.1 Informed Consent 
The requirements of the informed consent form are described in Section 15.2. 
Informed consent must be obtained prior to the subject entering into the trial, and before any 
protocol-directed procedures are performed. 
A unique subject number will be assigned to each subject after informed consent is obtained. If 
all eligibility criteria are fulfilled, this subject number will be used throughout the trial. Subject 
numbers assigned to subjects who fail screening should not be reused (Section 9.1.13). 
9.1.2 Demographics, Medical History and Prior Medications 
Demographic information to be obtained will include age, sex, race, and ethnicity as described 
by the subject. 
Medical history will also be collected, including but not limited to any medical history that may 
be relevant to subject eligibility for trial participation such as prior vaccinations, concomitant 
medications, and previous and ongoing illnesses and/or injuries. Relevant medical history can 
also include any medical history that contributes to the understanding of an AE that occurs 
during trial participation, if it represents an exacerbation of an underlying disease/preexisting 
problem. 
Medical history (including corresponding medication) to be obtained will include any significant 
conditions or diseases that have disappeared or resolved at or prior to signing of the informed 
consent form. 
Adverse medical occurrences emerging during the time between signing of the informed consent 
form and the first administration of trial vaccine will be recorded in the “Medical History” eCRF. 
If such an adverse medical occurrence is assessed as related to a trial procedure this should be 
recorded in the eCRF as an AE related to trial procedure. 
All medications, vaccines and blood products taken or received by the subjects are to be 
collected as “Prior and Concomitant Medications” and recorded on the “Prior and Concomitant 
Medications” eCRF and in the subject’s source document: 
a) Medications: from 1 month (minimum 28 days) prior to administration of each trial 
vaccine dose up to 1 month (minimum 28 days) thereafter, 
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b) Vaccines: from 1 month (minimum 28 days) prior to administration of each trial vaccine 
dose up to 1 month (minimum 28 days) thereafter, 
c) Steroids and immunostimulants within 60 days prior to Day 1 (M0), 
d) Immunoglobulins and blood products within 3 months prior to Day 1 (M0), 
e) Immunosuppressive therapy within 6 months prior to Day 1 (M0). 
The use of antipyretics and/or analgesic medications within 24 hours prior to vaccination must be 
identified and the reason for their use (prophylaxis versus treatment) must be documented in the 
subject’s source documents and the eCRF. Trial vaccine administration should be delayed if 
subjects have used antipyretics and/or analgesic medications within 24 hours prior to vaccine 
administration (see Section 7.3). 
Medications taken for prophylaxis are those intended to prevent the onset of AEs following 
vaccination. Medications taken for treatment are intended to reduce or eliminate the presence of 
symptoms that are present. 
Prohibited therapies (see also Section 7.2): 
a) Parenteral immunoglobulin preparation, blood products, and/or blood-derived products 
within 3 months prior to Day 1 (M0), 
b) Immunosuppressive therapy within 6 months or systemic (eg, oral or parenteral) 
corticosteroid treatment within 60 days prior to Day 1 (M0) or immunostimulants within 
60 days prior to Day 1 (M0), 
c) Any other vaccines within 14 days (for inactivated vaccines) or 28 days (for live 
vaccines) prior to Day 1 (M0) and Day 90 (M3), and 28 days after each trial vaccination, 
d) Any other dengue vaccines (investigational or licensed) for the entire trial period, 
e) Receipt of any other clinical trial product within 30 days prior to Day 1 (M0). 
These data must be written in the subject’s source documents. 
9.1.3 Documentation of Trial Entrance/Randomization 
Only subjects who have a signed informed consent form, meet all of the inclusion criteria and 
none of the exclusion criteria are eligible for entrance into the active phase. 
If the subject is found to be ineligible for entrance into the active phase, the investigator should 
record the primary reason for failure on the subject’s screening and enrollment log. 
Randomization is not applicable for the trial. 
9.1.4 Physical Examination 
Physical examinations must be performed by a qualified health professional in accordance with 
local regulations and as listed within the Site Responsibility Delegation Log. A complete 
physical examination will be performed at Screening (if applicable) and prior to vaccination on 
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Day 1 (M0) and Day 90 (M3). A complete physical examination includes but is not limited to: 
auscultation of heart and lungs, palpation of the abdomen, inspection of extremities (including 
skin over intended vaccination site), a check of general appearance and the measurement of 
weight and height; BMI will be calculated. Measurement of height is only required at Screening 
or Day 1 (M0), as applicable. Additional physical examinations may be performed if indicated 
by review of the subject’s medical history. The findings should be documented in the subject’s 
source document. 
Targeted symptom-directed physical examination including but not limited to measurement of 
vital signs (see Section 9.1.5) may be performed on Day 30 (M1), Day 120 (M4) and 
Day 360 (M12). Clinically significant changes from the Baseline examination should be 
recorded in the subject’s source documents and the eCRF. 
9.1.5 Vital Signs 
During the physical examination, a subject should have their vital signs measured. These will 
include (but are not limited to) the measurement of systolic blood pressure/diastolic blood 
pressure, heart rate, and body temperature at all trial visits subsequent to Visit 1 (Day 1 [M0]). 
9.1.6 Screening 
A blood sample (2.5 mL) will be collected at the screening visit (up to 70 days [10 weeks] prior 
to Day 1 [M0)]). A screening visit is only applicable if serological testing with regard to the 
inclusion criteria (see Section 7.1) is performed within the scope of this trial. 
All blood samples will be processed, labeled and stored according to the Laboratory Manual or 
other appropriate guideline provided to the trial site. 
9.1.7 Immunogenicity Assessments 
 Humoral immune responses (dengue neutralizing antibodies and anti-dengue NS1 antibodies 
for all subjects, and T cell epitope mapping in a subset of subjects with IFN- γ ELISpot 
responses >500 SFC/106 cells and availability of sufficient cells) will be measured using 
blood samples (20 mL) collected from all subjects pre first vaccination (Day 1 [M0]) and on 
Days 15, 30 (M1), and 60 (M2); pre second vaccination (Day 90 [M3]) and on Days 105, 
120 (M4), 150 (M5), 180 (M6), 270 (M9), and 360 (M12). 
 T cell-mediated immune response (60 mL) will be measured using blood samples collected 
from all subjects pre first vaccination (Day 1 [M0]) and on Days 15, 30 (M1), and 60 (M2); 
pre second vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 150 (M5), 180 (M6), 
270 (M9), and 360 (M12). 
 B cell-mediated immune response will be measured using blood samples (30 mL) collected 
from all subjects pre first vaccination (Day 1 [M0]) and on Day 30 (M1); pre second 
vaccination (Day 90 [M3]) and on Days 105, 120 (M4), 180 (M6), 270 (M9), and 360 (M12). 
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 The innate immune response will be measured using blood samples (5 mL) collected from all 
subjects pre first vaccination (Day 1 [M0]) and on Days 4 and 6; pre second vaccination 
(Day 90 [M3]) and on Days 94 and 96. 
All blood samples will be processed, labeled and stored according to the Laboratory Manual or 
other appropriate guideline provided to the trial site. 
9.1.8 Processing, Labeling and Storage of Biological Samples 
PBMC will be collected, processed, labeled and stored according to trial site Standard Operating 
Procedures (SOP). Refer to the SOP for detailed instructions. 
All biological samples will be processed, labeled and stored according to the Laboratory Manual 
or other appropriate guideline provided to the site. 
9.1.9 Safety Assessments 
Safety assessments will include collection and recording of solicited local (injection site) 
reactions and systemic events, unsolicited AEs (serious and non-serious), and MAAEs. Refer to 
Section 10.1 for safety definitions. Details on collection and reporting of AEs are in 
Section 10.4. 
9.1.10 Clinical Safety Laboratory Variables 
Vaccine viremia will be measured using blood samples (5 mL) collected from all subjects 
pre first vaccination (Day 1 [M0]) and on Days 6, 9, 12, 15, and 30 (M1); pre second vaccination 
(Day 90 [M3]) and on Days 96, 99, 102, 105, and 120 (M4). 
All blood samples will be processed, labeled and stored according to the Laboratory Manual or 
other appropriate guideline provided to the trial site. 
9.1.11 Contraception and Pregnancy Avoidance Procedure 
For female subjects of childbearing potential, serum or urine pregnancy testing will be performed 
at Screening (if applicable) and prior to each trial dose administration (Day 1 [M0] and 
Day 90 [M3]). Results must be confirmed and documented as negative prior to each trial dose 
administration. Additional pregnancy tests may be performed during the trial if deemed 
necessary by the investigator. Females of childbearing potential who are sexually active will be 
reminded during trial visits to adhere to acceptable contraceptive methods up to 6 weeks after the 
last dose of TDV (Day 90 [M3] + 6 weeks). Subjects will be provided with information on 
acceptable methods of contraception as part of the subject informed consent process and will be 
asked to sign a consent form stating that they understand the requirements for avoidance of 
pregnancy and donation of ova. During the course of the trial, subjects of childbearing potential 
will receive continued guidance with respect to the avoidance of pregnancy as part of the trial 
procedures (see Section 2.1). Refer also to Section 7.2. 
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9.1.12 Pregnancy 
To ensure subject safety and the safety of the unborn child, each pregnancy in a subject having 
received a trial vaccine must be reported to the sponsor within 24 hours of the site learning of its 
occurrence. If the subject becomes pregnant during the trial, she will not receive any further 
doses of any trial vaccine. The pregnancy must be followed to determine outcome, including 
spontaneous or voluntary termination, details of birth, and the presence or absence of any birth 
defects, congenital abnormalities, or maternal and/or newborn complications. This follow-up 
should occur even if the intended duration of safety follow-up for the trial has ended. 
Any pregnancy occurring following trial vaccine administration should be reported immediately, 
using a “Clinical Trial Pregnancy Form”, to the contact listed in the Investigator Site File. 
9.1.13 Documentation of Subjects Who are Not Randomized 
Investigators must account for all subjects who sign an informed consent. If the subject is found 
to be not eligible at Day 1 (M0), the investigator should complete the eCRF. Randomization is 
not applicable for the trial. 
The primary reason for not receiving the trial vaccine is to be recorded in the eCRF using the 
following categories: 
 Adverse medical occurrence prior to receipt of investigational vaccine. 
 Screen failure (did not meet one or more inclusion criteria or did meet one or more exclusion 
criteria). 
 Withdrawal by subject. 
 Site terminated by sponsor. 
 Trial terminated by sponsor. 
 Other (specify reason). 
Subject numbers assigned to enrolled subjects who are not vaccinated should not be reused. 
9.2 
Monitoring Subject Compliance 
The investigator must record all injections of trial vaccine given to the subject in the subject’s 
source document and the eCRF. 
9.3 
Schedule of Observations and Procedures 
The schedule for all trial-related procedures for all evaluations is shown in Section 2.1. 
Assessments should be completed at the designated visit(s)/time point(s). 
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9.3.1 Procedures at the Screening Visit (up to 70 days [10 weeks] prior to Day 1 [M0]) 
Note: A screening visit is only applicable if serological testing with regard to the inclusion 
criteria (see Section 7.1) is performed within the scope of this trial. 
1. Before performing any trial procedure, the signed informed consent form needs to be 
obtained. Refer to Section 9.1.1. 
2. Check inclusion and exclusion criteria. Refer to Section 7.1 and Section 7.2, respectively. 
3. Collect demographic data, medical history, and prior medication/vaccination. Refer to 
Section 9.1.2. 
4. Collect concomitant medications/vaccinations. Refer to Section 9.1.2. 
5. Perform a complete physical examination. Refer to Section 9.1.4. 
6. Perform pregnancy testing (serum or urine) for females of childbearing potential. Refer to 
Section 9.1.11. 
7. Collect blood sample. Refer to Section 9.1.6. 
Blood should be taken from the subject using an aseptic venipuncture technique. 
The site should schedule the next trial visit with the subject. 
The subject will receive a written reminder of the next trial visit. 
9.3.2 Pre-Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) 
1. Before performing any trial procedure, the signed informed consent form needs to be 
obtained (Day 1 [M0] unless obtained at Screening). Refer to Section 9.1.1. 
2. Check inclusion and exclusion criteria (Day 1 [M0]). Refer to Section 7.1 and Section 7.2, 
respectively. 
3. Assign subject to trial group (Day 1 [M0]). Refer to Section 6.1. 
4. Collect demographic data, medical history, and prior medication/vaccination (Day 1 [M0]). 
Refer to Section 9.1.2. 
5. Collect concomitant medications/vaccinations. Refer to Section 9.1.2. 
6. Review of systems: Review of systems is a structured interview that queries the subject as to 
any complaints the subject has experienced across each organ system. 
7. Perform a complete physical examination. Refer to Section 9.1.4. 
8. Check vital signs. Refer to Section 9.1.5. 
9. Perform pregnancy testing (serum or urine) for females of childbearing potential. Refer to 
Section 9.1.11. 
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10. Provide guidance with respect to the avoidance of pregnancy for females of childbearing 
potential who are sexually active. Refer to Section 9.1.11. 
11. Collect pre-vaccination blood samples. Refer to Sections 9.1.7, 9.1.8, and 9.1.10. 
Blood should be taken from the subject using an aseptic venipuncture technique. 
9.3.3 Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) 
1. Check criteria for delay of trial vaccine administration. Refer to Section 7.3. 
2. Check contraindications for trial vaccine administration. Refer to Section 7.5. 
3. Administer the trial vaccine. Refer to Section 8.1.3. 
9.3.4 Post Vaccination Procedures (Day 1 [M0] and Day 90 [M3]) 
 Careful training of the subject on how to measure solicited local (injection site) reactions and 
body temperature, how to complete the diary card and how often to complete the diary card. 
Training should be directed at the individual(s) who will perform the measurements of 
solicited local (injection site) reactions and those who will enter the information into the 
diary card. This individual may or may not be the subject, but if a person other than the 
subject enters information into the diary card, this person’s identity must be documented in 
the subject’s source document and this person must receive training on the diary card. 
Training of the subject on how to measure an injection site reaction and how to take their 
temperature, as well as how to record the information in the diary card, should be performed 
while the subject is under observation after vaccination. 
Diary card instructions must include the following: 
 The individual(s) who will enter the information into the diary card must understand that 
timely completion of the diary card on a daily basis is a critical component of trial 
participation. This individual should also be instructed to write clearly and to complete the 
diary card in pen. Any corrections to the diary card that are performed by the individual(s) 
completing the diary card should include a single strikethrough line with a brief explanation 
for any change and be initialed and dated. 
Please note: 
Diary cards will be the only source document allowed for remote collection of solicited local 
(injection site) reactions, solicited systemic events (including body temperature measurements), 
and unsolicited (non-serious) AEs. The following additional rules apply to the documentation of 
safety information collected by diary card: 
 The diary card should be reviewed with the subject. 
 No corrections or additions to the diary card will be allowed after it is reviewed with the 
investigator/designee. 
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 Any data that is identified as implausible or incorrect, and confirmed by the subject to be a 
transcription error, should be corrected by the subject on the diary card (the correction should 
include a single strikethrough line and should be initialed and dated by the subject). 
 Any blank or illegible fields on the diary card not otherwise corrected as above will be 
missing in the eCRF. 
 The site must enter all readable entries on the diary card into the eCRF. 
 Any newly described solicited and unsolicited (non-serious) safety information should be 
added to the diary card by the subject, initialed, and dated. 
 Starting on the day of vaccination (Day 1 [M0] and Day 90 [M3]), the subject will check for 
specific types of events at the injection site (ie, solicited local [injection site] reactions), any 
specific generalized symptoms (solicited systemic events), body temperature, unsolicited 
(non-serious) AEs, any other symptoms or change in the subject’s health status, any 
medications taken (excluding vitamins and minerals), and any vaccination. Solicited local 
(injection site) reactions, solicited systemic events, body temperature, and unsolicited 
(non-serious) AEs will be recorded in the diary. Assessments should preferably take place in 
the evening at day’s end. 
 Body temperature measurement is to be performed using the thermometer provided by the 
site. Assessments should take place at approximately the same time of day, preferably in the 
evening at day’s end. If the subject feels unusually hot or cold during the day, the subject 
should check their temperature. If the subject has fever, the highest body temperature 
observed that day should be recorded on the diary card. 
 The measurements of solicited local (injection site) reactions are to be performed using the 
ruler provided by the site. 
 The collection on the diary card of solicited local (injection site) reactions, solicited systemic 
events (including body temperature measurement), and unsolicited (non-serious) AEs will 
continue for a total of 7 days, 14 days, and 28 days; respectively, following administration of 
each trial vaccine dose (including the day of administration). Any solicited local (injection 
site) reaction or solicited systemic event observed as continuing on Day 8 or 15, respectively, 
following each trial vaccination will be recorded as an AE on the “Adverse Event” eCRF for 
follow-up. Any solicited local (injection site) reaction or systemic event that resolves before 
8 or 15 days, respectively, following each trial vaccination, but recurs at a later time (ie, if 
discontinues), should be recorded as an unsolicited AE on the “Adverse Event” eCRF (see 
Section 10.4.1 and Section 10.4.2). 
 Collect and record MAAEs. Refer to Section 10.4.3. 
 Collect and report SAEs. Refer to Section 10.4.4. 
After each trial vaccination, the subject will be observed for at least 30 minutes including 
observation for solicited local (injection site) reactions, solicited systemic events (including body 
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temperature measurement), and unsolicited AEs. Information should be recorded in the eCRF. 
The investigator or delegate will take the opportunity to remind the subject how to measure 
solicited local (injection site) reactions and body temperature as part of this observation period. 
All safety data will be collected in the subject’s source documents. 
The site should schedule the next trial visit with the subject. 
The subject will receive a written reminder of the next trial visit. 
The subject will be reminded to complete the diary card daily, to contact the site if there are any 
questions, and to contact the site immediately (or as soon as the subject is medically stable) if the 
subject has a medical condition that leads to a hospitalization or an emergency room visit or is 
otherwise perceived as serious. All contact details will be provided to the subject. 
9.3.5 Site Visits after Vaccination (Post First Vaccination: Days 4, 6, 9, 12, 15, 30 [M1], 
and 60 [M2]; Post Second Vaccination: Days 94, 96, 99, 102, 105, 120 [M4], 
150 [M5], 180 [M6], 270 [M9], and 360 [M12]) 
Note: Refer to Section 2.1 for visit windows(days) between trial visits that do NOT include a 
vaccination. 
1. Review the diary card with the subject (Days 4, 6, 9, 12, 15, 30 [M1], 94, 96, 99, 102, 105, 
and 120 [M4]). Refer to Section 9.3.4. 
2. Collect and record persistent/prolonged solicited local (injection site) reactions (Days 9, 12, 
15, 30 [M1], 60 [M2], 99, 102, 105, 120 [M4], 150 [M5], 180 [M6], 270 [M9], and 
360 [M12]). Refer to Section 9.3.4 and Section 10.4.2. 
3. Collect and record persistent/prolonged solicited systemic events (Days 15, 30[M1], 60 [M2], 
120 [M4], 150 [M5], 180 [M6], 270 [M9], and 360 [M12]). Refer to Section 9.3.4 and 
Section 10.4.2. 
4. Collect concomitant medications/vaccinations. Refer to Section 9.1.2. 
5. Perform a targeted physical examination (Days 30 [M1], 120 [M4], and 360 [M12]). Refer to 
Section 9.1.4 and Section 9.1.5. 
6. Check vital signs. Refer to Section 9.1.5. 
7. Provide guidance with respect to the avoidance of pregnancy for females of childbearing 
potential who are sexually active (Days 4, 6, 9, 12, 15, 30 [M1], 60 [M2], 94, 96, 99, 102, 
105, and 120 [M4]). Refer to Section 9.1.11. 
8. Collect and record MAAEs. Refer to Section 10.4.3. 
9. Collect and report SAEs. Refer to Section 10.4.4. 
10. Collect blood samples. Refer to Section 9.1.7 and Section 9.1.10. 
Blood should be taken from the subject using an aseptic venipuncture technique. 
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The investigator or delegate should schedule the next trial visit with the subject, as applicable. 
The subject will receive a written reminder of the next trial visit, as applicable. The subject will 
be reminded to complete the diary card daily (Days 4, 6, 9, 12, 15, 94, 96, 99, 102, and 105), to 
contact the site if there are any questions, and to contact the site immediately (or as soon as the 
subject is medically stable) if the subject has a medical condition that leads to a hospitalization or 
an emergency room visit or is otherwise perceived as serious. All contact details will be provided 
to the subject. 
9.3.6 Phone Contacts - Reminder Calls 
Not applicable. 
9.3.7 Phone Contacts – Safety Call 
When a site visit cannot be carried out due to the COVID-19 pandemic, telephone contacts will 
be made for subjects who are still under monitoring for safety reporting. Refer also to Section 
14.1. 
9.3.8 Final (End of Trial) Visit 
The final (end of trial) visit will be performed on Day 360 (M12). If a subject terminates earlier, 
the final (end of trial) visit procedures should be performed at their last trial visit, if possible. For 
all subjects receiving trial vaccine, the investigator must complete the “End of Trial” eCRF. 
9.3.9 Post-Trial Care 
No post-trial care will be provided. 
9.4 
Biological Sample Retention and Destruction 
In this trial, specimens will be collected for serological testing as described in Section 9.1.6, 
immune response testing as described in Section 9.1.7. and Section 9.1.8, and for clinical safety 
as described in Section 9.1.10. All blood samples will be processed, labeled and stored according 
to the Laboratory Manual or other appropriate guideline provided to the site. Samples will be 
preserved and retained at a central laboratory that was contracted by the sponsor for this purpose 
for up to but no longer than 20 years or as required by applicable law. The sponsor has put into 
place a system to protect the subject’s personal information to ensure optimal confidentiality and 
defined standard processes for sample and data collection, storage, analysis, and destruction. 
Serum samples and PBMC will be used for the analyses defined in this protocol, but can also, 
with permission from the subject, be used to assess, improve or develop tests related to dengue or 
other disease(s) or the investigational vaccine that will allow more reliable measurement of the 
response to the investigational vaccine. 
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10.0 
ADVERSE EVENTS 
10.1 
Definitions 
10.1.1 Adverse Events 
An AE is defined as any untoward medical occurrence in a clinical investigation subject 
administered a trial vaccine; it does not necessarily have to have a causal relationship with trial 
vaccine administration. 
An AE can therefore be any unfavorable and unintended sign (eg, a clinically significant 
abnormal laboratory finding), symptom, or disease temporally associated with the administration 
of a trial vaccine whether or not it is considered related to the trial vaccine. 
AEs will be graded by the investigator in the following manner: 
Mild 
Grade 1 
 
Awareness of symptoms that are easily tolerated, causing minimal discomfort and 
not interfering with everyday activities. Relieved with or without symptomatic 
treatment. 
Moderate 
Grade 2 
 
Sufficient discomfort is present to cause interference with normal activity. Only 
partially relieved with symptomatic treatment. 
Severe 
Grade 3 
 
Extreme distress, causing significant impairment of functioning or incapacitation. 
Prevents normal everyday activities. Not relieved with symptomatic treatment. 
10.1.2 Solicited Adverse Events 
The occurrence of selected indicators of safety will be measured/collected for 7 days (solicited 
local [injection site] reactions) and 14 days (solicited systemic events) following administration 
of each trial vaccine dose (including the day of administration) and will be recorded on the 
“Local and Systemic Reactions” eCRF, as applicable, and as listed in Table 10.a. 
Any solicited local (injection site) reaction or systemic event observed as continuing on Day 8 or 
Day 15, respectively, following each trial vaccination will be recorded as an AE on the “Adverse 
Event” eCRF for follow-up. For these persistent/prolonged solicited AEs, the end date will be 
captured on the “Adverse Event” eCRF to permit a separate analysis from the unsolicited AEs 
(see Section 10.4.1 and Section 10.4.2). 
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Table 10.a 
Solicited Local (Injection Site) Reactions and Systemic Events 
Local (injection site) reactions: 
Pain 
Erythema 
Swelling 
Systemic events: 
Fever (a) 
Headache 
Asthenia 
Malaise 
Myalgia 
(a) Fever is defined as body temperature greater than or equal to 38°C (100.4°F) regardless of method taken [19]. 
The severity of solicited safety parameters will be assessed as described in Table 10.b. 
Table 10.b 
Severity Scales for Solicited Safety Parameters 
AE 
Severity grade 
Severity 
Pain at injection site 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents daily activity with or without treatment 
Erythema at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm 
2 
Moderate: >50 – ≤100 mm 
3 
Severe: >100 mm 
Swelling at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm 
2 
Moderate: >50 – ≤100 mm 
3 
Severe: >100 mm 
Headache 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents normal activity with or without treatment 
Asthenia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Malaise  
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Myalgia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Fever (b) 
Record body temperature in °C/°F 
(a) Subjects are to record greatest surface diameter in mm on the diary card. 
(b)  Fever is defined as body temperature greater than or equal to 38°C (100.4°F) regardless of method taken [19]. 
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10.1.3 Adverse Events of Special Interest 
Not applicable. 
10.1.4 Medically Attended Adverse Events 
MAAEs are defined as AEs leading to an unscheduled medical visit to or by a healthcare 
professional including visits to an emergency department, but not fulfilling seriousness criteria. 
10.1.5 Serious Adverse Events 
An SAE is defined as any untoward medical occurrence that: 
1. Results in DEATH. 
2. Is LIFE THREATENING. 
 The term “life threatening” refers to an event in which the subject was at risk of death at 
the time of the event; it does not refer to an event that hypothetically might have caused 
death if it were more severe. 
3. Requires inpatient HOSPITALIZATION or prolongation of existing hospitalization. 
4. Results in persistent or significant DISABILITY/INCAPACITY. 
5. Leads to a CONGENITAL ANOMALY/BIRTH DEFECT in the offspring of a subject. 
6. Is an IMPORTANT MEDICAL EVENT that satisfies any of the following: 
 May require intervention to prevent items 1 through 5 above. 
 May expose the subject to danger, even though the event is not immediately life 
threatening or fatal or does not result in hospitalization. 
10.2 
Causality of Adverse Events 
Relationship (causality) to the trial vaccine will also be assessed by the investigator. The 
relationship of each AE to the trial vaccine, including solicited systemic events (solicited local 
[injection site] reactions are considered as related by default) will be assessed using the 
following categories: 
Related: 
There is suspicion that there is a relationship between the trial vaccine and 
the AE (without determining the extent of probability); there is a reasonable 
possibility that the trial vaccine contributed to the AE. 
Not Related: 
There is no suspicion that there is a relationship between the trial vaccine and 
the AE; there are other more likely causes and administration of the trial 
vaccine is not suspected to have contributed to the AE. 
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10.2.1 Relationship to Trial Procedures 
Relationship (causality) to trial procedures should be determined for all AEs. 
The relationship should be assessed as “Yes” if the investigator considers that there is a 
reasonable possibility that an event is due to a trial procedure. Otherwise, the relationship should 
be assessed as “No”. 
10.2.2 Outcome of Adverse Events 
Resolved: 
The subject has fully recovered from the event or the condition has 
returned to the level observed at Baseline. 
Resolving: 
The event is improving but the subject is still not fully recovered. 
Not resolved: 
The event is ongoing at the time of reporting and the subject has still 
not recovered. 
Resolved with 
sequelae: 
As a result of the AE, the subject suffered persistent and significant 
disability/incapacity (eg, became blind, deaf or paralysed). 
Fatal: 
The subject died due to the event. If the subject died due to other 
circumstances than the event, the outcome of the event per se should 
be stated otherwise (eg, not resolved or resolving). 
Unknown: 
If outcome is not known or not reported. 
10.3 
Additional Points to Consider for Adverse Events 
An untoward occurrence generally may: 
 Indicate a new diagnosis or unexpected worsening of a pre-existing condition. Intermittent 
events for pre-existing conditions or underlying disease should not be considered as AEs. 
 Necessitate therapeutic intervention. 
 Require an invasive diagnostic procedure. 
 Require trial vaccine discontinuation or a change in concomitant medication. 
 Be considered unfavorable by the investigator for any reason. 
Diagnoses vs signs and symptoms: 
 Each event should be recorded to represent a single diagnosis. Accompanying signs 
(including abnormal laboratory values) or symptoms should NOT be recorded as additional 
AEs. If a diagnosis is unknown, signs or symptoms should be recorded appropriately as AEs. 
Worsening of AEs: 
 If the subject experiences a worsening or complication of an AE after administration of the 
trial vaccine, the worsening or complication should be recorded as a new AE. Investigators 
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should ensure that the AE term recorded captures the change in the condition (eg, “worsening 
of…”). 
 If the subject experiences a worsening or complication of an AE, the worsening or 
complication should be recorded as a new AE. Investigators should ensure that the AE term 
recorded captures the change in the condition (eg, “worsening of…”). 
Changes in severity of AEs: 
 If the subject experiences changes in severity of an AE, the event should be captured once 
with the maximum severity recorded. 
Preplanned surgeries or procedures: 
 Preplanned procedures (surgeries or therapies) that were scheduled prior to signing of 
informed consent form are not considered AEs. Complications resulting from any planned 
surgery should be reported as AEs. 
Elective surgeries or procedures: 
 Elective procedures performed where there is no change in the subject’s medical condition 
should not be recorded as AEs but should be documented in the subject’s source documents. 
Complications resulting from an elective surgery should be reported as AEs. 
Trial procedures: 
 Adverse occurrences related to trial procedures after signing of the informed consent form 
are considered as AEs and should be reported as AEs. 
10.4 
Procedures 
10.4.1 Collection and Reporting of Adverse Events 
All AEs, whether considered related to the use of the trial vaccine or not, must be monitored 
until symptoms subside and any abnormal laboratory values have returned to Baseline, or until 
there is a satisfactory explanation for the changes observed, or until death, in which case a full 
autopsy report should be supplied, if possible. All findings must be reported on the “Adverse 
Event” eCRF and on the SAE form*, if necessary (see Section 10.4.4). All findings in subjects 
experiencing AEs must also be documented in the subject’s source documents. Any unsolicited 
AEs will be collected on diary cards by the subject for 28 days following administration of each 
trial vaccine dose (including the day of administration). AEs leading to discontinuation (from the 
trial or from the vaccination regimen) are collected throughout the trial. Even if the subject is 
deemed ineligible to receive further doses of trial vaccine, all efforts should be made to continue 
the collection of safety data according to protocol. 
 Reported term for the AE. 
 Start and end date. 
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 Serious (Yes/No). 
 Severity. 
 Investigator’s opinion of the causality (relationship) between the event and administration of 
trial vaccine (“related” or “not related”). 
 Investigator’s opinion of the causality (relationship) to trial procedure(s), including the 
details of the suspected procedure. 
 Action taken with the trial vaccine. 
 Outcome of event. 
*SAE reporting will be done by eCRF. If the eCRF system is unavailable, a paper sponsor SAE 
form/paper CRF should be completed and the event must be entered into the eCRF once access 
is restored. 
10.4.2 Collection and Reporting of Solicited Adverse Events 
The occurrence of selected indicators of safety will be collected on diary cards by the subject for 
7 days (solicited local [injection site] reactions) and 14 days (solicited systemic events) 
following administration of each trial vaccine dose (including the day of administration) and will 
be recorded on the “Local and Systemic Reactions” eCRF, as applicable. These will be 
summarized in the final report under the category “solicited AEs” to differentiate them from 
unsolicited AEs. 
Any solicited local (injection site) reaction or solicited systemic event observed as continuing on 
Day 8 or 15, respectively, following each trial vaccination will be recorded as an AE on the 
“Adverse Event” eCRF for follow-up. For these persistent/prolonged solicited AEs, the end date 
will be captured on the “Adverse Event” eCRF to permit a separate analysis from the unsolicited 
AEs. Any solicited local (injection site) reaction or systemic event that resolves before 8 or 
15 days, respectively, following each trial vaccination, but recurs at a later time (ie, if 
discontinues), should be recorded as an unsolicited AE on the “Adverse Event” eCRF (see 
Section 10.4.1). 
Any solicited AE that meets any of the following criteria must be entered as an AE on the 
“Adverse Event” eCRF: 
 Solicited local (injection site) reactions or systemic events that lead the subject to withdraw 
from the trial. 
 Solicited local (injection site) reactions or systemic events that lead to the subject being 
withdrawn from the trial by the investigator. 
 Solicited local (injection site) reactions and systemic events that otherwise meet the 
definition of an SAE (see Section 10.1.5). 
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10.4.3 Collection and Reporting of Adverse Events of Special Interest/Medically Attended 
Adverse Events 
AEs of special interest will not be collected for this trial. 
MAAEs occurring from first trial vaccination at Day 1 (M0) until the end of the trial 
(Day 360 [M12]) will be collected during site visits by interview and must be recorded as AEs 
on the “Adverse Event” eCRF. 
10.4.4 Collection and Reporting of Serious Adverse Events 
Collection of SAEs will commence from the time that the subject is first administered the trial 
vaccine (Day 1 [M0]). Routine collection of SAEs will continue until the end of the trial 
(Day 360 [M12]). 
SAEs should be reported according to the following procedure: 
A sponsor SAE form must be completed, in English, and signed by the investigator immediately 
or within 24 hours of first onset or notification of the event. The information should be 
completed as fully as possible but contain, at a minimum: 
 A short description of the event and the reason why the event is categorized as serious. 
 Causality assessment. 
 Protocol number. 
 Subject identification number. 
 Investigator’s name. 
 Name of trial vaccine. 
The SAE form should be transmitted within 24 hours to for the attention of the contact(s) in the 
list provided to each site. 
10.5 
Follow-up Procedures 
10.5.1 Adverse Events 
All AEs will be monitored until resolution or a stable status is reached or until a formal diagnosis 
can be made or until the end of the trial, whichever occurs first. 
10.5.2 Serious Adverse Events 
If information not available at the time of the first report becomes available later, the investigator 
should complete a follow-up SAE form or provide other written documentation immediately. 
Copies of any relevant data from the hospital notes (eg, laboratory tests, discharge summary, 
postmortem results) should be sent to the sponsor. 
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All SAEs should be followed up until resolution, permanent outcome of the event, or is 
otherwise explained. The timelines and procedure for follow-up reports are the same as those for 
the initial report. 
10.5.3 Safety Reporting to Investigators, Investigational Review Boards or Independent 
Ethics Committees, and Regulatory Authorities 
The sponsor or designee will be responsible for the reporting of all Suspected Unexpected 
Serious Adverse Reactions (SUSAR) and any other SAEs to regulatory authorities, investigators 
and IRBs or IECs, as applicable, in accordance with national regulations in the countries where 
the trial is conducted. Relative to the first awareness of the event by/or further provision to the 
sponsor or designee, SUSARs will be submitted within 7 days for fatal and life-threatening 
events and 15 days for other SUSARs, unless otherwise required by national regulations. The 
sponsor will also prepare an expedited report for other safety issues where these might materially 
alter the current benefit-risk assessment of an investigational vaccine or that would be sufficient 
to consider changes in the trial vaccine administration or in the overall conduct of the trial. The 
investigational site will also forward a copy of all expedited reports to their IRB or IEC in 
accordance with national regulations. 
10.5.4 Post-Trial Events 
Any SAE that occurs outside of the protocol-specified observation period or after the end of the 
trial but is considered to be caused by the trial vaccine must be reported to the sponsor. These 
SAEs will be processed by the sponsor’s Pharmacovigilance Department. Instructions for how to 
submit these SAEs will be provided in a handout in the Investigator Site File. 
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11.0 
TRIAL-SPECIFIC REQUIREMENT(S) 
11.1 
Trial-Specific Committees 
11.1.1 Data Monitoring Committee 
A DMC will have oversight of this trial. The DMC functions at a program level and further 
information is available in the DMC Charter. 
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12.0 
DATA HANDLING AND RECORD KEEPING 
The full details of procedures for data handling will be documented in the Data Management 
Plan. AEs, medical history, and concurrent medical conditions will be coded using the Medical 
Dictionary for Regulatory Activities (MedDRA). Drugs will be coded using the WHO Drug 
Dictionary [20]. 
12.1 
CRFs (Electronic) 
Completed eCRFs are required for each subject who provides a signed informed form. 
The sponsor or designee will supply the investigative site with access to eCRFs. The sponsor will 
make arrangements to train appropriate site staff in the use of the eCRF. Electronic CRFs must 
be completed in English. 
After completion of the entry process, computer logic checks will be run to identify items, such 
as inconsistent dates, missing data, and questionable values. Queries may be issued by sponsor 
personnel (or designee[s]) and will be answered by the site. 
The principal investigator must review the eCRFs for completeness and accuracy and must sign 
and date the appropriate eCRFs as indicated. Furthermore, the investigator must retain full 
responsibility for the accuracy and authenticity of all data entered on the eCRFs. 
Electronic CRFs will be reviewed for completeness and acceptability at the trial site during 
periodic visits by trial monitors. The sponsor or designee will be permitted to review the 
subject’s medical and hospital records pertinent to the trial to ensure accuracy of the eCRFs. The 
completed eCRFs are the sole property of the sponsor and should not be made available in any 
form to third parties, except for authorized representatives of appropriate governmental health or 
regulatory authorities, without written permission of the sponsor. 
When a site visit cannot be carried out due to the COVID-19 pandemic, telephone contacts will 
be made for subjects for whom monitoring for safety reporting is still ongoing. Refer also to 
Section 14.1. 
12.2 
Record Retention 
The investigator agrees to keep the records stipulated in Appendix A and those documents that 
include (but are not limited to) the trial-specific documents, the identification log of all 
participating subjects, medical records. Temporary media such as thermal sensitive paper should 
be copied and certified, source worksheets, all original signed and dated informed consent forms, 
subject authorization forms regarding the use of personal health information (if separate from the 
informed consent forms), electronic copy of CRFs, including the audit trail, and detailed records 
of vaccine disposition to enable evaluations or audits from regulatory authorities, the sponsor or 
designee. Furthermore, ICH E6 Section 4.9.5 requires the investigator to retain essential 
documents specified in ICH E6 (Section 8) until at least 2 years after the last approval of a 
marketing application for a specified vaccine indication being investigated or, if an application is 
not approved, until at least 2 years after the investigation is discontinued and regulatory 
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authorities are notified. In addition, ICH E6 Section 4.9.5 states that the trial records should be 
retained until an amount of time specified by applicable regulatory requirements or for a time 
specified in the Clinical Study Site Agreement between the investigator and sponsor. 
Refer to the Clinical Study Site Agreement for the sponsor’s requirements on record retention. 
The investigator should contact and receive written approval from the sponsor before disposing 
of any such documents. 
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13.0 
STATISTICAL METHODS 
13.1 
Statistical and Analytical Plans 
A Statistical Analysis Plan (SAP) will be prepared and finalized prior to the database lock. This 
document will provide further details regarding the definition of analysis variables and analysis 
methodology to address all trial objectives. 
Data reviews will be conducted prior to the database lock. These reviews will assess the accuracy 
and completeness of the trial database and subject evaluability. 
This trial is designed for a descriptive evaluation of the endpoints that does not require 
hypotheses testing. Further details will be provided in the SAP. 
Summaries will be provided by trial group (Group 1 and Group 2) and further for Group 2 by 
subgroups (DENV-1 or DENV-3 seropositive). 
As trial laboratory procedures are not carried out during the COVID-19 pandemic there is a 
possibility that not all pre-defined test results will be available prior to database lock. Thus, there 
is a risk for database unlock/relock and additional data may be added in CSR amendments or 
addenda to the final CSR. 
13.1.1 Analysis Sets 
Safety Set: The Safety Set will consist of all subjects who received at least 1 dose of trial 
vaccine. 
Full Analysis Set (FAS): The FAS will include all subjects who received at least 1 dose of trial 
vaccine and for whom a valid pre-dose and at least one valid post-dose blood sample is taken. 
Per–Protocol Set (PPS): The PPS will include all subjects in the FAS who have no major 
protocol violations. The major protocol violation criteria will be defined as part of the data 
review prior to database lock. The categories of major protocol violations include: (1) not 
meeting selected entry criteria, (2) receiving prohibited therapies, (3) not receiving 2 doses of 
TDV, or receiving the second vaccination inadmissibly outside of the visit window, and (4) other 
major protocol violations that may be identified during data review prior to database lock. 
13.1.2 Analysis of Demographics and Other Baseline Characteristics 
Age, gender, race, and other Baseline characteristics will be summarized descriptively. 
Summaries will be provided by trial group (Group 1 and Group 2) and further for Group 2 by 
subgroups (DENV-1 or DENV-3 seropositive). Unless specified otherwise, number of subjects 
with non-missing observations, mean, SD, median, minimum and maximum will be presented 
for continuous data; frequency and percent will be presented for categorical data. 
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13.1.3 Immunogenicity Analysis 
For the primary immunogenicity endpoint, descriptive statistics and 95% CIs will be provided 
for each applicable visit. Summaries will be provided by trial group (Group 1 and Group 2) and 
further for Group 2 by subgroups (DENV-1 or DENV-3 seropositive). 
The primary immunogenicity analyses will be based on the PPS; supportive analyses may be 
provided based on the FAS. 
Similar descriptive analyses as for the primary immunogenicity endpoint will be provided for the 
secondary and exploratory endpoints for each applicable assay at all relevant time points, based 
on the PPS. Supportive analyses based on the FAS may also be provided for selected endpoints. 
Antibody titers will be analyzed under log-normal distribution assumption. GMT will be 
calculated for each relevant time point as anti-logarithm of arithmetic mean of natural log 
transformed titers. The 95% CI for GMT will be calculated as the anti-log transformation of 
upper and lower limits for a 2-sided CI of the mean of the log-transformed titers (based on 
Student’s t-distribution). 
Other quantitative data will be summarized descriptively on the original scale. 
Seropositivity rates will be accompanied with 95% CIs calculated by exact (Clopper-Pearson) 
method [21]. 
Relationship between immunogenicity measures will be assessed using non-parametric 
correlation coefficient, as appropriate [22]. Graphical methods will also be explored. Further 
details will be provided in the SAP. 
Handling of missing data will be described in the SAP. 
13.1.4 Safety Analysis 
All safety data will be summarized descriptively using the Safety Set, by trial group (Group 1 
and Group 2) and further for Group 2 by subgroups (DENV-1 or DENV-3 seropositive). 
Solicited AEs 
For each solicited AE, the number and percentage of subjects with local (injection site) reactions 
and systemic events will be summarized by event severity for each day after each vaccination 
(ie, Day 1 [M0] through Day 7 for local [injection site] reactions and Day 1 [M0] through 
Day 14 for systemic events), and overall. Summaries of first onset of each event and the number 
of days subjects reported experiencing each event will also be provided. For subjects with more 
than 1 episode of the same event, the maximum severity will be used for tabulations. 
Persistent/prolonged solicited local (injection site) reactions or systemic events continuing on 
Day 8 or Day 15, respectively, following each trial vaccination will be assessed separately. 
Unless otherwise specified, these events will not be included in the analyses/tabulations of 
unsolicited AEs and will have separate listings. 
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Unsolicited AEs 
Unsolicited AEs will be coded using MedDRA, and summarized by System Organ Class (SOC) 
and Preferred Term (PT). Unsolicited AEs will be summarized for 28 days following each 
vaccination (day of vaccination + 27 days) as follows: by PT including events with frequency 
greater than a pre-defined threshold (the percentage will be specified in the SAP); by SOC and 
PT; by SOC, PT, and severity; and by SOC, PT, and relationship to TDV. Subjects reporting 
more than 1 occurrence for the term (level) being summarized will be counted only once. 
AEs leading to trial vaccine withdrawal or trial discontinuation will be collected and summarized 
for the entire trial period. 
SAEs and MAAEs 
SAEs and MAAEs will be coded using MedDRA, and summarized by SOC and PT for the entire 
trial period. 
Other Safety Observations 
Vaccine viremia (incidence, duration, and level) will be evaluated descriptively at all applicable 
time points. 
Relationship between vaccine viremia and immunogenicity measures will be assessed using non-
parametric correlation coefficient, as appropriate [21]. Graphical methods will also be explored. 
Further details will be provided in the SAP. 
13.2 
Interim Analysis and Criteria for Early Termination 
No interim analysis is planned. 
13.3 
Determination of Sample Size 
This trial is designed for a descriptive evaluation of the endpoints that does not require 
hypotheses testing. Therefore, the sample size was not determined based on formal statistical 
power calculations. 
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14.0 
QUALITY CONTROL AND QUALITY ASSURANCE 
14.1 
Trial-Site Monitoring Visits 
Monitoring visits to the trial site will be made periodically during the trial to ensure that all 
aspects of the protocol are followed. Source documents will be reviewed for verification of data 
recorded on the eCRFs. Source documents are defined as original documents, data, and records. 
The investigator and institution guarantee access to source documents by the sponsor or designee 
(Clinical Research Organization [CRO]) and by the IRB or IEC. 
All aspects of the trial and its documentation will be subject to review by the sponsor or designee 
(as long as blinding is not jeopardized), including but not limited to the Investigator Site File, 
trial vaccine records, subject medical records, informed consent form documentation, 
documentation of subject authorization to use personal health information (if separate from the 
informed consent forms), and review of eCRFs and associated source documents. It is important 
that the investigator and other trial personnel are available during the monitoring visits and that 
sufficient time is devoted to the process. 
In the event a monitor cannot visit the site in a timely manner due to the COVID-19 pandemic, 
alternative monitoring approaches such as remote source data verification (SDV) or telephone 
contact may be used to ensure data quality and integrity and maintain subject safety. Alternative 
monitoring approaches should be used only where allowed by the local Health Authority and 
when approved by the IRB/IEC. During remote monitoring, the monitor should focus on trial 
activities that are essential to the safety of trial subjects and/or data reliability. 
14.2 
Protocol Deviations 
The investigator should not deviate from the protocol, except where necessary to eliminate an 
immediate hazard to trial subjects. Should other unexpected circumstances arise that will require 
deviation from protocol-specified procedures, the investigator should consult with the medical 
monitor (and IRB or IEC, as required) to determine the appropriate course of action. There will 
be no exemptions (a prospective approved deviation) from the inclusion or exclusion criteria. 
14.3 
Quality Assurance Audits and Regulatory Agency Inspections 
The trial site also may be subject to quality assurance audits by the sponsor or designee. In this 
circumstance, the sponsor-designated auditor will contact the site in advance to arrange an 
auditing visit. The auditor may ask to visit the facilities where laboratory samples are collected, 
where the vaccine is stored and prepared, and any other facility used during the trial. In addition, 
there is the possibility that this trial may be inspected by regulatory agencies, including those of 
foreign governments (eg, the Food and Drug Administration [FDA], the Medicines and 
Healthcare Products Regulatory Agency of the United Kingdom [MHRA], the Pharmaceuticals 
and Medical Devices Agency of Japan [PMDA]). If the trial site is contacted for an inspection by 
a regulatory body, the sponsor should be notified immediately. The investigator and institution 
guarantee access for quality assurance auditors to all trial documents as described in 
Section 14.1. 
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14.4 
Trial Risk Management 
The ICH E6 addendum (R2) guidance encourages a risk-based approach to the management of 
clinical trials and includes requirements for risk control and risk reporting. Takeda or designee 
established Quality Tolerance Limits (QTL) taking into consideration the medical and statistical 
characteristics of the variables and the statistical design of the trial. This process was performed 
according to Takeda internal procedures. 
At the end of the trial, the quality management approach implemented will be described in the 
CSR. If applicable, the CSR will summarize important deviations from the predefined QTL and 
the remedial actions taken. 
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15.0 
ETHICAL ASPECTS OF THE TRIAL 
This trial will be conducted with the highest respect for the trial subjects according to the 
protocol, the ethical principles that have their origin in the Declaration of Helsinki [1], and the 
ICH Harmonized Tripartite Guideline for GCP [2]. Each investigator will conduct the trial 
according to applicable local or regional regulatory requirements and align his or her conduct in 
accordance with the “Responsibilities of the Investigator” that are listed in Appendix A. The 
principles of Helsinki are addressed through the protocol and through appendices containing 
requirements for informed consent and investigator responsibilities. 
15.1 
Institutional Review Board and/or Independent Ethics Committee Approval 
IRBs and IECs must be constituted according to the applicable federal requirements of each 
participating region. The sponsor or designee will require documentation noting all names and 
titles of members who make up the respective IRB or IEC. If any member of the IRB or IEC has 
direct participation in this trial, written notification regarding his or her abstinence from voting 
must also be obtained. If the US site is unwilling to provide names and titles of all members due 
to privacy and conflict of interest concerns, the site should instead provide a Federal Wide 
Assurance Number or comparable number assigned by the Department of Health and Human 
Services. 
The sponsor or designee will supply relevant documents for submission to the respective IRB or 
IEC for the protocol’s review and approval. This protocol, the IB, a copy of the informed consent 
form, and, if applicable, subject recruitment materials and/or advertisements and other 
documents required by all applicable laws and regulations, must be submitted to a central or 
local IRB or IEC for approval. The IRB’s or IEC’s written approval of the protocol and subject 
informed consent form must be obtained and submitted to the sponsor or designee before 
commencement of the trial (ie, before shipment of the trial vaccine(s) or trial specific screening 
activity). The IRB or IEC approval must refer to the trial by exact protocol title, number, and 
version date; identify versions of other documents (eg, informed consent form) reviewed; and 
state the approval date. The sponsor will notify the site once the sponsor has confirmed the 
adequacy of site regulatory documentation and, when applicable, the sponsor has received 
permission from the competent authority to begin the trial. Until the site receives notification, no 
protocol activities, including screening may occur. 
The site must adhere to all requirements stipulated by their respective IRB or IEC. This may 
include notification to the IRB or IEC regarding protocol amendments, updates to the informed 
consent form, recruitment materials intended for viewing by subjects, local safety reporting 
requirements, reports and updates regarding the ongoing review of the trial at intervals specified 
by the respective IRB or IEC, and submission of the investigator’s final status report to IRB or 
IEC. All IRB and IEC approvals and relevant documentation for these items must be provided to 
the sponsor or designee. 
Incentives should not be used to exert undue influence on subjects for participation. Payments to 
subjects must be approved by the IRB or IEC and sponsor. 
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15.2 
Subject Information, Informed Consent, and Subject Authorization 
Written consent documents will embody the elements of informed consent as described in the 
Declaration of Helsinki [1] and the ICH Guidelines for GCP [2] and will be in accordance with 
all applicable laws and regulations. The informed consent form, subject authorization form (if 
applicable), and subject information sheet describe the planned and permitted uses, transfers, and 
disclosures of the subject’s personal and personal health information for the purpose of 
conducting the trial. The informed consent form and the subject information sheet further explain 
the nature of the trial, its objectives, and potential risks and benefits, as well as the date informed 
consent is given. The informed consent form will detail the requirements of the subject and the 
fact that the subject is free to withdraw at any time without giving a reason and without prejudice 
to the subject’s further medical care. 
The investigator is responsible for the preparation, content, and IRB or IEC approval of the 
informed consent form and if applicable, the subject authorization form. The informed consent 
form, subject authorization form (if applicable), and subject information sheet must be approved 
by both the IRB or IEC and the sponsor prior to use. 
The informed consent form, subject authorization form (if applicable), and subject information 
sheet must be written in a language fully comprehensible to the prospective subject. It is the 
responsibility of the investigator to explain the detailed elements of the informed consent form, 
subject authorization form (if applicable), and subject information sheet (if applicable) to the 
subject. Information should be given in both oral and written form whenever possible and in the 
manner deemed appropriate by the IRB or IEC. 
The subject must be given ample opportunity to: (1) inquire about details of the trial and 
(2) decide whether or not to participate in the trial. If the subject determines he or she will 
participate in the trial, then the informed consent form and subject authorization form (if 
applicable) must be signed and dated by the subject, at the time of consent and prior to the 
subject entering into the trial. The subject should be instructed to sign using their legal names, 
not nicknames, using blue or black ballpoint ink. The investigator must also sign and date the 
informed consent form and subject authorization (if applicable) at the time of consent and prior 
to the subject entering into the trial; however, the sponsor may allow a designee of the 
investigator to sign to the extent permitted by applicable law. 
Once signed, the original informed consent, subject authorization form (if applicable), and 
subject information sheet will be stored in the investigator’s site file. The investigator must 
document the date the subject signs the informed consent form in the subject’s medical record 
and eCRF. Copies of the signed informed consent form, the signed subject authorization form (if 
applicable), and subject information sheet (if applicable) shall be given to the subject. 
All revised informed consent forms must be reviewed and signed by the subject in the same 
manner as the original informed consent form. The date the revised consent was obtained should 
be recorded in the subject’s medical record and eCRF, and the subject should receive a copy of 
the revised informed consent form. 
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15.3 
Subject Confidentiality 
The sponsor and designee affirm and uphold the principle of the subject’s right to protection 
against invasion of privacy. Throughout this trial, a subject’s source data will only be linked to 
the sponsor’s clinical trial database or documentation via a unique identification number. As 
permitted by all applicable laws and regulations, limited subject attributes, such as sex, age, or 
date of birth, and subject initials may be used to verify the subject and accuracy of the subject’s 
unique identification number. 
To comply with ICH Guidelines for GCP and to verify compliance with this protocol, the 
sponsor requires the investigator to permit its monitor or designee, representatives from any 
regulatory authority (eg, FDA, MHRA, PMDA), the sponsor’s designated auditors, and the 
appropriate IRBs and IECs to review the subject’s original medical records (source data or 
documents), including, but not limited to, laboratory test result reports, electrocardiogram (ECG) 
reports, admission and discharge summaries for hospital admissions occurring during a subject’s 
trial participation, and autopsy reports. Access to a subject’s original medical records requires 
the specific authorization of the subject as part of the informed consent form process (see 
Section 15.2). 
Copies of any subject source documents that are provided to the sponsor must have certain 
personally identifiable information removed (ie, subject name, address, and other identifier fields 
not collected on the subject’s eCRF). 
15.4 
Clinical Trial Registration, Publication and Disclosure Policy 
15.4.1 Clinical Trial Registration 
In order to ensure that information on clinical trials reaches the public in a timely manner and to 
comply with applicable law, regulation and guidance, the sponsor will, as a minimum register all 
clinical trials conducted in subjects that it sponsors anywhere in the world, on publicly accessible 
websites such as ClinicalTrials.gov and EudraCT, according to local requirements, before trial 
initiation. The sponsor contact information, along with the investigator’s city, country, and 
recruiting status will be registered and available for public viewing. 
15.4.2 Clinical Trial Results Disclosure 
The sponsor will post the results of this clinical trial regardless of outcome, on publicly 
accessible websites such as ClinicalTrials.gov and/or EudraCT, as required by applicable laws 
and/or regulations. 
Trial completion corresponds to the date on which the final subject was examined or received an 
intervention for the purposes of final collection of data (Last Subject Last Visit). 
15.4.3 Publication of Trial Results 
The results of this trial are expected to be published in a scientific journal. It is anticipated that 
clinical and laboratory co-investigators will participate in authorship. The authorship, order of 
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authors and choice of journal will be proposed by the sponsor to the Principal Investigator, to be 
eventually agreed upon by all authors themselves. The data analysis center for this trial will 
provide the analyses needed for publication. Information regarding this trial will be posted on 
ClinicalTrials.gov and/or other registries according to the local requirements. 
15.5 
Insurance and Compensation for Injury 
Each subject in the trial must be insured in accordance with the regulations applicable to the site 
where the subject is participating. If a local underwriter is required, then the sponsor or sponsor’s 
designee will obtain clinical trial insurance against the risk of injury to clinical trial subjects. 
Refer to the Clinical Study Site Agreement regarding the sponsor’s policy on subject 
compensation and treatment for injury. If the investigator has questions regarding this policy, he 
or she should contact the sponsor or sponsor’s designee. 
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16.0 
REFERENCES 
1. World Medical Association Declaration of Helsinki: ethical principals for medical research 
involving human subjects. JAMA 2013;310(20):2191-4. 
2. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for 
Human Use (ICH). ICH harmonised guideline. Integrated Addendum to ICH E6 (R1): 
Guideline for Good Clinical Practice E6 (R2). Available at https://database.ich.org/sites/ 
default/ files/E6_R2_Addendum.pdf. [Accessed 18 December 2020]. 
3. Simmons CP, Farrar JJ, Nguyen vV, Wills B. Dengue. N Engl J Med 2012;366(15):1423-32. 
4. Gubler DJ. Epidemic dengue/dengue hemorrhagic fever as a public health, social and 
economic problem in the 21st century. Trends Microbiol 2002;10(2):100-3. 
5. Guzman M, Kouri G. Dengue: an update. Lancet Infect Dis 2002;2(1):33-42. 
6. World Health Organization, 2018. Dengue and severe dengue. Fact Sheet. Available at 
https://www.who.int/en/news-room/fact-sheets/detail/dengue-and-severe-dengue [Accessed 
18 December 2020]. 
7. World Health Organization, 1997. Dengue hemorrhagic fever: diagnosis, treatment, 
prevention and control, 2nd Edition. Geneva. Available at 
https://www.who.int/csr/resources/publications/dengue/Denguepublication/en/ [Accessed 18 
December 2020]. 
8. World Health Organization, 2009. Dengue guidelines for diagnosis, treatment, prevention 
and control. Available at http://www.who.int/tdr/publications/documents/dengue-
diagnosis.pdf. [Accessed 18 December 2020]. 
9. Halstead S, Deen J. The future of dengue vaccines. Lancet 2002;360(9341):1243-5. 
10. Wichmann O, Vannice K, Asturias EJ, de Albuquerque Luna EJ, Longini I, Lopez AL, et al. 
Live-attenuated tetravalent dengue vaccines: The needs and challenges of post-licensure 
evaluation of vaccine safety and effectiveness. Vaccine. 2017. 35(42):5535-42. 
11. Hadinegoro SR, Arredondo-García JL, Capeding MR, Deseda C, Chotpitayasunondh T, 
Dietze R, et al. Efficacy and long-term safety of a dengue vaccine in regions of endemic 
disease. N Engl J Med 2015;373(13):1195-206. 
12. Sridhar S, Luedtke A, Langevin E, Zhu M, Bonaparte M, Machabert T, et al. Effect of 
dengue serostatus on dengue vaccine safety and efficacy. N Engl J Med. 2018. 379(4):327-
40. 
13. World Health Organization. Weekly Epidemiological Record. 2018. 93:329-44. Available at 
http://www.who.int/wer [Accessed 18 December 2020]. 
14. Yoksan S, Bhamarapravati N, Halstead SB. Dengue virus vaccine development: study on 
biological markers of uncloned dengue 1-4 viruses serially passaged in primary kidney cells. 
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In: TD St. George, BH Kay and J Blok (ed.), Arbovirus research in Australia. Proceedings of 
the 4th Symposium. CSIRO/QIMR, Brisbane, Australia, 1986:35-8. 
15. Huang CY, Butrapet S, Tsuchiya KR, Bhamarapravati N, Gubler DJ, Kinney RM. Dengue 2 
PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development. J. Virol 
2003;77(21):11436-47. 
16. Huang CY, Butrapet S, Pierro DJ, Chang GJ, Hunt AR, Bhamarapravati N, et al. Chimeric 
dengue type 2 (vaccine strain PDK-53)/dengue type 1 virus as a potential candidate dengue 
type 1 virus vaccine. J Virol. 2000;74(7):3020-8. 
17. Policy and communication bulletin – The clinical center. Guidelines for limits of blood 
drawn for research purposes in the clinical center. Manual transmittal sheet, no. M95-9 (rev.), 
2009. Available at https://irb.research.chop.edu/.../files/documents/g_nih_blooddraws 
[Accessed 18 December 2020]. 
18. World Health Organization. Reducing pain at the time of vaccination: WHO position paper – 
September 2015.Wkly Epidemiol Rec 2015;90(39):505-10. 
19. Marcy SM, Kohl KS, Dagan R, Nalin D, Blum M, Jones MC, et al. Brighton Collaboration 
Fever Working Group. Fever as an adverse event following immunization: case definition 
and guidelines of data collection, analysis and presentation. Vaccine 2004;22(5-6):551-6. 
20. WHO Drug Dictionary. Available at http://www.who.int/medicines/publications/ 
druginformation/en/ [Accessed 18 December 2020]. 
21. Clopper CJ, Pearson ES. The use of confidence or fiducial limits illustrated in the case of the 
binomial. Biometrika 1934;26(4):404-13. 
22. Kruskal WH. Ordinal Measures of Association. JASA 1958;53(284):814-61. 
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Appendix A Responsibilities of the Investigator 
Clinical research studies sponsored by the sponsor are subject to ICH GCP and all the applicable 
local laws and regulations. 
The investigator agrees to assume the following responsibilities: 
1. Conduct the trial in accordance with the protocol. 
2. Personally conduct or supervise the staff that will assist in the protocol. 
3. Ensure that trial related procedures, including trial specific (non-routine/non-standard panel) 
screening assessments, are NOT performed on potential subjects prior to the receipt of 
written approval from relevant governing bodies/authorities. 
4. Ensure that all colleagues and employees assisting in the conduct of the trial are informed of 
these obligations. 
5. Secure prior approval of the trial and any changes by an appropriate IRB/IEC that conform to 
21 Code of Federal Regulations (CFR) Part 56, ICH, and local regulatory requirements. 
6. Ensure that the IRB/IEC will be responsible for initial review, continuing review, and 
approval of the protocol. Promptly report to the IRB/IEC all changes in research activity and 
all anticipated risks to subjects. Make at least yearly reports on the progress of the trial to the 
IRB/IEC, and issue a final report within 3 months of trial completion. 
7. Ensure that requirements for informed consent, as outlined in 21 CFR Part 50, ICH and local 
regulations, are met. 
8. Obtain valid informed consent from each subject who participates in the trial, and document 
the date of consent in the subject’s medical chart. Valid informed consent form is the most 
current version approved by the IRB/IEC. Each informed consent form should contain a 
subject authorization section that describes the uses and disclosures of a subject’s personal 
information (including personal health information) that will take place in connection with 
the trial. If an informed consent form does not include such a subject authorization, then the 
investigator must obtain a separate subject authorization form from each subject. 
9. Prepare and maintain adequate case histories of all persons entered into the trial, including 
eCRFs, hospital records, laboratory results, etc, and maintain these data for a minimum of 
2 years following notification by the sponsor that all investigations have been discontinued or 
that the regulatory authority has approved the marketing application. The investigator should 
contact and receive written approval from the sponsor before disposing of any such 
documents. 
10. Allow possible inspection and copying by the regulatory authority of GCP-specified essential 
documents. 
11. Maintain current records of the receipt, administration, and disposition of sponsor-supplied 
vaccines, and return all unused sponsor-supplied vaccines to the sponsor. 
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12. Report AEs to the sponsor promptly. In the event of an SAE, notify the sponsor within 
24 hours. 
13. Review and provide a signature as approval of the content of the CSR. 
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Appendix B Investigator Consent to Use of Personal Information 
Takeda will collect and retain personal information of the investigator, including his or her name, 
address, and other personally identifiable information. In addition, investigator’s personal 
information may be transferred to other parties located in countries throughout the world (eg, the 
United Kingdom, United States, and Japan), including the following: 
 Takeda, its affiliates, and licensing partners. 
 Business partners assisting Takeda, its affiliates, and licensing partners. 
 Regulatory agencies and other health authorities. 
 IRBs and IECs. 
Investigator’s personal information may be retained, processed, and transferred by Takeda and 
these other parties for research purposes including the following: 
 Assessment of the suitability of investigator for the trial and/or other clinical studies. 
 Management, monitoring, inspection, and audit of the trial. 
 Analysis, review, and verification of the trial results. 
 Safety reporting and pharmacovigilance relating to the trial. 
 Preparation and submission of regulatory filings, correspondence, and communications to 
regulatory agencies relating to the trial. 
 Preparation and submission of regulatory filings, correspondence, and communications to 
regulatory agencies relating to other vaccines used in other clinical studies that may contain 
the same chemical compound present in the investigational vaccine. 
 Inspections and investigations by regulatory authorities relating to the trial. 
 Self-inspection and internal audit within Takeda, its affiliates, and licensing partners. 
 Archiving and audit of trial records. 
 Posting investigator site contact information, trial details and results on publicly accessible 
clinical trial registries, databases, and websites. 
Investigator’s personal information may be transferred to other countries that do not have data 
protection laws that offer the same level of protection as data protection laws in investigator’s 
own country. Investigator acknowledges and consents to the use of his or her personal 
information by Takeda and other parties for the purposes described above. 
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Signature Page for DEN-210 Protocol Amendment 4, Version 5.0, 18 December 2020
Title: An Open-Label, Phase 2 Trial to Investigate the Humoral and Cell-Mediated
Document Number: TAK-003-02389 v5.0
Approval
Clinical
12-Jan-2021 13:09:42 GMT+0000
Approval
Clinical
12-Jan-2021 13:17:19 GMT+0000
Approval
Pharmacovigilance
12-Jan-2021 16:58:40 GMT+0000
Approval
Statistics
12-Jan-2021 22:41:18 GMT+0000
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