Conditions: Cytomegalovirus (CMV) Infections
Keywords: Prevention of cytomegalovirus infection (CMVi)
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 2200
Healthy volunteers: yes

Brief summary:
This study evaluated the safety, tolerability, and efficacy of the cytomegalovirus (CMV) vaccine (V160) administered in a 2-dose or 3-dose regimen to healthy seronegative women 16 to 35 years of age. Participants received blinded V160 on Day 1, Month 2, and Month 6 (3-dose regimen), V160 on Day 1 and Month 6 and placebo at Month 2 (2-dose regimen), or placebo on Day 1, Month 2, and Month 6, and were followed to approximately Month 24. The primary hypothesis of the study was that administration of a 3-dose regimen of V160 will reduce the incidence of primary CMV infection compared to placebo.

Detailed description:

Arms:
arm 1: Participants received 3 doses of vaccine V160 (100 Units/0.5 mL dose with Merck aluminum phosphate adjuvant \[MAPA\], 4°C stable formulation) administered by intramuscular (IM) injection on Day 1, Month 2, and Month 6. arm 2: Participants received 2 doses of vaccine V160 (100 Units/0.5 mL dose with MAPA, 4°C stable formulation) administered IM on Day 1 and Month 6 and a placebo-saline solution at Month 2. arm 3: Participants received placebo (saline solution) by IM injection on Day 1, Month 2, and Month 6.

Interventions: intervention 1: V160 intervention 2: Placebo
Intervention descriptions: intervention 1: V160 was administered as a 0.5 mL (100 Units/0.5 mL dose with Merck aluminum phosphate adjuvant \[MAPA\], 4°C stable formulation) IM injection. intervention 2: Saline solution administered as a 0.5 mL IM injection

Protocol document:
Prot_SAP_000.pdf: 
Official Protocol Title: 
NCT number: 
NCT03486834  
Document Date: 
07-MAY-2019
Double-Blind, Randomized, Placebo-Controlled Phase 2b, Multi-center
Study to Evaluate the Safety, Tolerability, Efficacy and Immunogenicity of a 
2-Dose and a 3-Dose Regimen of V160 (Cytomegalovirus [CMV] Vaccine) in 
Healthy Seronegative Women, 16 to 35 Years of Age


Product:  V160 
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Title Page
THIS PROTOCOL AMENDMENT AND ALL OF THE INFORMATION RELATING 
TO IT ARE CONFIDENTIAL AND PROPRIETARY PROPERTY OF MERCK 
SHARP & DOHME CORP., A SUBSIDIARY OF MERCK & CO., INC., 
WHITEHOUSE STATION, NJ, U.S.A.
Protocol Title: Double-Blind, Randomized, Placebo-Controlled Phase 2b, Multi-center 
Study to Evaluate the Safety, Tolerability, Efficacy and Immunogenicity of a 2-Dose and a 3-
Dose Regimen of V160 (Cytomegalovirus [CMV] Vaccine) in Healthy Seronegative 
Women, 16 to 35 Years of Age
Protocol Number: 002-02
Compound Number: V160
Sponsor Name and Legal Registered Address: 
Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc. 
(hereafter referred to as the Sponsor or Merck)
One Merck Drive
P.O. Box 100
Whitehouse Station, New Jersey, 08889-0100, U.S.A.
Regulatory Agency Identifying Number(s):
IND NUMBER: [15778]
EudraCT NUMBER: [2017-004233-86]
Approval Date:  07 May 2019
 
 
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Sponsor Signatory
Typed Name:
Title:
Date
Protocol-specific Sponsor Contact information can be found in the Investigator Trial 
File Binder (or equivalent). 
Investigator Signatory
I agree to conduct this clinical trial in accordance with the design outlined in this protocol 
and to abide by all provisions of this protocol.
Typed Name:
Title:
Date
 
 
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DOCUMENT HISTORY
Document
Date of Issue
Overall Rationale
V160-002-02
07-May-2019
Updated cCMVi Case Definition, clarified the infant sample collection 
strategy, and adjusted visit schedules to be more accommodating and 
maintain more contact with study participants
V160-002-01
06-Aug-2018
Updated protocol procedures for clarification
V160-002-00
31-Oct-2017
Original Protocol
 
 
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PROTOCOL AMENDMENT SUMMARY OF CHANGES
Amendment [02]
Overall Rationale for the Amendment:
Updated the cCMVi Case Definition, clarified the infant sample collection strategy and adjusted visit schedules to be more 
accommodating and maintain more contact with study participants
Summary of Changes Table:
Section # and 
Name
Description of Change
Brief Rationale
2.1 Schedule of 
Activities: 
Screening 
Through 
Treatment Phase
Changed Visit 3/Month 2 visit window to 
21 days; Changed Visit 4/Month 6 visit 
window to 28 days.
The visit windows for the vaccination visits at Month 2 and Month 
6 were widened to allow more flexibility with scheduling these 
visits.
2.2 Schedule of 
Activities: 
Follow-up Period
Changed Month 18 telephone contact to 
Study Visit; Added Month 30 Study Visit.
Two in-clinic study visits are being added to maintain participant-
site contact and promote retention of study participants.  The 
additional visits will also allow for more efficient management of 
urine/saliva self-collection kits. One visit/row was added to SoA 
2.2, and the other to SoA 2.3.
 
 
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Section # and 
Name
Description of Change
Brief Rationale
2.3 Schedule of 
Activities for 
Congenital 
Cytomegalovirus 
Infection 
Assessment

Clarified the visit window for cCMVi 
Visit 2.

Added a row for an activity to record 
any anti-CMV treatments received by 
participants during pregnancy.

Added a column for “Additional 
cCMVi visit (if necessary)”

Added a Notes column and moved 
saliva collection information to 
column

cCMVi Visit 2 should be scheduled between the day of birth 
and up to 14 days after birth. A window of +7 days is specified 
to ensure infant samples are collected within 21 days of birth
(a requirement of the cCMVi case definition).

Collection of anti-CMV treatments received by participants 
during pregnancy is needed for comprehensive documentation 
of pregnancy outcomes.

The optional additional visit was added to allow for the 
collection of a 2nd set of infant urine and saliva samples, if 
they need to be collected on a separate day.  The visit window 
on this visit is also specified to ensure that the 2nd set of infant 
samples is collected within 21 days of birth (a requirement of 
the cCMVi case definition)

Clarification of information
5.1.1 Study 
Diagram
Updated the Study Diagram to include 
on−site visits at Months 18 and 30.
The updates were added for consistency with the Schedule of 
Activities (SoA).
6.2.1 Participant 
Deferment 
Criteria before 
Vaccination
Deleted language stating the participant 
would only be permitted to continue with 
a delayed vaccination if the vaccine can be 
administered within the allowed window
The requirement to only allow delayed vaccination if the vaccine 
can be administered within the allowed window is unnecessarily 
restrictive.
 
 
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Section # and 
Name
Description of Change
Brief Rationale
7.3 Method of 
Treatment 
Assignment
Clarified that approximately 360 
participants (or as many as operationally 
feasible) will be included in the Detailed 
Immunogenicity sub-study
Due to the high screen failure rate, it may not be logistically 
feasible to enroll 360 participants in the Detailed Immunogenicity 
Sub-Study as initially planned for this exploratory endpoint.
8.1 
Discontinuation 
of Study 
Treatment

Clarified that the self-collection of 
urine and saliva samples by study 
participants should continue regardless 
of whether they complete the entire 
vaccine regimen

Removed last bullet point stating that a
participant who has received any 
prohibited medications or vaccinations 
since Dose 1 administration must be 
discontinued and added a bullet for 
“Received a non-study CMV vaccine.”

Statement added to emphasize that self-collection of urine and 
saliva samples by study participants should continue 
regardless of whether they complete the entire vaccine 
regimen.

The requirement for participants to be discontinued from study 
treatment if they received any prohibited medications or 
vaccination since Dose 1 is unnecessarily restrictive. However, 
participants who receive a non-study CMV vaccine must be 
discontinued from study treatment.
Section 9 Study 
Assessments and 
Procedures
Updated visit numbers for the Month 24 
and 36 sample collections in Table 3.
Visit numbers in Table 3 were updated for consistency with the 
new visit numbers for the Month 24 and Month 30 visits in the 
SoA.
9.1.8 Treatment 
Administration
Updated first sentence to expand 
information regarding qualified study 
personnel
Conformation with current template language
 
 
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Section # and 
Name
Description of Change
Brief Rationale
9.2.1.1.2 CMVi 
Case Detection
Clarified that the monthly urine and saliva 
samples may be collected outside of the 
study site or at scheduled site visits (when 
applicable), as indicated in the SoA
Urine and saliva samples are self-collected by the participant at 
monthly intervals. For intervals that coincide with scheduled study 
visits, it is permissible for the samples to be collected during the 
on-site visit.
9.2.1.2.1 cCMVi 
Case Definition
Updated cCMVi Case Definition
With input from external experts and regulatory agencies, it has
been determined that only cases of cCMVi which have been 
virologically confirmed will be consider as endpoints for analysis. 
For this reason, “Pregnancy impacted by CMV but CMV not 
virologically confirmed” has been removed from the cCMVi case 
definition. Language was also revised to provide further 
clarification.
9.2.1.2.2 cCMVi 
Case Detection
Updated cCMVi case detection procedures

Clarified that collection of anti-
CMV treatments received by 
participants during pregnancy will 
be collected for the duration of the 
study.  Added statement that “all 
relevant medical records should be 
placed in the participant’s trial file.”

Clarified that 2 sets of urine and 
saliva will be collected from the 
infant within 21 days of birth.

Revised the trigger for adjudication 
to be suspicion of congenital CMV 
during pregnancy of a study 
participant or in the infant.

Collection of anti-CMV treatments received by participants 
during pregnancy is needed for comprehensive documentation 
of pregnancy outcomes.  Clarification of information.

2 sets of infant urine and saliva samples collected within 21 
days of birth are needed to support the updated cCMVi Case 
Definition.

Because the cCMVi Case Definition was revised, the trigger 
for adjudication was revised accordingly.
 
 
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Section # and 
Name
Description of Change
Brief Rationale
9.3.1 Time Period 
and Frequency for 
Collecting AE, 
SAE and Other 
Reportable Safety 
Event 
Information
In Table 4, in the row for “Lactation 
Exposure” added the following 
information:

Reporting Time Period: After the 
Protocol Specified Follow-up 
Period 2 column: “Previously 
reported – follow to completion”.

Timeframe to Report Event and 
Follow-up Information to 
SPONSOR column: “Within 24 
hours of learning of event.”
In V160-002-01, the “Reporting Time Period: After the Protocol 
Specified Follow-up Period 2” and “Timeframe to Report Event 
and Follow-up Information to SPONSOR” columns were 
inadvertently left blank in the Lactation Exposure row.
9.3.3 Follow-up 
of AE, SAE and 
Other Reportable 
Safety Event 
Information
Clarified that participants who 
inadvertently become pregnant before 
receiving all 3 doses of study vaccine or 
placebo, and any participant who becomes 
pregnant after the treatment phase will 
remain in the trial for follow-up of 
pregnancy and infant outcome
Previous text was ambiguous and clarifies that all pregnancies will 
be followed for outcome regardless of the vaccination status of the 
participant.
10.5.1 Efficacy 
Analysis 
Populations
Changed PPE Analysis population 
definition for all arms to include all 3 
doses
The definition was revised to minimize potential bias in efficacy 
analyses.
 
 
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Section # and 
Name
Description of Change
Brief Rationale
12.3 Appendix 3 
Contraceptive 
Guidance and 
Pregnancy 
Testing
Removed bilateral tubal ligation occlusion 
as allowable contraceptive.  Added 
statement: “As WOCBP are the target 
population of this study and pregnancy is 
permissible after the treatment phase, 
permanent contraception methods at study 
entry are not allowed.”
This study is designed to enroll women of childbearing potential 
(WOCBP). Bilateral tubal ligation occlusion is a permanent 
procedure therefore it is not an allowable contraceptive method.
12.4 Appendix 4 
Adverse Events: 
Definitions and 
Procedures for 
Recording, 
Evaluating, 
Follow-up, and 
Reporting
Updated text in Assessment of Intensity
section.
Conformation with current template language
Additional non-substantive editorial/grammatical/typographical changes were made as appropriate.
 
 
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Table of Contents
DOCUMENT HISTORY ........................................................................................................3
PROTOCOL AMENDMENT SUMMARY OF CHANGES...............................................4
1.
Synopsis.......................................................................................................................19
2.
Schedule of Activities (SoA)......................................................................................22
2.1
Schedule of Activities: Screening Through Treatment Phase ............................22
2.2
Schedule of Activities: Follow-up Period Through Approximately 
Month 36 (Approximate Time Point to Reach Cases Required for 
Primary Endpoint Assessment) .............................................................................25
2.3
Schedule of Activities for Congenital Cytomegalovirus Infection 
Assessment...............................................................................................................26
3.
Introduction................................................................................................................27
3.1
Study Rationale .......................................................................................................27
3.2
Background..............................................................................................................27
Pharmaceutical and Therapeutic Background .....................................................29
Pre-clinical and Clinical Studies..........................................................................29
3.2.2.1
Pre-Clinical ...................................................................................................29
3.2.2.2
Clinical Studies.............................................................................................30
3.2.2.3
V160 Replication ..........................................................................................31
3.3
Benefit/Risk Assessment.........................................................................................31
4.
Objectives/Hypotheses and Endpoints.....................................................................32
5.
Study Design...............................................................................................................34
5.1
Overall Design .........................................................................................................34
Study Diagram .....................................................................................................35
5.2
Number of Participants ..........................................................................................35
5.3
Beginning and End of Study Definition ................................................................36
Clinical Criteria for Early Study Termination .....................................................36
5.4
Scientific Rationale for Study Design....................................................................36
Rationale for Endpoints .......................................................................................37
5.4.1.1
Rationale for Efficacy Endpoint ...................................................................37
 
 
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5.4.1.2
Rationale for Immunogenicity Endpoints.....................................................37
5.4.1.3
Rationale for Safety Endpoints .....................................................................38
5.4.1.4
Rationale for Congenital CMVi Exploratory Endpoint................................38
5.4.1.5
Planned Exploratory Biomarker Research....................................................39
5.4.1.5.1
Future Biomedical Research..................................................................39
5.4.1.6
Rationale for the Use of Comparator/Placebo ..............................................39
5.4.1.7
Rationale for Adjuvant..................................................................................39
5.5
Justification for Dose ..............................................................................................40
Rationale for Dose Interval and Trial Design ......................................................40
6.
Study Population........................................................................................................40
6.1
Inclusion Criteria....................................................................................................40
6.2
Exclusion Criteria ...................................................................................................41
Participant Deferment Criteria before Vaccination .............................................43
Criteria to be Checked for Excluding Participants from Receiving 
Subsequent Doses ................................................................................................43
6.3
Lifestyle Restrictions...............................................................................................43
Meals and Dietary Restrictions............................................................................43
Caffeine Alcohol, and Tobacco Restrictions .......................................................44
Activity ................................................................................................................44
6.4
Screen Failures ........................................................................................................44
6.5
Participant Replacement Strategy.........................................................................44
7.
Treatments..................................................................................................................44
7.1
Treatments Administered.......................................................................................44
7.2
Dose Modification ...................................................................................................45
7.3
Method of Treatment Assignment.........................................................................45
Stratification.........................................................................................................47
7.4
Blinding....................................................................................................................47
7.5
Preparation/Handling/Storage/Accountability ....................................................47
Dose Preparation..................................................................................................47
Handling, Storage and Accountability.................................................................48
7.6
Treatment Compliance...........................................................................................48
7.7
Concomitant Therapy.............................................................................................49
 
 
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Rescue Medications and Supportive Care ...........................................................49
7.8
Treatment After the End of the Study ..................................................................49
7.9
Clinical Supplies Disclosure...................................................................................49
8.
Discontinuation/Withdrawal Criteria......................................................................50
8.1
Discontinuation of Study Treatment.....................................................................50
8.2
Withdrawal from the Study ...................................................................................51
8.3
Lost to Follow Up....................................................................................................51
9.
Study Assessments and Procedures..........................................................................51
9.1
Administrative and General Procedures ..............................................................53
Informed Consent/Assent.....................................................................................53
9.1.1.1
General Informed Consent/Assent................................................................53
9.1.1.2
Consent/Assent and Collection of Specimens for Future
Biomedical Research ....................................................................................53
Inclusion/Exclusion Criteria ................................................................................54
Participant Identification Card.............................................................................54
Medical History ...................................................................................................54
Prior and Concomitant Medications Review .......................................................54
9.1.5.1
Prior Medications..........................................................................................54
9.1.5.2
Concomitant Medications.............................................................................54
Assignment of Screening Number.......................................................................54
Assignment of Treatment/Randomization Number.............................................55
Treatment Administration....................................................................................55
9.1.8.1
Timing of Dose Administration....................................................................56
Withdrawal/Discontinuation................................................................................56
9.1.9.1
Withdrawal From Future Biomedical Research ...........................................56
Participant Blinding/Unblinding..........................................................................56
Distribution of Electronic Vaccination Report Card............................................57
Calibration of Equipment.....................................................................................57
9.2
Efficacy and Immunogenicity Assessments ..........................................................58
Efficacy Assessment ............................................................................................58
9.2.1.1
CMVi ............................................................................................................58
9.2.1.1.1
CMVi Case Definition...........................................................................58
 
 
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9.2.1.1.2
CMVi Case Detection............................................................................58
9.2.1.1.3
CMV PCR Testing.................................................................................58
9.2.1.2
cCMVi...........................................................................................................59
9.2.1.2.1
cCMVi Case Definition .........................................................................59
9.2.1.2.2
cCMVi Case Detection ..........................................................................59
9.2.1.2.3
Health Follow-Up of Infants Diagnosed with cCMVi...........................61
Immunogenicity Assessments..............................................................................61
9.2.2.1
Viral Neutralizing Antibody Assay ..............................................................61
9.2.2.2
IgG ELISA Assay .........................................................................................61
9.2.2.3
CMV IFN-γ ELISPOTs Assay......................................................................61
9.3
Adverse Events (AE), Serious Adverse Events (SAE) and Other 
Reportable Safety Events .......................................................................................62
Time Period and Frequency for Collecting AE, SAE and Other 
Reportable Safety Event Information ..................................................................62
Method of Detecting AE, SAE and Other Reportable Safety Events..................64
Follow-up of AE, SAE and Other Reportable Safety Event Information............64
Regulatory Reporting Requirements for SAE .....................................................64
Disease-Related Events and/or Disease-Related Outcomes Not 
Qualifying as AEs or SAEs..................................................................................64
Pregnancy and Exposure During Breastfeeding ..................................................65
Events of Clinical Interest (ECIs) ........................................................................65
9.4
Treatment of Overdose...........................................................................................65
9.5
Safety........................................................................................................................65
Physical Examinations (Full and Targeted Examinations)..................................66
Vital Signs and Body Temperature......................................................................66
Laboratory Procedures and Assessments.............................................................66
9.5.3.1
Pregnancy Test..............................................................................................67
Safety Assessments Using the eVRC ..................................................................67
9.6
Pharmacokinetics....................................................................................................67
9.7
Pharmacodynamics.................................................................................................67
9.8
Biomarkers ..............................................................................................................68
9.9
Future Biomedical Research Sample Collection..................................................68
9.10
Visit Requirements..................................................................................................68
 
 
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10.
Statistical Analysis Plan ............................................................................................68
10.1
Statistical Analysis Plan Summary........................................................................69
10.2
Responsibility for Analyses/In-House Blinding....................................................71
10.3
Hypotheses/Estimation ...........................................................................................73
10.4
Analysis Endpoints..................................................................................................73
Efficacy Endpoints...............................................................................................73
Immunogenicity Endpoints..................................................................................73
Safety Endpoints ..................................................................................................73
10.5
Analysis Populations...............................................................................................74
Efficacy Analysis Populations .............................................................................74
Immunogenicity Analysis Populations ................................................................74
Safety Analysis Population..................................................................................74
10.6
Statistical Methods..................................................................................................75
Statistical Methods for Efficacy Analyses ...........................................................75
Statistical Methods for Immunogenicity Analyses ..............................................76
Statistical Methods for Safety Analyses ..............................................................76
10.7
Interim Analyses .....................................................................................................78
Efficacy Interim Analysis ....................................................................................78
Safety Interim Analysis........................................................................................79
10.8
Multiplicity ..............................................................................................................80
10.9
Sample Size and Power Calculations ....................................................................80
Sample Size and Power for Efficacy Analyses....................................................80
10.9.1.1 Sample Size and Power for Testing the Primary Efficacy 
Hypothesis ....................................................................................................80
10.9.1.2 Sample Size and Power for Testing the Secondary Efficacy 
Hypothesis ....................................................................................................83
Sample Size and Power for Safety Analyses .......................................................85
10.10 Subgroup Analyses and Effect of Baseline Factors..............................................86
10.11 Compliance ..............................................................................................................86
Adherence to Scheduled Vaccinations .........................................................86
Adherence to Scheduled Self-Collection of Urine and Saliva 
Samples................................................................................................................86
10.12 Extent of Exposure..................................................................................................87
 
 
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11.
References...................................................................................................................88
12.
Appendices..................................................................................................................92
12.1
Appendix 1: Study Governance Considerations ..................................................92
Merck Code of Conduct for Clinical Trials .....................................................................92
Financial Disclosure.........................................................................................................94
Data Protection.................................................................................................................94
Confidentiality of Data ....................................................................................................94
Confidentiality of Participant Records.............................................................................94
Confidentiality of IRB/IEC Information..........................................................................95
Committees Structure.......................................................................................................95
Scientific Advisory Committee....................................................................................95
Executive Oversight Committee..................................................................................95
Data Monitoring Committee........................................................................................95
Clinical Adjudication Committee ................................................................................96
Publication Policy............................................................................................................96
Compliance with Trial Registration and Results Posting Requirements.........................96
Compliance with Law, Audit and Debarment .................................................................96
Data Quality Assurance ...................................................................................................97
Source Documents ...........................................................................................................98
Study and Site Closure.....................................................................................................98
12.2
Appendix 2: Collection and Management of Specimens for Future 
Biomedical Research...............................................................................................99
12.3
Appendix 3: Contraceptive Guidance and Pregnancy Testing.........................103
Contraception Requirements..........................................................................................103
Pregnancy Testing..........................................................................................................105
12.4
Appendix 4: Adverse Events: Definitions and Procedures for 
Recording, Evaluating, Follow-up, and Reporting............................................106
Definition of AE ............................................................................................................106
Definition of SAE ..........................................................................................................107
Additional Events Reported...........................................................................................108
Recording AE and SAE .................................................................................................108
Reporting of AE, SAE, and Other Reportable Safety Events to the Sponsor................113
12.5
Appendix 5: Laboratory Assays ..........................................................................115
 
 
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12.6
Appendix 6: Abbreviations and Trademarks.....................................................118
 
 
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LIST OF TABLES
    
Table 1
Study Treatments..................................................................................................45
Table 2
Allocation Ratio across Treatment Arms and Detailed Immunogenicity 
Sub-Study .............................................................................................................46
Table 3
Approximate Blood/Tissue Volumes Drawn/Collected by Trial Visit 
and by Sample Types............................................................................................52
Table 4
Reporting Time Periods and Timeframes for Adverse Events and Other 
Reportable Safety Events......................................................................................63
Table 5
Analysis Strategy for Safety Parameters ..............................................................78
Table 6
Decision Rule and Corresponding Type 1 Error and Power for Testing 
the Primary Efficacy Hypothesis of VE >0% Against CMVi (3-Dose 
Regimen vs. Placebo) ...........................................................................................82
Table 7
Decision Rule and Corresponding Type 1 Error and Power for Testing 
the Secondary Efficacy Hypothesis of VE >0% Against CMVi (2-Dose 
Regimen vs. Placebo) ...........................................................................................84
Table 8
Critical Counts of Participants with Adverse Events in V160 and 
Placebo Arms Resulting in the Lower Bound of 95% CI Estimate of 
Risk Difference to be >0% ...................................................................................85
Table 9
Highly Effective Contraception Methods...........................................................104
 
 
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LIST OF FIGURES
          
Figure 1
Trial Design Diagram ...........................................................................................35
Figure 2
Plan for Testing the Primary and Secondary Efficacy Hypotheses......................79
 
 
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1. Synopsis
Protocol Title:  
Double-Blind, Randomized, Placebo-Controlled Phase 2b, Multi-center Study to 
Evaluate the Safety, Tolerability, Efficacy and Immunogenicity of a 2-Dose and a 3-
Dose Regimen of V160 (Cytomegalovirus [CMV] Vaccine) in Healthy Seronegative 
Women, 16 to 35 Years of Age
Short Title: 
V160 2-Dose and 3-Dose Regimens vs. Placebo in Healthy CMV Seronegative Females
Objectives/Hypotheses and Endpoints:
In healthy females between 16 and 35 years of age:
Objective/Hypothesis
Endpoint
Primary

Objective: To demonstrate the 
efficacy of a 3-dose regimen of V160 
in reducing the incidence of primary 
CMVi during the follow-up period 
after the last dose of the vaccine
Hypothesis 1: Administration of a 3-
dose regimen of V160 will reduce the 
incidence of primary CMVi 
compared to placebo. (The statistical 
criterion for success requires the 
lower limit of the 95% confidence 
interval [CI] of vaccine efficacy [VE]
to be greater than 0%).

Incidence of CMVi in the 3-dose 
vaccine and placebo groups during the 
follow-up period after the last dose of 
a 3-dose regimen of the vaccine

Objective: To assess the safety and 
tolerability of a 2-dose and a 3-dose 
regimen of V160

Number of participants in the 2-dose 
and 3-dose vaccine groups 
experiencing solicited injection site 
adverse events (AEs) within Days 1 
through 5 after each vaccination visit; 
solicited systemic AEs; and vaccine-
related serious AEs (SAEs) within 
Days 1 through 14 after each 
vaccination visit
 
 
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Secondary 

Objective: To demonstrate the 
efficacy of a 2-dose regimen of V160 
in reducing the incidence of primary 
CMVi during the follow-up period 
after the last dose of the vaccine
Hypothesis 2: Administration of a 2-
dose regimen of V160 will reduce the 
incidence of primary CMVi 
compared to placebo. (The statistical 
criterion for success requires the 
lower limit of the 95% CI of VE to be 
greater than 0%).

Incidence of CMVi in the 2-dose 
vaccine and placebo groups during the 
follow-up period after the last dose of 
a 2-dose regimen of the vaccine
Overall Design:
Study Phase
Phase 2b
Clinical Indication
Prevention of CMVi
Population
Participants will be healthy, CMV seronegative, non-
pregnant, female, age 16-35 years 
Infants of participants will be followed for congenital 
CMVi
Study Type
Interventional
Type of Design
Parallel, controlled, multi-center
Type of Control
Placebo
Study Blinding
Double-blind
Estimated Duration 
of Trial
The Sponsor estimates that the trial will require 
approximately 3 years from the time the first participant 
signs the informed consent/assent until the last 
participant’s last study-related phone call or visit.  For 
purposes of analysis and reporting, the overall study ends 
when the Sponsor receives the last laboratory result or at 
the time of final contact with the last participant, 
whichever comes last.
 
 
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Number of Participants:
Approximately 2100 participants will be enrolled.
Treatment Groups and Duration:
Treatment Groups
3 treatment groups, 1:1:1 randomization as follows:

Active treatment group will receive 3 doses of vaccine 
V160 (100 Units/0.5 mL dose with Merck aluminum 
phosphate adjuvant [MAPA], 4°C stable formulation)
administered intramuscularly (IM) according to a Day 1, 
Month 2, Month 6 schedule

Active treatment group will receive 2 doses of vaccine 
V160 (100 Units/0.5 mL dose with MAPA, 4°C stable 
formulation) administered IM according to a Day 1 
(V160), Month 2 (placebo-saline solution), Month 6 
(V160) schedule

A control group will receive 3 doses of placebo (saline 
solution) administered IM according to a Day 1, Month
2, Month 6 schedule
Duration of 
Participation
A participant may be in the trial for up to approximately 3 
years, depending on when they are enrolled. Because this is an 
endpoint driven study, participation times will vary depending 
on when the participant is enrolled (subjects will be enrolled 
over an 18- month period and the study is planned to be a 3-
year trial overall, based on reaching the primary endpoint). 
Study governance considerations are outlined in Appendix 1. A list of abbreviations used in 
this document can be found in Appendix 6.
 
 
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2. Schedule of Activities (SoA)
2.1
Schedule of Activities: Screening Through Treatment Phase
Trial Period
Screening
Visit Number/Title
Visit 1 
Screening
Visit 2
Day 1
Vacc 1
Visit 3
Month 2
Vacc 2
Visit 4
Month 6
Vacc 3
Visit 5
Month 7
Reminder 
Contacts
(Monthly)
Notes
Scheduled Day
-21 to -1
1
61
183
213
Scheduling Window
-21 to -1
Day 1
21 days
28 days
21 to 49 
days after 
Vacc 3
Informed consent
X
Informed consent for future biomedical 
research
X
Check inclusion/exclusion criteria
X
X
Medical history 
X
Prior/concomitant medication review
X
X
X
X
X
Full physical examination 
X
Includes height, weight, vital signs
(temperature, heart rate, 
respiratory rate, and blood 
pressure), and other assessments 
specified in Section 9.5.1  
Screening for CMV serostatus by IgG
X
Serum pregnancy test (-HCG)
X
Participant identification card
X
Card provided after informed 
consent. Site to add randomization 
number at Visit 2
Treatment randomization 
X
Urine and saliva sample self-collection 
training
X
Will be performed prior to 
vaccination
Directed physical examination
X
X
X
Includes vital signs (temperature, 
heart rate, respiratory rate, and 
blood pressure), and limited 
assessments specified in Section 
9.5.1
Urine pregnancy test
X
X
X
X
Will be performed prior to 
vaccination
 
 
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Trial Period
Screening
Visit Number/Title
Visit 1 
Screening
Visit 2
Day 1
Vacc 1
Visit 3
Month 2
Vacc 2
Visit 4
Month 6
Vacc 3
Visit 5
Month 7
Reminder 
Contacts
(Monthly)
Notes
Scheduled Day
-21 to -1
1
61
183
213
Scheduling Window
-21 to -1
Day 1
21 days
28 days
21 to 49 
days after 
Vacc 3
Participants NOT included in the Detailed 
Immunogenicity sub-study: 
Serum for NAbs and IgG ELISA 
antibodies
X
X
Baseline sample to be collected 
prior to vaccination
Detailed Immunogenicity sub-study 
participants only: Blood for CMV IFN-γ
ELISPOT (PBMCs) and serum for NAbs 
and IgG ELISA antibodies
X
X
X
Baseline and Month 6 samples to 
be collected prior to vaccination
Blood (DNA) for future biomedical 
research
X
Will be performed prior to 
vaccination
V160/Placebo administration
X
X
X
30-minute postdose safety observation 
X
X
X
Distribution of eVRC device
X
Check postdose 1 exclusion criteria (and 
any contraindication to vaccination)
X
X
Will be performed prior to 
vaccination
 
 
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Trial Period
Screening
Visit Number/Title
Visit 1 
Screening
Visit 2
Day 1
Vacc 1
Visit 3
Month 2
Vacc 2
Visit 4
Month 6
Vacc 3
Visit 5
Month 7
Reminder 
Contacts
(Monthly)
Notes
Scheduled Day
-21 to -1
1
61
183
213
Scheduling Window
-21 to -1
Day 1
21 days
28 days
21 to 49 
days after 
Vacc 3
Adverse event monitoring
X
X
X
X
X
Screening to randomization: All 
AEs, SAEs, and other reportable 
events that cause the participant to 
be excluded from the trial or are 
the result of a protocol-specified 
intervention are to be reported. 
Randomization through 14 days 
following Dose 3: AEs (serious 
and non-serious) are to be reported 
from Days 1 through 14 following 
each vaccination. SAEs, deaths, 
other reportable events (ie, cancer, 
overdose, pregnancy, and infant 
SAEs) are to be reported 
throughout the duration of an 
individual’s study participation. 
Exposure during breastfeeding is 
to be reported from Day 1 through 
Month 7.
Review of eVRC data
X
X
X 
Reminder contact for urine/saliva self-
collection and pregnancy status
X
Urine and saliva for CMV
X
Collected monthly
Self-collected by participant 
starting at Day 1 (prior to 
vaccination) and then continue 
monthly through ≈Month 36 or 
beyond, depending on event 
accrual
Vacc = Vaccine; eVRC = Electronic vaccination report card; NAbs = neutralizing antibodies; -hCG = Human chorionic gonadotropin; IgG = immunoglobin G; 
IFN-γ = Interferon gamma; ELISA = Enzyme-linked immunosorbent assay; PBMC = peripheral blood mononuclear cell; 
DNA = Deoxyribonucleic acid; CMV = Cytomegalovirus; AE = Adverse event; SAE = Serious adverse event
 
 
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2.2
Schedule of Activities: Follow-up Period Through Approximately Month 36 (Approximate Time Point to Reach Cases 
Required for Primary Endpoint Assessment)
Trial period
Follow-up period
Notes
Visit number/title
Visit 6
Month 12
Year 1
Visit 7
Month 18
Year 1.5
Visit 8
Month 24
Year 2
Visit 9
Month 30
Year 2.5
Visit 10
Month 36
Year 3
Reminder 
contacts
(monthly)
Study closeout activities 
will be conducted at the 
participant’s closest visit 
to the end of the study. 
Close out activities must 
be completed in the 
event of early 
discontinuation.
Since the study 
completion is driven by 
the primary endpoint
(i.e., number of events), 
the number of visits will 
depend on the timing of
event accrual. 
Scheduled Day
365
548
731
914
1096
Scheduling Window
28
days
28
days
28
days
28
days
90
days
Reminder contact for urine/saliva self-collection 
and pregnancy status 
X
Deaths and SAEs monitoring
X
X
X
X
X
Participants NOT included in the Detailed 
Immunogenicity sub-study: 
Serum for NAbs and IgG ELISA antibodies
X
X
X
Detailed Immunogenicity sub-study participants
only: 
Serum for NAbs and IgG ELISA antibodies
Blood for CMV IFN-γ ELISPOT (PBMCs)
X
X
X
Urine and saliva sample for CMV
collected monthly
Self-collected by 
participant starting at 
Day 1 (prior to 
vaccination) and then 
continue monthly 
through ≈Month 36 or 
beyond, depending on 
event accrual
SAE = serious adverse event; NAbs = neutralizing antibodies; IgG = immunoglobin G; ELISA = Enzyme-linked immunosorbent assay; ELISPOT = Enzyme-
linked immunosorbent spot; PBMC = peripheral blood mononuclear cell; IFN-γ = Interferon gamma
 
 
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2.3
Schedule of Activities for Congenital Cytomegalovirus Infection Assessment
Trial period visit number/title
cCMVi
visit 1
Monthly 
pregnancy 
contacts
cCMVi
visit 2
Additional 
cCMVi 
visit (if 
necessary)
Reminder 
contacts
(monthly)
cCMVi infant
follow-up, yearly 
contacts for 3 years
Notes
Scheduled day
Pregnancy
reported
Once per 
month
Up to 14 
days
after 
birth
of the 
infant
Up to 14 
days after 
birth
of the 
infant
Scheduling window
-
-
+ 7 days + 7 days
-
Reminder contact for urine/saliva self-collection
and pregnancy status
X
Provide instruction on pregnancy and infant follow-
up 
X
Contact from site to assess progress of pregnancy
including any suspected congenital CMV infection
X
Record anti-CMV treatments received by 
participants during pregnancy
X
Pregnancy/infant outcome assessment
X
X
X    
Informed consent for infant urine and saliva sample
if live birth 
X
Infant urine and saliva sample collected for CMV 
PCR
X
X
Saliva sample should 
not be collected within 
2 hours of 
breastfeeding. Two sets 
of samples (saliva and 
urine) must be collected 
at 2 different time 
points separated by at 
least 3 hours within 14 
(+7) days after birth
SAE monitoring and collection for live birth infants 
X
X
X
X
Yearly follow-up for hearing, vision, and other 
neurologic outcomes (for infants with cCMVi only)  
X
cCMVi = congenital cytomegalovirus infection; PCR = Polymerase chain reaction
 
 
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3. Introduction
3.1
Study Rationale 
Merck is developing an investigational CMV vaccine (referred to as V160) for the prevention 
of infection and disease caused by CMV in vulnerable populations. Although CMVi is 
asymptomatic in healthy individuals, CMV is an important pathogen for congenitally 
infected infants and immunosuppressed individuals, such as solid organ transplant or
hematopoietic stem cell transplant recipients.  
Congenital cytomegalovirus infection (cCMVi) is the most frequent infectious cause of 
newborn malformation in developed countries. Congenital CMVi, which can lead to 
congenital CMV disease (cCMVd), presents as sensorineural hearing loss (responsible for 
almost one-third of all cases), and neurodevelopmental abnormalities. As many affected 
children require significant ongoing care and special therapeutic and educational services, the 
economic burden associated with congenital CMV infection is substantial. In the United 
States (US) in 2000, the estimated annual cost of congenital CMV infection is up to $2 
billion in 1992 dollars, which correlates with in excess of $3 billion in 2017 [Marsico, C. 
2017]. In a study of economic burden in Germany in 2011, the total cost of CMV infection in 
Germany per child and for the society as a whole, amounted to 242.91 million euros (940.77 
million euros, undiscounted) annually [Walter, E,, et al 2011].
The impact of cCMVd on public health is likely significantly under-appreciated with very 
low public awareness, largely because relatively few infected infants are severely ill at birth 
and symptoms may not be recognized in the newborn. When the later symptoms of cCMVd, 
such as visual and hearing impairment, do appear, the diagnosis of CMV can no longer be 
confirmed. Congenital CMVd occurs more frequently than Down's syndrome, Spina Bifida, 
Fetal Alcohol Syndrome and, prior to the availability of a vaccine, non-epidemic Congenital 
Rubella Syndrome. Development of a vaccine for prevention of cCMVi has been rated a high 
priority by the National Academy of Medicine [Griffiths, P. D., et al 2011]. Refer to Section 
3.2 for more details regarding burden of disease.
The V160 development strategy focuses initially on women of childbearing age. The vaccine 
was developed based on the principle that an effective CMV vaccine should be designed to 
parallel the immune response commonly seen in healthy CMV seropositive participants and 
should induce both humoral and cellular immunity.
3.2
Background
Disease 
Human CMV, also known as human herpesvirus 5 (HHV-5) is a double-stranded 
deoxyribonucleic acid (DNA) virus belonging to the beta-herpesvirus family of 
Herpesviridae. Acquisition appears to require close or intimate contact with persons who are 
excreting CMV in their urine, saliva or other secretions. CMV can also be transmitted via 
blood transfusion, breast milk, sexual intercourse, and transplanted organs. 
In most healthy individuals, CMVi is asymptomatic. When symptoms are present, they are 
often mild, can be confused with other illnesses, and include fever, sore throat, fatigue, 
and/or swollen glands. After infection, the virus establishes lifelong latency, the evidence of 
 
 
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which is the detection of anti-CMV antibodies in the blood (CMV seropositivity). Healthy 
individuals with latent CMV infection can reactivate to shed the virus in their saliva or urine, 
which is also predominantly asymptomatic.
It is known that CMV can cause serious disease in congenitally infected newborns and 
immunocompromised individuals. The spectrum of disease in newborns with cCMVi ranges 
from fetal/infant death to neurological and sensory impairments which are diagnosed later in 
childhood [Hahn, G., et al 1998] [Just-Nubling, G., et al 2003] [Sinclair, J. 2008] [Lanzieri, 
T. M., et al 2014].
Diagnosis
Incident CMVi can be diagnosed by finding new anti-CMV antibodies in the blood (in a 
previously CMV seronegative individual) or by detection of CMV DNA by real time 
quantitative polymerase chain reaction (PCR) in the blood, saliva, or urine.
In immunocompromised individuals, CMV PCR in the blood is the diagnostic method of 
choice. In newborns and immunocompetent adults, detection of CMV DNA in the saliva and 
urine is the preferred method of diagnosis. After a new CMVi, CMV DNA can be detected in 
the saliva or urine for 6-8 weeks in adults (and perhaps longer). Latently CMV infected 
individuals (CMV seropositive subjects) can also have intermittent reactivation evidenced by 
detection of CMV DNA in saliva and urine; the triggers of this “viral shedding” are 
unknown.
The focus of this study will be CMV detection in saliva and urine. Refer to Section 5.4.1.1 
regarding the rationale for the CMVi endpoint. 
Epidemiology and Disease Burden
In developed countries, maternal seropositivity/seroprevalence (IgG positivity) for CMV is 
approximately 50% and the prevalence of cCMVi among infants is estimated to be 
approximately 0.7% among live births. In developing countries, maternal seroprevalence is 
usually higher than in developed countries, and ranges from 84 to 100%. The prevalence of 
cCMVi in developing countries varies substantially, both within and between countries, and 
has been reported to range from 0.6% to 6.1% [Dollard, S. C., et al 2007] [Kenneson, A. and 
Cannon, M. J. 2007] [Lanzieri, T. M., et al 2014] [Wang, S., et al 2017]. The rate of CMVi 
during pregnancy is assumed to be similar to the rate in the general population 
(approximately 1% to 4% depending on risk factors), although data are limited.
The CMV seropositivity rate in a population (a surrogate marker for CMVi) is affected by 
several factors. CMV seroprevalence varies by age (increase with age), race/ethnicity, and 
region. African Americans and Hispanics are infected with CMV at a younger age than non-
Hispanic white individuals in the US, and approximately 90% are CMV seropositive by 40 
years of age. Low socioeconomic status, residence in an urban area, and a history of sexually 
transmitted diseases are also risk factors for CMV seropositivity [Bate, S. L., et al 2010]. In 
studies conducted in Europe, CMV seroprevalence was similar to the US and was highest 
among non-Western, immigrant groups.
In the 1999 to 2004 NHANES survey of US general population, the annualized rates of CMV 
seroconversion were about 1.6% in the US overall, and were 5.7%, 5.1% and 1.4% among 
black, Mexican-American, and non-Hispanic white participants aged 12-49 years, 
 
 
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respectively [Colugnati, F. A., et al 2007]. The strongest risk factor for CMV seroconversion 
is exposure to young children, particularly those attending daycare [Hyde, T. B., et al 2010].
It is estimated that of the approximately 4 million live births each year in the US, 
approximately 0.5%-2.0% (20,000-80,000) will have cCMVi [Dollard, S. C., et al 2007] 
[Sung, H. and Schleiss, M. R. 2010]. In other developed countries (in Europe, Japan), rates 
have been reported to be similar, 0.65 per 100 births.  At least 15% (3,000 to 9,000 in the 
US) will develop symptoms of CNS damage either at birth or progressively in the first few 
years of life, with clinical manifestations including sensorineural hearing loss, mental 
retardation, and cerebral palsy. 
Refer to the investigator’s brochure (IB) for additional details.
Pharmaceutical and Therapeutic Background
There is no effective CMV vaccine to protect women from acquiring CMVi during 
pregnancy or any recognized intervention that can effectively reduce transmission of CMV 
from newly infected pregnant women to the fetus.
Immune responses to natural CMVi can protect individuals against re-infection [Plotkin, S. 
A., et al 1989] [Adler, S. P., et al 1995] [Wang, D., et al 2016]. Maternal immunity from 
naturally acquired infection can protect a fetus from cCMVi [Fowler, K. B., et al 2003]. In 
addition, CMV-seropositivity is associated with a lower incidence of severe CMV disease 
post-transplantation [Opelz, G., et al 2004]. An effective CMV vaccine should therefore be 
designed to parallel the immune response commonly seen in healthy CMV seropositive 
individuals and should induce both humoral and cellular immunity. Such a vaccine could 
protect women from acquisition of CMVi, and thereby prevent vertical transmission to their 
baby, and would be a major medical advance that addresses a critical unmet public health 
need.
Pre-clinical and Clinical Studies
3.2.2.1
Pre-Clinical 
The AD169 strain of CMV was originally isolated at the National Institutes of Health from 
the adenoids of a 7-year-old girl. The isolate was attenuated through multiple passages in 
human fibroblasts. This live attenuated virus was tested as a vaccine in a Phase-1 study in 24
CMV seronegative male participants by Neff, et.al., [Weibel, R. E., et al 1980]. The vaccine 
was well tolerated and did not cause any SAEs. No virus could be recovered either from 
peripheral lymphocytes or urine of the vaccinated participants.
The V160 vaccine was created using the Merck variant of AD169 virus (MAD169). The 
vaccine virus was modified to be conditionally replication defective in the absence of a 
synthetic molecule called Shld-1 through the ddFKBP-mediated destabilization of two viral 
proteins critical for viral replication. With Shld-1 present, viral stocks for V160 can be 
grown, but in vivo with no Shld-1, replication of V160 is prevented. In order to facilitate 
broad immune responses, the pentameric complex (gH/gL/pUL128-131) was restored on the 
V160 virus. This restores elicitation of antibodies against the gH antigens, as well as viral 
tropism of V160 for epithelial and endothelial cells, permitting entry and some protein 
 
 
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production without replication (in the absence of Shld-1). Cell-mediated and humoral 
antibody responses are thereby enhanced. 
Since there is no relevant animal model available to assess CMV vaccine efficacy (VE), the 
key immunological parameters for preclinical evaluations were based on neutralizing 
antibodies (NAbs) and T-cell responses in animals. V160 has been extensively evaluated in 
rhesus macaques and can elicit potent neutralizing antibodies superior to those seen with the 
gB/MF59 candidate CMV vaccine as well as balanced CD4 and CD8 T cell responses. V160 
was well tolerated in toxicology studies in animals supporting the administration of this 
vaccine to humans. 
Refer to the IB for further details regarding viral replication, virological characteristics of the 
vaccine virus, as well as detailed background information on animal studies. 
3.2.2.2
Clinical Studies
Phase 1 Clinical Study
The first-in-human study was a Phase 1 dose ranging study to assess the safety and 
immunogenicity of various antigen dose levels, formulations, and routes of administration in 
healthy volunteers 18 years of age and older. In this study, the safety and tolerability as well 
as the immunogenicity of V160 was evaluated in both CMV seronegative and seropositive 
participants. The Phase 1 study is briefly summarized below. Refer to the IB for more 
detailed information.
Participants in the Phase 1 study received a 3-dose vaccination regimen (0-1-6-month 
schedule) at various doses (10-250 units) administered intramuscularly or intradermally, with 
and without MAPA adjuvant. A saline placebo was used as the control. 
In initially CMV seronegative participants, all vaccination groups receiving V160 with and 
without MAPA exhibited statistically significant higher CMV-specific neutralizing antibody 
titers compared with the placebo group. The neutralizing antibody geometric mean titer 
(GMT) at Month 7 was highest in recipients of V160 100-unit dose + MAPA. Robust cell-
mediated immune (CMI) responses were also noted among CMV seronegative participants 
but no differences across dose groups of V160 were observed. Overall, CMI responses at all 
doses, and neutralizing antibody responses at some doses (including 100U+MAPA) in CMV 
seronegative participants approximated those observed in CMV seropositive participants.
All tested V160 formulations were safe and generally well-tolerated. No SAEs and no events 
of clinical interest were reported. Overall, adjuvant-containing formulations were more 
reactogenic than unadjuvanted irrespective of dose. Across the different vaccination groups, 
the majority of AEs were the solicited injection site AEs (pain/tenderness, erythema/redness, 
and swelling) and the solicited systemic AEs (headache, fatigue, muscle pain, and joint pain); 
all of these events were transient, and most were mild to moderate in intensity. No vaccine-
strain CMV was detected by PCR in the saliva or urine of vaccine recipients.
Phase 2 clinical study
The Phase 2b study protocol described herein will be conducted to further evaluate the V160 
formulation containing 100 units/dose + MAPA, which elicited high CMV-specific 
neutralizing antibody in the Phase 1 study. Both humoral and cellular immune responses as 
well as protection against CMVi in vaccine recipients will be assessed.
 
 
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The targeted population in this trial is women of child-bearing age and thus, healthy females 
between 16 and 35 years of age will be included in the study. Infant outcomes of any study 
participant who becomes pregnant will be assessed to evaluate protection against cCMVi as 
well. 
Refer to Section 5.4 for the scientific rationale for the study endpoints and use of placebo and 
to Section 5.5 for dose justification. 
3.2.2.3
V160 Replication
As described, the V160 vaccine virus is built on a live-attenuated virus backbone with a 
designed block of V160 replication by a chemical genetic switch. Because of this novel 
design, V160 is a conditionally replication defective virus and its replication requires the 
presence of Shld-1. 
There is a theoretical possibility that certain medications taken by participants could mimic 
Shld-1 to stabilize ddFKBP and achieve sufficient plasma concentrations to inadvertently 
sustain replication in vivo, at local injection sites. Based on results of a cell-based assay with 
high-through-put capacity for screening of medicinal agents that can mimic Shld-1, only 
tacrolimus was found to be able to stabilize ddFKBP. As a precaution, participants taking 
tacrolimus will be excluded. Refer to the IB for details.
In this Phase 2b study, urine and saliva samples will be collected and analyzed for CMV by 
PCR during the vaccination period and thereafter through Month 36. Any detected virus will 
be typed to distinguish non-vaccine strain CMV (causing a case of CMVi) from vaccine 
strain. Refer to Section 5.4.1.1 for additional details.
3.3
Benefit/Risk Assessment
Congenital CMVi can lead to life-long sequelae in the infected newborns. A focus on 
prevention of primary CMVi in women of childbearing age is therefore needed. V160 has 
demonstrated an acceptable safety profile, with immunogenicity that approaches natural 
infection with CMV (known to be protective against cCMVi). These results warrant further 
evaluation of the safety, immunogenicity, and efficacy of this investigational vaccine.
It cannot be guaranteed that participants in clinical trials will directly benefit from treatment 
during participation, as clinical vaccine trials are generally designed to provide information 
about the safety, immunogenicity, and/or efficacy of an investigational vaccine.
Additional information regarding specific benefits and risks for participants in this clinical 
trial may be found in the accompanying IB and informed consent documents.
 
 
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4. Objectives/Hypotheses and Endpoints
In healthy females between 16 and 35 years of age:
Objective/Hypothesis
Endpoint
Primary

Objective: To demonstrate the efficacy 
of a 3-dose regimen of V160 in reducing 
the incidence of primary CMVi during 
the follow-up period after the last dose of 
the vaccine

Hypothesis 1: Administration of a 3-
dose regimen of V160 will reduce the 
incidence of primary CMVi 
compared to placebo. (The statistical 
criterion for success requires the 
lower limit of the 95% CI of VE to be 
greater than 0%)

Incidence of CMVi in the 3-dose 
vaccine and placebo groups during the 
follow-up period after the last dose of a 
3-dose regimen of the vaccine

Objective: To assess the safety and 
tolerability of a 2-dose and a 3-dose 
regimen of V160

Number of participants in the 2-dose 
and 3-dose vaccine groups
experiencing solicited injection site 
AEs within Days 1 through 5 after each 
vaccination visit; solicited systemic 
AEs; and vaccine-related SAEs within 
Days 1 through 14 after each 
vaccination visit
Secondary

Objective: To demonstrate the efficacy 
of a 2-dose regimen of V160 in reducing 
the incidence of primary CMVi during 
the follow-up period after the last dose of
the vaccine

Hypothesis 2: Administration of a 2-
dose regimen of V160 will reduce the 
incidence of primary CMVi 
compared to placebo. (The statistical 
criterion for success requires the 
lower limit of the 95% CI of VE to be 
greater than 0%).

Incidence of CMVi in the 2-dose 
vaccine and placebo groups during the 
follow-up period after the last dose of a 
2-dose regimen of the vaccine
 
 
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Objective/Hypothesis
Endpoint
Exploratory 

To assess and compare the kinetics of 
humoral immune response induced by 2-
dose and 3-dose regimens of V160

Percent of participants who seroconvert 
based on the NAb and IgG ELISA 
assays from CMV seronegative at Day 
1 to CMV seropositive at Month 7a

Seropositivity based on the NAb and 
IgG ELISA assays at Day 1, Months 6, 
7a, 12, 24, and 36

GMT based on the NAb and the IgG 
ELISA assays at Day 1, Months 6, 7a, 
12, 24, and 36

To assess and compare the kinetics of 
CMI response induced by 2-dose and 3-
dose regimens of V160 in a subset of
study participantsb

Geometric mean count (GMc) of spot 
forming cells (SFC) per million 
PBMCs (SFC/106 PBMCs) based on 
the ELISPOT assay at Day 1, Months 
6, 7a, 12, 24 and 36

To evaluate the incidence of cCMVi in 
infants born to recipients of 2-dose and 
3-dose regimens of V160 compared to 
placebo, during the follow-up period 
after the last dose of vaccine or placebo

To follow the outcomes of infants 
diagnosed with cCMVi in the study

Incidence of cCMVi cases in the V160 
2-dose, V160 3-dose, and placebo 
groups during the follow-up period 
after the last dose of vaccine or placebo

Hearing, vision, and neurological 
outcomes of infants diagnosed with 
cCMVi over 3 years

To explore potential correlations between 
humoral and CMI response to 2-dose and 
3-dose regimens of V160 and efficacy of 
2-dose and 3-dose regimens of V160 
against CMVi

Immune responses in participants in the 
2-dose and 3-dose vaccine groups with 
and without CMVi during the follow-
up period after the last dose of V160
a Month 7 is the time point corresponding to 1 month following last dose of the 2-dose and 3-dose 
regimen of V160
b In Detailed Immunogenicity sub-study participants only (N≈360).
cCMVi = Congenital CMV infection; CMV = Cytomegalovirus; CMVi = CMV infection; 
DNA = Deoxyribonucleic acid; ELISA = Enzyme-linked immunosorbent assay
ELISPOT = Enzyme-linked immunosorbent spot; IgG = Immunoglobulin G 
NAb = Neutralizing antibody
PCR = Polymerase chain reaction; PBMC = Peripheral blood mononuclear cells
SFC = Spot forming cells
 
 
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5. Study Design 
5.1
Overall Design 
This study is a randomized, placebo-controlled, parallel-group, multi-center, double-blind
trial of V160, in healthy CMV seronegative adolescent and adult women 16 to 35 years of 
age. Refer to Section 6 for details regarding the study population. 
This is a proof-of-concept study intended to evaluate the safety and efficacy against CMVi of 
a 3-dose and 2-dose regimen of V160 administered over 6 months compared to placebo. The 
time period for the treatment phase of the study is Day 1 through Month 7. The study is 
event-driven, requiring a fixed number of CMVi cases to be accumulated in order to test the 
hypotheses relating to the efficacy of 3-dose and 2-dose regimens of V160. The study is 
designed to accumulate the required number of CMVi cases in ≈3 years with ≈2100 study 
participants. Since the study duration is driven by the primary endpoint (ie, number of 
events), the study duration will depend on the timing of event accrual. The study will also 
evaluate the immunogenicity of a 3-dose and 2-dose regimen of V160 (humoral immunity in 
the whole study and detailed immunogenicity including CMI in an approximately 360-
participant sub-study) as well as the impact of administration of V160 on the incidence of 
cCMVi in participants who get pregnant during the study.
Specific procedures to be performed during the trial, as well as their prescribed times and 
associated visit windows, are outlined in the Trial SoA - Section 2. Details of each procedure 
are provided in Section 9 – Study Assessments and Procedures.
There will be 1 formal interim analysis (IA) for testing the primary hypothesis addressing the 
efficacy of a 3-dose regimen of V160 compared to placebo, to be conducted upon 
accumulation of 15 total cases of CMVi in the combined V160 3-dose and placebo arms of 
the study. The interim analysis will also assess futility. It is anticipated that the required 15 
cases will be accumulated at ≈18 months after enrollment of the first study participant.
An external data monitoring committee (eDMC) will be formed to conduct an interim 
analysis of the efficacy data, and a review of the safety data, collected during this study. 
Results of the IA will be reviewed by the eDMC, who will make recommendations to the 
Sponsor’s Executive Oversight Committee. Additional details regarding the responsibilities 
of the eDMC will be provided in an eDMC charter.
Details relating to the planned IA are provided in Section 10.7. There will be no IA for the 
testing of the secondary hypothesis addressing the efficacy of a 2-dose regimen of V160 
compared to placebo.
All pregnancies that occur during the study will be monitored for evidence of cCMVi in the 
fetus/infant. Determinations of cCMVi will be conducted by an independent, blinded 
adjudication committee. Refer to Section 9.2.1.2.2 for further details.
Refer to Section 10.9 for sample size calculations.
 
 
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Study Diagram
The trial design is depicted in Figure 1.
M= month; V1, V2, etc, = Visit 1, Visit 2, etc; Vacc = vaccination visit
SAEs = serious adverse events; NAbs= neutralizing antibodies, CMI= cell-mediated immunity; cCMVi = congenital CMV 
infection; ELISPOT = Enzyme-linked immunosorbent spot
Monthly contacts through M36 will be conducted for self-collection reminder (saliva and urine) and to assess pregnancy 
status. During pregnancy participants will have another monthly contact to assess pregnancy and infant outcome.
a Participants in the V160 2-dose regimen arm will receive V160 at Vacc 1, Placebo at Vacc 2, and V160 at Vacc 3.
b Any time a pregnancy is reported the participant will be asked to return to the clinic for 2 or 3 visits - one to provide 
information on pregnancy monitoring and infant follow-up and 1 or 2 others to bring in the infant for collection of infant 
saliva and urine sample for CMV PCR testing. Infants will be screened for cCMVi by collecting saliva and urine for CMV 
PCR within 2 weeks of birth. cCMVi positive infants will be monitored for up to 3 years after birth.
Figure 1
Trial Design Diagram
5.2
Number of Participants
Approximately 2100 participants will be randomized in order to accumulate the required 
number of CMVi cases to provide sufficient power for the primary and secondary efficacy 
hypotheses.
 
 
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5.3
Beginning and End of Study Definition
The overall study begins when the first participant signs the informed consent/assent form 
(ICF).  The overall study ends when the last participant completes the last study-related 
contact or visit, withdraws from the study or is lost to follow-up (ie, the participant is unable 
to be contacted by the investigator).  For purposes of analysis and reporting, the overall study 
ends when the Sponsor receives the last laboratory result or at the time of final contact with 
the last participant, whichever comes last.
Clinical Criteria for Early Study Termination
An eDMC will be formed to monitor accumulating safety data from this study. They will also 
review the results of the IA of the primary endpoint. At the IA for testing the primary 
efficacy hypothesis, the eDMC may recommend termination of the study for demonstration 
of futility (as described in Section 10.7). The eDMC may also recommend termination of the 
study for safety if concerning safety trends are detected. Guidance relating to 
recommendations for temporary pause or termination of the study will be provided in the 
eDMC charter that will be written for this study.
5.4
Scientific Rationale for Study Design 
This study is designed to evaluate the efficacy of V160 in preventing CMVi in women of 
childbearing age who are seronegative for CMV prior to vaccination. The primary reason to 
vaccinate women against CMV is to prevent cCMVi and cCMVd in infants of women who 
become infected with CMV during pregnancy. A CMV seronegative woman infected during 
pregnancy has a high probability of transmitting CMV to her fetus (~40%). Congenital 
CMVi and cCMVd are rare endpoints which are difficult to assess in clinical studies.
Therefore, the primary efficacy endpoint in this study is prevention of CMVi in CMV 
seronegative women. It is postulated that if a woman is durably protected against CMV 
infection, she should continue to be protected during any future pregnancies thereby 
eliminating the potential to transmit CMV to her fetus and cases of cCMVi/cCMVd. 
Although a CMV seropositive woman can also transmit CMV to her fetus, the probability of 
transmission is much lower (~1-2%), and the sequelae in the infected infant, less severe. 
Whether this transmission, in CMV seropositive women, occurs as a result of acquisition of a 
new CMVi or reactivation of a latent infection is unknown.  
Since persons who are CMV seropositive (latently infected with the virus) are known to shed 
the virus intermittently, this study will include only CMV seronegative women in order to 
optimize demonstration of vaccine efficacy against CMVi. Additional studies in the V160 
development program will evaluate the safety and immunogenicity of V160 among CMV 
seropositive women.
Additionally, women who have exposure to a child/children ≤5 years old at home or 
occupationally will be included in the study to increase the risk of incident CMVi. As women 
with young children may be breastfeeding, breastfeeding a child will not be an exclusion 
criterion for vaccination in the study. Breastfeeding is not a contraindication for other live 
virus vaccines - Measles, Mumps, Rubella vaccine (M-M-R™II) or Varicella virus vaccine 
(Varivax™). The position of the Advisory Committee on Immunization Practices (ACIP)
regarding vaccines and breastfeeding is, “Neither inactivated nor live virus vaccines 
 
 
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administered to a lactating woman affect the safety of breast-feeding for mothers of infants.” 
and “Although live vaccines multiply within the mother's body, the majority have not been 
demonstrated to be excreted in human milk”[Centers for Disease Control and Prevention 
2002]. Similarly, for V160, as replication of the virus within the mother is not expected, and 
shedding has not been detected in saliva or urine, presence of V160 in breast milk is unlikely.
This relatively large Phase 2b study is designed to allow a rapid and robust demonstration of 
clinical efficacy of V160, with a high probability of success at the IA when the VE ≥85%. 
The rigor of the analysis criteria is intended to reduce the risk of a false-negative study in the 
case that the vaccine is truly efficacious.
The 2-dose arm is included to allow for evaluation of a schedule that would optimally 
coincide with the current adolescent vaccination schedule, allowing maximum uptake when 
licensed.
Rationale for Endpoints
5.4.1.1
Rationale for Efficacy Endpoint
CMVi (the primary endpoint for this study) is largely asymptomatic in adults, but the virus 
can be shed in saliva or urine for 6-8 weeks (or perhaps longer) in persons who are infected. 
Therefore, detection of CMV in the saliva or urine of previously uninfected women will be 
used to identify cases of incident CMV infection. Sampling of saliva and urine will be 
conducted monthly, and self-collection will be used in order to encourage compliance. 
Adequacy of sampling during self-collection will be assessed by evaluation of the samples 
for a human housekeeping gene, beta-globin, by polymerase chain reaction (PCR) for a 
period of time after the start of the study (eg, 6 months).  
Prior CMV vaccine studies identified incident CMVi by detecting new immunity to CMV 
and confirming by urine or saliva viral detection. In these studies, serum, urine, and saliva 
sampling was conducted every 3 months and urine/saliva detection identified nearly all cases 
of infection [Pass, R. F., et al 2009]. As immunity to V160 is designed to mimic natural 
immune responses, postdose immune responses cannot be used to identify incident CMVi. 
Therefore, the focus of this study will be CMV detection in saliva and urine by PCR and the 
frequency of assessment has been increased from every 3 months to monthly. 
If CMV is identified by PCR in a participant, the virus will be typed to determine whether it 
is vaccine or non-vaccine CMV strain. Since this is the first study to use monthly sampling of 
saliva and urine to identify incident CMVi, the performance of the sampling methodology 
will be carefully monitored. If it is determined that reduced sampling frequency or use of a 
single matrix (saliva or urine) could adequately identify endpoint CMVi cases, modifications 
to the sampling methodology can be implemented in future studies.
5.4.1.2
Rationale for Immunogenicity Endpoints
The rationale for the immunogenicity endpoint is to characterize the humoral and CMI 
responses to V160 and explore the relationship between immune responses to V160 and 
efficacy of the vaccine.
 
 
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Robust and balanced humoral and cellular CMV-specific immune responses have been found 
to play an important role in the protection against cCMVi [Plotkin, S. A., et al 1989] [Adler, 
S. P., et al 1995] [Wang, D., et al 2016].
The purpose of the CMV-specific NAb assay is to assess humoral immunity by quantifying
neutralizing antibody titers to CMV before and after vaccination. This assay provided the 
primary serological readout in Phase 1, allowed quantitative comparison of responses to 
different doses, and provides a functional characterization of the vaccine response.
For an additional measure of the humoral response, an IgG ELISA assay being will be used 
to measure the total IgG response to V160 as a secondary serological read-out. This assay 
will also be evaluated as a surrogate of the functional neutralization assay.
The purpose of the CMV ELISPOT assay is to assess CMI responses before and after 
vaccination. This assay was also used in Phase 1 and while robust responses were 
demonstrated, it did not allow quantitative comparison between groups. It will be performed 
on only a subset of participants to confirm CMI responses to V160.
5.4.1.3
Rationale for Safety Endpoints
The safety endpoints evaluated in this study were selected based on the product’s safety 
profile demonstrated in the Phase 1 study, and on feedback received from regulatory agencies 
during product development.
In this study, as in the Phase I study, pre-specified solicited systemic AEs include tiredness 
(fatigue), muscle pain (myalgia), headache, and joint pain (arthralgia) and will be collected 
Days 1 to 14 following each dose of V160/placebo. The vaccine has limited potential for 
replication, therefore, an extended collection time of safety follow-up typically used for live 
virus vaccines is not warranted in this study.
The timepoints chosen for safety assessment are based on peak occurrence of AEs generally 
observed following vaccination in clinical trials with multi-dose vaccines.
AEs will be assessed and graded for intensity/toxicity per Guidance for Industry. Although
death is not included in the Toxicity Grade Guidance for Industry, for this protocol, any 
death will be assessed as Toxicity Grade 4.
Details on AEs, including definitions and reporting requirements, can be found in Appendix 
4.
5.4.1.4
Rationale for Congenital CMVi Exploratory Endpoint
V160 is being developed to prevent CMVi in women of childbearing potential and thereby 
cCMVi in their infants. Evaluation of infant outcomes of any women who become pregnant 
is an exploratory objective in this trial. As there is no routine screening for CMVi in 
pregnancy or cCMVi at birth, the study will collect a woman’s prenatal care records for 
evidence of suspicion of congenital infection (eg, ultrasounds with increased placental 
thickness, fetal growth retardation, amniocentesis procedures looking for CMV, etc.). When 
a pregnancy is not carried to term, data on spontaneous abortions and terminations will be 
collected. Additionally, suspicion for congenital infections at birth will be determined from 
review of delivery records. Infants will be screened for cCMVi in the study by collecting 
 
 
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saliva and urine for CMV PCR within 3 weeks of birth. When there is a suspicion or 
diagnosis of cCMVi outside the study, or infant samples are collected outside the study 
window, endpoint determination for cCMVi will be performed by an independent, blinded, 
clinical adjudication committee.
Refer to Section 9.2.1.2 for cCMVi endpoint assessment including case definition and 
detection. Details of the cCMVi exploratory endpoint analysis will be provided in the
supplementary statistical analysis plan (sSAP).
5.4.1.5
Planned Exploratory Biomarker Research
5.4.1.5.1 Future Biomedical Research
The Sponsor will conduct Future Biomedical Research on DNA specimens for which consent 
was provided during this clinical study. This research may include genetic analyses (DNA), 
gene expression profiling (ribonucleic acid [RNA]), proteomics, metabolomics (serum, 
plasma) and/or the measurement of other analytes, depending on which specimens are 
consented for future biomedical research.
Such research is for biomarker testing to address emergent questions not described elsewhere 
in the protocol (as part of the main study) and will only be conducted on specimens from 
appropriately consented participants. The objective of collecting/retaining specimens for 
Future Biomedical Research is to explore and identify biomarkers that inform the scientific 
understanding of diseases and/or their therapeutic treatments. The overarching goal is to use 
such information to develop safer, more effective drugs/vaccines, and/or to ensure that 
participants receive the correct dose of the correct drug/vaccine at the correct time. The 
details of this Future Biomedical Research sub-study are presented in Appendix 2.
5.4.1.6
Rationale for the Use of Comparator/Placebo
A concurrent placebo arm (sterile saline) is included in this study in order to provide CMV 
infection rates for determination of vaccine efficacy. 
There is currently no active comparator licensed for the prevention of CMV infection
5.4.1.7
Rationale for Adjuvant
The Merck Aluminum Phosphate Adjuvant (MAPA) was assessed in the Phase 1 trial in 
which formulations containing MAPA tended to be more immunogenic than those without, 
particularly at the 100-unit dose. Therefore, MAPA is included in the V160 formulation for 
this Phase 2 trial. The dose level of MAPA in the V160 formulation to be used in this trial is 
consistent with the amount of aluminum phosphate adjuvant (125 µg/dose to 500 µg/dose) 
used in many other licensed vaccines.
 
 
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5.5
Justification for Dose 
The V160 formulation selected for this trial is 100 Units/0.5 mL dose +225 µg of MAPA as 
this formulation provided the greatest immunogenicity (measured by the NAb response) in 
the Phase 1 trial. All participants randomized to either of the two vaccine groups will be 
given either 2 or 3 injections (depending on randomization) of the V160 100 units/0.5 mL 
+MAPA. The maximum dose at each dosing time point is the same as the starting dose.
Rationale for Dose Interval and Trial Design
The proposed dosing interval is intended to provide early induction of CMV-specific immune 
responses (with 2 “priming” doses given within 2 months) and establishment of long-term
immunogenicity by providing a third “boost” dose at a later time (6 months). A 3-dose 
schedule of V160 administered at Day 1, Month 1, and Month 6 was used for the Phase 1 
study. For this Phase 2b study, a Day 1, Month 2, and Month 6 schedule and a Day 1, Month 
6 schedule were selected. Since the 0, 1, 6 month and 0, 2, 6-month regimens are designed 
with 2 priming doses followed by a boost dose, it is not anticipated that the change from 1 to 
2 months for the second dose in the 3-dose regimen will impact immunogenicity. The 2-dose 
regimen with a single priming dose followed by a boost dose is being evaluated because it 
would allow greater compliance. Both these regimens would allow alignment with other 
adolescent vaccines such as Gardasil™ and Gardasil™9. 
6. Study Population
Participants will be healthy, CMV seronegative, non-pregnant, female participants of child-
bearing potential, age 16-35 years.
Prospective approval of protocol deviations to recruitment and enrollment criteria, also 
known as protocol waivers or exemptions, is not permitted.
6.1
Inclusion Criteria
Participants are eligible to be included in the study only if all of the following criteria apply:
1. Healthy, based on medical history and physical examination
2. Female, 16 to 35 years of age
3. Serologically confirmed as CMV seronegative (based on CMV IgG) prior to 
receiving the first dose of V160/placebo
4. Have direct exposure to young children (≤5 years of age) at home or occupationally 
5. Childbearing potential (see definition of women of childbearing potential [WOCBP]
in Appendix 3)
6. Agrees to avoid becoming pregnant during the 6-month treatment phase and for 4 
weeks after the last dose of V160/placebo (from Day 1 through Month 7) by either 
practicing abstinence from heterosexual activity OR by using allowable methods of 
birth control during heterosexual activity. Allowable methods of birth control are
provided in the Appendix 3.
 
 
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Informed Consent/Assent
7. Voluntarily agrees to participate (or legally acceptable representative, for minors, if 
applicable); and provide written informed consent/assent for the trial. The participant 
may also provide consent/assent for Future Biomedical Research. However, the 
participant may participate in the main trial without participating in Future 
Biomedical Research.
Protocol Adherence
8. Be able to complete all scheduled visits and to comply with the study procedures
9. Agrees to provide study personnel with a primary telephone number as well as an 
alternate form of contact, if available, for follow-up purposes
6.2
Exclusion Criteria 
Participants are excluded from the study if any of the following criteria apply:
Medical Conditions
1. Has a history or current evidence of any condition, therapy, laboratory abnormality or 
other circumstance that might expose the participant to risk by participating in the 
trial, confound the results of the trial, or interfere with participation for the full 
duration of the trial, as assessed by the investigator
2. Has a history of any allergic reaction or an anaphylactic/anaphylactoid reaction to any 
vaccination that required medical intervention, or of any severe allergic reaction (eg, 
swelling of the mouth and throat, difficulty breathing, hypotension, or shock), to any 
vaccine component that required medical intervention
3. Has a recent (<72 hours) history of febrile illness (temperature ≥100.4°F/38.0°C, oral 
equivalent)
4. Is currently immunocompromised or has been diagnosed as having a congenital or 
acquired immunodeficiency, human immunodeficiency virus (HIV) infection, 
lymphoma, or leukemia. Participants with autoimmune conditions that require 
immunosuppressive medications (eg, systemic lupus erythematosus, rheumatoid 
arthritis, juvenile rheumatoid arthritis, inflammatory bowel disease) are also to be 
excluded. No testing for HIV will be required in the study.
5. Has a condition in which repeated venipuncture or injections pose more than minimal 
risk for the participant, such as hemophilia, thrombocytopenia, other severe 
coagulation disorders, or significantly impaired venous access
6. A woman of childbearing potential (WOCBP) (see Appendix 3) who has a positive 
pregnancy test at screening or within 24 hours before the first dose of study treatment.
If the urine test on the day of vaccination cannot be confirmed as negative, a serum 
pregnancy test is required. In such cases, the participant must be excluded from 
participation if the serum pregnancy result is positive. If pregnancy is diagnosed any 
time after the first dose but before the completion of the vaccination series, the 
participant is excluded from any remaining vaccinations but is included in the study 
for safety follow-up.
 
 
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Prior/Concomitant Therapy
7. Has previously received a CMV vaccine
8. Had any live virus vaccine administered or scheduled to be administered in the period 
from 4 weeks prior to, and 4 weeks following receipt of any dose of trial vaccine
9. Had any inactivated vaccine administered or scheduled within the period from 14 
days prior to, through 14 days following, any dose of trial vaccine
10. Had administration of any immune globulin or blood product within 90 days prior to 
injection with V160/placebo or scheduled within 30 days thereafter
11. Has received systemic corticosteroids (equivalent of ≥2 mg/kg total daily dose of 
prednisone or ≥20 mg/d for persons weighing >10 kg) for ≥14 consecutive days and 
has not completed treatment at least 30 days prior to trial entry
12. Has received systemic corticosteroids exceeding physiologic replacement doses 
(≈5 mg/d prednisone equivalent) within 14 days prior to the first vaccination
(participants using inhaled, nasal, or topical steroids are considered eligible for the 
trial)
13. Has received any anti-viral agent with proven or potential activity against CMV two 
weeks prior to vaccination or is likely to receive such an agent within 2 weeks after 
vaccination. Anti-viral agents prohibited include letermovir, ganciclovir, 
valganciclovir, foscarnet, cidofovir, and valacyclovir.
14. Is receiving or has received in the year prior to enrollment immunosuppressive 
therapies including but not limited to rapamycin (also sirolimus), tacrolimus (also 
FK-506 or Fujimycin), or other therapies used for solid organ/cell transplant, 
radiation therapy, immunosuppressive/cytotoxic immunotherapy, chemotherapy and 
other immunosuppressive therapies known to interfere with the immune response.
Topical tacrolimus is allowed provided that it is not used within 2 weeks prior to, or 2 
weeks following a V160 dose.
Prior/Concurrent Clinical Study Experience
15. Has participated in another clinical trial in the past 4 weeks or plans to participate in a 
treatment-based trial or a trial in which an invasive procedure is to be performed 
while enrolled in this trial. (Participation in a safety surveillance or non-interventional 
trial is acceptable).
Other Exclusions
16. Planned donation of eggs at any time from signing the informed consent through 1 
month after receiving the last dose of the trial V160/placebo
17. Is or has an immediate family member (eg, spouse, parent/legal guardian, sibling or 
child) who is investigational site or sponsor staff directly involved with this study
18. Any other reason that in the opinion of the investigator might interfere with the 
evaluation required by the study
 
 
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Participant Deferment Criteria before Vaccination
These deferment criteria must be reviewed for each participant before each vaccination to 
ensure that none of the criteria apply to the participant and or that vaccination is deferred for 
an appropriate period of time. The participant should be deferred if:
1. Has a recent (<72 hours) history of febrile illness (temperature ≥100.4ºF/38.0°C, oral 
equivalent)
2. Had any live virus vaccine administered or scheduled to be administered in the period 
from 4 weeks prior to, through 1 month following receipt of any dose of study 
vaccine
3. Had any inactivated vaccine administered or scheduled within the period from 14
days prior to, through 14 days following, any dose of trial vaccine
4. Has received systemic corticosteroids (equivalent of ≥2 mg/kg total daily dose of 
prednisone or ≥20 mg/d for persons weighing >10 kg) for ≥14 consecutive days and 
has not completed treatment at least 30 days prior to any dose of trial vaccine
5. Has received systemic corticosteroids exceeding physiologic replacement doses (≈5 
mg/d prednisone equivalent) within 14 days prior to the any dose of trial vaccine (use 
of inhaled, nasal, or topical steroids is allowed for the trial)
6. Had administration of any immune globulin or blood product within 90 days prior to 
injection with V160/placebo or scheduled within 30 days thereafter
7. Has received any anti-viral agent with proven or potential activity against CMV 2
weeks prior to vaccination or is likely to receive such an agent within 2 weeks after 
vaccination. 
Anti-viral 
agents 
prohibited 
include 
letermovir, 
ganciclovir, 
valganciclovir, foscarnet, cidofovir, and valacyclovir
8. Has received or is scheduled to receive topical tacrolimus within 2 weeks prior to or 2 
weeks following a V160 dose.
Criteria to be Checked for Excluding Participants from Receiving Subsequent 
Doses
Specific exclusion criteria must be reviewed for each participant before administration of any 
subsequent dose of V160/placebo to ensure that none of the criteria apply to the participant. 
If any apply, the participant will be excluded from receiving the scheduled vaccination (and 
therefore be excluded from the per protocol analysis) but will continue in the study for all 
other follow-up procedures. A complete list of criteria to be checked prior to Dose 2 and 
Dose 3 are provided in Section 8.1.
6.3
Lifestyle Restrictions
No lifestyle restrictions are required.
Meals and Dietary Restrictions
No specific meal and/or dietary restrictions are required.
 
 
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Caffeine Alcohol, and Tobacco Restrictions
No specific caffeine, alcohol, or tobacco restrictions are required.
Activity
No specific activity restrictions are required.
6.4
Screen Failures
Screen failures are defined as participants who consent/assent to participate in the clinical 
study but are not subsequently randomized in the study.  A minimal set of screen failure 
information is required to ensure transparent reporting of screen failure participants to meet 
the Consolidated Standards of Reporting Trials (CONSORT) publishing requirements and to 
respond to queries from regulatory authorities. Minimal information includes demography, 
screen failure details, eligibility criteria, and any adverse events or serious adverse events 
(SAE) meeting reporting requirements as outlined in the data entry guidelines.
6.5
Participant Replacement Strategy
A participant who discontinues from trial treatment/vaccination or withdraws from the trial
will not be replaced.
7. Treatments
Study treatment is defined as any investigational treatment(s), marketed product(s), placebo,
or medical device(s) intended to be administered to a study participant according to the study 
protocol.
Clinical supplies [study treatment(s) provided by the Sponsor] will be packaged to support 
enrollment.  Clinical supplies will be affixed with a clinical label in accordance with 
regulatory requirements.
7.1
Treatments Administered
The study treatments to be used in this trial are outlined below in Table 1.  
 
 
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Table 1
Study Treatments
Study Treatment 
Name:
V160
MAPA Diluent
Sterile Saline Placebo
Dosage 
Formulation:
Lyophilized 
Cake
Sterile Suspension for 
reconstitution
Sterile Solution
Unit Dose Strength:
200 Units/mL
450 µg/mL
Not applicable
Dosage Level:
100 Units with 225 µg aluminum per 
0.5mL dose
0.5mL
Route of 
Administration:
IM injection
IM injection
Sourcing:
Sponsor
Sponsor
Sponsor
MAPA Diluent= V160 Merck Aluminum Phosphate Adjuvant (MAPA) Diluent
Sterile Saline Placebo = V160 Sterile Saline Placebo 
Unit dose strength= strength of the V160 and MAPA products after reconstitution
Dosage level = post reconstitution per 0.5mL dose
All supplies indicated in Table 1 will be provided per the ‘Sourcing’ row depending upon 
local country operational requirements.  Every attempt should be made to source these 
supplies from a single lot/batch number. The trial site is responsible for recording the lot 
number, manufacturer, and expiry date for any locally purchased product as per local 
guidelines unless otherwise instructed by the Sponsor.
Refer to section 9.1.8 for details regarding administration of the study treatment.
7.2
Dose Modification  
All vaccine doses will be 0.5 mL (100 Units+MAPA): no modifications will be made during 
the trial.
7.3
Method of Treatment Assignment 
All eligible participants will be randomly allocated and will receive a 
treatment/randomization number. The treatment/randomization number identifies the 
participant for all procedures occurring after treatment allocation/randomization.  Once a 
treatment/randomization number is assigned to a participant, it can never be re-assigned to 
another participant. A single participant cannot be assigned more than 1 
treatment/randomization number.
Treatment allocation/randomization will occur centrally using interactive response 
technology (IRT). There are 3 study treatment arms. Participants will be assigned randomly 
in a 1:1:1 ratio.
Furthermore, a subset of study participants will be enrolled to participate in a Detailed 
Immunogenicity sub-study. Sub-study participants will be asked to provide an additional 
blood sample for NAbs and IgG ELISA antibodies at Month 6, prior to the last vaccination, 
 
 
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and blood samples for PBMC isolation to allow for assessment of CMI response (CMV IFN-
γ ELISPOTs) at timepoints shown in the SoA.
Detail Immunogenicity sub-study participants will be recruited from sub-study sites, which 
are sites that are near a validated facility that can process blood samples for PBMCs. Non-
sub-study sites are clinical study sites that are not near a validated facility that can process 
blood samples for PBMCs. Study participants enrolled in non-sub-study clinical sites are 
ineligible to participate in the Detailed Immunogenicity sub-study. 
The randomization ratios relating to allocation of study participants into the 3 treatment arms 
and the sub-study are shown in Table 2.
Table 2
Allocation Ratio across Treatment Arms and Detailed Immunogenicity Sub-Study
Allocation Ratio (N)
V160 3-Dose
(Group A)
V160 2-Dose
(Group B)
Placebo
(Group C)
All Participants
1 (700)
1 (700)
1 (700)
Sub-study participants
1 (120)
1 (120)
1 (120)
Sub-study non-participants
1 (580)
1 (580)
1 (580)
N = Sample size
Approximately 360 study participants (or as many as are operationally feasible) will be 
enrolled in the sub-study, to be allocated in a 1:1:1 ratio in each of the 3-dose regimen, 2-
dose regimen, and placebo arms. For operational simplicity, the participants of the sub-study 
will be selected as follows: 

Starting at the time when the facility that can process blood samples for PBMCs near 
a particular sub-study site is validated, study sites near the facility become a sub-
study site subject to the capacity of the facility. Study participants randomized into 
the study by this particular sub-study site will become participants in the Detailed 
Immunogenicity sub-study. A site may choose to withdraw from participation in the 
sub-study after approval from the Sponsor. Participants of the Detailed 
Immunogenicity sub-study will be recruited in this manner until approximately 360 
sub-study participants (or as many as are operationally feasible) across all sub-study 
sites have been enrolled.
o
Sites may randomize participants into the study prior to the time the PBMC 
blood sample processing facility is validated; however, these participants will 
not be included in the Detailed Immunogenicity sub-study.

Additional study participants enrolled by sub-study sites over and above 360 sub-
study participants will not be enrolled in the sub-study.
 
 
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In order to increase the likelihood that approximately 360 participants enrolled into the study 
by sub-study sites are allocated approximately equally into the 3 treatment arms, two 
allocation schedules will be generated; one for use by non-sub-study sites and another for use 
by sub-study sites. 
Randomization will stop when ≈2100 participants have been randomized into the study.
Stratification
No stratification based on age, sex, or other characteristics will be used in this trial.
7.4
Blinding 
A double-blinding technique with in-house blinding will be used.  V160 and placebo will be 
prepared and administered in a blinded fashion by an unblinded pharmacist or qualified trial 
site personnel because the two products are visually different (the vaccine is opaque, and the 
placebo is clear).  The participant and the investigator who is involved in the clinical 
evaluation of the participants are unaware of the group assignments.
In order to avoid bias, the unblinded individual/s will not be involved in any postdose safety 
assessment procedures. The unblinded individual/s also must not disclose any information 
regarding the allocation of the clinical supplies or the appearance of the V160/placebo to any 
blinded member of the site staff or to the participant. No blinded member of the site staff 
should have contact with the clinical supplies at any point during the course of the study.
Note that participants must be discontinued from vaccination if they become unblinded 
(Section 8.1).
See Section 9.1.10 for a description of the method of unblinding a participant during the trial, 
should such action be warranted.
7.5
Preparation/Handling/Storage/Accountability
Dose Preparation
As described in Section 7.4, unblinded personnel will be responsible for reconstitution of
study vaccine in order to produce the final study vaccine formulation that will be 
administered to the participant. Blinded personnel will not be present during vaccine 
administration.
Reconstitution of V160 Lyophilized Cake:
Reconstitution of the V160 lyophilized cake should be completed within 90 minutes after 
removal from the refrigerated storage unit.
1. Remove V160 and V160 MAPA diluent vials from the refrigerated storage unit.
2. Mix the contents of the V160 MAPA diluent vial by shaking thoroughly to get a 
homogeneous mixture.
3. Using a syringe and needle, withdraw 0.7 mL of V160 MAPA diluent from the 
diluent vial, and inject immediately into the V160 lyophilized cake vial. 
 
 
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4. Gently swirl the V160 vial and examine for undissolved material. Continue swirling
until the lyophilized cake has completely dissolved and the appearance is opalescent. 
Disregard any bubbles that may be present. 
5. Withdraw and administer 0.5 mL of the reconstituted V160 vial.
Study vaccine should be administered immediately following dose preparation. If the 
reconstituted product is not used immediately, the vial/syringe contents should be mixed by 
inverting several times to get a homogeneous mixture of the product immediately prior to 
use. If the study vaccine or placebo is not administered within 60 minutes, it should be 
disposed of as a biohazardous medical waste and a new study vaccine should be prepared.
Preparation of Placebo
The sterile saline placebo will be used as supplied.
Handling, Storage and Accountability
The investigator or designee must confirm appropriate temperature conditions have been 
maintained during transit for all study treatment received and any discrepancies are reported 
and resolved before use of the study treatment.
Only participants enrolled in the study may receive study treatment and only authorized site 
staff may supply or administer study treatment. All study treatments must be stored in a 
secure, environmentally controlled, and monitored (manual or automated) area in accordance 
with the labeled storage conditions with access limited to the investigator and authorized site 
staff.
The investigator, institution, or the head of the medical institution (where applicable) is 
responsible for study treatment accountability, reconciliation, and record maintenance (ie, 
receipt, reconciliation, and final disposition records).
For all trial sites, the local country Sponsor personnel or designee will provide appropriate 
documentation that must be completed for drug accountability and return, or local discard 
and destruction if appropriate.  Where local discard and destruction is appropriate, the 
investigator is responsible for ensuring that a local discard/destruction procedure is 
documented.
The trial site is responsible for recording the lot number, manufacturer, and expiry date for 
any locally purchased product as per local guidelines unless otherwise instructed by the 
Sponsor.
The investigator shall take responsibility for and shall take all steps to maintain appropriate 
records and ensure appropriate supply, storage, handling, distribution and usage of study 
treatments in accordance with the protocol and any applicable laws and regulations.
7.6
Treatment Compliance
Trial compliance is defined in this study as participants who received all 3 doses of
scheduled study vaccinations/placebo.
 
 
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Interruptions from the protocol specified vaccination plan require consultation between the 
investigator and the Sponsor and written documentation of the collaborative decision on 
participant management.
7.7
Concomitant Therapy
Medications or vaccinations specifically prohibited in the exclusion criteria are not allowed 
during time periods specified by this protocol for the particular medication or vaccination.
If there is a clinical indication for any medication or vaccination specifically prohibited, 
discontinuation from trial therapy or vaccination may be required. The investigator should 
discuss any questions regarding this with the Sponsor Clinical Director. The final decision on 
any supportive therapy or vaccination rests with the investigator and/or the participant's 
primary physician. However, the decision to continue the participant on study treatment 
requires the mutual agreement of the investigator, the Sponsor and the participant.
See the exclusion criteria for specific restrictions for prior and concomitant medications at 
Day 1 (Section 6.2) and prerequisites for other vaccination visits (Section 6.2.2). 
Participants may receive allergen desensitization therapy and tuberculin skin testing while 
participating in the study.
Topical tacrolimus is allowed provided that it is not used 2 weeks prior to, or 2 weeks 
following a V160 dose.
Use of prior and concomitant medications/vaccination should be recorded as described in 
Section 9.1.5. 
Rescue Medications and Supportive Care
No rescue or supportive medications are specified to be used in this trial.
7.8
Treatment After the End of the Study 
There is no study-specified treatment following the end of the study.
7.9
Clinical Supplies Disclosure
This trial is blinded but supplies are provided open label; therefore, an unblinded pharmacist 
or qualified trial site personnel will be used to blind supplies.  Study treatment identity 
(name, strength or potency) is included in the label text; random code/disclosure envelopes or 
lists are not provided.
The emergency unblinding call center will use the treatment/randomization schedule for the 
trial to unblind participants and to unmask study treatment identity. The emergency 
unblinding call center should only be used in cases of emergency (see Section 9.1.10). In the 
event that the emergency unblinding call center is not available for a given site in this trial, 
the interactive response technology (IRT) should be used in order to unblind participants and 
to unmask study treatment identity.  The Sponsor will not provide random code/disclosure 
envelopes or lists with the clinical supplies.
 
 
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See Section 9.1.10, Participant Blinding/Unblinding, for a description of the method of 
unblinding a participant during the trial, should such action be warranted.
8. Discontinuation/Withdrawal Criteria 
8.1
Discontinuation of Study Treatment 
Discontinuation of study treatment does not represent withdrawal from the study.
As certain data on clinical events beyond study treatment discontinuation may be important 
to the study, they must be collected through the participant’s last scheduled follow-up, even 
if the participant has discontinued study treatment.  Therefore, all participants who 
discontinue study treatment prior to completion of the vaccination regimen will still continue 
to participate in the study as specified in the SoA. This includes the self-collection of urine 
and saliva samples by study participants.
Participants may discontinue study treatment at any time for any reason or be dropped from 
the study treatment at the discretion of the investigator should any untoward effect occur.  In 
addition, a participant may be discontinued from study treatment by the investigator or the 
Sponsor if study treatment is inappropriate, the trial plan is violated, or for administrative 
and/or other safety reasons.  Specific details regarding procedures to be performed at study 
treatment discontinuation are provided in Section 9.1.9. 
A participant must be discontinued from study treatment but continue to be monitored in the 
study for any of the following reasons:
○
The participant or participant’s legally acceptable representative requests to discontinue 
study treatment.
o
The participant’s treatment assignment has been unblinded by the investigator, Merck 
subsidiary or through the emergency unblinding call center. 
o
The participant has a medical condition or personal circumstance which, in the opinion of 
the investigator and/or sponsor, places the participant at unnecessary risk from continued 
administration of study treatment.
o
The participant has a confirmed positive pregnancy test.
o
The participant had an allergic reaction or an anaphylactic/anaphylactoid reaction that 
required medical intervention following Dose 1 administration.
o
The participant enrolled in another interventional clinical trial following Dose 1 
administration.
o
Received a non-study CMV vaccine.
For participants who are discontinued from study treatment but continue to be monitored in 
the trial, all visits and procedures, as outlined in the SoA, should be completed.
Discontinuation from study treatment is “permanent.” Once a participant is discontinued, she 
shall not be allowed to restart study treatment.
 
 
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8.2
Withdrawal from the Study
A participant must be withdrawn from the study if the participant or participant’s legally 
acceptable representative withdraws consent from the study.
If a participant withdraws from the study, they will no longer receive study treatment or be 
followed at scheduled protocol visits.
Specific details regarding procedures to be performed at the time of withdrawal from the 
study, as well as specific details regarding withdrawal from Future Biomedical Research are 
outlined in Section 9.1.9. The procedures to be performed should a participant repeatedly fail 
to return for scheduled visits and/or if the study site is unable to contact the participant are 
outlined in Section 8.3.
8.3
Lost to Follow Up
If a participant fails to return to the clinic for a required study visit and/or if the site is unable 
to contact the participant, the following procedures are to be performed:
o
The site must attempt to contact the participant and reschedule the missed visit.  If the 
participant is contacted, the participant should be counseled on the importance of 
maintaining the protocol-specified visit schedule.
o
The investigator or designee must make every effort to regain contact with the 
participant at each missed visit (eg, phone calls and/or a certified letter to the 
participant’s last known mailing address or locally equivalent methods). These 
contact attempts should be documented in the participant’s medical record.
o
Note:  A participant is not considered lost to follow-up until the last scheduled visit 
for the individual participant.  The amount of missing data for the participant will be 
managed via the pre-specified data handling and analysis guidelines.
9. Study Assessments and Procedures
●
Study procedures and their timing are summarized in the SoA. 
●
Adherence to the study design requirements, including those specified in the SoA, is 
essential and required for study conduct.
●
The Investigator is responsible for assuring procedures are conducted by staff 
appropriately qualified by education, training or experience.  Delegation of study site 
personnel responsibilities will be documented in the Investigator Trial File Binder (or 
equivalent).
●
All screening evaluations must be completed and reviewed to confirm that potential 
participants meet all eligibility criteria. The investigator will maintain a screening log 
to record details of all participants screened and to confirm eligibility or record 
reasons for screening failure, as applicable. 
●
Procedures conducted as part of the participant’s routine clinical management (eg, 
blood count) and obtained before signing of ICF may be utilized for screening or 
baseline purposes provided the procedure met the protocol-specified criteria and were 
performed within the time frame defined in the SoA.
 
 
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●
Additional evaluations/testing may be deemed necessary by the investigator and or 
the Sponsor for reasons related to participant safety. In some cases, such 
evaluation/testing may be potentially sensitive in nature (eg, HIV, Hepatitis C, etc.), 
and thus local regulations may require that additional informed consent, and assent if 
applicable, be obtained from the participant.  In these cases, such evaluations/testing 
will be performed in accordance with those regulations.
The maximum amount of blood collected from each participant over the duration of the 
study, including any extra assessments that may be required is shown in
Table 3.
Repeat or unscheduled samples may be taken for safety reasons or for technical issues with 
the samples.
Table 3
Approximate Blood/Tissue Volumes Drawn/Collected by Trial Visit and by 
Sample Types
Population
Sample
Visit Number, 
Time point
Approximate 
Volume Collected
All participants
Serum for CMV serostatus by IgG
Visit 1, Screening
3.5 mL
Serum -Human Chorionic Gonadotropin 
(-hCG)
Visit 1, Screening
3.5 mL
Blood (DNA) for Future Biomedical 
Research
Visit 2, Day 1
8.5 mL
Serum for Neutralizing Antibody 
Serum for IgG ELISA antibodies
Visit 2, Day1
15 mL
Visit 5, Month 7
15 mL
Visit 6, Month 12
15 mL
Visit 8, Month 24
15 mL
Visit 10, Month 36
15 mL
Non-sub Study Participants Expected Total Volume Collected
90.5 mL
Detailed 
Immunogenicity 
sub-study 
participants 
onlya
Serum for Neutralizing Antibody 
Serum for IgG ELISA antibodies
Visit 4, Month 6
15 mL
Blood for ELISPOT (PBMCs)
Visit 2, Day1
50 mL
Visit 4, Month 6
50 mL
Visit 5, Month 7
50 mL
Visit 6, Month 12
50 mL
Visit 8, Month 24
50 mL
Visit 10, Month 36
50 mL
Additional Sub-Study Expected Total Volume Collected
315 mL 
Sub-study Participants Expected Total Volume Collected (all 
visits included)
405.5 mL
a Note that these samples are in addition to samples collected for all participants.
 
 
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9.1
Administrative and General Procedures
Informed Consent/Assent
The investigator or qualified designee must obtain documented consent, and assent if 
applicable, from each potential participant or each participant’s legally acceptable 
representative prior to participating in a clinical trial or Future Biomedical Research. If there 
are changes to the participant’s status during the trial (eg, health or age of majority 
requirements), the investigator or qualified designee must ensure the appropriate 
consent/assent is in place. 
9.1.1.1
General Informed Consent/Assent
Consent/assent must be documented by the participant’s dated signature or by the 
participant’s legally acceptable representative’s dated signature on a consent/assent form 
along with the dated signature of the person conducting the consent discussion. 
A copy of the signed and dated consent/assent form should be given to the participant before 
participation in the trial.
The initial informed consent/assent form, any subsequent revised written informed 
consent/assent form and any written information provided to the participant must receive the 
IRB/IEC’s approval/favorable opinion in advance of use.  The participant or his/her legally 
acceptable representative should be informed in a timely manner if new information becomes 
available that may be relevant to the participant’s willingness to continue participation in the 
trial.  The communication of this information will be provided and documented via a revised 
consent/assent form or addendum to the original consent/assent form that captures the 
participant’s dated signature or by the participant’s legally acceptable representative’s dated 
signature.
Specifics about a trial and the trial population will be added to the consent/assent form 
template at the protocol level.  
The informed consent will adhere to IRB/IEC requirements, applicable laws and regulations 
and Sponsor requirements. The assent, as applicable will adhere to IRB/IEC requirements, 
applicable laws and regulations and Sponsor requirements.
Pregnancy follow-up will be included in the general informed consent form in the event that 
any participant becomes pregnant, and will allow the site to monitor the pregnancy, and have 
access to medical records pertaining to the pregnancy and the infant outcome. 
A separate informed consent will be requested for any participant who becomes pregnant 
during the study. This consent will be obtained when the pregnancy is reported and a visit is 
scheduled for pregnancy and infant follow-up (at cCMVi Visit 1).
9.1.1.2
Consent/Assent and Collection of Specimens for Future Biomedical Research
The investigator or qualified designee will explain the Future Biomedical Research 
consent/assent to the participant, answer all of his/her questions, and obtain written informed 
consent/assent before performing any procedure related to the Future Biomedical Research 
sub-trial.  A copy of the informed consent/assent will be given to the participant.
 
 
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Inclusion/Exclusion Criteria
All inclusion and exclusion criteria will be reviewed by the investigator or qualified designee 
to ensure that the participant qualifies for the study. An additional review of 
exclusion/discontinuation criteria will be done at subsequent doses after Dose 1. These 
criteria are listed in Section 8.1.
Participant Identification Card
All participants will be given a Participant Identification Card identifying them as 
participants in a research study. The card will contain trial site contact information (including 
direct telephone numbers) to be utilized in the event of an emergency. The investigator or 
qualified designee will provide the participant with a Participant Identification Card 
immediately after the participant provides written informed consent/assent. At the time of 
treatment allocation/randomization, site personnel will add the treatment/randomization 
number to the Participant Identification Card.
The participant identification card also contains contact information for the emergency 
unblinding call center so that a health care provider can obtain information about study 
treatment in emergency situations where the investigator is not available.
Medical History
A medical history will be obtained by the investigator or qualified designee at the
screening visit in order to ensure that the participant satisfies the inclusion and exclusion
criteria of the study. The participant's medical conditions will be reported on the
appropriate eCRF, including any chronic or serious conditions.
Prior and Concomitant Medications Review
9.1.5.1
Prior Medications
The investigator or qualified designee will review prior medication/vaccination use and 
record prior medication taken by the participant to assess inclusion and exclusion criteria 
including time windows for medication/vaccination use.
9.1.5.2
Concomitant Medications
The investigator or qualified designee will record medication, if any, taken by the participant 
during the trial. As outlined in the exclusion criteria, there are specific restrictions for prior 
medications and vaccines, as well as concomitant medications and vaccines that are 
anticipated throughout the duration of the study. Investigators should make every attempt to
adhere to the entry criteria concerning prior, concomitant, and anticipated medications and
vaccines.
Assignment of Screening Number
All consented participants will be given a unique screening number that will be used to 
identify the participant for all procedures that occur prior to randomization.  Each participant 
will be assigned only one screening number.  Screening numbers must not be re-used for 
different participants.
 
 
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Any participant who is screened multiple times will retain the original screening number 
assigned at the initial screening visit.
Note that participants who meet screening eligibility may be randomized outside of the 21-
day window; however, all screening procedures must be repeated when a participant is not 
randomized within 21 days of Visit 1 (screening). Participants who are re-screened must be 
randomized within 21 days of the re-screening visit.
Assignment of Treatment/Randomization Number
All eligible participants will be randomly allocated and will receive a 
treatment/randomization number.  The treatment/randomization number identifies the 
participant for all procedures occurring after treatment allocation/randomization.  Once a 
treatment/randomization number is assigned to a participant, it can never be re-assigned to 
another participant.
A single participant cannot be assigned more than 1 treatment/randomization number.
Treatment Administration
V160/placebo should be prepared and administered by appropriately qualified members of 
the study personnel (eg, physician, nurse, physician’s assistant, nurse practitioner, 
pharmacist, or medical assistant) as allowed by local/state, country, and institutional 
guidance. The study personnel responsible for vaccine preparation/administration will be 
unblinded and will have no further contact with the participants or data.  Open-label, single-
dose vials will be supplied to the unblinded individual(s) at the clinical site. Supplies will be 
affixed with a clinical label in accordance with regulatory requirements.
Note that the V160/placebo should be administered within 60 minutes of preparation. Refer 
to Section 7.5.1 for details.
The V160/placebo will be administered intramuscularly in the deltoid muscle using the 
syringe that was prepared by the unblinded individual/s. Injections should be administered at
a 90° angle into the muscle tissue using a needle long enough to ensure intramuscular 
deposition of V160/placebo. A separate, sterile disposable unit should be used for the 
administration of V160/placebo to each participant. Needles should not be recapped. Safe 
disposal procedures should be followed. Study participants will be observed for 30 minutes 
following each vaccination for any immediate adverse events and longer if necessary. 
Adequate treatment provision, including epinephrine, should be available for immediate use 
should an anaphylactic or anaphylactoid reaction occur.
Participants who meet any of the deferred exclusion criteria (Section 6.2.1) may subsequently 
return to the clinic to determine eligibility status prior to vaccination. 
Prior to each subsequent vaccination, exclusion criteria for subsequent vaccination (Section 
6.2.2 and Section 8.1) will be assessed.
Any participant who has a positive pregnancy test prior to vaccination, will not be 
vaccinated.
 
 
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9.1.8.1
Timing of Dose Administration
Dose 1 of study treatment for all treatment groups will begin on the day of treatment 
randomization at Visit 2 (Day 1). 
For participants randomized to the 3-dose regimen (Day 1, Month 2, and Month 6), a second 
dose of study treatment will be given at Visit 3 (Month 2) and a third dose will be given at 
Visit 4 (Month 6). 
For participants randomized to the 2-dose regimen (Day 1 and Month 6), placebo will be 
given at Visit 3 (Month 2), and the second dose will be given at Visit 4 (Month 6).
For participants randomized to the placebo arm, placebo will be given at Visit 2 (Day 1), 
Visit 3 (Month 2), and Visit 4 (Month 6).
Withdrawal/Discontinuation
When a participant withdraws from participation in the trial, all applicable activities 
scheduled for the Month 7 visit (if not completed) or the final trial visit should be performed 
at the time of withdrawal.  Any adverse events which are present at the time of withdrawal 
should be followed in accordance with the safety requirements outlined in Section 9.3 -
Adverse Events.
9.1.9.1
Withdrawal From Future Biomedical Research
A Participant’s consent for Future Biomedical Research may be withdrawn by the participant 
or the participant’s legally acceptable representative (as appropriate) and their specimens and 
all derivatives destroyed. A participant’s consent may be withdrawn at any time by 
contacting the principal investigator for the main trial.  If medical records for the main trial 
are still available, the investigator will contact the Sponsor using the designated mailbox 
(clinical.specimen.management@merck.com).  Subsequently, the participant's consent for 
Future Biomedical Research will be withdrawn.  A letter will be sent from the Sponsor to the 
investigator confirming the withdrawal. It is the responsibility of the investigator to inform 
the participant of completion of withdrawal.  Any analyses in progress at the time of request 
for withdrawal or already performed prior to the request being received by the Sponsor will 
continue to be used as part of the overall research trial data and results.  No new analyses 
would be generated after the request is received.
In the event that the medical records for the main trial are no longer available (eg, if the 
investigator is no longer required by regulatory authorities to retain the main trial records) or 
the specimens have been completely anonymized, there will no longer be a link between the 
participant’s personal information and their specimens.  In this situation, the request for 
specimen withdrawal cannot be processed.
Participant Blinding/Unblinding
STUDY TREATMENT IDENTIFICATION INFORMATION IS TO BE UNMASKED 
ONLY IF NECESSARY FOR THE WELFARE OF THE PARTICIPANT. EVERY 
EFFORT SHOULD BE MADE NOT TO UNBLIND THE PARTICIPANT UNLESS 
NECESSARY.
 
 
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For emergency situations where the investigator or delegate needs to identify the treatment
used by a participant and the dosage administered, he/she will contact the emergency 
unblinding call center by telephone and make a request for emergency unblinding.  As 
requested by the investigator or delegate the emergency unblinding call center will provide 
the information to him/her promptly and report unblinding to the sponsor.  Prior to contacting 
the emergency unblinding call center to request unblinding of a participant’s treatment 
assignment, the investigator or delegate must enter the toxicity/intensity grade of the adverse 
events observed, the relation to study treatment, the reason thereof, etc., in the medical chart.
Participants whose treatment assignment has been unblinded by the investigator/delegate 
and/or non-study treating physician must be discontinued from study treatment, but should 
continue to be monitored in the trial.
Additionally, the investigator must go into the IRT system and perform the unblind in the 
IRT system to update treatment disposition.  In the event that the emergency unblinding call 
center is not available for a given site in this trial, IRT should be used for emergency 
unblinding in the event that this is required for participant safety.
Study treatment identification information is to be unmasked ONLY if necessary for the 
welfare of the participant.  Every effort should be made not to unblind the participant unless 
necessary. 
In the event that unblinding has occurred, the circumstances around the unblinding (eg, date, 
reason and person performing the unblinding) must be documented promptly, and the 
Sponsor Clinical Director notified as soon as possible. Once an emergency unblinding has 
taken place, the principal investigator, site personnel, and Sponsor personnel may be 
unblinded so that the appropriate follow-up medical care can be provided to the participant.
At the end of the trial, unblinding logs are to be returned to the Sponsor or designee.
Distribution of Electronic Vaccination Report Card
The electronic vaccination report card (eVRC) was developed to be administered 
electronically via a hand-held device. This item was structured as recommended in the final 
FDA Patient Reported Outcome (PRO) Guidance. The investigator or delegate will train the 
participant in the use of the eVRC prior to dispensing it to the participant at Visit 2 (Day 1).
Body temperatures, injection site reactions, vaccine specific complaints, other complaints or
illnesses and medications/vaccinations will be recorded on the eVRC following each 
vaccination (refer to Section 9.5.4 for safety assessments using the eVRC).
Calibration of Equipment
The investigator or qualified designee has the responsibility to ensure that any device or 
instrument used for a clinical evaluation/test during a clinical study that provides information 
about inclusion/exclusion criteria and/or safety or efficacy parameters shall be suitably 
calibrated and/or maintained to ensure that the data obtained is reliable and/or reproducible. 
Documentation of equipment calibration must be retained as source documentation at the 
study site.
 
 
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9.2
Efficacy and Immunogenicity Assessments
Efficacy Assessment
Efficacy of the V160 vaccine will be assessed by monitoring for CMVi among study 
participants (primary endpoint) and cCMVi in pregnancies and/or infants of study 
participants (exploratory endpoint). This will be done by evaluating self-collected urine and 
saliva samples from study participants by PCR for CMV and pregnancies and/or infants of 
study participants for evidence of cCMVi.
9.2.1.1
CMVi
9.2.1.1.1 CMVi Case Definition
A case of CMVi is defined as the detection of CMV (non-vaccine type) by PCR from a 
single saliva or urine sample in a previously CMV uninfected participant (CMV seronegative 
and no prior non-vaccine CMV detected) in the per protocol population.
9.2.1.1.2 CMVi Case Detection
At Visit 2 (Day 1), participants will be trained on proper saliva and urine sample collection 
procedures using the self-collection kits provided. The first set of samples will be collected 
on site by the participant. All other samples will be collected monthly, from Month 1 to 
approximately Month 36, by the participant outside of the study site (eg. at home or place of 
employment) or at scheduled site visits (when applicable), as indicated in the SoA.
In addition to on-site training, instructional materials will be provided to the participant to 
use as a reference. Participants will contact the site with any questions or concerns they have 
regarding the self-collection process. Reminder contacts will be made monthly to the 
participants, to prompt their collection of samples. Receipt of the samples will be tracked in 
real-time in order to provide feedback to the site. The site will contact and re-train 
participants who are missing samples or submitting samples which are unacceptable for 
testing. Detailed instructions will be provided to the site for monitoring self-collection and 
sample handling.
9.2.1.1.3 CMV PCR Testing 
DNA will be extracted from the self-collected urine and saliva samples and assayed for the 
presence of CMV by a PCR assay. If CMV is detected, distinction between V160 vaccine 
virus and non-vaccine virus will be performed in a separate PCR assay.  Additionally, in 
order to assess the adequacy of the self-collection, extracted DNA will be assayed for a 
human housekeeping gene, beta-globin by PCR. Beta-globin testing will stop after 6 months, 
once samples are demonstrated to be consistently positive. If self-collected samples are not 
consistently positive or there is concern about declining quality/quantity of DNA isolated, 
beta-globin testing may be reinstituted.
Refer to Appendix 5 for a summary of the CMV PCR assay. 
 
 
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9.2.1.2
cCMVi
9.2.1.2.1 cCMVi Case Definition
Virologically-confirmed cases of cCMVi will be used to test the efficacy of V160 against 
cCMVi. A virologically-confirmed case of cCMVi is defined as the occurrence of any of the 
following events: 
1. PCR-positive result by Merck-validated CMV PCR assay in at least 1 urine sample 
obtained within 21 days of birth from an infant born to a study participant.

PCR-negative results by Merck-validated CMV PCR assay in all available urine 
samples obtained within 21 days of birth from an infant born to a study participant are 
considered definitive evidence of absence of CMV infection, and therefore constitute 
a non-case of cCMVi.
2. In the absence of any urine sample obtained within 21 days of birth from an infant born 
to a study participant and tested by Merck-validated CMV PCR assay, a PCR-positive 
result by Merck-validated CMV PCR assay in at least 2 saliva samples obtained within 
21 days of birth from an infant born to a study participant.

The cCMVi case definition based on saliva samples will be applied only in the 
absence of results from urine samples. PCR-negative results by Merck-validated 
CMV PCR assay in all available urine samples obtained within 21 days of birth from 
an infant born to a study participant are considered definitive evidence of absence of 
CMV infection, and therefore constitute a non-case of cCMVi, regardless of the PCR 
results in saliva samples.
3. A consensus diagnosis by the Clinical Adjudication Committee (CAC) of a case of 
cCMVi in the fetus/infant of a study participant based on detection of CMV by PCR or 
other virologic methods outside the context of the study (ie, based on methods not 
validated by Merck) from:
a.
saliva, urine, or other body fluid samples (including the neonatal dried blood spot) 
obtained within 21 days of birth from an infant born to a study participant;
OR
b. amniocentesis fluid obtained from a study participant, or samples from still-birth or 
products of conception obtained from a study participant.
9.2.1.2.2 cCMVi Case Detection
After a pregnancy is reported to the site, the participant will be asked to come to the site for a 
study visit (referred to as cCMVi Visit 1). 
At this visit, the site staff will explain that the participant’s pregnancy will be monitored via 
monthly contacts from the site until the outcome of the pregnancy is established and 
recorded. Instructions on when to call the site for any changes in pregnancy status will be 
provided and an explanation and informed consent/assent regarding follow-up of infant/live 
birth will also be provided. 
 
 
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For the pregnancy follow-up, medical records of the pregnant study participant will be 
collected and reviewed by the site personnel to:

confirm date of conception and expected delivery date

obtain information on the outcome of the pregnancy

obtain information that could indicate a suspicion/diagnosis of congenital CMV 
infection and whether the participant had any related interventions
o
Any anti-CMV treatments received by participants during pregnancy or by
infants will be collected for the duration of the trial.

obtain information that could indicate suspected congenital infection in the infant
All relevant medical records should be placed in the participant’s trial file. Infant outcomes 
will be recorded in the eCRF for all pregnant participants within 24 hours of the delivery or 
as soon as possible. All pregnancies will be followed to outcome. Refer to Section 9.3.6 for 
additional requirements for pregnancy follow-up and reporting.
For any live birth occurring during the study, the participant will be asked to return to the site 
with their infant for an additional visit(s) (referred to as cCMVi Visit 2) within 21 days after 
birth of the infant to collect urine and saliva samples from the infant. During this time, 2 sets 
of urine and saliva samples at 2 different time points will be collected from the infant to 
determine the infant’s CMV status by PCR testing as described in Section 9.2.1.1.3. Each set 
of samples can be collected on different days or on the same day, at least 3 hours apart. The 
saliva sample should not be collected within 2 hours of breastfeeding (if the woman is 
breastfeeding). Infant sample collection kits will be provided to the sites and site personnel 
should collect the samples according to the instructions provided. It is critical that both sets 
of samples are collected within 21 days of the birth of the infant.
If it is not possible for the participant to return to the clinic with their infant to obtain urine 
and saliva samples, additional efforts will be made to obtain the sample (eg, study personnel 
may be dispatched to a hospital or the infant’s home).
Medical review and adjudication by the CAC will be triggered by suspicion of congenital 
CMV during pregnancy of a study participant or in the infant.

The CAC will review all relevant information captured in V160 case report forms.

Suspicion of congenital CMV during pregnancy of a study participant will be 
documented and captured in the V160 fetus-infant details (FID) case report form.

Detection of CMV by PCR or other virologic methods outside the context of the 
study will also be documented and captured in the FID form.

Detection of CMV by PCR or other virologic methods outside the context of the
study is a necessary requirement for the CAC to arrive at a diagnosis of cCMVi case 
attributable to the study participant.
Data to be used to determine cases and details regarding the adjudication process will be 
provided in the Adjudication Charter. Refer to Section 9.2.1.2.1 for the cCMVi case 
definition and Appendix 1 for information regarding the CAC.
 
 
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9.2.1.2.3 Health Follow-Up of Infants Diagnosed with cCMVi
Infants who are diagnosed with cCMVi will be followed for up to years 3 after birth to 
collect hearing, vision, and/ or neurological outcomes as well as any other manifestations of 
cCMVi. These infant health follow-up visits will be conducted on a yearly basis as outlined 
in the SoA.
Immunogenicity Assessments 
Sera collected from all participants at time points specified in the SoA will be used to 
measure humoral immune responses to V160 using both viral NAbs and IgG ELISA assays. 
Blood collected from a subset of subjects at time points specified in the SoA will be used to 
measure cell-mediated immunity using the CMV ELIPSOT assay.
Blood collection, storage and shipment instructions for serum and blood samples will be 
provided in the operations/laboratory manual.
Note that all blood samples collected on the day of vaccination will be drawn prior to
V160/placebo administration.
Refer to Table 3 for a listing of assays to be done at each visit and volume of blood collected.
9.2.2.1
Viral Neutralizing Antibody Assay 
The viral NAb assay measures the functional antibodies blocking viral entry and is 
considered the primary assay for immunogenicity. 
Refer to Appendix 5 for additional details of the assay.
9.2.2.2
IgG ELISA Assay 
IgG ELISA measuring total IgG against V160 virus will be validated as a surrogate 
immunogenicity assay. This assay does not provide a functional read-out of immunogenicity 
of V160 but is easier to perform. If it is found to correlate with immunogenicity measured by 
the NAb, it may replace the NAb assay in future studies.  
Refer to Appendix 5 for additional details of the assay.
9.2.2.3
CMV IFN-γ ELISPOTs Assay
The CMV interferon gamma (IFN-γ) ELISPOTs (CMV ELISPOT) assay will be used to 
assess CMI responses by detecting (IFN-secreting CMV-specific T cells from PBMCs in 
response to stimulation of three different CMV antigens (pp65 and IE1 CMV proteins, and 
purified CMV).
This assay will be performed on a subset of participants as shown in the SoA.
Refer to Appendix 5 for additional details of the assay.
 
 
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9.3
Adverse Events (AE), Serious Adverse Events (SAE) and Other Reportable Safety 
Events 
The definitions of an adverse event (AE) or serious adverse event (SAE), as well as the 
method of recording, evaluating, and assessing causality of AE and SAE and the procedures 
for completing and transmitting AE, SAE and other reportable safety event reports can be 
found in Appendix 4.
AE, SAEs, and other reportable safety events will be reported by the participant (or, when 
appropriate, by a caregiver, surrogate, or the participant's legally authorized representative). 
The investigator, who is a qualified physician, and any designees are responsible for 
detecting, assessing, documenting, and reporting events that meet the definition of an AE or 
SAE as well as other reportable safety events. Investigators remain responsible for following 
up AE, SAEs and other reportable safety events for outcome according to Section 9.3.3.
Time Period and Frequency for Collecting AE, SAE and Other Reportable 
Safety Event Information 
All AEs, SAEs and other reportable safety events that occur after the consent form is signed 
but before allocation/randomization must be reported by the investigator if they cause the 
participant to be excluded from the trial, or are the result of a protocol-specified intervention, 
including but not limited to washout or discontinuation of usual therapy, diet, placebo 
treatment or a procedure. 
From the time of allocation/randomization through 14 days following the first vaccination(s) 
and from the time of any subsequent vaccination(s) through 14 days thereafter, all AEs, 
SAEs and other reportable safety events must be reported by the investigator. SAEs 
(regardless of causality), other reportable safety events (ie, Cancer, Overdose, pregnancy, and 
infant SAEs) are reportable throughout the duration of an individual’s study participation. 
Exposure during breast feeding is reportable from Day 1 through Month 7.
Additionally, any SAE brought to the attention of an investigator at any time outside of the 
time period specified in the previous paragraph also must be reported immediately to the 
Sponsor if the event is either:
1. A death that occurs prior to the participant completing the trial, but outside the time 
period specified in the previous paragraph.
or
2. A serious adverse event that is considered by an investigator who is a qualified physician 
to be vaccine related.
Investigators are not obligated to actively seek AE or SAE or other reportable safety events 
in former study participants. However, if the investigator learns of any SAE, including a 
death, at any time after a participant has been discharged from the study, and he/she 
considers the event to be reasonably related to the study treatment or study participation, the 
investigator must promptly notify the sponsor.
All initial and follow-up AEs, SAEs and other reportable safety events will be recorded and 
reported to the sponsor or designee within the timeframes as indicated in Table 4.
 
 
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Table 4
Reporting Time Periods and Timeframes for Adverse Events and Other 
Reportable Safety Events
Type of Event
Reporting Time 
Period:
Consent to 
Randomization/ 
Allocation
Reporting Time 
Period:
Randomization/ 
Allocation 
through 
Protocol-
Specified Follow-
up Period
Reporting Time 
Period:
After the Protocol 
Specified Follow-
up Period
Timeframe to 
Report Event 
and Follow-up 
Information to 
SPONSOR:
Non-Serious 
Adverse Event 
(NSAE)
Report if:
- due to protocol-
specified 
intervention
- causes exclusion
- participant is 
receiving placebo 
run-in or other run-
in treatment
Report all from 
the time of each 
vaccination 
through 14 days 
thereafter.
Not required
Per data entry 
guidelines
Serious Adverse 
Event (SAE)
Report if:
- due to protocol-
specified 
intervention
- causes exclusion
- participant is 
receiving placebo 
run-in or other run-
in treatment
Report all for the 
duration of an 
individual’s study 
participation.
Report if:
- drug/vaccine 
related.
- any death until 
participant
completion of trial
(Follow ongoing to 
outcome)
Within 24 hours 
of learning of 
event
Pregnancy
Report if:
- due to intervention
- causes exclusion
Report all –
Follow to 
completion/termin
ation and report 
outcome
Previously reported 
– Follow to 
completion/termina
tion; report 
outcome
Within 24 hours 
of learning of 
event
Lactation 
Exposure
Report if:
- due to intervention
- causes exclusion
Report all - Day 1 
through Month 7
Previously reported 
– follow to 
completion
Within 24 hours 
of learning of 
event
Cancer
Report if:
- due to intervention
- causes exclusion
Report all for the 
duration of an 
individual’s study 
participation.
Not required
Within 5 
calendar days of 
learning of 
event
Overdose
Report if:
- receiving placebo 
run-in or other run-
in medication 
Report all for the 
duration of an 
individual’s study 
participation.
Not required
Within 5 
calendar days of 
learning of 
event
 
 
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Method of Detecting AE, SAE and Other Reportable Safety Events
Care will be taken not to introduce bias when detecting AE and/or SAE and other reportable 
safety events. Open-ended and non-leading verbal questioning of the participant is the 
preferred method to inquire about AE occurrence.
Follow-up of AE, SAE and Other Reportable Safety Event Information 
After the initial AE/SAE report, the investigator is required to proactively follow each 
participant at subsequent visits/contacts. All AE, SAE and other reportable safety events 
including pregnancy and exposure during breastfeeding, Cancer and Overdose will be 
followed until resolution, stabilization, until the event is otherwise explained, or the 
participant is lost to follow-up (as defined in Section 8.3). In addition, the investigator will 
make every attempt to follow all non-serious AEs that occur in randomized participants for 
outcome.  Further information on follow-up procedures is given in Appendix 4. Although 
participants are to agree not to become pregnant during the vaccination period through 
Month 7, if a pregnancy is reported before the last planned V160/placebo dose, the 
participant will be discontinued from further planned vaccination regimen per exclusion 
criterion but will not be discontinued from the study. Participants who inadvertently become 
pregnant before receiving all 3 doses of study vaccine or placebo, and any participant who 
becomes pregnant after the treatment phase will remain in the trial for follow-up of 
pregnancy and outcome.
Regulatory Reporting Requirements for SAE

Prompt notification (within 24 hours) by the investigator to the sponsor of SAE is 
essential so that legal obligations and ethical responsibilities towards the safety of 
participants and the safety of a study treatment under clinical investigation are met. 

The sponsor has a legal responsibility to notify both the local regulatory authority and 
other regulatory agencies about the safety of a study treatment under clinical 
investigation. All Adverse Events will be reported to regulatory authorities, IRB/IECs 
and investigators in accordance with all applicable global laws and regulations, ie, per 
ICH Topic E6 (R1) Guidelines for Good Clinical Practice.

Investigator safety reports must be prepared for suspected unexpected serious adverse 
reactions (SUSAR) according to local regulatory requirements and sponsor policy and 
forwarded to investigators as necessary.

An investigator who receives an investigator safety report describing an SAE or other 
specific safety information (eg, summary or listing of SAE) from the sponsor will file it 
along with the Investigator’s Brochure and will notify the IRB/IEC, if appropriate 
according to local requirements.
Disease-Related Events and/or Disease-Related Outcomes Not Qualifying as AEs 
or SAEs
There are no disease-related events and/or disease related outcomes not qualifying as AEs or 
SAEs for this study.
 
 
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Pregnancy and Exposure During Breastfeeding
Although pregnancy and infant exposure during breastfeeding are not considered adverse 
events, any pregnancy or infant exposure during breastfeeding in a participant (spontaneously 
reported to the investigator or their designee) that occurs during the trial are reportable to the 
Sponsor. A detailed previous pregnancy history will be collected from all randomized 
participants who report a pregnancy during the study.
All female participants will undergo pregnancy testing based on serum analysis for β-hCG at 
screening, and urine sample analysis prior to each vaccination. Participants found to be 
pregnant at Day 1 are not eligible to participate in the trial and participants are instructed to 
use effective contraception through Month 7, one month after the treatment phase. All 
pregnancies that occur during the trial are reportable to the Sponsor.
Participants who inadvertently become pregnant before receiving all 3 doses of study vaccine
or placebo, and any participant who becomes pregnant after the treatment phase will remain 
in the trial for follow-up of pregnancy and infant outcome. Infant SAEs for all infants born to 
participants who received the study vaccine or placebo or who were breastfed during the 
treatment phase of the trial must be reported for the duration of the trial.
Infant exposure to vaccination during the treatment phase refers to any participant who is 
breastfeeding an infant and receives a vaccination; all cases of infant exposure must be 
reported during the trial. Lactation events do not need to be reported if breastfeeding begins 
after the treatment phase.
All reported pregnancies in randomized participants must be followed to the 
completion/termination of the pregnancy. Pregnancy outcomes of spontaneous abortion, 
missed abortion, benign hydatidiform mole, blighted ovum, fetal death, intrauterine death, 
miscarriage and stillbirth must be reported as serious events (Important Medical Events). If 
the pregnancy continues to term, the outcome (health of infant) must also be reported. Refer 
to Section 9.2.1.2.3 for additional requirements regarding infant outcomes for confirmed 
cases of cCMVi.
Events of Clinical Interest (ECIs)
Selected non-serious and serious adverse events are also known as Events of Clinical Interest 
(ECIs) and must be reported to the Sponsor. There are no ECIs for this study.
9.4
Treatment of Overdose
In this trial, an overdose is defined as receipt of more than 1 dose of study vaccine in a 24-
hour period.
Sponsor does not recommend specific treatment for an overdose.
9.5
Safety 
Details regarding specific safety procedures/assessments to be performed in this trial are 
provided below. The total amount of blood/tissue to be drawn/collected over the course of 
the trial (from pre-trial to post-trial visits), including approximate blood/tissue volumes 
drawn/collected by visit and by sample type per participant can be found in Table 3.
 
 
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Planned time points for all safety assessments are provided in the SoA.
Physical Examinations (Full and Targeted Examinations)
A full physical examination should be performed prior to vaccination at Visit 2. It should 
include vital signs (heart rate, respiratory rate, seated blood pressure, and oral temperature), 
height, weight, heart and lungs abdomen, as well as head, eyes, ears, nose, and throat 
(HEENT), skin, lymph nodes, neurological system, and musculoskeletal system. Any 
clinically significant abnormality should be recorded on the appropriate eCRF. Special 
attention should be paid to clinical signs related to a previous serious illness.
A directed physical examination should be performed prior to each vaccination at Visit 3 
(Month 2) and Visit 4 (Month 6), and at Visit 5 (Month 7). This should include vital signs, 
heart, lungs, and abdomen.
Vital Signs and Body Temperature
Vital signs and body temperature recordings are part of the physical examination and include
oral temperature, heart rate, respiratory rate, and blood pressure. Blood pressure and heart 
rate measurements will be assessed after the participant has at least 5 minutes of rest in a 
quiet setting without distractions, and from a sitting position with a completely automated 
device. Manual techniques will be used only if an automated device is not available.
Oral body temperatures will also be documented by participants using their eVRC during the 
eVRC-specified postvaccination follow-up period.
Laboratory Procedures and Assessments
Refer to Table 3 for the list of clinical laboratory tests to be performed and to the SoA for the 
timing and frequency. 
●
The investigator must review the laboratory report, document this review, and record 
any clinically relevant changes occurring during the study in the AE section of the 
CRF. The laboratory reports must be filed with the source documents. Clinically 
significant abnormal laboratory findings are those which are not associated with the 
underlying disease, unless judged by the investigator to be more severe than expected 
for the participant's condition.
●
All protocol-required laboratory assessments, as defined in Table 3, must be 
conducted in accordance with the laboratory manual and the SoA.
●
If laboratory values from non-protocol specified laboratory assessments performed at 
the institution’s local laboratory require a change in study participant management or 
are considered clinically significant by the investigator (eg, SAE or AE or dose 
modification), then the results must be recorded in the appropriate CRF (eg, SLAB).
●
For any laboratory tests with values considered clinically significantly abnormal 
during participation in the study or within 30 days after the last dose of study 
treatment, every attempt should be made to perform repeat assessments until the 
values return to normal or baseline or if a new baseline is established as determined 
by the investigator. 
 
 
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9.5.3.1
Pregnancy Test
A serum β-hCG pregnancy test (which must be sensitive enough to detect β-hCG at 
concentrations of 25 IU/L) will be performed for women of childbearing potential during 
screening. 
In addition, a urine β-hCG pregnancy test (which must be sensitive to detect β-hCG at 
concentrations of 25 IU/L) will be performed on each vaccination day prior to any 
V160/placebo dose, and at Month 7 (one month after the last dose). A negative urine β-hCG 
pregnancy test must be documented on the day of vaccination before the administration of
V160/placebo. Additional pregnancy tests may be performed at the discretion of the 
investigator at any time during the study.
From Month 7 through the end of the study, study pregnancy tests will not be performed, 
however, participants will be asked to report a pregnancy as soon as they become aware of 
the pregnancy. Refer to Section 9.3.6 for details.
Safety Assessments Using the eVRC
All participants will be observed for 30 minutes after each vaccination for any immediate 
reactions. If any immediate AEs are observed during this period, the time at which the event 
occurred within this timeframe, as well as the event itself, any concomitant medications that 
were administered, and resolution of the event, must be recorded on the appropriate eCRF.
Participants will use the eVRC (Section 9.1.11) to document the following information:

Oral body temperatures measured from Day 1 (day of vaccination) through Day 14
postdose; any temperature ≥100.4°F (≥38.0ºC) oral or equivalent, will be considered an 
AE of fever. 

Solicited injection-site AEs (pain/tenderness, erythema/redness, and swelling) from Day 
1 through Day 5 postdose;

Solicited systemic AEs (headache, fatigue, muscle pain, and joint pain) from Day 1 
through Day 14 postdose;

Any other injection-site or systemic AEs from Day 1 through Day 14 postdose; and

Concomitant medications from Day 1 to Day 14 postdose.
The investigator or delegate will review the data captured on the eVRC with the participant 
at the subsequent visit after each vaccination (refer to the SoA).
For AEs reported on the eVRC, the investigator will use the information provided by the 
participant both on the eVRC, and verbally at the time of eVRC review, to apply the 
appropriate intensity/toxicity grade as described in Appendix 4.
9.6
Pharmacokinetics
PK parameters will not be evaluated in this study.
9.7
Pharmacodynamics
Pharmacodynamic parameters will not be evaluated in this study.
 
 
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9.8
Biomarkers
Biomarkers are not evaluated in this study.
9.9
Future Biomedical Research Sample Collection 
The following specimens are to be obtained as part of Future Biomedical Research:
(and will be stored for future biomedical research)

DNA for future research

Leftover main study saliva which test positive from CMV DNA assay stored for 
future research 

Leftover main study urine which test positive from CMV DNA assay stored for future 
research

Leftover main study serum from immunologic testing stored for future research and 
assay development

Leftover sub-study PBMC from immunologic testing stored for future research and 
assay development
9.10 Visit Requirements
Visit requirements are outlined in Section 2 – Schedule of Activities (SoA).  Specific 
procedure-related details are provided above in Section 9 – Study Assessments and 
Procedures.
10. Statistical Analysis Plan 
This section outlines the statistical analysis strategy and procedures for the study. If, after the 
study has begun, but prior to any unblinding/final database lock, changes made to primary 
and/or key secondary hypotheses, or the statistical methods related to those hypotheses, then 
the protocol will be amended (consistent with ICH Guideline E-9). Changes to exploratory or 
other non-confirmatory analyses made after the protocol has been finalized, but prior to 
unblinding/final database lock, will be documented in a sSAP and referenced in the Clinical 
Study Report (CSR) for the study. Post hoc exploratory analyses will be clearly identified in 
the CSR.  
 
 
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10.1 Statistical Analysis Plan Summary
Key elements of the statistical analysis plan are summarized below; the comprehensive plan 
is provided in Sections 10.2 to 10.12. For the purposes of analysis and reporting, the overall 
study ends when all study efficacy, immunogenicity, and safety data are available and ready 
for database lock execution.
Study Design Overview
Double-Blind, Randomized, Placebo-Controlled Phase 2b, 
Multi-center Study to Evaluate the Safety, Tolerability, 
Efficacy and Immunogenicity of a 2-Dose and a 3-Dose 
Regimen of V160 (Cytomegalovirus [CMV] Vaccine) in 
Healthy Seronegative Women, 16 to 35 Years of Age
Treatment Assignment
Participants will be randomized in a 1:1:1 allocation to one of 
the following 3 treatment arms:

V160 3-dose regimen: will receive 3 doses of V160 
administered IM according to a Day 1, Month 2, Month 6 
schedule;

V160 2-dose regimen: will receive 2 doses of V160 
administered IM according to a Day 1 (V160), Month 2 
(placebo), Month 6 (V160) schedule;

Placebo regimen: will receive 3 doses of saline solution 
placebo administered IM according to a Day 1, Month 2, 
Month 6 schedule.
No stratification based on age, sex or other characteristics will 
be used in this trial.
Analysis Populations
Efficacy: Per-Protocol Efficacy (PPE)
Safety: All Participants as Treated (APaT)
Primary Endpoint(s)
1. Efficacy: number of participants with CMVi in the V160 
3-dose regimen and placebo arms.
2. Safety: number of participants experiencing solicited 
injection site AEs within Days 1 through 5 after each 
vaccination visit; solicited systemic AEs and vaccine-
related serious AEs within Days 1 through 14 after each 
vaccination visit.
Key Secondary 
Endpoints
Efficacy: number of participants with CMVi in the V160 2-
dose regimen and placebo arms.
 
 
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Statistical Methods for 
Key Efficacy Analyses
The primary and secondary efficacy hypotheses will be 
evaluated by calculating the efficacy of V160 compared to 
placebo with respect to the CMVi endpoint. The p-value for 
testing the hypothesis that VE>0% as well as estimation of the 
95% CI of VE will be based on the exact binomial method
proposed by Chan and Bohidar [Chan, I. S. F. and Bohidar, N. 
R. 1998].
Statistical Methods for 
Key Safety Analyses
There are no a priori clinical events identified in this trial as 
Tier 1 events. Tier 2 events identified in this trial include 
solicited injection site and systemic AEs; vaccine-related AEs 
(systemic or serious); any serious AEs (in trial participants or 
infants born to trial participants); discontinuations due to AEs, 
AEs by system organ class observed in 7 participants in at 
least one vaccination group and elevated temperatures.
Estimates of 95% confidence intervals for between-treatment 
differences in the percentage of participants with events will 
be calculated using the Miettinen and Nurminen method
[Miettinen, O. and Nurminen, M. 1985].
Interim Analyses
One IA will be performed in this trial relating to the testing of 
the primary efficacy hypothesis. Results will be reviewed by 
an eDMC. A summary of the IA is provided below. Further 
details are provided in Section 10.7. 
IA:

Timing: To be conducted upon accumulation of 15 CMVi 
cases in the combined V160 3-dose regimen and placebo arms, 
expected at ≈18 months from trial start.

Testing: The primary efficacy hypothesis will be tested.
Final analysis

Timing: If the result of testing the primary efficacy hypothesis 
during the IA was inconclusive, a final analysis will be 
conducted upon accumulation of 24 CMVi cases in the 
combined V160 3-dose regimen and placebo arms, expected at 
≈3 years from trial start.
Multiplicity
The primary efficacy hypothesis will be tested once at an IA, 
and if necessary, a second time at a final analysis. The 
secondary efficacy hypothesis will be tested in one final 
analysis, without an IA, only if the primary efficacy 
hypothesis is successfully demonstrated. The overall one-
sided trial Type 1 error associated with testing the primary and 
secondary efficacy hypotheses is 2.5% based on a 
gatekeeping procedure described in Section 10.8.
 
 
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Sample Size and Power 
The power of the decision rules that will be implemented in 
this trial for testing of the primary efficacy hypothesis is based 
on a fixed-event design.

Corresponding to the decision rules shown in Figure 2 in 
Section 10.7, the trial has >90% power to demonstrate 
that the efficacy of a V160 3-dose regimen compared to 
placebo to prevent CMVi is >0% if the underlying 
vaccine efficacy is 80% at an overall one-sided Type 1
error equal to 2.3% (see Table 6).
The trial will randomize 2,100 participants equally allocated 
into one of the 3 treatment arms as described in the
Treatment Assignment panel of this section. Under study 
design assumptions described in Section 10.9.1.1, the IA of 
the primary efficacy hypothesis is expected to be conducted at 
≈18 months from trial start; and the final analysis if needed is 
expected to be conducted at ≈3 years from trial start, as 
mentioned in the Interim Analyses panel of this section.
The power of the decision rules that will be implemented in 
this trial for testing of the secondary efficacy hypothesis is 
also based on a fixed-event design.

Corresponding to the decision rules shown in Figure 2 in 
Section 10.7, if the secondary efficacy hypothesis is 
tested based on a total of 15 observed CMVi cases in the 
combined V160 2-dose and placebo regimens, then the 
trial has power >85% at an overall one-sided Type 1 error 
≤2.5% to demonstrate that the efficacy of a V160 2-dose 
regimen compared to placebo to prevent CMVi is >0% if 
the underlying vaccine efficacy is 85% (see Table 7).
10.2 Responsibility for Analyses/In-House Blinding
This trial will be conducted as a double-blind trial under in-house blinding procedures. 
The statistical analysis of the data obtained from this trial will be the responsibility of the 
Clinical Biostatistics department of the Sponsor.
The Clinical Biostatistics department will generate the randomized allocation schedules for 
assignment of study treatment and selection of immunogenicity sub-study participants as 
described in Section 7.3. Randomization will be implemented using IRT.
An external statistician and scientific programmer will be identified and will comprise the 
unblinded team. This team will be unblinded on a participant-level. They will not be involved 
in any discussions regarding modifications to the protocol, statistical methods, identification 
of protocol deviations, or data validation efforts at any time during the trial.
 
 
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This trial has one planned IA, and its key aspects are described in Section 10.7. The eDMC 
will serve as the primary reviewer of the results of the IA. Treatment-level results of the IA 
will be provided by the external unblinded statistician to the eDMC. The eDMC will make 
recommendations relating to trial discontinuation or modification to an Executive Oversight
Committee (EOC) (see Appendix 1) of the Sponsor. If the eDMC recommends modifications 
to the design of the protocol or discontinuation of the trial, the EOC may be unblinded to 
results at the treatment level in order to act on these recommendations. If necessary, limited 
additional Sponsor personnel may be unblinded to the treatment level results of the IA in 
order to act on the recommendations of the eDMC. The extent to which individuals are 
unblinded with respect to results of IA will be documented by the external unblinded 
statistician. Additional logistical details will be provided in the eDMC Charter.
Upon reaching the milestone when definitive disposition of the primary and secondary 
efficacy hypotheses has been reached:

A database lock will be executed.

All Sponsor personnel who are members of the study team and the V160 program team 
will be unblinded and a clinical study report will be written.

If this milestone is reached at or beyond study Year 3, or if the outcome in this 
milestone is failure of the test of the primary efficacy hypothesis, then the study 
investigators and participants will be unblinded upon completion of the clinical study 
report.
Definitive disposition of the primary and secondary hypotheses is reached when one of the 
following study outcomes occur:
1. Test of the primary efficacy hypothesis fails.
2. Tests of both the primary and secondary efficacy hypotheses succeed.
3.
Test of the primary efficacy hypothesis succeeds but the test of the secondary 
efficacy hypothesis fails.
If at least the primary efficacy hypothesis is successfully demonstrated prior to reaching 
study Year 3, the study may continue to follow participants up to study Year 3 in order to 
continue to accumulate more information relating to vaccine efficacy against CMVi and 
cCMVi. Under this scenario, study investigators and participants will remain blinded during 
the continuing follow-up for CMVi and cCMVi information. Upon reaching the third year 
of the study:

A database lock will be executed.

An end-of-study clinical report will be written.

The study investigators and participants will be unblinded upon completion of the end-
of-study clinical report.
For the purpose of external reporting of study results in clinical trial registries (eg, EudraCT 
and Clinicaltrials.gov), the end-of-study milestone corresponds to the last data generated 
date, which is the date when all study efficacy, immunogenicity, and safety data are available 
and ready for database lock execution.
 
 
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10.3 Hypotheses/Estimation
Objectives and hypotheses of the study are stated in Section 4.
10.4 Analysis Endpoints
The efficacy, immunogenicity, and safety endpoints that will be evaluated are listed below.
Efficacy Endpoints
The efficacy endpoint that will be used to evaluate VE and test the primary and secondary 
efficacy hypotheses is the number of participants with primary CMVi, described in Section 4 
and Section 5.4.1.
CMVi is defined as detection of CMV DNA by PCR in urine or saliva samples.
As stated in Section 4, in the evaluation of the primary and secondary efficacy hypotheses, 
cases of CMVi will be counted starting after the last dose of the assigned vaccination 
regimen. As stated in Section 10.5.1, the primary and secondary efficacy hypothesis will be 
evaluated on the PPE population, where eligibility includes being PCR negative post Day 1 
through Month 7. Thus, CMVi cases that will be included in the PPE testing of the primary 
and secondary efficacy hypothesis are those that occurred after Month 7.
Immunogenicity Endpoints
All immunogenicity endpoints in this study are exploratory endpoints and are listed below. 
These will be further described and defined in a sSAP that will be written for this study.

Seroconversion based on the NAb and IgG ELISA assays, defined as transition from 
CMV seronegative at Day 1 to CMV seropositive at Month 7.

Seropositivity based on the NAb and IgG ELISA assays at Day 1, Months, 7, 12, 24, 
and 36.

GMTs based on the NAb and the IgG ELISA assays at Day 1, Months 7, 12, 24, and 
36.

GMc of SFC/106 PBMCs based on the ELISPOT assay Day 1, Months 6, 7, 12, and 
24. 
Safety Endpoints
As specified in Section 4, Section 5.4.1.3, and Section 10.6.3, the endpoints that will be used 
to assess the safety and tolerability of 2-dose and 3-dose regimens of V160 include the 
following:

Number of participants experiencing solicited injection site AEs within Days 1 through 5
after each vaccination visit; solicited systemic AEs (including tiredness, muscle pain, 
headache, and joint pain); and vaccine-related systemic AEs, SAEs, and elevated 
temperatures within Days 1 through 14 after each vaccination visit 

Incidence of any SAE at any time during the study
 
 
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
Incidence of vaccine-type viral shedding

Outcomes of pregnancies.
10.5 Analysis Populations
Efficacy Analysis Populations
Per Protocol Efficacy (PPE)
The PPE population will serve as the primary population for the evaluation of VE and for 
testing the primary and secondary efficacy hypotheses. To be eligible for inclusion in the 
PPE population, study participants must satisfy the following criteria:

CMV seronegative at Day 1 and CMV negative by PCR for non-vaccine strain virus
from post Day 1 through Month 7

Have received all 3 injections/vaccinations within the vaccination visit window 
specified in the SoA (Section 2)

At any time from Day 1 through Month 7, did not have events, deviations from 
protocol, or engaged in activities that are deemed by the study Clinical Director to 
potentially interfere with the evaluation of efficacy or immune response to injections 
of V160.
Participants whose CMV status by PCR post Day 1 through Month 7 is not evaluable will be 
ineligible for inclusion in the PPE population. The categories of events, deviations from 
protocol, or activities that are deemed by the study Clinical Director to potentially interfere 
with the evaluation of efficacy or immune response to injections of V160 will be identified 
and documented in a memo to study file prior to the time of the IA for efficacy assessment.
Full Analysis Set (FAS)
The FAS will serve as a supportive analysis population for the evaluation of VE. The FAS 
population consists of all randomized participants who received at least 1 dose of the correct 
clinical material corresponding to the treatment regimen the participants were randomized 
into.
Immunogenicity Analysis Populations
The immunogenicity analysis populations that will be used in exploratory analyses of 
immunogenicity will be described in the sSAP that will be written for this study.
Safety Analysis Population
All Participants as Treated (APaT)
The APaT population will serve as the primary population for evaluation of safety. The 
APaT population includes all randomized participants who received at least 1 injection of 
V160 or placebo, have safety follow-up data, and assigns participants to the treatment arm 
corresponding to the actual clinical material received. For participants who received 
injections of V160 or placebo corresponding to a sequence that does not correspond to any of 
 
 
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the protocol-defined treatment arms (ie, cross-treated participants), a safety profile listing 
will be created separate from the safety reports that will be provided for the protocol-defined 
treatment arms.
10.6 Statistical Methods
Statistical testing and inference relating to efficacy are described in Section 10.6.1. Statistical 
methods relating to safety analyses are described in Section 10.6.3. Efficacy results that will 
be deemed to be statistically significant after consideration of the Type 1 error control 
strategy are described in Section 10.8. Nominal p-values may be computed for other efficacy 
analyses, but should be interpreted with caution due to potential issues of multiplicity, 
sample size, etc. Unless otherwise stated, all statistical tests will be conducted at the α=0.025 
(1-sided) level.
Statistical Methods for Efficacy Analyses
To address the primary and secondary efficacy hypotheses, one-sided tests of the null 
hypothesis that the vaccine efficacy is 0% versus the alternative that vaccine efficacy is >0% 
will be conducted separately for each of the primary and secondary hypotheses. Each of the 
primary and secondary efficacy hypotheses will be tested at one-sided α=0.025 level (ie,
2.5% Type 1 error). Vaccine efficacy is defined as:
VE = 100%*{1-(Rv/Rp)}
where Rv and Rp are the incidence rates CMVi in the V160 dose regimen (3-dose for the 
primary efficacy hypothesis testing; 2-dose for the secondary efficacy hypothesis testing) and 
placebo groups, respectively. The incidence rate Rv is defined as Rv = Cv/Tv, where Cv = 
the count of CMVi cases in the vaccine group and Tv = total person-years of follow-up for 
efficacy in the vaccine group. The incidence rate Rp is defined similarly. The null hypothesis 
that vaccine is not efficacious (ie, VE=0%) will be tested by constructing a 100*(1α)%
confidence interval (CI) for VE, denoted as (VEL, VEU), based on study data at a particular 
analysis time point (eg, at IA or at final analysis). The statistical criterion for success with 
respect to the primary efficacy hypothesis (and likewise for the secondary efficacy 
hypothesis) will be met if VEL >0%. 
The 100*(1α) % CI for VE, (VEL, VEU), is constructed as follows. Under the assumption 
that Rv and Rp are the means of independent Poisson processes, and given that there is a total 
of n = Cv + Cp efficacy endpoint cases observed in the combined V160 dose regimen and 
placebo groups, then the number of primary efficacy cases Cv in the V160 dose regimen 
group is distributed as Binomial(n,), where the binomial probability is defined as = 
TvRv/(TvRv+TpRp). The probability is a person-years-adjusted estimate of the probability 
that a particular study participant who became a case of CMVi belongs to the V160 dose 
regimen group. The lower bound of the 100*(1α) % exact CI for the probability is 
obtained by searching for the proportion L such that the probability of observing Cv or more 
efficacy endpoint cases out of n total efficacy cases is ≤α/2. Similarly, the upper bound of the 
100*(1α)% exact CI for the probability is obtained by searching for the proportion U 
such that the probability of observing Cv or fewer efficacy endpoint cases out of n total 
 
 
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efficacy cases is ≤α/2 [Chan, I. S. F. and Bohidar, N. R. 1998]. VEL and VE are then 
calculated from L and U as follows:
VEL = 100%*{1((U, (1+()}/(1U);
VEU = 100%*{1((L, (1+()}/(1L);
where = Tp/Tv is the ratio of the total person-years of follow-up for efficacy in the placebo 
group over the V160 dose regimen group.
There is only one final analysis of the secondary efficacy endpoint. However, the testing of 
the secondary efficacy hypothesis may occur at the time of the IA or the final analysis of the 
primary efficacy hypothesis. The secondary efficacy hypothesis will only be tested if the 
primary efficacy hypothesis is successfully demonstrated. In addition, the secondary efficacy 
hypothesis will be tested contingent on accumulation of 15 total CMVi cases in the V160 2-
dose and placebo regimens (see Figure 2 in Section 10.7, and Section 10.9.1.2). Thus, the 
possible timing for testing the secondary hypothesis is as follows.

If the primary hypothesis test is successful at the IA and there are 15 total CMVi 
cases in the V160 2-dose and placebo regimens at the time of IA, then the secondary 
efficacy hypothesis will be tested immediately after the testing of the primary efficacy 
hypothesis at the time of the IA.

If the primary hypothesis test is successful at the IA and there are <15 total CMVi 
cases in the V160 2-dose and placebo regimens at the time of IA, the secondary 
efficacy hypothesis will not be tested at the IA. In this case, follow-up will continue 
until at least 15 total CMVi cases in the V160 2-dose and placebo regimens groups 
are accrued. Upon accrual of 15 total CMVi cases in the V160 2-dose and placebo 
regimens, the secondary efficacy hypothesis will be tested.

If the primary hypothesis test is not successful at the IA, the secondary efficacy 
hypothesis will not be tested at the time of the IA.
Statistical Methods for Immunogenicity Analyses
Statistical methods for analysis of exploratory immunogenicity endpoints will be provided in 
the sSAP that will be written for this study.
Statistical Methods for Safety Analyses
Safety and tolerability will be assessed by clinical review of all relevant parameters including 
AEs and outcomes of pregnancies.
The analysis of safety results will follow a tiered approach (Table 5). The tiers differ with 
respect to the analyses that will be performed. Safety parameters or adverse events of special 
interest that are identified a priori constitute “Tier 1” safety endpoints that will be subject to 
inferential testing for statistical significance with p-values and 95% confidence intervals 
provided for between-group comparisons. Other safety parameters will be considered Tier 2 
or Tier 3. Tier 2 parameters will be assessed via point estimates with 95% confidence 
intervals provided for V160 dose regimen versus placebo comparisons; only point estimates 
by treatment group are provided for Tier 3 safety parameters. Safety results will be 
summarized for each vaccination dose and across all three vaccination doses.
 
 
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There were no SAEs nor vaccine-type viral shedding (which is an event of interest for V160) 
observed in the V160 Phase I study. While vaccine-type viral shedding is an event of interest 
that can occur only in V160-vaccinated subjects, barring false-positive tests in placebo 
recipients, the 0% incidence of vaccine-type viral shedding observed in the Phase I study 
provides no informed, data-driven, non-zero incidence level around which a safety 
hypothesis can be formulated and tested. Therefore, there is no a priori identified AE or ECI
for which a test of hypothesis will be conducted. Thus, no Tier 1 event is identified in this 
study. Categories of events considered as Tier 2 and Tier 3 in this study are shown in 
Table 5. In addition, AEs observed in at least 7 participants (representing 1% of N=700 
participants per group) in any vaccination group will be categorized as a Tier 2 event.
Incidences of the categories of events identified in Table 5 will be reported as percentages, 
generally calculated as 100% x (number of participants who experienced the event/total 
number of participants with safety follow-up), except for outcomes of pregnancies, SAEs 
throughout the study duration, and vaccine-type viral shedding. The 95% CIs will be 
provided for vaccine versus placebo percentage point differences (percent in V160 dose 
regimen group minus percent in placebo) will be calculated using the Miettinen and 
Nurminen method [Miettinen, O. and Nurminen, M. 1985], an unconditional, asymptotic 
method. No stratification factor will be used in the calculation of 95% CI.
Percentages of outcomes of pregnancies will be generally calculated as 100% x (total number 
of specific pregnancy outcome being reported/total number of all relevant pregnancy 
outcomes). For example, the percent associated with the pregnancy outcome of live births
will be calculated as 100% x (total number of live births/total number of fetuses/infants with 
known outcomes). The numerator and denominator account for pregnancies with multiple 
fetuses (ie, twins, triplets, etc.) and do not treat such occurrences as a single pregnancy 
outcome or event. Further details will be provided in the sSAP that will be written for this 
study.
Vaccine-type viral shedding will be assessed in V160-vaccinated participants. In addition to 
the analysis strategy shown in Table 5, point and corresponding 95% CI estimates of 
incidence of vaccine-type viral shedding (number of participants observed with vaccine-type 
viral shedding/total person-years of follow-up) will be calculated separately for the V160 3-
dose, V160 2-dose, and placebo arms. No treatment group comparisons (V160 vs. placebo or 
V160 3-dose vs. 2-dose arms) of the percent of participants detected with vaccine-type viral 
shedding will be conducted. 
Summaries of SAEs (throughout the study duration) by system organ class will be provided 
as incidence rates (number of participants [or infants] observed with a specific SAE/total 
person-years of follow-up) rather than percentages.
 
 
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Table 5
Analysis Strategy for Safety Parameters
Safety 
Tier
Safety Endpoint
p-value
95% CI for 
Comparison
of V160 to 
Placebo
Descriptive 
Statistics
Tier 2
Solicited injection site AEsa
X
X
Solicited systemic AEs
X
X
Vaccine-related systemic AEs
X
X
Any serious AEs (in study participants or infants 
born to study participants)
X
Xb
Vaccine-related serious AEs within Days 1 
through 14 after vaccination
X
X
Discontinuations due to AEs
X
X
Elevated temperaturec
X
X
AEs by SOC observed in 7 participants in at 
least one vaccination group
X
X
Tier 3
AEs by SOC
X
Outcomes of pregnancies
X
Vaccine-type viral shedding
X‡
a All injection site AEs are considered vaccine-related.
b Incidence rates (cases/total person-years follow-up) will be provided instead of percentages.
c Defined as ≥38.0ºC [≥100.4°] oral or equivalent
AE = Adverse event; CI = Confidence interval; SOC = System organ class; X = indicated summary statistic 
will be provided.
10.7 Interim Analyses
Efficacy Interim Analysis
An IA for testing the primary efficacy hypothesis will be conducted upon accrual of a total of 
15 CMVi endpoint cases in the combined V160 3-dose and placebo regimens. Possible 
outcomes of this IA and consequent study pathways are as described below and shown in 
schematic form in Figure 2.
1. 7 of 15 CMVi cases observed in the V160 3-dose arm

Study fails to demonstrate the primary efficacy hypothesis at the IA and has no 
chance of successfully demonstrating the primary efficacy hypothesis in a final 
analysis when a total of 24 CMVi cases are accrued in the combined V160 3-dose 
regimen and placebo groups. 

eDMC may recommend the study to be terminated for futility.
2. (4, 5, or 6) of 15 CMVi cases observed in the V160 3-dose arm.

Result of testing of the primary efficacy hypothesis is inconclusive.

Study fails to demonstrate the primary efficacy hypothesis at the IA but still has a 
chance of demonstrating the primary efficacy hypothesis in a final analysis when a 
 
 
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total of 24 CMVi cases are accrued in the combined V160 3-dose regimen and 
placebo groups. 

Study (which is still blinded) will continue until a total of 24 CMVi cases are 
accrued in the combined V160 3-dose regimen and placebo groups, at which time, 
the final testing of the primary efficacy hypothesis will be conducted.
3. ≤3 of 15 CMVi cases observed in the V160 3-dose arm

Study successfully demonstrates the primary efficacy hypothesis.

The appropriate timing for the testing the secondary efficacy hypothesis is evaluated 
(see Section 10.6.1).
Figure 2
Plan for Testing the Primary and Secondary Efficacy Hypotheses
Safety Interim Analysis
At the time of IA for efficacy, the eDMC will also conduct a review of the cumulative study 
safety data recorded in the study database upon which the 15 CMVi cases was counted and 
confirmed to have been accumulated. No hypothesis relating to safety will be tested. 
Summaries of safety parameters identified in Section 10.6.3 Statistical Methods for Safety 
Analyses, will be prepared for review by the eDMC.
 
 
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10.8 Multiplicity
The overall 1-sided Type 1 error in testing the efficacy hypotheses is controlled to not exceed 
2.5% based on gatekeeping procedure. 
The primary efficacy hypothesis will be tested at an overall (over the IA and final analysis) 
1-sided Type 1 error <2.5%, as discussed in Section 10.9.1.1. The secondary efficacy 
hypothesis will be tested at 1-sided Type 1 error 2.5% only if the primary efficacy 
hypothesis is successfully demonstrated, as discussed in Section 10.9.1.2. The primary 
efficacy hypothesis is a reasonable gatekeeper to the secondary efficacy hypothesis because 
if a V160 3-dose regimen with last dose at Month 6 is not efficacious, then a V160 2-dose 
regimen with last dose also at Month 6 is highly unlikely to be efficacious.
10.9 Sample Size and Power Calculations
Sample Size and Power for Efficacy Analyses
10.9.1.1 Sample Size and Power for Testing the Primary Efficacy Hypothesis
The power corresponding to the decision rule that will be implemented in testing the primary 
efficacy hypothesis is based on a fixed-event design. For testing the primary efficacy 
hypothesis, participants will be followed until 15 total CMVi cases in the combined V160 3-
dose and placebo regimens have been accumulated, at which point, an IA will be conducted. 
If the testing of the primary efficacy hypothesis is inconclusive at the IA, follow-up will 
continue until 24 total CMVi cases have been observed in the combined V160 3-dose and 
placebo regimens. The decision rules to be implemented that will determine success or 
failure to demonstrate the primary efficacy hypothesis are as shown in Figure 2 in Section 
10.7 and in Table 6. Corresponding to these decision rules, the study has >90% power to 
demonstrate that the efficacy of a V160 3-dose regimen compared to placebo to prevent 
CMVi is >0% if the underlying vaccine efficacy is 80% at an overall one-sided Type 1 error 
equal to 2.3%. The operating characteristics of the decision rules with one IA and a final 
analysis if necessary are evaluated by a 5 million-replication simulation using SAS v9.4. In 
the simulation conducted, the underlying probability distribution of accumulation of CMVi 
cases in the V160 3-dose regimen conditional on the total cases in the combined V160 and 
placebo regimens is based on an exact method under a large sample Poisson distribution 
assumption proposed by Chan and Bohidar (1998) [Chan, I. S. F. and Bohidar, N. R. 1998].
This study will randomize 700 participants into the V160 3-dose regimen and 700 into the 
placebo regimen. The 15 total CMVi cases in the combined V160 3-dose and placebo 
regimens are projected to be accumulated in ≈18 to 19 months under the following 
assumptions: 1) all 1,400 participants are enrolled within 18 months; 2) incidence of CMVi 
in the placebo group is 2% per year; 2) VE=85%, so that the incidence of CMVi in the V160 
arm is 0.3% per year; 3) loss-to-follow-up is ≈5% to 10% per year; and 4) participants 
ineligible for the PPE analysis is ≈2% to 5% of participants randomized. Under the same 
assumptions, the total of 24 CMVi cases in the combined V160 3-dose and placebo regimens 
is projected to be accumulated in ≈3 years.
 
 
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It is possible that at the time when CMVi cases are counted to determine whether a total of 
15 cases have been accrued, the actual CMVi cases accumulated will already be greater than 
15. In such an eventuality, the IA will be conducted based on the actual number of cases 
accrued (eg, 16, or 17, or some other count). The operating characteristics shown in Table 6
will be recalculated, which will involve recalculating the total number of events needed at the 
final analysis in order to maintain an overall (ie, cumulative over the IA and final analysis) 1-
sided Type 1 error ≤2.5% and overall power >90% when the underlying vaccine efficacy is 
80%.
 
 
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Table 6
Decision Rule and Corresponding Type 1 Error and Power for Testing the Primary Efficacy Hypothesis of VE >0% 
Against CMVi (3-Dose Regimen vs. Placebo)
Analysis
Time
Point
Total 
Cases to 
Accrue
Stop for 
Futility
(Vacc Cases)
Case Split
for Success
(Vacc:Pbo)
VE (95% CI)
Cumulative
Type I 
Error
True Vaccine Efficacy
70%
75%
80%
85%
90%
95%
Cumulative Power
IA
15
7
3 : 12
75.0 (7.4, 95.5)
1.8%
53.2%
64.8%
76.8%
87.8%
95.9%
99.5%
Final†
24
7
6 : 18
66.7 (12.3, 89.2)
2.3%
73.0%
83.7%
92.3%
97.5%
99.6%
100%
† Type I error and power shown are cumulative over the IA and final analysis.
CI = Confidence interval; IA = Interim analysis; VE = Vaccine efficacy.
 
 
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10.9.1.2 Sample Size and Power for Testing the Secondary Efficacy Hypothesis
The power corresponding to the decision rule that will be implemented in testing the 
secondary efficacy hypothesis is based on a fixed-event design. The secondary efficacy 
hypothesis will be tested when a total of 15 CMVi cases in the combined V160 2-dose and 
placebo arms have been observed (see Figure 2 in Section 10.7). When the secondary 
efficacy hypothesis is tested based on a total of 15 CMVi cases, the study has >85% power 
to demonstrate that the efficacy of a V160 2-dose regimen compared to placebo to prevent 
CMVi is >0% if the underlying vaccine efficacy is 85% at an overall one-sided Type 1 error 
2.5% (see Table 7). The calculations are based on an exact method under a large sample 
Poisson distribution assumption proposed by Chan and Bohidar (1998) [Chan, I. S. F. and 
Bohidar, N. R. 1998] and carried out using the R software.
This study will randomize 700 participants into the V160 2-dose regimen and 700 into the 
placebo regimen. Under the same assumptions governing the accumulation of CMVi cases in 
the V160 3-dose and placebo arms, a total of 15 CMVi cases in the combined V160 2-dose 
and placebo arms are projected to be accumulated in ≈18 to 19 months; and a total of 24 in 
≈3 years. 
 
 
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Table 7
Decision Rule and Corresponding Type 1 Error and Power for Testing the Secondary Efficacy Hypothesis of VE >0% 
Against CMVi (2-Dose Regimen vs. Placebo)
Analysis
Time
Point
Total 
Cases 
Accrued
Case Split
for Success
(Vacc:Pbo)
VE (95% CI)
Type I 
Error
True Vaccine Efficacy
70%
75%
80%
85%
90%
95%
Power
Final
15
3 : 12
75.0 (7.4, 95.5)
1.8%
53.2%
64.8%
76.8%
87.8%
95.9%
99.5%
16
3 : 13
76.9 (16.0, 95.8)
1.1%
47.7%
59.8%
72.9%
85.4%
94.9%
99.4%
17
4 : 13
79.2 (0.4, 92.7)
2.5%
64.7%
75.8%
86.0%
93.9%
98.5%
99.9%
18
4 : 14
71.4 (9.0, 93.2)
1.5%
59.6%
71.6%
83.2%
92.5%
98.1%
99.9%
19
4 : 15
73.3 (16.3, 93.6)
1.0%
54.5%
67.3%
80.1%
90.9%
97.6%
99.8%
20
5 : 15
66.7 (3.5, 90.5)
2.1%
69.3%
80.4%
89.8%
96.2%
99.3%
100%
21
5 : 16
68.8 (10.7, 91.0)
1.3%
64.8%
76.9%
87.7%
95.3%
99.1%
100%
22
5 : 17
70.6 (16.9, 91.5)
0.8%
60.2%
73.3%
85.3%
94.2%
98.8%
100%
23
6 : 17
64.7 (6.2, 88.6)
1.7%
73.2%
84.0%
92.5%
97.7%
99.7%
100%
24
6 : 18
66.7 (12.3, 89.2)
1.1%
69.1%
81.1%
90.9%
97.1%
99.6%
100%
CI = Confidence interval; VE = Vaccine efficacy.
 
 
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Sample Size and Power for Safety Analyses
Testing of hypotheses relating to safety will not be conducted in this study. Thus, power and 
Type 1 error considerations relating to safety analyses are not applicable in this study.
The probability of observing a specific AE in specific treatment arms of this study depends 
on the number of vaccinated participants with safety follow-up and the underlying incidence 
of that specific AE in the study population. In each treatment arm, assuming that all 700 
participants randomized into the study will have safety follow-up, then there is a >99% 
chance of observing at least one specific AE if the incidence of that AE is at least 1 in every 
150 participants; >90% chance if the incidence is at least 1 in every 300 participants. If no 
vaccine-related SAEs are observed among the 700 participants randomized in a specific 
V160 treatment arm, this study will provide 97.5% confidence that the underlying percentage 
of participants with vaccine-related SAE is 0.5% (1 in every 200 participants) among V160 
vaccinated participants in that V160 arm. This is based on calculation of a 1-sided 97.5% 
upper confidence limit of a binomial proportion using the exact binomial method of Clopper 
and Pearson [Clopper, C. J. and Pearson, E. S. 1934] and was done using PASS 2008. For a 
sample size of 700 per treatment arm, Table 8 provides the critical split of participants with 
AEs between V160 and placebo arms for which it will be possible to state with 95% 
confidence that the risk difference (defined as percent in V160 dose regimen group minus 
percent in placebo) excludes 0%. For a specific AE, given a sample size of 700 per arm, the 
minimum total number of participants with AEs for which it is possible to state with 95% 
confidence that the risk difference excludes 0% is 5. As stated in Section 10.6.3, AEs 
observed in at least 7 participants (representing 1% of N=700 participants per group) in any 
vaccination group will be categorized as a Tier 2 event. The 95% CIs of the percentage point 
differences were calculated using the Miettinen and Nurminen method [Miettinen, O. and 
Nurminen, M. 1985].
Table 8
Critical Counts of Participants with Adverse Events in V160 and Placebo Arms 
Resulting in the Lower Bound of 95% CI Estimate of Risk Difference to be >0%
Observed Count of Participants
with Adverse Events
Risk Difference† (V160 – Placebo)
Total
V160
(N=700)
Placebo
(N=700)
Percentage Points‡
95% CI§
5
5
0
0.7%
(0.1%, 1.7%)
6
6
0
0.8%
(0.3%, 1.8%)
7
7
0
1.0%
(0.4%, 2.0%)
8
7
1
0.9%
(0.1%, 1.9%)
9
8
1
1.0%
(0.2%, 2.1%)
10
9
1
1.1%
(0.3%, 2.3%)
11
9
2
1.0%
(0.1%, 2.2%)
12
10
2
1.1%
(0.2%, 2.4%)
13
11
2
1.3%
(0.4%, 2.5%)
14
11
3
1.1%
(0.1%, 2.4%)
15
12
3
1.3%
(0.3%, 2.6%)
16
13
3
1.4%
(0.4%, 2.8%)
17
13
4
1.3%
(0.2%, 2.6%)
18
14
4
1.4%
(0.3%, 2.8%)
 
 
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Observed Count of Participants
with Adverse Events
Risk Difference† (V160 – Placebo)
Total
V160
(N=700)
Placebo
(N=700)
Percentage Points‡
95% CI§
19
15
4
1.6%
(0.4%, 3.0%)
14
5
1.3%
(0.1%, 2.7%)
20
15
5
1.4%
(0.2%, 2.9%)
† Defined as percent of participants in the V160 group with adverse events minus the corresponding 
percent in the placebo group.
‡ Calculated as 100%×(V160 – Placebo)÷700.
§ Calculated based on the Miettinen and Nurminen method.
N = Number of participants randomized.
CI = Confidence interval
10.10 Subgroup Analyses and Effect of Baseline Factors
To determine whether VE is consistent across various subgroups associated with specific 
baseline participant characteristics, the estimate of the VE (with a nominal 95% CI) for the 
primary efficacy endpoint (comparing 3-dose regimen versus placebo) and for the secondary 
efficacy endpoint (comparing 2-dose regimen versus placebo) will be estimated and plotted 
within each subgroup. Subgroup assessments of VE will be conducted for the following 
baseline participant characteristics:

Age category (≤21 vs. >21 years)

Race (white, non-white)

Region (US, Ex-US)
For each of the primary and secondary efficacy endpoints, a Forest plot will be produced, 
which provides the estimated point and 95% CI estimates of VE across the subgroups 
associated with specific baseline participant characteristics listed above. The consistency of 
VE across subgroups will be assessed descriptively based on these Forest plots. 
10.11 Compliance
Adherence to Scheduled Vaccinations
Compliance to scheduled vaccinations in this study is defined as receipt of all 3 vaccinations 
of either V160 or placebo. To summarize compliance, the numbers of participants who 
receive each vaccination will be tabulated. For each of vaccination visits 2 and 3, histograms 
of the time (in days) of administration of the vaccine or placebo relative to the target 
vaccination visit will be provided.
Adherence to Scheduled Self-Collection of Urine and Saliva Samples
Accurate and reliable ascertainment of the primary and secondary efficacy endpoints (ie,
incidence of CMVi) will depend on compliance of study participants to the self-collection of 
urine and saliva samples based on the schedule indicated in Section 2. Thus, compliance of 
 
 
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study participants to the self-collection of urine and saliva samples will be assessed as 
follows.
On a participant-level (say the i-th participant among the N=2100 study participants), percent 
compliance (denoted by Pi) of a study participant to the self-collection of urine and/or saliva 
samples will be calculated as:
where 
ni = number of visits for which either a urine or saliva sample was received from the study 
participant;
Mi = total number of study visits throughout the study for which a urine or saliva sample is 
expected to be received from the study participant. This number will vary based on the 
time when the study participant enters the study and the time when the study ends.
For each of the V160 3-dose, V160 2-dose, and placebo arms, point and 95% CI estimates of 
the geometric mean of the participant-level percent compliance Pi will be calculated to 
summarize the vaccination group-level compliance of study participants to the self-collection 
of urine and saliva samples.
This assessment of compliance of study participants to the self-collection of urine and saliva 
samples will be conducted in both the PPE and FAS analysis populations.
10.12 Extent of Exposure
As indicated in Section 5.5, participants who will receive an injection of V160 at a 
vaccination visit will receive a 0.5mL dose containing 100 units of V160. Thus:

Participants randomized to the 3-dose regimen of V160 are expected to be 
administered a cumulative total of 1.5 mL containing 300 units of V160 over a 6-
months duration;

Participants randomized to the 2-dose regimen of V160 are expected to be 
administered a cumulative total of 1.0 mL containing 200 units of V160 over a 6-
months duration;

Participants randomized to placebo are not expected to receive V160.
 
 
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11. References
[Adler, S. P., et al 1995]
Adler SP, Starr SE, Plotkin SA, Hempfling 
SH, Buis J, Manning ML, et al. Immunity 
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[Bate, S. L., et al 2010]
Bate SL, 
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Cannon MJ. 
Cytomegalovirus seroprevalence in the 
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Clin Infect Dis. 2010 Jun 1;50(11):1439-47.
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the 
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03R303
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N. R. 1998]
Chan ISF, Bohidar NR. Exact power and 
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03PRMD
[Clopper, C. J. and Pearson, 
E. S. 1934]
Clopper CJ, Pearson ES. The use of 
confidence or fiducial limits illustrated in 
the case of the binomial. Biometrika 
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[Colugnati, F. A., et al 2007]
Colugnati FA, Staras SA, Dollard SC, 
Cannon MJ. Incidence of cytomegalovirus 
infection among the general population and 
pregnant women in the United States. BMC 
Infect Dis. 2007 Jul 2;7:71.
04PCNN
[Dollard, S. C., et al 2007]
Dollard SC, Grosse SD, Ross DS. New 
estimates of the prevalence of neurological 
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infection. Rev Med Virol 2007;17:355-63.
03RMFN
 
 
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[Fowler, K. B., et al 2003]
Fowler KB, Stagno S, Pass RF. Maternal 
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[Griffiths, P. D., et al 2011]
Griffiths PD, Stanton A, McCarrell E, 
Smith C, Osman M, Harber M, et al. 
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controlled trial. Lancet 2011;377:1256-63.
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[Hahn, G., et al 1998]
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G, 
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ES. 
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[Hyde, T. B., et al 2010]
Hyde TB, Schmid DS, Cannon MJ. 
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risk factors: implications for congenital 
CMV. Rev Med Virol. 2010 Sep;20(5):311-
26.
04PCRS
[Just-Nubling, G., et al 2003]
Just-Nubling G, Korn S, Ludwig B, 
Stephan C, Doerr HW, Preiser W. Primary 
cytomegalovirus infection in an outpatient 
setting--laboratory markers and clinical 
aspects. Infection. 2003 Oct;31(5):318-23.
04GCTX
[Kenneson, A. and Cannon, 
M. J. 2007]
Kenneson A, Cannon MJ. Review and 
meta-analysis of the epidemiology of 
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infection . Rev Med Virol 2007;17:253-76.
03RMF4
[Lanzieri, T. M., et al 2014]
Lanzieri TM, Dollard SC, Bialek SR, 
Grosse SD. Systematic review of the birth 
prevalence of congenital cytomegalovirus 
infection in developing countries. Int J 
Infect Dis. 2014 May;22:44-8.
04PCPQ
[Marsico, C. 2017]
Marsico C, Kimberlin DW. Congenital 
Cytomegalovirus infection: advances and 
challenges in diagnosis, prevention and 
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Ital 
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2017 
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[Miettinen, O. and Nurminen, 
M. 1985]
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[Opelz, G., et al 2004]
Opelz G, Döhler B, Ruhenstroth A. 
Cytomegalovirus prophylaxis and graft 
outcome in solid organ transplantation: A 
collaborative transplant study report. Am J 
Transplant 2004;4:928-36.
03RV0M
[Pass, R. F., et al 2009]
Pass RF, Zhang C, Evans A, Simpson T, 
Andrews W, Huang M-L, et al. Vaccine 
prevention of maternal cytomegalovirus 
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[Plotkin, S. A., et al 1989]
Plotkin SA, Starr SE, Friedman HM, 
Gonczol E, Weibel RE. Protective effects 
of towne cytomegalovirus vaccine against 
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[Sinclair, J. 2008]
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[Sung, H. and Schleiss, M. R. 
2010]
Sung H, Schleiss MR. Update on the 
current status of cytomegalovirus vaccines. 
Expert Rev Vaccines 2010;9(11):1303-14.
03RM43
[Walter, E,, et al 2011]
Walter E., Brennig C., Schollbauer V. 
Congenital Cytomegalovirus Infection. 1st 
ed. Germany: Springer, Vienna; c2011. 
Chapter 7, How to save money: congenital 
CMV infection and the economy; P. 121-
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04R7G4
[Wang, D., et al 2016]
Wang D, Freed DC, He X, Li F, Tang A, 
Cox KS, et al. A replication-defective 
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[Wang, S., et al 2017]
Wang S, Wang T, Zhang W, Liu X, Wang 
X, Wang H, et al. Cohort study on maternal 
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(Baltimore). 2017 Feb;96(5):e6007.
04PCQ0
[Weibel, R. E., et al 1980]
Weibel RE, Villarejos VM, Klein EB, 
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12. Appendices
12.1 Appendix 1: Study Governance Considerations
Merck Code of Conduct for Clinical Trials
Merck*
Code of Conduct for Clinical Trials
I.
Introduction
A. Purpose
Merck, through its subsidiaries, conducts clinical trials worldwide to evaluate the safety and effectiveness of our 
products. As such, we are committed to designing, implementing, conducting, analyzing and reporting these trials in 
compliance with the highest ethical and scientific standards. Protection of participant safety is the overriding concern 
in the design of clinical trials. In all cases, Merck clinical trials will be conducted in compliance with local and/or 
national regulations and in accordance with the ethical principles that have their origin in the Declaration of Helsinki. 
B. Scope
Such standards shall be endorsed for all clinical interventional investigations sponsored by Merck irrespective of the 
party (parties) employed for their execution (eg, contract research organizations, collaborative research efforts). This 
Code is not intended to apply to trials which are observational in nature, or which are retrospective. Further, this Code 
does not apply to investigator-initiated trials which are not under the control of Merck.
II. Scientific Issues
A. Trial Conduct
1. Trial Design
Except for pilot or estimation trials, clinical trial protocols will be hypothesis-driven to assess safety, efficacy 
and/or pharmacokinetic or pharmacodynamic indices of Merck or comparator products.  Alternatively, Merck may 
conduct outcomes research trials, trials to assess or validate various endpoint measures, or trials to determine 
participant preferences, etc.  
The design (ie, participant population, duration, statistical power) must be adequate to address the specific purpose 
of the trial.  Research participants must meet protocol entry criteria to be enrolled in the trial. 
2. Site Selection
Merck selects investigative sites based on medical expertise, access to appropriate participants, adequacy of 
facilities and staff, previous performance in Merck trials, as well as budgetary considerations.  Prior to trial 
initiation, sites are evaluated by Merck personnel to assess the ability to successfully conduct the trial.
3. Site Monitoring/Scientific Integrity
Trial sites are monitored to assess compliance with the trial protocol and general principles of Good Clinical 
Practice.  Merck reviews clinical data for accuracy, completeness and consistency. Data are verified versus source 
documentation according to standard operating procedures.  Per Merck policies and procedures, if fraud, 
misconduct or serious GCP-non-Compliance are suspected, the issues are promptly investigated. When necessary, 
the clinical site will be closed, the responsible regulatory authorities and ethics review committees notified and 
data disclosed accordingly. 
B. Publication and Authorship
To the extent scientifically appropriate, Merck seeks to publish the results of trials it conducts.  Some early phase or 
pilot trials are intended to be hypothesis-generating rather than hypothesis testing.  In such cases, publication of 
results may not be appropriate since the trial may be underpowered and the analyses complicated by statistical issues 
of multiplicity.
Merck’s policy on authorship is consistent with the requirements outlined in the ICH-Good Clinical Practice 
guidelines. In summary, authorship should reflect significant contribution to the design and conduct of the trial, 
performance or interpretation of the analysis, and/or writing of the manuscript.  All named authors must be able to 
defend the trial results and conclusions.  Merck funding of a trial will be acknowledged in publications. 
 
 
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III. Participant Protection
A. IRB/IEC review
All clinical trials will be reviewed and approved by an independent IRB/IEC before being initiated at each site.  
Significant changes or revisions to the protocol will be approved by the IRB/IEC prior to implementation, except that 
changes required urgently to protect participant safety and well-being may be enacted in anticipation of IRB/IEC 
approval. For each site, the IRB/IEC and Merck will approve the participant informed consent form. 
B. Safety
The guiding principle in decision-making in clinical trials is that participant welfare is of primary importance.  
Potential participants will be informed of the risks and benefits of, as well as alternatives to, trial participation. At a 
minimum, trial designs will take into account the local standard of care. Participants are never denied access to 
appropriate medical care based on participation in a Merck clinical trial. 
All participation in Merck clinical trials is voluntary.  Participants are enrolled only after providing informed consent 
for participation.  Participants may withdraw from a Merck trial at any time, without any influence on their access to, 
or receipt of, medical care that may otherwise be available to them.
C. Confidentiality
Merck is committed to safeguarding participant confidentiality, to the greatest extent possible.  Unless required by 
law, only the investigator, sponsor (or representative) and/or regulatory authorities will have access to confidential 
medical records that might identify the research participant by name.  
D. Genomic Research
Genomic Research will only be conducted in accordance with informed consent and/or as specifically authorized by 
an Ethics Committee.
IV. Financial Considerations
A. Payments to Investigators
Clinical trials are time- and labor-intensive.  It is Merck’s policy to compensate investigators (or the sponsoring 
institution) in a fair manner for the work performed in support of Merck trials.  Merck does not pay incentives to 
enroll participants in its trials.  However, when enrollment is particularly challenging, additional payments may be 
made to compensate for the time spent in extra recruiting efforts.
Merck does not pay for participant referrals.  However, Merck may compensate referring physicians for time spent 
on chart review to identify potentially eligible participants.
B. Clinical Research Funding 
Informed consent forms will disclose that the trial is sponsored by Merck, and that the investigator or sponsoring 
institution is being paid or provided a grant for performing the trial.  However, the local IRB/IEC may wish to alter 
the wording of the disclosure statement to be consistent with financial practices at that institution.  As noted above, 
publications resulting from Merck trials will indicate Merck as a source of funding.
C. Funding for Travel and Other Requests
Funding of travel by investigators and support staff (eg, to scientific meetings, investigator meetings, etc.) will be 
consistent with local guidelines and practices including, in the U.S., those established by the American Medical 
Association (AMA). 
V. Investigator Commitment
Investigators will be expected to review Merck’s Code of Conduct as an appendix to the trial protocol, and in signing 
the protocol, agree to support these ethical and scientific standards.
* In this document, "Merck" refers to Merck Sharp & Dohme Corp. and Schering Corporation, each of which is a 
subsidiary of Merck & Co., Inc.  Merck is known as MSD outside of the United States and Canada.  As warranted by 
context, Merck also includes affiliates and subsidiaries of Merck & Co., Inc."
 
 
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Financial Disclosure
Financial Disclosure requirements are outlined in the US Food and Drug Administration 
Regulations, Financial Disclosure by Clinical Investigators (21 CFR Part 54).  It is the 
Sponsor's responsibility to determine, based on these regulations, whether a request for 
Financial Disclosure information is required.  It is the investigator's/subinvestigator's 
responsibility to comply with any such request.  
The investigator/subinvestigator(s) agree, if requested by the Sponsor in accordance with 21 
CFR Part 54, to provide his/her financial interests in and/or arrangements with the Sponsor to 
allow for the submission of complete and accurate certification and disclosure statements.  
The investigator/subinvestigator(s) further agree to provide this information on a 
Certification/Disclosure Form, commonly known as a financial disclosure form, provided by 
the Sponsor. The investigator/subinvestigator(s) also consent to the transmission of this 
information to the Sponsor in the United States for these purposes.  This may involve the 
transmission of information to countries that do not have laws protecting personal data.
Data Protection
Participants will be assigned a unique identifier by the sponsor. Any participant records or 
datasets that are transferred to the sponsor will contain the identifier only; participant names 
or any information which would make the participant identifiable will not be transferred. 
The participant must be informed that his/her personal study-related data will be used by the 
sponsor in accordance with local data protection law. The level of disclosure must also be 
explained to the participant. 
The participant must be informed that his/her medical records may be examined by Clinical 
Quality Assurance auditors or other authorized personnel appointed by the sponsor, by 
appropriate IRB/IEC members, and by inspectors from regulatory authorities.
Confidentiality of Data
By signing this protocol, the investigator affirms to the Sponsor that information furnished to 
the investigator by the Sponsor will be maintained in confidence, and such information will 
be divulged to the institutional review board, ethics review committee (IRB/IEC) or similar 
or expert committee; affiliated institution and employees, only under an appropriate 
understanding of confidentiality with such board or committee, affiliated institution and 
employees.  Data generated by this trial will be considered confidential by the investigator, 
except to the extent that it is included in a publication as provided in the Publications section 
of this protocol.
Confidentiality of Participant Records
By signing this protocol, the investigator agrees that the Sponsor (or Sponsor representative), 
IRB/IEC, or regulatory authority representatives may consult and/or copy trial documents in 
order to verify worksheet/case report form data.  By signing the consent form, the participant 
agrees to this process.  If trial documents will be photocopied during the process of verifying 
 
 
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worksheet/case report form information, the participant will be identified by unique code 
only; full names/initials will be masked prior to transmission to the Sponsor.
By signing this protocol, the investigator agrees to treat all participant data used and 
disclosed in connection with this trial in accordance with all applicable privacy laws, rules 
and regulations.
Confidentiality of IRB/IEC Information
The Sponsor is required to record the name and address of each IRB/IEC that reviews and 
approves this trial.  The Sponsor is also required to document that each IRB/IEC meets 
regulatory and ICH GCP requirements by requesting and maintaining records of the names 
and qualifications of the IRB/IEC members and to make these records available for 
regulatory agency review upon request by those agencies.
Committees Structure
Scientific Advisory Committee
This trial was developed in collaboration with a Scientific Advisory Committee (SAC).  The 
SAC comprises both Sponsor and non-Sponsor scientific experts who provide input with 
respect to trial design, interpretation of trial results and subsequent peer-reviewed scientific 
publications.
Executive Oversight Committee
The Executive Oversight Committee (EOC) comprises members of Sponsor Senior 
Management.  The EOC will receive and decide upon any recommendations made by the 
Data Monitoring Committee (DMC) regarding the trial.
Data Monitoring Committee
To supplement the routine trial monitoring outlined in this protocol, an external Data 
Monitoring Committee (DMC) will monitor the interim data from this trial.  The voting 
members of the committee are external to the Sponsor.  The members of the DMC must not 
be involved with the trial in any other way (eg, they cannot be trial investigators) and must 
have no competing interests that could affect their roles with respect to the trial. 
The DMC will make recommendations to the EOC regarding steps to ensure both participant 
safety and the continued ethical integrity of the trial.  Also, the DMC will review interim trial 
results, consider the overall risk and benefit to trial participants (see Section 10.7 - Interim 
Analyses) and recommend to the EOC if the trial should continue in accordance with the 
protocol.
Specific details regarding composition, responsibilities, and governance, including the roles 
and responsibilities of the various members and the Sponsor protocol team; meeting 
facilitation; the trial governance structure; and requirements for and proper documentation of 
DMC reports, minutes, and recommendations will be described in the DMC charter that is 
reviewed and approved by all the DMC members.
 
 
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Clinical Adjudication Committee
A Clinical Adjudication Committee (CAC) will evaluate the potential cCMVi cases for the 
purposes of confirming them according to the criteria in Section 9.2.1.2.1 – cCMVi Case 
Definition. Cases confirmed as cCMVi by the CAC will be used in the analysis of the 
cCMVi endpoint described in the sSAP.
All personnel involved in the adjudication process will remain blinded to study treatment 
allocation throughout the trial.
Publication Policy
The results of this study may be published or presented at scientific meetings. The sponsor 
will comply with the requirements for publication of study results. In accordance with 
standard editorial and ethical practice, the sponsor will generally support publication of 
multicenter studies only in their entirety and not as individual site data. In this case, a 
coordinating investigator will be designated by mutual agreement.
If publication activity is not directed by the sponsor, the investigator agrees to submit all 
manuscripts or abstracts to the sponsor before submission.  This allows the sponsor to protect 
proprietary information and to provide comments.  
Authorship will be determined by mutual agreement and in line with International Committee 
of Medical Journal Editors authorship requirements.
Compliance with Trial Registration and Results Posting Requirements
Under the terms of the Food and Drug Administration Amendments Act (FDAAA) of 2007 
and the European Medicines Agency (EMA) clinical trial Directive 2001/20/EC, the Sponsor 
of the trial is solely responsible for determining whether the trial and its results are subject to 
the requirements for submission to http://www.clinicaltrials.gov, 
www.clinicaltrialsregister.eu or other local registries.  Merck, as Sponsor of this trial, will 
review this protocol and submit the information necessary to fulfill these requirements. 
Merck entries are not limited to FDAAA or the EMA clinical trial directive mandated trials.  
Information posted will allow participants to identify potentially appropriate trials for their 
disease conditions and pursue participation by calling a central contact number for further 
information on appropriate trial locations and trial site contact information.  
By signing this protocol, the investigator acknowledges that the statutory obligations under 
FDAAA, the EMA clinical trials directive or other locally mandated registries are that of the 
Sponsor and agrees not to submit any information about this trial or its results to those 
registries.
Compliance with Law, Audit and Debarment
By signing this protocol, the investigator agrees to conduct the trial in an efficient and 
diligent manner and in conformance with this protocol; generally accepted standards of Good 
Clinical Practice (eg, International Conference on Harmonization of Technical Requirements 
for Registration of Pharmaceuticals for Human Use Good Clinical Practice: Consolidated 
 
 
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Guideline and other generally accepted standards of good clinical practice); and all 
applicable federal, state and local laws, rules and regulations relating to the conduct of the 
clinical trial.
The Code of Conduct, a collection of goals and considerations that govern the ethical and 
scientific conduct of clinical investigations sponsored by Merck, is provided in this appendix 
under the Merck Code of Conduct for Clinical Trials.
The investigator agrees not to seek reimbursement from participants, their insurance 
providers or from government programs for procedures included as part of the trial 
reimbursed to the investigator by the Sponsor.
The investigator will promptly inform the Sponsor of any regulatory authority inspection 
conducted for this trial.
The Investigator agrees to provide the Sponsor with relevant information from inspection 
observations/findings to allow the Sponsor to assist in responding to any citations resulting 
from regulatory authority inspection, and will provide the Sponsor with a copy of the 
proposed response for consultation before submission to the regulatory authority. 
Persons debarred from conducting or working on clinical trials by any court or regulatory 
authority will not be allowed to conduct or work on this Sponsor’s trials. The investigator 
will immediately disclose in writing to the Sponsor if any person who is involved in 
conducting the trial is debarred or if any proceeding for debarment is pending or, to the best 
of the investigator’s knowledge, threatened.
Data Quality Assurance
All participant data relating to the study will be recorded on printed or electronic CRF unless 
transmitted to the sponsor or designee electronically (eg, laboratory data). The investigator or 
qualified designee is responsible for verifying that data entries are accurate and correct by 
physically or electronically signing the CRF. 
Detailed information regarding Data Management procedures for this protocol will be 
provided separately.
The investigator must maintain accurate documentation (source data) that supports the 
information entered in the CRF. 
The investigator must permit study-related monitoring, audits, IRB/IEC review, and 
regulatory agency inspections and provide direct access to source data documents. 
Trial documentation will be promptly and fully disclosed to the Sponsor by the investigator 
upon request and also shall be made available at the trial site upon request for inspection, 
copying, review and audit at reasonable times by representatives of the Sponsor or any 
regulatory authorities.  The investigator agrees to promptly take any reasonable steps that are 
requested by the Sponsor or regulatory authority as a result of an audit or inspection to cure 
deficiencies in the trial documentation and worksheets/case report forms.
The sponsor or designee is responsible for the data management of this study including 
quality checking of the data. 
 
 
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Study monitors will perform ongoing source data review and verification to confirm that data 
entered into the CRF by authorized site personnel are accurate, complete, and verifiable from 
source documents; that the safety and rights of participants are being protected; and that the 
study is being conducted in accordance with the currently approved protocol and any other 
study agreements, ICH GCP, and all applicable regulatory requirements. 
Records and documents, including signed ICF, pertaining to the conduct of this study must 
be retained by the investigator for 15 years after study completion unless local regulations or 
institutional policies require a longer retention period. No records may be destroyed during 
the retention period without the written approval of the sponsor. No records may be 
transferred to another location or party without written notification to the sponsor. 
Source Documents
Source documents provide evidence for the existence of the participant and substantiate the 
integrity of the data collected. Source documents are filed at the investigator’s site.
Data reported on the CRF or entered in the eCRF that are transcribed from source documents 
must be consistent with the source documents or the discrepancies must be explained. The 
investigator may need to request previous medical records or transfer records, depending on 
the study. Also, current medical records must be available.
Study and Site Closure
The sponsor or its designee may stop the study or study site participation in the study for 
medical, safety, regulatory, administrative, or other reasons consistent with applicable laws, 
regulations, and GCP. 
In the event the Sponsor prematurely terminates a particular trial site, the Sponsor will 
promptly notify that trial site’s IRB/IEC.
 
 
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12.2 Appendix 2: Collection and Management of Specimens for Future Biomedical 
Research
1. Definitions
a.
Biomarker:  A biological molecule found in blood, other body fluids, or tissues that is a 
sign of a normal or abnormal process or of a condition or disease.  A biomarker may be 
used to see how well the body responds to a treatment for a disease or condition.1
b. Pharmacogenomics:  The investigation of variations of DNA and RNA characteristics as 
related to drug/vaccine response.2
c.
Pharmacogenetics:  A subset of pharmacogenomics, pharmacogenetics is the influence of 
variations in DNA sequence on drug/vaccine response.2
d. DNA:  Deoxyribonucleic acid.
e.
RNA:  Ribonucleic acid.
2. Scope of Future Biomedical Research
The specimens consented and/or collected in this trial as outlined in Section 9.9 – Future 
Biomedical Research Sample Collection9.9 – Future Biomedical Research Sample 
Collection will be used in various experiments to understand:
o
The biology of how drugs/vaccines work
o
Biomarkers responsible for how a drug/vaccine enters and is removed by the body
o
Other pathways drugs/vaccines may interact with
o
The biology of disease
The specimen(s) may be used for future assay development and/or drug/vaccine 
development.
It is now well recognized that information obtained from studying and testing clinical 
specimens offers unique opportunities to enhance our understanding of how individuals 
respond to drugs/vaccines, enhance our understanding of human disease and ultimately 
improve public health through development of novel treatments targeted to populations 
with the greatest need.  All specimens will be used by the Sponsor or those working for 
or with the Sponsor.
3. Summary of Procedures for Future Biomedical Research
a.
Participants for Enrollment
All participants enrolled in the clinical trial will be considered for enrollment in the 
Future Biomedical Research sub-trial.
b. Informed Consent
Informed consent for specimens (ie, DNA, RNA, protein, etc.) will be obtained 
during screening for protocol enrollment from all participants or legal guardians, at a 
trial visit by the investigator or his or her designate.  Informed consent for Future 
Biomedical Research should be presented to the participants on the visit designated in 
the SoA.  If delayed, present consent at next possible Participant Visit.  Consent 
 
 
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forms signed by the participant will be kept at the clinical trial site under secure 
storage for regulatory reasons.  
A template of each trial site’s approved informed consent will be stored in the 
Sponsor’s clinical document repository.
c.
eCRF Documentation for Future Biomedical Research Specimens
Documentation of participant consent for Future Biomedical Research will be 
captured in the electronic Case Report Forms (eCRFs). Any specimens for which 
such an informed consent cannot be verified will be destroyed.
d. Future Biomedical Research Specimen(s)
Collection of specimens for Future Biomedical Research will be performed as 
outlined in the SoA. In general, if additional blood specimens are being collected for 
Future Biomedical Research, these will usually be obtained at a time when the 
participant is having blood drawn for other trial purposes.
4. Confidential Participant Information for Future Biomedical Research 
In order to optimize the research that can be conducted with Future Biomedical Research 
specimens, it is critical to link participant' clinical information with future test results. In 
fact little or no research can be conducted without connecting the clinical trial data to the 
specimen. The clinical data allow specific analyses to be conducted. Knowing participant 
characteristics like gender, age, medical history and treatment outcomes are critical to 
understanding clinical context of analytical results.
To maintain privacy of information collected from specimens obtained for Future 
Biomedical Research, the Sponsor has developed secure policies and procedures. All 
specimens will be single-coded per ICH E15 guidelines as described below. 
At the clinical trial site, unique codes will be placed on the Future Biomedical Research 
specimens.  This code is a random number which does not contain any personally 
identifying information embedded within it. The link (or key) between participant 
identifiers and this unique code will be held at the trial site.  No personal identifiers will 
appear on the specimen tube.  
5. Biorepository Specimen Usage
Specimens obtained for the Sponsor will be used for analyses using good scientific 
practices.  Analyses utilizing the Future Biomedical Research specimens may be 
performed by the Sponsor, or an additional third party (eg, a university investigator) 
designated by the Sponsor. The investigator conducting the analysis will follow the 
Sponsor’s privacy and confidentiality requirements. Any contracted third party analyses 
will conform to the specific scope of analysis outlined in this sub-trial. Future Biomedical 
Research specimens remaining with the third party after specific analysis is performed 
will be reported to the Sponsor.
6. Withdrawal From Future Biomedical Research 
Participants may withdraw their consent for Future Biomedical Research and ask that 
their biospecimens not be used for Future Biomedical Research.  Participants may 
withdraw consent at any time by contacting the principal investigator for the main trial. If 
 
 
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medical records for the main trial are still available, the investigator will contact the 
Sponsor using the designated mailbox (clinical.specimen.management@merck.com). 
Subsequently, the participant's specimens will be flagged in the biorepository and 
restricted to main study use only. If specimens were collected from study participants 
specifically for Future Biomedical Research, these specimens will be removed from the 
biorepository and destroyed. Documentation will be sent to the investigator confirming 
withdrawal and/or destruction, if applicable. It is the responsibility of the investigator to 
inform the participant of completion of the withdrawal and/or destruction, if applicable.  
Any analyses in progress at the time of request for withdrawal/destruction or already 
performed prior to the request being received by the Sponsor will continue to be used as 
part of the overall research trial data and results. No new analyses would be generated 
after the request is received.
In the event that the medical records for the main trial are no longer available (eg, if the 
investigator is no longer required by regulatory authorities to retain the main trial 
records) or the specimens have been completely anonymized, there will no longer be a 
link between the participant’s personal information and their specimens. In this situation, 
the request for withdrawal of consent and/or destruction cannot be processed. 
7. Retention of Specimens 
Future Biomedical Research specimens will be stored in the biorepository for potential 
analysis for up to 20 years from the end of the main study. Specimens may be stored for 
longer if a regulatory or governmental authority has active questions that are being 
answered. In this special circumstance, specimens will be stored until these questions 
have been adequately addressed.
Specimens from the trial site will be shipped to a central laboratory and then shipped to 
the Sponsor-designated biorepository. If a central laboratory is not utilized in a particular 
trial, the trial site will ship directly to the Sponsor-designated biorepository. The 
specimens will be stored under strict supervision in a limited access facility which 
operates to assure the integrity of the specimens.  Specimens will be destroyed according 
to Sponsor policies and procedures and this destruction will be documented in the 
biorepository database.
8. Data Security
Databases containing specimen information and test results are accessible only to the 
authorized Sponsor representatives and the designated trial administrator research 
personnel and/or collaborators. Database user authentication is highly secure, and is 
accomplished using network security policies and practices based on international 
standards to protect against unauthorized access.
9. Reporting of Future Biomedical Research Data to Participants
No information obtained from exploratory laboratory studies will be reported to the 
participant, family, or physicians. Principle reasons not to inform or return results to the 
participant include: Lack of relevance to participant health, limitations of predictive 
capability, and concerns regarding misinterpretation.
 
 
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If important research findings are discovered, the Sponsor may publish results, present 
results in national meetings, and make results accessible on a public website in order to 
rapidly report this information to doctors and participants.  Participants will not be 
identified by name in any published reports about this study or in any other scientific 
publication or presentation.
10. Future Biomedical Research Study Population
Every effort will be made to recruit all participants diagnosed and treated on Sponsor 
clinical trials for Future Biomedical Research.
11. Risks Versus Benefits of Future Biomedical Research 
For future biomedical research, risks to the participant have been minimized and are 
described in the Future Biomedical Research informed consent.
The Sponsor has developed strict security, policies and procedures to address participant 
data privacy concerns.  Data privacy risks are largely limited to rare situations involving 
possible breach of confidentiality. In this highly unlikely situation there is risk that the 
information, like all medical information, may be misused.
12. Questions
Any questions related to the future biomedical research should be e-mailed directly to 
clinical.specimen.management@merck.com.
13. References
1. National Cancer Institute: http://www.cancer.gov/dictionary/?searchTxt=biomarker
2. International Conference on Harmonization: DEFINITIONS FOR GENOMIC 
BIOMARKERS, PHARMACOGENOMICS, PHARMACOGENETICS, GENOMIC 
DATA AND SAMPLE CODING CATEGORIES - E15; 
http://www.ich.org/LOB/media/MEDIA3383.pdf
3. Industry Pharmacogenomics Working Group. Understanding the Intent, Scope and 
Public Health Benefits of Exploratory Biomarker Research: A Guide for IRBs/IECs 
and Investigational Site Staff. Available at http://i-pwg.org/
4. Industry Pharmacogenomics Working Group. Pharmacogenomics Informational 
Brochure for IRBs/IECs and Investigational Site Staff. Available at http://i-pwg.org/
 
 
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12.3 Appendix 3: Contraceptive Guidance and Pregnancy Testing
Only women of childbearing potential (WOCBP) 16 to 35 years of age are to be enrolled in 
this study.
A woman is considered fertile following menarche and until becoming post-menopausal 
unless permanently sterile (see below) 
Women in the following categories are NOT considered WOCBP, and therefore will NOT be 
enrolled:
●
Premenopausal female with 1 or more of the following:
o
Documented hysterectomy
o
Documented bilateral salpingectomy
o
Documented bilateral oophorectomy
o
Documented bilateral tubal ligation
Note:  Documentation can come from the site personnel’s review of the participant’s 
medical records, medical examination, or medical history interview.
●
Postmenopausal female
○
A postmenopausal state is defined as no menses for 12 months without an alternative 
medical cause. 
▪
A high follicle stimulating hormone (FSH) level in the postmenopausal range may 
be used to confirm a postmenopausal state in women not using hormonal 
contraception or hormonal replacement therapy (HRT). However, in the absence 
of 12 months of amenorrhea, confirmation with two FSH measurements in the 
postmenopausal range is required. 
○
Females on HRT and whose menopausal status is in doubt will be required to use one 
of the non-hormonal highly effective contraception methods if they wish to continue 
their HRT during the study. Otherwise, they must discontinue HRT to allow 
confirmation of postmenopausal status before study enrollment.
Contraception Requirements
Female participants of childbearing potential are eligible to participate if they agree to use a 
highly effective method of contraception consistently and correctly as described in Table 9
during the protocol-defined time frame in Section 6.1. 
 
 
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Table 9
Highly Effective Contraception Methods
Highly Effective Contraceptive Methods That Are User Dependent a
Failure rate of <1% per year when used consistently and correctly.

Combined (estrogen- and progestogen- containing) hormonal contraception b
o
Oral
o
Intravaginal
o
Transdermal
o
Injectable

Progestogen-only hormonal contraception b
o
Oral
o
Injectable
Highly Effective Methods That Have Low User Dependency
Failure rate of <1% per year when used consistently and correctly.

Progestogen- only contraceptive implant b

Intrauterine hormone-releasing system (IUS)

Intrauterine device (IUD)

Vasectomized partner
A vasectomized partner is a highly effective contraception method provided that the 
partner is the sole male sexual partner of the WOCBP and the absence of sperm has been 
confirmed. If not, an additional highly effective method of contraception should be used.

Sexual abstinence
Sexual abstinence is considered a highly effective method only if defined as refraining 
from heterosexual intercourse during the entire period of risk associated with the study 
treatment. The reliability of sexual abstinence needs to be evaluated in relation to the 
duration of the study and the preferred and usual lifestyle of the participant.
Notes:
Use should be consistent with local regulations regarding the use of contraceptive 
methods for participants of clinical studies.
As WOCBP are the target population of this study and pregnancy is permissible after the 
treatment phase, permanent contraception methods at study entry are not allowed.
a Typical use failure rates are lower than perfect-use failure rates (ie, when used 
consistently and correctly).
b If locally required, in accordance with Clinical Trial Facilitation Group (CTFG) 
guidelines, acceptable hormonal contraceptives are limited to those which inhibit 
ovulation.
 
 
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Pregnancy Testing
Women of childbearing potential are to be included after a negative highly sensitive urine or 
serum pregnancy test and in accordance with local requirements. This test should be repeated 
a maximum of 24-hours before the first dose/vaccination.
 
 
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12.4 Appendix 4: Adverse Events: Definitions and Procedures for Recording, 
Evaluating, Follow-up, and Reporting
Definition of AE
AE Definition
●
An AE is any untoward medical occurrence in a patient or clinical study participant, 
temporally associated with the use of study treatment, whether or not considered 
related to the study treatment.
●
NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a study treatment.
●
NOTE: for purposes of AE definition, study treatment (also referred to as Sponsor’s 
product) includes any pharmaceutical product, biological product, vaccine, device, 
diagnostic agent or protocol specified procedure whether investigational (including 
placebo or active comparator product) or marketed, manufactured by, licensed by, 
provided by or distributed by the sponsor for human use in this study.
Events Meeting the AE Definition 
●
Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) or 
other safety assessments (eg, ECG, radiological scans, vital signs measurements), 
including those that worsen from baseline, or are considered clinically significant in 
the medical and scientific judgment of the investigator.
●
Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.
●
New conditions detected or diagnosed after study treatment administration even 
though it may have been present before the start of the study.
●
Signs, symptoms, or the clinical sequelae of a suspected drug-drug interaction.
●
Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication. 
●
For all reports of overdose (whether accidental or intentional) with an associated 
adverse event, the AE term should reflect the clinical symptoms or abnormal test 
result.  An overdose without any associated clinical symptoms or abnormal laboratory 
results is reported using the terminology “accidental or intentional overdose without 
adverse effect.”
●
Any new cancer or progression of existing cancer.
 
 
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Events NOT Meeting the AE Definition 
●
Medical or surgical procedure (eg, endoscopy, appendectomy): the condition that 
leads to the procedure is the AE.
●
Situations in which an untoward medical occurrence did not occur (social and/or 
convenience admission to a hospital).
●
Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen.
●
Surgery planned prior to informed consent to treat a pre-existing condition that has 
not worsened.
●
Refer to section 9.3.5 for protocol specific exceptions
Definition of SAE
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met 
A SAE is defined as any untoward medical occurrence that, at any dose:
a.
Results in death
b. Is life-threatening
●
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
participant was at risk of death at the time of the event. It does not refer to an event, 
which hypothetically might have caused death, if it were more severe.
c.
Requires inpatient hospitalization or prolongation of existing hospitalization
●
Hospitalization is defined as an inpatient admission, regardless of length of stay, even 
if the hospitalization is a precautionary measure for continued observation.  (Note: 
Hospitalization for an elective procedure to treat a pre-existing condition that has not 
worsened is not a serious adverse event.  A pre-existing condition is a clinical condition 
that is diagnosed prior to the use of a Merck product and is documented in the patient’s 
medical history.
d. Results in persistent or significant disability/incapacity
●
The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.
●
This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, and 
accidental trauma (eg, sprained ankle) which may interfere with or prevent everyday life 
functions but do not constitute a substantial disruption.
 
 
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e.
Is a congenital anomaly/birth defect
●
in offspring of participant taking the product regardless of time to diagnosis
f.
Other important medical events:
●
Medical or scientific judgment should be exercised in deciding whether SAE 
reporting is appropriate in other situations such as important medical events that may not 
be immediately life-threatening or result in death or hospitalization but may jeopardize 
the participant or may require medical or surgical intervention to prevent one of the other 
outcomes listed in the above definition. These events should usually be considered 
serious.
Examples of such events include invasive or malignant cancers, intensive treatment 
in an emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse.
Additional Events Reported
Additional Events which require reporting 
In addition to the above criteria, adverse events meeting either of the below criteria, 
although not serious per ICH definition, are reportable to the Sponsor.

Is a cancer;

Is associated with an overdose
Recording AE and SAE
AE and SAE Recording
●
When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (eg, hospital progress notes, laboratory, and diagnostics reports) 
related to the event.
●
The investigator will record all relevant AE/SAE information on the Adverse Event 
case report forms/worksheets at each examination.
●
It is not acceptable for the investigator to send photocopies of the participant’s 
medical records to the Sponsor in lieu of completion of the AE CRF page.
●
There may be instances when copies of medical records for certain cases are 
requested by the Sponsor. In this case, all participant identifiers, with the exception of 
the participant number, will be blinded on the copies of the medical records before 
submission to the Sponsor.
●
The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information. In such cases, the diagnosis (not the 
individual signs/symptoms) will be documented as the AE/SAE.
 
 
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Assessment of Intensity
●
An event is defined as ‘serious’ when it meets at least 1 of the predefined outcomes as 
described in the definition of an SAE, NOT when it is rated as severe.
●
The investigator will make an assessment of intensity for each AE and SAE (and 
other reportable safety event) reported during the study and assign it to 1 of the 
following categories: 
●
Mild: An event that is easily tolerated by the participant, causing minimal 
discomfort and not interfering with everyday activities.  (for pediatric trials, 
awareness of symptoms, but easily tolerated)
●
Moderate: An event that causes sufficiently discomfort and interferes with normal 
everyday activities. (for pediatric trials, definitely acting like something is wrong)
●
Severe: An event that prevents normal everyday activities. An AE that is assessed 
as severe should not be confused with an SAE. Severe is a category utilized for 
rating the intensity of an event; and both AE and SAE can be assessed as severe 
(for pediatric trials, extremely distressed or unable to do usual activities).

Injection site erythema/redness or induration/swelling from the day of vaccination 
through Day 5 postvaccination will be evaluated by maximum size.

The investigator will assess toxicity for each AE and SAE (and other reportable 
event) reported during the study. A toxicity grade will be assigned to injection-
site AEs, specific systemic AEs, other systemic AEs, and vital sign (temperature) 
as shown in the following tables. The toxicity grading scales used in this study are 
adapted from the “FDA Guidance for Industry: Toxicity Grading Scale for 
Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical 
Trials, September 2007.”
Injection Site Reactions Intensity/Toxicity Grading Scale
Injection Site 
Reaction to 
V160/Placeboa
Grade 1
Grade 2
Grade 3
Grade 4
Injection-site AEs occurring Days 1 through 5 following receipt of V160/placebo
Pain/Tenderness
Does not 
interfere 
with activity
Repeated use of non-
narcotic pain reliever 
>24 hours or interferes 
with activity
Any use of narcotic 
pain reliever or 
prevents daily 
activity
ER visit or 
hospitalization
Erythema/Redness
Size 
measured as
B
Size measured as C or 
D
Size measured as
E→
Necrosis or 
exfoliative 
dermatitis or 
results in ER 
visit or 
hospitalization
Induration/Swelling
Size 
measured as 
B
Size measured as C or 
D
Size measured as
E→
Necrosis or ER 
visit or 
hospitalization
 
 
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Other 
Does not 
interfere 
with activity
Repeated use of non-
narcotic pain reliever 
>24 hours or interferes 
with activity
Any use of narcotic 
pain reliever or 
prevents daily 
activity
ER visit or 
hospitalization
Any injection-site reaction that begins ≥6 days after receipt of V160/placebo
Pain/tenderness
Erythema/Redness
Induration/Swelling
Other
Does not 
interfere 
with 
activity
Repeated use of non-
narcotic pain reliever 
>24 hours or interferes 
with activity
Any use of narcotic 
pain reliever or 
prevents daily 
activity
ER visit or 
hospitalization
Abbreviations: AE = adverse event; ER = emergency room; eVRC = electronic Vaccine Report Card; SAE 
= serious adverse event
a Based upon information provided by the patient on the eVRC and verbally during VRC review.  
Erythema/Redness/Induration and Swelling are specific injection-site AEs with size designations of letters 
A through E→, based upon a graphic in the eVRC. Size A is not assigned a toxicity grade; however, 
injection-site AEs that measure size A should be reported as adverse experiences. If the participant has an 
ER visit or is hospitalized for any injection-site AE, that AE is to be assigned a toxicity grade of 4, 
regardless of the size measured.
Specific Systemic AE Intensity/Toxicity Grading Scale
Systemic 
(General)
Mild 
(Grade 1)
Moderate 
(Grade 2)
Severe 
(Grade 3)
Potentially Life 
Threatening
(Grade 4)
Headache
No interference 
with activity
Repeated use of 
non-narcotic pain 
reliever >24 hours 
or some 
interference with 
activity
Significant; any 
use of narcotic 
pain reliever or 
prevents daily 
activity
ER visit or 
hospitalization
Fatigue
No interference 
with activity
Some interference 
with activity
Significant; 
prevents daily 
activity
ER visit or 
hospitalization
Myalgia
No interference 
with activity
Some interference 
with activity
Significant; 
prevents daily 
activity
ER visit or 
hospitalization
Abbreviations: ER = emergency room
Other Systemic AEs Intensity/Toxicity Grading Scale
Systemic Illnessa
Mild 
(Grade 1)
Moderate 
(Grade 2)
Severe 
(Grade 3)
Potentially Life 
Threatening
(Grade 4)b
Illness or clinical 
AE (as defined 
according to 
applicable 
regulations)
No interference 
with activity
Some interference 
with activity not 
requiring medical 
intervention
Prevents daily 
activity and 
required medical 
intervention
ER visit or 
hospitalization
Abbreviations: ER = emergency room; eVRC = electronic Vaccine Report Card; SAE = serious adverse event
a Based upon information provided by the patient on the eVRC and verbally during the eVRC review during the 
primary safety follow-up period. For SAEs reported beyond the primary safety follow-up period, grading will be based 
upon the initial report and/or follow-up of the event.
b AEs resulting in death will be assessed as Grade 4
 
 
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Vital Sign (Temperature) Toxicity Grading Scale
Vital Signsa
Mild 
(Grade 1)
Moderate 
(Grade 2)
Severe 
(Grade 3)
Potentially Life 
Threatening
(Grade 4)
Fever 
(°C) b
              (°F) b
38.0° to 38.4°
100.4° to 101.1°
38.5° to 38.9°
101.2° to 102.0°
39.0° to 40.0°
102.1° to 104.0°
>40.0°
>104.0°
a Participant should be at rest for all vital sign requirements
b Oral temperature; no recent hot or cold beverages or smoking
Assessment of Causality
●
Did the Sponsor's product cause the adverse event?
●
The determination of the likelihood that the Sponsor's product caused the adverse 
event will be provided by an investigator who is a qualified physician.  The 
investigator’s signed/dated initials on the source document or worksheet that 
supports the causality noted on the AE form, ensures that a medically qualified 
assessment of causality was done.  This initialed document must be retained for 
the required regulatory time frame.  The criteria below are intended as reference 
guidelines to assist the investigator in assessing the likelihood of a relationship 
between the test product and the adverse event based upon the available 
information
●
The following components are to be used to assess the relationship between 
the Sponsor's product and the AE; the greater the correlation with the 
components and their respective elements (in number and/or intensity), the more 
likely the Sponsor's product caused the adverse event:
●
Exposure:  Is there evidence that the participant was actually exposed to the 
Sponsor's product such as:  reliable history, acceptable compliance assessment 
(diary, etc.), seroconversion or identification of vaccine virus in bodily 
specimen?
●
Time Course:  Did the AE follow in a reasonable temporal sequence from 
administration of the Sponsor's product? Is the time of onset of the AE 
compatible with a vaccine-induced effect?
●
Likely Cause:  Is the AE not reasonably explained by another etiology such 
as underlying disease, other drug(s)/vaccine(s), or other host or environmental 
factors
●
Rechallenge: Was the participant re-exposed to the Sponsor's product in this 
trial?
●
If yes, did the AE recur or worsen?
●
If yes, this is a positive rechallenge. 
●
If no, this is a negative rechallenge.
(Note:  This criterion is not applicable if:  (1) the initial AE resulted in death 
or permanent disability, or (2) the trial is a single-dose vaccine trial); or (3) 
Sponsor's product(s) is/are used only one time.)
 
 
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NOTE:  IF A RECHALLENGE IS PLANNED FOR AN ADVERSE EVENT WHICH 
WAS SERIOUS AND WHICH MAY HAVE BEEN CAUSED BY THE SPONSOR'S 
PRODUCT, OR IF RE-EXPOSURE TO THE SPONSOR'S PRODUCT POSES 
ADDITIONAL POTENTIAL SIGNIFICANT RISK TO THE PARTICIPANT THEN 
THE RECHALLENGE MUST BE APPROVED IN ADVANCE BY THE SPONSOR 
CLINICAL DIRECTOR, AND IF REQUIRED, THE INSTITUTIONAL REVIEW 
BOARD/INDEPENDENT ETHICS COMMITTEE.  
●
Consistency with Study treatment Profile:
Is the clinical/pathological 
presentation of the AE consistent with previous knowledge regarding the 
Sponsor's product or drug class pharmacology or toxicology?
●
The assessment of relationship will be reported on the case report forms /worksheets 
by an investigator who is a qualified physician according to his/her best clinical 
judgment, including consideration of the above elements.
●
Use the following scale of criteria as guidance (not all criteria must be present to be 
indicative of a Sponsor's product relationship).
●
Yes, there is a reasonable possibility of Sponsor's product relationship:
There is evidence of exposure to the Sponsor's product.  The temporal sequence 
of the AE onset relative to the administration of the Sponsor's product is 
reasonable.  The AE is more likely explained by the Sponsor's product than by 
another cause. 
●
No, there is not a reasonable possibility of Sponsor's product relationship:
Participant did not receive the Sponsor's product OR temporal sequence of the AE 
onset relative to administration of the Sponsor's product is not reasonable OR the 
AE is more likely explained by another cause than the Sponsor’s product.  (Also 
entered for a participant with overdose without an associated AE.)
●
For each AE/SAE, the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.
●
There may be situations in which an SAE has occurred and the investigator has 
minimal information to include in the initial report to the Sponsor. However, it is very 
important that the investigator always make an assessment of causality for every event 
before the initial transmission of the SAE data to the Sponsor.
●
The investigator may change his/her opinion of causality in light of follow-up 
information and send an SAE follow-up report with the updated causality assessment.
●
The causality assessment is one of the criteria used when determining regulatory 
reporting requirements
 
 
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Follow-up of AE and SAE
●
The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as medically indicated or as requested by Sponsor 
to elucidate the nature and/or causality of the AE or SAE as fully as possible. This 
may include additional laboratory tests or investigations, histopathological 
examinations, or consultation with other health care professionals.
●
New or updated information will be recorded in the CRF.
●
The investigator will submit any updated SAE data to the Sponsor within 24 hours of 
receipt of the information.
Reporting of AE, SAE, and Other Reportable Safety Events to the Sponsor 
AE, SAE, and Other Reportable Safety Event Reporting to Sponsor via Electronic 
Data Collection Tool
●
The primary mechanism for reporting to the Sponsor will be the electronic data 
collection (EDC) tool.  
●
Electronic reporting procedures can be found in the EDC data entry guidelines (or 
equivalent).
●
If the electronic system is unavailable for more than 24 hours, then the site will 
use the paper AE Reporting form.  
●
Reference section 9.3.1 – Time Period and Frequency for Collecting AE and 
SAE and Other Reportable Safety Event Information for reporting time 
requirements 
●
The site will enter the SAE data into the electronic system as soon as it becomes 
available.
●
After the study is completed at a given site, the electronic data collection tool will be 
taken off-line to prevent the entry of new data or changes to existing data.
●
If a site receives a report of a new SAE from a study participant or receives updated 
data on a previously reported SAE after the electronic data collection tool has been 
taken off-line, then the site can report this information on a paper SAE form or by 
telephone (see next section).
●
Contacts for SAE reporting can be found in the Investigator Trial File Binder (or 
equivalent).
 
 
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SAE Reporting to the Sponsor via Paper CRF
●
If the electronic data collection tool is not operational, facsimile transmission or 
secure e-mail of the SAE paper CRF is the preferred method to transmit this 
information to the Sponsor.
●
In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable with a copy of the SAE data collection tool sent by overnight 
mail or courier service.
●
Initial notification via telephone does not replace the need for the investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames.
●
Contacts and instructions for SAE reporting and paper reporting procedures can be 
found in the Investigator Trial File Binder (or equivalent).
 
 
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12.5 Appendix 5: Laboratory Assays
Safety laboratory assessments (ie, blood chemistry, hematology and urinalysis) are not 
planned for this study. 
A serum pregnancy test will be performed at screening as indicated in the SoA (protocol-
specific requirements for inclusion or exclusion of participants are detailed in Sections 6.1
and 6.2) and a urine pregnancy test will be performed prior to each vaccine dose and one 
month after the last dose. Additional tests may be performed at any time during the study as 
determined necessary by the investigator or required by local regulations. Investigators must 
document their review of any laboratory safety report.
Any laboratory results that could unblind the study will not be reported to investigative sites 
or other blinded personnel (until the study has been unblinded).
Efficacy and Immunogenicity Laboratory Assays
CMV PCR Assays
The purpose of the CMV polymerase chain reaction (PCR) assay is to detect CMV both non-
vaccine type and vaccine-type strain deoxyribonucleic acid DNA in specimens obtained from 
participants. 
The specificity of the PCR assay has been evaluated against a panel of viral and non-viral 
targets to confirm 1) specific amplification of CMV and 2) absence of cross-reactivity to the 
other microorganisms tested. PCR sensitivity for urine and saliva detection will be 
determined prior to initiation of sample testing, with a target goal to achieve sensitivity of at 
least 200 IU/mL for both sample types. This will help assess whether V160 is able to 
establish latency, shed, and potentially serve as a source of transmission to close contacts 
(unexpected).
This is the primary assay for the detection of vaccine-type viral shedding in clinical samples. 
The assay is planned to be performed by an outside vendor on behalf of Merck Research 
Laboratories.
The assay uses virus-specific primers and probes to detect and discriminate among wild type 
CMV and those of the vaccine strain-specific construct. 
The CMV PCR assay system is designed as a two-part assay to 1) detect and quantify low 
levels of CMV DNA and 2) discriminate Merck vaccine virus from non-vaccine type viral 
strains. 
Viral DNA from urine and saliva clinical samples will be extracted using a commercially 
available DNA purification kit and initially PCR tested using UL54 and UL55 as the viral 
DNA target in a quantitative PCR (qPCR) assay to determine the presence and quantity of 
any CMV.
Samples identified as positive for CMV DNA using the initial assay will then be tested using 
a multiplex CMV real time qPCR (RT-PCR) assay to detect 3 specific targets (US28/29 
junction region; loxP; and UL51-FKBP fusion) for the discrimination of vaccine-derived 
viral DNA from wild type viral DNA. In the vaccine discrimination assay, the US28/29 
junction region is found in both vaccine and wild type strains. The loxP sequence is a 
bacterial recombination sequence and is only found in bacterial artificial chromosome 
 
 
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(BAC)- derived CMV strains (ie, the vaccine strain). The UL51-FKBP target is a genetically 
engineered gene fusion specific to the Merck vaccine strain.
For both components of the CMV PCR assay system, analysis of the raw fluorescence data is 
generated and captured using RT-PCR instrumentation. 
For CMV qPCR, the number of DNA copies in each sample is quantified compared to a 
standard curve which is calibrated to an international standard.
For RT-PCR the qualitative assessment of the presence of wild type vs vaccine strain CMV 
will be performed.
In addition to the CMV PCRs, DNA extracted from the clinical samples will be used to test 
for a human housekeeping gene, beta-globin, to confirm the integrity of sample collection.  
Beta-globin testing will stop after 6 months, once samples are demonstrated to be 
consistently positive. If self-collected samples are not consistently positive or there is 
concern about declining quality/quantity of DNA isolated, beta-globin testing may be 
reinstituted.
Viral Neutralization Antibody (NAb) Assay 
The purpose of the viral NAb assay is to quantify neutralizing antibody titers to CMV before 
and after vaccination with CMV vaccine. Functional antibodies will be measured by viral 
neutralization assay to assess the ability of vaccine-induced immune sera to inhibit viral 
infection of cells.
This is one of the primary assays used by the Merck Research Laboratories to evaluate the 
serological response to the vaccine and is currently being validated by an outside vendor to 
be performed on behalf of Merck Research Laboratories.
A CMV neutralization assay was developed using a report CMV with green-fluorescent 
protein (GFP) to provide consistent results and flexibility for the screening of large numbers 
of clinical samples, as needed. In this assay, a virus with a GFP gene inserted into its genome 
is used to infect ARPE-19cells. GFP levels are measured as an indication of viral infection. 
Neutralizing antibodies present in test serum prevent the entry of CMV into target cells, 
prevent expression of the reporter GFP protein and, thus, reduce the signal intensity. 
In the viral NAb assay, serum samples are serially diluted and mixed with an epithelial cell 
tropic HCMV before being added into ARPE-19 cells. Cells are fixed after 48-hour 
incubation with the serum/virus mixture and then subsequently scanned using an EiSight 
imager. Data are presented as number of GFP-positive cells per well. The neutralizing 
activity in the test sera is presented as NT50, which is interpolated dilution corresponding to 
the 50% of the maximum (median of the no serum control wells) and the minimum (median 
of the no virus control wells).
Preliminary results using clinical samples from V160-001 showed comparable results to the 
In-Cell Western neutralization assay used in Phase 1 trial, where the IE-1 protein was used as 
a read-out by immune-staining on the Li-Cor reader.
IgG ELISA Assay
Viral neutralization assay measures the functional antibodies blocking viral entry and is 
considered the primary assay for immunogenicity. However, there are certain degrees of 
 
 
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technical challenges to implement the neutralization assay to support a large Phase 3 trial 
which may enroll over 24,000 subjects. Thus, a viral antigen ELISA measuring surrogate 
antibody responses is desired.  
An ELISA assay measuring the serological responses to purified V160 virus was developed 
by Merck Research Laboratories and is planned to be validated by an outside vendor to be 
performed on behalf of Merck Research Laboratories. In this assay, purified V160 virus is 
coated on microtiter plates. Clinical samples are incubated on the plate and antibodies against 
HCMV virus will be captured. An HRP-labeled anti-human IgG antibody is used to detect 
IgG antibodies that are bound to the virus. 
Data generated during Phase 2 determine if an ELISA assay is suitable to replace viral 
neutralization assay in Phase 3.
CMV Cell-Mediated Immunity Assays, CMV IFN-γ ELISPOTs
The purpose of the CMV enzyme-linked immunospot (ELISPOT) assay is to detect 
interferon gamma (IFN-γ) secreting HCMV-specific cells from peripheral blood 
mononuclear cells (PBMCs) before and after vaccination with V160. This assay is planned to 
be performed by an outside vendor on behalf of Merck Research Laboratories to evaluate the 
cellular immune response to the vaccine. Results for the assay are expressed as the frequency 
of spot forming cells (SFCs) per million PBMCs. 
The IFN-γ ELISPOT assay utilizes 2 high-affinity IFN-γ specific monoclonal antibodies that 
are directed against different epitopes on the IFN-γ molecule. In the CMV-specific IFN-γ 
ELISPOT assay, PBMCs are stimulated by the particular CMV antigen in wells of cell 
culture plates that have been pre-coated with one mouse monoclonal antibody to human 
IFN-γ. Three different antigens are used to stimulate CMV-specific IFN-γ responses: 
overlapping 15-mer peptide pools of pp65 and IE1 CMV proteins, and purified CMV virion 
stocks. 
Mitogen and mock-stimulated assay controls will also be performed for each subject. IFN-γ 
released by the CMV-specific T-cells then binds to the first antibody present in close 
proximity to the cells that produced it. After ≈18 hours in culture, the cells are washed away, 
a biotinylated form of the second antibody to IFN-γ is added to the wells of the plate and the 
plate is incubated overnight at 4ºC. The assay plates are washed, and then alkaline 
phosphatase-streptavidin is added to each well of the plate. After 2 hours of incubation at 
room temperature, the plates are washed again and a chromogenic substrate (NBT/BCIP) is 
added to react with the alkaline phosphatase. As a result, dark blue spots develop against the 
white background of the plates. The IFN-γ produced by each cell results in the formation of a 
spot on the culture plate and the number of spots approximates the number of cells that 
produced IFN-γ in response to CMV antigen. The frequency of SFCs is usually expressed per 
million input PBMCs. This reflects the T-cell precursor frequency specific to CMV 
circulating in the blood at a defined time point.
 
 
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12.6 Appendix 6: Abbreviations and Trademarks
AE
Adverse event/experience
APaT
All participants as treated
CAC
Clinical Adjudication Committee
CI
Confidence intervals
cCMVd
Congenital cytomegalovirus disease
cCMVi
Congenital cytomegalovirus infection 
CMVi
Cytomegalovirus infection
CRF
Case report form
CSR
Clinical Study Report 
DMC
Data Monitoring Committee
DNA 
Deoxyribonucleic acid
ECI
Event of clinical interest
eCRF
Electronic case report form
eDMC
External Data Monitoring Committee
EMA
European Medicines Agency
eVRC
Electronic vaccine report card 
ELISA
Enzyme-linked immunosorbent assay
ELISPOT
Enzyme-linked immunosorbent spot
EOC
Executive Oversight Committee
ER
Emergency room
ERC
Ethics Committee
FAS
Full Analysis Set 
FDAAA 
Food and Drug Administration Amendments Act
FK-506
Tacrolimus
GCP
Good Clinical Practice
GFP
Green-fluorescent protein
GMc
Geometric mean count 
GMT
Geometric mean titer
-hCG
Beta-human chorionic gonadotropin
HEENT 
Head, eyes, ears, nose, and throat  
HIV
Human immunodeficiency virus
 
 
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HHV-5
Human herpesvirus 5
HRT
Hormone replacement therapy
IB
Investigators Brochure   
ICF
Informed Consent Form 
ICH
International Conference on Harmonization
ICW
In-cell Western
IEC
Independent Ethics Committee
IFN-γ
Interferon gamma
IgG
Immunoglobin G
IM
Intramuscular
IRT
Interactive response technology
IRB
Institutional Review Boards
LLN
Lower limits of normal
MAPA
Merck aluminum phosphate adjuvant
NAb
Neutralizing antibody
PPE
Per-protocol efficacy
PRO
Patient reported outcome 
PBMCs
Peripheral blood mononuclear cell
PCR
Polymerase chain reaction  
RNA
Ribonucleic acid
SAC
Scientific Advisory Committee
SAE
Serious adverse event
SFC
Spot forming cells
SAP
Statistical Analysis Plan 
sSAP
Supplemental Statistical Analysis Plan
TC
Telephone contact
US
United States
V160
Merck’s candidate CMV vaccine
VE
Vaccine Efficacy
WOCBP
Women of childbearing potential
 
 
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PAGE 1
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Supplemental Statistical Analysis Plan (sSAP)
Double-Blind, Randomized, Placebo-Controlled Phase 2b, Multi-center Study to Evaluate the 
Safety, Tolerability, Efficacy and Immunogenicity of a 2-Dose and a 3-Dose Regimen of 
V160 (Cytomegalovirus [CMV] Vaccine) in Healthy Seronegative Women, 16 to 35 Years 
of Age
(Version 2.0)
Prepared by
.
CONFIDENTIAL, PROPRIETARY PROPERTY OF 
MERCK & CO., INC.
 
 
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PAGE 2
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Table of Contents
Listing of Tables ....................................................................................................................... 4
Listing of Figures...................................................................................................................... 5
1.
Introduction ....................................................................................................................... 6
2.
Summary of Changes......................................................................................................... 6
2.1
Version 1.0 ................................................................................................................. 6
2.2
Version 2.0 ................................................................................................................. 6
3.
Analytical and Methodological Details ............................................................................. 7
3.1
Statistical Analysis Plan Summary ............................................................................ 7
3.2
Responsibility for Analyses/In-House Blinding ........................................................ 9
3.3
Hypotheses/Estimation............................................................................................. 11
3.4
Analysis Endpoints................................................................................................... 11
Efficacy Endpoints............................................................................................ 11
Immunogenicity Endpoints............................................................................... 11
Safety Endpoints............................................................................................... 11
3.5
Analysis Populations................................................................................................ 12
Efficacy Analysis Populations.......................................................................... 12
Immunogenicity Analysis Populations ............................................................. 13
Safety Analysis Population............................................................................... 14
3.6
Statistical Methods................................................................................................... 14
Statistical Methods for Efficacy Analyses........................................................ 14
Statistical Methods for Immunogenicity Analyses........................................... 15
Statistical Methods for Safety Analyses ........................................................... 16
3.7
Interim Analyses ...................................................................................................... 17
Efficacy Interim Analysis ................................................................................. 17
Safety Interim Analysis..................................................................................... 18
3.8
Multiplicity............................................................................................................... 19
3.9
Sample Size and Power Calculations....................................................................... 19
Sample Size and Power for Efficacy Analyses................................................. 19
3.9.1.1
Sample Size and Power for Testing the Primary Efficacy Hypothesis......... 19
3.9.1.2
Sample Size and Power for Testing the Secondary Efficacy Hypothesis..... 24
Sample Size and Power for Safety Analyses .................................................... 26
 
 
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3.10
Subgroup Analyses and Effect of Baseline Factors ................................................. 27
3.11
Compliance............................................................................................................... 27
Adherence to Scheduled Vaccinations.............................................................. 27
Adherence to Scheduled Self-Collection of Urine and Saliva Samples ........... 27
3.12
Extent of Exposure................................................................................................... 28
References............................................................................................................................... 29
 
 
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Listing of Tables
Table 1
Analysis Strategy for Safety Parameters ..............................................................17
Table 2
Decision Rule and Corresponding Type 1 Error and Power for Testing 
the Primary Efficacy Hypothesis of VE >0% Against CMVi (3-Dose 
Regimen vs. Placebo) ...........................................................................................21
Table 3
Decision Rule and Corresponding Type 1 Error and Power for Testing 
the Primary Efficacy Hypothesis of VE >0% Against CMVi,
Conditional on the IA Being Conducted at 17 Total Cases (3-Dose 
Regimen vs. Placebo) ...........................................................................................23
Table 4
Decision Rule and Corresponding Type 1 Error and Power for Testing 
the Secondary Efficacy Hypothesis of VE >0% Against CMVi (2-Dose 
Regimen vs. Placebo) ...........................................................................................25
Table 5
Critical Counts of Participants with Adverse Events in V160 and 
Placebo Arms Resulting in the Lower Bound of 95% CI Estimate of 
Risk Difference to be >0% ...................................................................................26
 
 
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Listing of Figures
Figure 1
Plan for Testing the Primary and Secondary Efficacy Hypotheses......................18
 
 
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1. Introduction
This document outlines the statistical analysis strategy and procedures for the V160-002 study. 
If, after the study has begun, but prior to any unblinding/final database lock, changes made to 
primary and/or key secondary hypotheses, or the statistical methods related to those 
hypotheses, then the protocol will be amended (consistent with ICH Guideline E-9). Changes 
to exploratory or other non-confirmatory analyses made after the protocol has been finalized, 
but prior to unblinding/final database lock, will be documented in this Supplementary 
Statistical Analysis Plan (sSAP) and referenced in the Clinical Study Report (CSR) for the 
study. Post hoc exploratory analyses will be clearly identified in the CSR.
2. Summary of Changes
2.1
Version 1.0
1. Table 2 in this sSAP, which corresponds to Table 6 in the V160-002 protocol, is 
expanded to show the decision rules that will be used when the interim analysis (IA) is 
conducted with more than 15 total CMVi cases in the V160 3-dose and placebo groups 
combined. Such an eventuality (i.e., over-running of the planned 15 primary efficacy 
endpoint cases needed for the IA) may happen if more than 15 cases are available in 
the study database when counting of cases is performed.

A counting of total cases of CMVi in the combined V160 3-dose and placebo 
groups was conducted on 22-November-2019. A total of 6 CMVi cases were 
obtained.

This total number of cases is indicative of an incidence rate that is higher than the 
anticipated CMVi incidence rate in the placebo group. As such, there is a potential 
to over-run at the next case counting the 15 CMVi cases needed for conducting the 
IA.

Table 2 in this sSAP is being expanded at this time in order to pre-specify the 
decision rules, prior to conducting the second counting of total cases of CMVi in 
the combined V160 3-dose and placebo groups, that will be implemented in an 
eventuality that the 15 CMVi cases needed for testing the primary efficacy 
hypothesis at the IA is over-run.
2. Section 3.5.1 enumerates the categories of events, deviations from protocol, or 
activities that are deemed by the study Clinical Director to potentially interfere with the 
evaluation of efficacy or immune response to injections of V160, and hence cause a 
study participant to be excluded from the per-protocol efficacy population.
2.2
Version 2.0
1. Section 3.4.2 no longer has the seroconversion and seropositivity immunogenicity 
endpoints. These endpoints cannot be defined because both the CMV-specific NAb 
assay and the IgG ELISA currently do not have identified seropositivity cutoffs that 
can be used to identify participants who are seropositive based on either of the two 
assays.
 
 
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2. Section 3.5.2 provides the definition of the per-protocol immunogenicity (PPI) analysis 
population.
3. Section 3.6.2 provides the method that will be used in calculating geometric mean titers 
(GMTs) and the corresponding 95% confidence interval (CI) based on the CMV-
specific neutralizing antibody (NAb) assay and IgG enzyme-linked immunosorbent 
assay (ELISA).
4. Section 3.9.1.1 has a new subsection (Final Testing of the Primary Efficacy 
Hypothesis) along with a new Table 3 that provides the decision rules that will be 
applied in the event that a final testing of the primary efficacy hypothesis needs to be 
conducted at total primary efficacy endpoint case count that is greater than 26 in the 
combined V160 3-dose and placebo groups (conditional on the IA being conducted at 
17 total cases in the combined V160 3-dose and placebo groups).
5. Table 4 in Section 3.9.1.2 has been expanded to cover situations when the secondary 
efficacy hypothesis is tested at 25 or more (through 35 cases) total cases in the V160 
2-dose and placebo groups.
3. Analytical and Methodological Details 
3.1
Statistical Analysis Plan Summary
Key elements of the statistical analysis plan are summarized below; the comprehensive plan 
is provided in Sections 3.2 to 3.12 of this sSAP. For the purposes of analysis and reporting, 
the overall study ends when all study efficacy, immunogenicity, and safety data are available 
and ready for database lock execution. 
Study Design Overview
Double-Blind, Randomized, Placebo-Controlled Phase 2b,
Multi-center Study to Evaluate the Safety, Tolerability, 
Efficacy and Immunogenicity of a 2-Dose and a 3-Dose 
Regimen of V160 (Cytomegalovirus [CMV] Vaccine) in 
Healthy Seronegative Women, 16 to 35 Years of Age
Treatment Assignment
Participants will be randomized in a 1:1:1 allocation to one of 
the following 3 treatment arms:

V160 3-dose regimen: will receive 3 doses of V160 
administered IM according to a Day 1, Month 2, Month 6 
schedule;

V160 2-dose regimen: will receive 2 doses of V160 
administered IM according to a Day 1 (V160), Month 2 
(placebo), Month 6 (V160) schedule;

Placebo regimen: will receive 3 doses of saline solution 
placebo administered IM according to a Day 1, Month 2, 
Month 6 schedule.
No stratification based on age, sex or other characteristics will 
be used in this trial.
 
 
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Analysis Populations
Efficacy: Per-Protocol Efficacy (PPE)
Safety: All Participants as Treated (APaT)
Primary Endpoint(s)
1. Efficacy: number of participants with CMVi in the V160 3-
dose regimen and placebo arms.
2. Safety: number of participants experiencing solicited 
injection site AEs within Days 1 through 5 after each 
vaccination visit; solicited systemic AEs and vaccine-
related serious AEs within Days 1 through 14 after each 
vaccination visit.
Key Secondary 
Endpoints
Efficacy: number of participants with CMVi in the V160 2-
dose regimen and placebo arms.
Statistical Methods for 
Key Efficacy Analyses
The primary and secondary efficacy hypotheses will be 
evaluated by calculating the efficacy of V160 compared to 
placebo with respect to the CMVi endpoint. The p-value for 
testing the hypothesis that VE>0% as well as estimation of the 
95% CI of VE will be based on the exact binomial method 
proposed by Chan and Bohidar [1].
Statistical Methods for 
Key Safety Analyses
There are no a priori clinical events identified in this trial as 
Tier 1 events. Tier 2 events identified in this trial include 
solicited injection site and systemic AEs; vaccine-related AEs 
(systemic or serious); any serious AEs (in trial participants or 
infants born to trial participants); discontinuations due to AEs, 
AEs by system organ class observed in 7 participants in at 
least one vaccination group and elevated temperatures.
Estimates of 95% confidence intervals for between-treatment 
differences in the percentage of participants with events will 
be calculated using the Miettinen and Nurminen method [2].
Interim Analyses
One IA will be performed in this trial relating to the testing of 
the primary efficacy hypothesis. Results will be reviewed by 
an eDMC. A summary of the IA is provided below. Further 
details are provided in Section 3.7. 
IA:

Timing: To be conducted upon accumulation of 15 CMVi cases 
in the combined V160 3-dose regimen and placebo arms, 
expected at ≈18 months from trial start.

Testing: The primary efficacy hypothesis will be tested.
Final analysis

Timing: If the result of testing the primary efficacy hypothesis 
during the IA was inconclusive, a final analysis will be 
conducted upon accumulation of 24 CMVi cases in the 
 
 
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combined V160 3-dose regimen and placebo arms, expected at 
≈3 years from trial start.
Multiplicity
The primary efficacy hypothesis will be tested once at an IA, 
and if necessary, a second time at a final analysis. The 
secondary efficacy hypothesis will be tested in one final 
analysis, without an IA, only if the primary efficacy 
hypothesis is successfully demonstrated. The overall one-
sided trial Type 1 error associated with testing the primary and 
secondary efficacy hypotheses is 2.5% based on a 
gatekeeping procedure described in Section 3.8.
Sample Size and Power 
The power of the decision rules that will be implemented in 
this trial for testing of the primary efficacy hypothesis is based 
on a fixed-event design.

Corresponding to the decision rules shown in Figure 1 in 
Section 3.7, the trial has >90% power to demonstrate that 
the efficacy of a V160 3-dose regimen compared to 
placebo to prevent CMVi is >0% if the underlying vaccine 
efficacy is 80% at an overall one-sided Type 1 error equal 
to 2.3% (see Table 2).
The trial will randomize 2,100 participants equally allocated 
into one of the 3 treatment arms as described in the 
Treatment Assignment panel of this section. Under study 
design assumptions described in Section 3.9.1, the IA of the 
primary efficacy hypothesis is expected to be conducted at 
≈18 months from trial start; and the final analysis if needed is 
expected to be conducted at ≈3 years from trial start, as 
mentioned in the Interim Analyses panel of this section.
The power of the decision rules that will be implemented in 
this trial for testing of the secondary efficacy hypothesis is 
also based on a fixed-event design. 

Corresponding to the decision rules shown in Figure 1 in 
Section 3.7, if the secondary efficacy hypothesis is tested 
based on a total of 15 observed CMVi cases in the 
combined V160 2-dose and placebo regimens, then the trial 
has power >85% at an overall one-sided Type 1 error 
≤2.5% to demonstrate that the efficacy of a V160 2-dose 
regimen compared to placebo to prevent CMVi is >0% if 
the underlying vaccine efficacy is 85% (see Table 3).
3.2
Responsibility for Analyses/In-House Blinding
This trial will be conducted as a double-blind trial under in-house blinding procedures. 
 
 
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The statistical analysis of the data obtained from this trial will be the responsibility of the 
Clinical Biostatistics department of the Sponsor.
The Clinical Biostatistics department will generate the randomized allocation schedules for 
assignment of study treatment and selection of immunogenicity sub-study participants as 
described in Section 7.3 of the V160-002 protocol. Randomization will be implemented 
using IRT.
An external statistician and scientific programmer will be identified and will comprise the 
unblinded team. This team will be unblinded on a participant-level. They will not be involved 
in any discussions regarding modifications to the protocol, statistical methods, identification 
of protocol deviations, or data validation efforts at any time during the trial.
This trial has one planned IA, and its key aspects are described in Section 3.7 of this sSAP. 
The eDMC will serve as the primary reviewer of the results of the IA. Treatment-level results 
of the IA will be provided by the external unblinded statistician to the eDMC. The eDMC 
will make recommendations relating to trial discontinuation or modification to an Executive 
Oversight Committee (EOC) (see Appendix 1) of the Sponsor. If the eDMC recommends 
modifications to the design of the protocol or discontinuation of the trial, the EOC may be 
unblinded to results at the treatment level in order to act on these recommendations. If 
necessary, limited additional Sponsor personnel may be unblinded to the treatment level 
results of the IA in order to act on the recommendations of the eDMC. The extent to which 
individuals are unblinded with respect to results of IA will be documented by the external 
unblinded statistician. Additional logistical details will be provided in the eDMC Charter.
Upon reaching the milestone when definitive disposition of the primary and secondary 
efficacy hypotheses has been reached:

A database lock will be executed.

All Sponsor personnel who are members of the study team and the V160 program team 
will be unblinded and a clinical study report will be written.

If this milestone is reached at or beyond study Year 3, or if the outcome in this milestone 
is failure of the test of the primary efficacy hypothesis, then the study investigators and 
participants will be unblinded upon completion of the clinical study report.
Definitive disposition of the primary and secondary hypotheses is reached when one of the 
following study outcomes occur:
1. Test of the primary efficacy hypothesis fails.
2. Tests of both the primary and secondary efficacy hypotheses succeed.
3.
Test of the primary efficacy hypothesis succeeds but the test of the secondary efficacy 
hypothesis fails.
If at least the primary efficacy hypothesis is successfully demonstrated prior to reaching 
study Year 3, the study may continue to follow participants up to study Year 3 in order to 
continue to accumulate more information relating to vaccine efficacy against CMVi and 
cCMVi. Under this scenario, study investigators and participants will remain blinded during 
the continuing follow-up for CMVi and cCMVi information. Upon reaching the third year 
of the study:
 
 
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
A database lock will be executed.

An end-of-study clinical report will be written.

The study investigators and participants will be unblinded upon completion of the end-
of-study clinical report.
For the purpose of external reporting of study results in clinical trial registries (eg, EudraCT 
and Clinicaltrials.gov), the end-of-study milestone corresponds to the last data generated 
date, which is the date when all study efficacy, immunogenicity, and safety data are available 
and ready for database lock execution.
3.3
Hypotheses/Estimation
Objectives and hypotheses of the study are stated in Section 4 of the V160-002 protocol.
3.4
Analysis Endpoints
The efficacy, immunogenicity, and safety endpoints that will be evaluated are listed below.
Efficacy Endpoints
The efficacy endpoint that will be used to evaluate VE and test the primary and secondary 
efficacy hypotheses is the number of participants with primary CMVi, described in Section 4 
and Section 5.4.1 of the V160-002 protocol.
CMVi is defined as detection of CMV DNA by PCR in urine or saliva samples.
As stated in Section 4 of the V160-002 protocol, in the evaluation of the primary and 
secondary efficacy hypotheses, cases of CMVi will be counted starting after the last dose of 
the assigned vaccination regimen. As stated in Section 3.5.1, the primary and secondary 
efficacy hypothesis will be evaluated on the PPE population, where eligibility includes being 
PCR negative post Day 1 through Month 7. Thus, CMVi cases that will be included in the 
PPE testing of the primary and secondary efficacy hypothesis are those that occurred after 
Month 7.
Immunogenicity Endpoints
All immunogenicity endpoints in this study are exploratory endpoints and are listed below. 

GMTs based on the NAb and the IgG ELISA assays at Day 1, Months 7, 12, 24, and 
36.

GMc of SFC/106 PBMCs based on the ELISPOT assay Day 1, Months 6, 7, 12, and 
24. 
Safety Endpoints
As specified in Sections 4 and 5.4.1.3 of the V160-002 protocol and Section 3.6.3 of this sSAP, 
the endpoints that will be used to assess the safety and tolerability of 2-dose and 3-dose 
regimens of V160 include the following:
 
 
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
Number of participants experiencing solicited injection site AEs within Days 1 through 5 
after each vaccination visit; solicited systemic AEs (including tiredness, muscle pain, 
headache, and joint pain); and vaccine-related systemic AEs, SAEs, and elevated
temperatures within Days 1 through 14 after each vaccination visit 

Incidence of any SAE at any time during the study

Incidence of vaccine-type viral shedding

Outcomes of pregnancies.
3.5
Analysis Populations
Efficacy Analysis Populations
Per Protocol Efficacy (PPE)
The PPE population will serve as the primary population for the evaluation of VE and for 
testing the primary and secondary efficacy hypotheses. To be eligible for inclusion in the 
PPE population, randomized study participants must satisfy the following criteria:

CMV seronegative at Day 1 and CMV negative by PCR for non-vaccine strain virus 
from post Day 1 through Month 7

Have received all 3 injections/vaccinations within the vaccination visit window 
specified in the SoA (Section 2)

At any time from Day 1 through Month 7, did not have events, deviations from 
protocol, or engaged in activities that are deemed by the study Clinical Director to 
potentially interfere with the evaluation of efficacy or immune response to injections 
of V160.
Participants whose CMV status by PCR post Day 1 through Month 7 is not evaluable will be 
ineligible for inclusion in the PPE population. The categories of events, deviations from 
protocol, or activities that are deemed by the study Clinical Director to potentially interfere 
with the evaluation of efficacy or immune response to injections of V160 include the 
following:
1. Participant was not serologically confirmed as CMV seronegative (based on CMV IgG) 
during screening;
2. Participant was immunocompromised or had been diagnosed as having a congenital or 
acquired immunodeficiency, human immunodeficiency virus (HIV) infection, 
lymphoma, leukemia, systemic lupus erythematosus, rheumatoid arthritis, juvenile 
rheumatoid arthritis, inflammatory bowel disease, or other autoimmune condition at 
any time between Day 1 to Month 7;
3. Participant had previously received a non-study CMV vaccine.
4. Participant received systemic corticosteroids (equivalent ≥2 mg/kg total daily dose of 
prednisone or ≥20 mg/d for persons weighing >10 kg) for ≥14 consecutive days and 
has not completed treatment at least 30 days prior to any vaccination dose;
 
 
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5. Participant received systemic corticosteroids exceeding physiologic replacement doses 
(approximately 5 mg/d prednisone equivalent) within 14 days prior to any vaccination 
dose;
6. Participant received any anti-viral agent with proven or potential activity against CMV 
(e.g. letermovir, ganciclovir, valganciclovir, foscarnet, cidofovir, valacyclovir) two 
weeks prior to any vaccination dose or likely to receive such an agent within two weeks 
after any vaccination dose.
7. Participant was receiving or has received in the year prior to randomization
immunosuppressive therapies including but not limited to rapamycin (also sirolimus), 
tacrolimus (also FK-506 or Fujimycin), or other therapies used for solid organ/cell 
transplant, 
radiation 
therapy, 
immunosuppressive/cytotoxic 
immunotherapy, 
chemotherapy and other immunosuppressive therapies known to interfere with the 
immune response. 
8. Participant used topical tacrolimus within 2 weeks before or after any vaccination dose;
9. Participant received study clinical material in a sequence that is different from the 
sequence corresponding to the vaccination group to which the participant was 
randomized into (i.e., incorrect vaccination sequence or cross-treatment).
10. Participant was administered improperly stored study clinical material that was deemed 
unacceptable for use.
11. Participant received improperly reconstituted study clinical material that may affect 
efficacy and/or safety.
Full Analysis Set (FAS)
The FAS will serve as a supportive analysis population for the evaluation of VE. The FAS 
population consists of all randomized participants who received at least 1 dose of the correct 
clinical material corresponding to the treatment regimen the participants were randomized into.
Immunogenicity Analysis Populations
Per Protocol Immunogenicity (PPI)
The PPI population will serve as the primary population for the evaluation of 
immunogenicity. To be eligible for inclusion in the PPI population, randomized study 
participants must satisfy the following criteria:

Eligible to be included in the PPE population as defined in Section 3.5.1;

Have provided serum sample for NAb and IgG ELISA evaluation at 1 month post last 
dose (Month 7) within the visit window specified in the SoA (i.e., 21 to 49 days after 
dose 3 of V160 or placebo).
 
 
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Safety Analysis Population
All Participants as Treated (APaT)
The APaT population will serve as the primary population for evaluation of safety. The APaT 
population includes all randomized participants who received at least 1 injection of V160 or 
placebo, have safety follow-up data, and assigns participants to the treatment arm 
corresponding to the actual clinical material received. For participants who received injections 
of V160 or placebo corresponding to a sequence that does not correspond to any of the 
protocol-defined treatment arms (ie, cross-treated participants), a safety profile listing will be 
created separate from the safety reports that will be provided for the protocol-defined treatment 
arms.
3.6
Statistical Methods
Statistical testing and inference relating to efficacy are described in Section 3.6.1. Statistical 
methods relating to safety analyses are described in Section 3.6.3. Efficacy results that will be 
deemed to be statistically significant after consideration of the Type 1 error control strategy 
are described in Section 3.8. Nominal p-values may be computed for other efficacy analyses, 
but should be interpreted with caution due to potential issues of multiplicity, sample size, etc. 
Unless otherwise stated, all statistical tests will be conducted at the α=0.025 (1-sided) level.
Statistical Methods for Efficacy Analyses
To address the primary and secondary efficacy hypotheses, one-sided tests of the null 
hypothesis that the vaccine efficacy is 0% versus the alternative that vaccine efficacy is >0% 
will be conducted separately for each of the primary and secondary hypotheses. Each of the 
primary and secondary efficacy hypotheses will be tested at one-sided α=0.025 level (ie, 2.5% 
Type 1 error). Vaccine efficacy is defined as: 
where Rv and Rp are the incidence rates CMVi in the V160 dose regimen (3-dose for the 
primary efficacy hypothesis testing; 2-dose for the secondary efficacy hypothesis testing) and 
placebo groups, respectively. The incidence rate Rv is defined as Rv = Cv/Tv, where Cv = the 
count of CMVi cases in the vaccine group and Tv = total person-years of follow-up for efficacy 
in the vaccine group. The incidence rate Rp is defined similarly. The null hypothesis that 
vaccine is not efficacious (ie, VE=0%) will be tested by constructing a 100*(1α)% confidence 
interval (CI) for VE, denoted as (VEL, VEU), based on study data at a particular analysis time 
point (eg, at IA or at final analysis). The statistical criterion for success with respect to the 
primary efficacy hypothesis (and likewise for the secondary efficacy hypothesis) will be met 
if VEL >0%. 
The 100*(1α) % CI for VE, (VEL, VEU), is constructed as follows. Under the assumption that 
Rv and Rp are the means of independent Poisson processes, and given that there is a total of n 
= Cv + Cp efficacy endpoint cases observed in the combined V160 dose regimen and placebo 
groups, then the number of primary efficacy cases Cv in the V160 dose regimen group is 
distributed as Binomial(n,), where the binomial probability is defined as = 
TvRv/(TvRv+TpRp). The probability is a person-years-adjusted estimate of the probability 
VE = 100%*{1 (Rv/Rp)}
 
 
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that a particular study participant who became a case of CMVi belongs to the V160 dose 
regimen group. The lower bound of the 100*(1α) % exact CI for the probability is obtained 
by searching for the proportion L such that the probability of observing Cv or more efficacy 
endpoint cases out of n total efficacy cases is ≤α/2. Similarly, the upper bound of the 
100*(1α)% exact CI for the probability is obtained by searching for the proportion U such 
that the probability of observing Cv or fewer efficacy endpoint cases out of n total efficacy 
cases is ≤α/2 [1]. VEL and VE are then calculated from L and U as follows:
where = Tp/Tv is the ratio of the total person-years of follow-up for efficacy in the placebo 
group over the V160 dose regimen group.
There is only one final analysis of the secondary efficacy endpoint. However, the testing of the 
secondary efficacy hypothesis may occur at the time of the IA or the final analysis of the 
primary efficacy hypothesis. The secondary efficacy hypothesis will only be tested if the 
primary efficacy hypothesis is successfully demonstrated. In addition, the secondary efficacy 
hypothesis will be tested contingent on accumulation of 15 total CMVi cases in the V160 2-
dose and placebo regimens (see Figure 1 in Section 3.7, and Section 3.9.2). Thus, the possible 
timing for testing the secondary hypothesis is as follows.

If the primary hypothesis test is successful at the IA and there are 15 total CMVi cases 
in the V160 2-dose and placebo regimens at the time of IA, then the secondary efficacy 
hypothesis will be tested immediately after the testing of the primary efficacy 
hypothesis at the time of the IA.

If the primary hypothesis test is successful at the IA and there are <15 total CMVi cases 
in the V160 2-dose and placebo regimens at the time of IA, the secondary efficacy 
hypothesis will not be tested at the IA. In this case, follow-up will continue until at least 
15 total CMVi cases in the V160 2-dose and placebo regimens groups are accrued. 
Upon accrual of 15 total CMVi cases in the V160 2-dose and placebo regimens, the 
secondary efficacy hypothesis will be tested. 

If the primary hypothesis test is not successful at the IA, the secondary efficacy 
hypothesis will not be tested at the time of the IA.
Statistical Methods for Immunogenicity Analyses
Within a vaccination group, GMT and the corresponding 95% CI will be derived by taking 
the anti-logarithm of the mean of the log-transformed antibody titers and its corresponding 
95% CI (calculated based on the t-distribution). Antibody titers reported as less than the
lower limit of quantitation (LLOQ) of the relevant assay will be replaced by the half of the 
LLOQ in the log-transformation of antibody titers.
Geometric mean concentration (GMc) of ELISPOT measures and the corresponding 95% CI 
will be calculated similar to the calculations of GMT and its corresponding 95% CI.
VEL = 100%*{1- U (1+)}/(1 - U);
VEU = 100%*{1- L (1+)}/(1 - L);
 
 
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Statistical Methods for Safety Analyses
Safety and tolerability will be assessed by clinical review of all relevant parameters including 
AEs and outcomes of pregnancies.
The analysis of safety results will follow a tiered approach (Table 1). The tiers differ with 
respect to the analyses that will be performed. Safety parameters or adverse events of special 
interest that are identified a priori constitute “Tier 1” safety endpoints that will be subject to 
inferential testing for statistical significance with p-values and 95% confidence intervals 
provided for between-group comparisons. Other safety parameters will be considered Tier 2 
or Tier 3. Tier 2 parameters will be assessed via point estimates with 95% confidence intervals 
provided for V160 dose regimen versus placebo comparisons; only point estimates by 
treatment group are provided for Tier 3 safety parameters. Safety results will be summarized 
for each vaccination dose and across all three vaccination doses.
There were no SAEs nor vaccine-type viral shedding (which is an event of interest for V160) 
observed in the V160 Phase I study. While vaccine-type viral shedding is an event of interest 
that can occur only in V160-vaccinated subjects, barring false-positive tests in placebo 
recipients, the 0% incidence of vaccine-type viral shedding observed in the Phase I study 
provides no informed, data-driven, non-zero incidence level around which a safety hypothesis 
can be formulated and tested. Therefore, there is no a priori identified AE or ECI for which a 
test of hypothesis will be conducted. Thus, no Tier 1 event is identified in this study. Categories 
of events considered as Tier 2 and Tier 3 in this study are shown in 
Table 1. In addition, AEs observed in at least 7 participants (representing 1% of N=700 
participants per group) in any vaccination group will be categorized as a Tier 2 event.
Incidences of the categories of events identified in Table 1 will be reported as percentages, 
generally calculated as 100% x (number of participants who experienced the event/total 
number of participants with safety follow-up), except for outcomes of pregnancies, SAEs 
throughout the study duration, and vaccine-type viral shedding. The 95% CIs will be provided 
for vaccine versus placebo percentage point differences (percent in V160 dose regimen group 
minus percent in placebo) will be calculated using the Miettinen and Nurminen method [2], an 
unconditional, asymptotic method. No stratification factor will be used in the calculation of 
95% CI.
Percentages of outcomes of pregnancies will be generally calculated as 100% x (total number 
of specific pregnancy outcome being reported/total number of all relevant pregnancy 
outcomes). For example, the percent associated with the pregnancy outcome of live births will 
be calculated as 100% x (total number of live births/total number of fetuses/infants with known
outcomes). The numerator and denominator account for pregnancies with multiple fetuses (ie, 
twins, triplets, etc.) and do not treat such occurrences as a single pregnancy outcome or event. 
Further details will be provided in the sSAP that will be written for this study.
Vaccine-type viral shedding will be assessed in V160-vaccinated participants. In addition to 
the analysis strategy shown in Table 1, point and corresponding 95% CI estimates of incidence 
of vaccine-type viral shedding (number of participants observed with vaccine-type viral 
shedding/total person-years of follow-up) will be calculated separately for the V160 3-dose, 
V160 2-dose, and placebo arms. No treatment group comparisons (V160 vs. placebo or V160 
 
 
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Protocol No. 02
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MAY.7, 2019 Amendment 02
3-dose vs. 2-dose arms) of the percent of participants detected with vaccine-type viral shedding 
will be conducted. 
Summaries of SAEs (throughout the study duration) by system organ class will be provided as 
incidence rates (number of participants [or infants] observed with a specific SAE/total person-
years of follow-up) rather than percentages.
Table 1
Analysis Strategy for Safety Parameters
Safety 
Tier
Safety Endpoint
p-value
95% CI for 
Comparison 
of V160 to 
Placebo
Descriptive 
Statistics
Tier 2
Solicited injection site AEsa
X
X
Solicited systemic AEs
X
X
Vaccine-related systemic AEs
X
X
Any serious AEs (in study participants or infants born 
to study participants)
X
Xb
Vaccine-related serious AEs within Days 1 through 
14 after vaccination
X
X
Discontinuations due to AEs
X
X
Elevated temperaturec
X
X
AEs by SOC observed in 7 participants in at least 
one vaccination group
X
X
Tier 3
AEs by SOC
X
Outcomes of pregnancies
X
Vaccine-type viral shedding
X‡
a All injection site AEs are considered vaccine-related.
b Incidence rates (cases/total person-years follow-up) will be provided instead of percentages.
c Defined as ≥38.0ºC [≥100.4°] oral or equivalent
AE = Adverse event; CI = Confidence interval; SOC = System organ class; X = indicated summary statistic 
will be provided.
3.7
Interim Analyses
Efficacy Interim Analysis
An IA for testing the primary efficacy hypothesis will be conducted upon accrual of a total of 
15 CMVi endpoint cases in the combined V160 3-dose and placebo regimens. Possible 
outcomes of this IA and consequent study pathways are as described below and shown in 
schematic form in Figure 1.
1. 7 of 15 CMVi cases observed in the V160 3-dose arm

Study fails to demonstrate the primary efficacy hypothesis at the IA and has no chance 
of successfully demonstrating the primary efficacy hypothesis in a final analysis when 
a total of 24 CMVi cases are accrued in the combined V160 3-dose regimen and 
placebo groups. 

eDMC may recommend the study to be terminated for futility.
2. (4, 5, or 6) of 15 CMVi cases observed in the V160 3-dose arm.
 
 
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Protocol No. 02
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MAY.7, 2019 Amendment 02

Result of testing of the primary efficacy hypothesis is inconclusive.

Study fails to demonstrate the primary efficacy hypothesis at the IA but still has a 
chance of demonstrating the primary efficacy hypothesis in a final analysis when a 
total of 24 CMVi cases are accrued in the combined V160 3-dose regimen and placebo 
groups. 

Study (which is still blinded) will continue until a total of 24 CMVi cases are accrued 
in the combined V160 3-dose regimen and placebo groups, at which time, the final 
testing of the primary efficacy hypothesis will be conducted.
3. ≤3 of 15 CMVi cases observed in the V160 3-dose arm

Study successfully demonstrates the primary efficacy hypothesis.

The appropriate timing for the testing the secondary efficacy hypothesis is evaluated 
(see Section 10.6.1).
Figure 1
Plan for Testing the Primary and Secondary Efficacy Hypotheses
Safety Interim Analysis
At the time of IA for efficacy, the eDMC will also conduct a review of the cumulative study 
safety data recorded in the study database upon which the 15 CMVi cases was counted and 
confirmed to have been accumulated. No hypothesis relating to safety will be tested. 
 
 
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Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Summaries of safety parameters identified in Section 10.6.3 Statistical Methods for Safety 
Analyses, will be prepared for review by the eDMC.
3.8
Multiplicity
The overall 1-sided Type 1 error in testing the efficacy hypotheses is controlled to not exceed 
2.5% based on gatekeeping procedure. 
The primary efficacy hypothesis will be tested at an overall (over the IA and final analysis) 1-
sided Type 1 error <2.5%, as discussed in Section 3.9.1. The secondary efficacy hypothesis 
will be tested at 1-sided Type 1 error 2.5% only if the primary efficacy hypothesis is 
successfully demonstrated, as discussed in Section 3.9.2. The primary efficacy hypothesis is a 
reasonable gatekeeper to the secondary efficacy hypothesis because if a V160 3-dose regimen 
with last dose at Month 6 is not efficacious, then a V160 2-dose regimen with last dose also at 
Month 6 is highly unlikely to be efficacious.
3.9
Sample Size and Power Calculations
Sample Size and Power for Efficacy Analyses
3.9.1.1
Sample Size and Power for Testing the Primary Efficacy Hypothesis
The power corresponding to the decision rule that will be implemented in testing the primary 
efficacy hypothesis is based on a fixed-event design. For testing the primary efficacy 
hypothesis, participants will be followed until 15 total CMVi cases in the combined V160 3-
dose and placebo regimens have been accumulated, at which point, an IA will be conducted. 
If the testing of the primary efficacy hypothesis is inconclusive at the IA, follow-up will 
continue until 24 total CMVi cases have been observed in the combined V160 3-dose and 
placebo regimens. The decision rules to be implemented that will determine success or failure 
to demonstrate the primary efficacy hypothesis are as shown in Figure 1 in Section 3.7 and in 
Table 2. Corresponding to these decision rules, the study has >90% power to demonstrate that 
the efficacy of a V160 3-dose regimen compared to placebo to prevent CMVi is >0% if the 
underlying vaccine efficacy is 80% at an overall one-sided Type 1 error equal to 2.3%. The 
operating characteristics of the decision rules with one IA and a final analysis if necessary are 
evaluated by a 5 million-replication simulation using SAS v9.4. In the simulation conducted, 
the underlying probability distribution of accumulation of CMVi cases in the V160 3-dose 
regimen conditional on the total cases in the combined V160 and placebo regimens is based 
on an exact method under a large sample Poisson distribution assumption proposed by Chan 
and Bohidar (1998) [1].
This study will randomize 700 participants into the V160 3-dose regimen and 700 into the 
placebo regimen. The 15 total CMVi cases in the combined V160 3-dose and placebo regimens 
are projected to be accumulated in ≈18 to 19 months under the following assumptions: 1) all 
1,400 participants are enrolled within 18 months; 2) incidence of CMVi in the placebo group 
is 2% per year; 2) VE=85%, so that the incidence of CMVi in the V160 arm is 0.3% per year; 
3) loss-to-follow-up is ≈5% to 10% per year; and 4) participants ineligible for the PPE analysis 
is ≈2% to 5% of participants randomized. Under the same assumptions, the total of 24 CMVi 
cases in the combined V160 3-dose and placebo regimens is projected to be accumulated in ≈3 
years.
 
 
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Protocol No. 02
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It is possible that at the time when CMVi cases are counted to determine whether a total of 15 
cases have been accrued, the actual CMVi cases accumulated will already be greater than 15. 
In such an eventuality, the IA will be conducted based on the actual number of cases accrued 
(eg, 16, or 17, or some other count). Table 2 also shows the operating characteristics of decision 
rules that will be used in the eventuality that the IA is conducted at a total case count greater 
than 15.
 
 
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Protocol No. 02
Supplemental SAP 2.0
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Table 2
Decision Rule and Corresponding Type 1 Error and Power for Testing the Primary Efficacy Hypothesis of VE >0% 
Against CMVi (3-Dose Regimen vs. Placebo)
Total 
Cases 
Available 
at IA
Analysis
Time
Point
Total 
Cases to 
Accrue
Stop for 
Futility
(Vacc 
Cases)
Case Split
for Success
(Vacc:Pbo)
VE (95% CI)a
Cumulative
Type I 
Errorb
True Vaccine Efficacy
70%
75%
80%
85%
90%
95%
Cumulative Power
15
IA
15
7
3 : 12
75.0 (7.4, 95.5)
1.8%
53.2%
64.8%
76.8%
87.8%
95.9%
99.5%
Final†
24
7
6 : 18
66.7 (12.3, 89.2)
2.3%
73.0%
83.7%
92.3%
97.5%
99.6%
100%
16
IA
16
8
3 : 13
76.9 (16.0, 95.8)
1.1%
47.7%
59.9%
72.9%
85.4%
94.9%
99.4%
Final†
26
8
7 : 19
63.2 (8.5, 86.9)
2.1%
78.0%
87.8%
94.9%
98.7%
99.9%
100%
17
IA
17
8
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
26
8
7 : 19
63.2 (8.5, 86.9)
1.7%
77.3%
87.4%
94.7%
98.6%
99.8%
100%
18
IA
18
8
4 : 14
71.4 (9.0, 93.2)
1.6%
59.6%
71.6%
83.2%
92.5%
98.1%
99.9%
Final†
26
8
7 : 19
63.2 (8.5, 86.9)
2.3%
78.8%
88.3%
95.1%
98.7%
99.9%
100%
19
IA
19
8
4 : 15
73.3 (16.3, 93.6)
1.0%
54.5%
67.3%
80.1%
90.9%
97.5%
99.8%
Final†
26
8
7 : 19
63.2 (8.5, 86.9)
1.9%
77.7%
87.7%
94.8%
98.6%
99.9%
100%
20
IA
20
8
5 : 15
66.7 (3.5, 90.5)
2.1%
69.4%
80.4%
89.8%
96.3%
99.3%
100%
Final†
27
8
7 : 20
65.0 (13.8, 87.5)
2.4%
78.1%
87.9%
94.9%
98.6%
99.9%
100%
21
IA
21
9
5 : 16
68.8 (10.7, 91.0)
1.3%
64.8%
76.9%
87.6%
95.3%
99.1%
100%
Final†
28
9
8 : 20
60.0 (5.2, 84.8)
2.3%
83.5%
91.7%
97.0%
99.3%
99.9%
100%
22
IA
22
9
5 : 17
70.6 (16.9, 91.5)
0.9%
60.2%
73.3%
85.3%
94.2%
98.8%
100%
Final†
29
9
8 : 21
61.9 (10.5, 85.4)
1.6%
80.4%
89.9%
96.2%
99.1%
99.9%
100%
a Upon completion of the analyses, a multiplicity-adjusted p-value and confidence interval will be calculated using the group-sequential methodology of Jennison and Turnbull 
(2000). 
b Type I error and power shown are cumulative over the IA and final analysis.
CI = Confidence interval; IA = Interim analysis; VE = Vaccine efficacy.
 
 
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Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Final Testing of the Primary Efficacy Hypothesis
The eDMC conducted the IA on March 17, 2020 based on a total of 17 primary endpoint 
cases in the combined V160 3-dose and placebo groups. After the IA was conducted, the 
eDMC recommended the study to continue unchanged. This eDMC recommendation 
implies:
1. The study did not meet the futility criterion at the IA.
2. The testing of the efficacy hypotheses (or hypothesis if only the secondary efficacy 
hypothesis needs to be tested at a future time) is paused.
3. It is unknown to the V160-002 blinded study team whether:
i. The test of the primary efficacy hypothesis succeeded but there are not 
enough primary efficacy endpoint cases available in the combined V160 
2-dose and placebo groups to test the secondary efficacy hypothesis (see 
Figure 1); OR
ii. The test of the primary efficacy hypothesis is inconclusive so that more 
cases of the primary efficacy endpoint in the combined V160 3-dose and 
placebo groups need to be accumulated (see Figure 1).
If the pause in efficacy hypotheses testing is due to 3.ii above, because the IA was conducted 
at 17 total cases of the primary endpoint in the combined V160 3-dose and placebo groups,
then the final testing of the primary efficacy hypothesis will be conducted when at least 26 
total cases of the primary endpoint have been accrued (see Table 2). In the event of over 
running the required 26 total cases of the primary endpoint in the combined V160 3-dose and 
placebo groups when the available efficacy endpoint cases are counted, Table 3 provides the 
decision rules that will be applied at the final testing of the primary efficacy hypothesis 
(conditional on the IA being conducted at 17 total cases in the combined V160 3-dose and 
placebo groups).
 
 
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PAGE 23
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Table 3
Decision Rule and Corresponding Type 1 Error and Power for Testing the Primary Efficacy Hypothesis of VE >0% 
Against CMVi, Conditional on the IA Being Conducted at 17 Total Cases (3-Dose Regimen vs. Placebo)
Cases 
Available 
at Final 
Analysis
Analysis
Time
Point
Total 
Cases to 
Accrue
Stop for 
Futility
(Vacc 
Cases)
Case Split
for Success
(Vacc:Pbo)
VE (95% CI)a
Cumulative
Type I 
Errorb
True Vaccine Efficacy
70%
75%
80%
85%
90%
95%
Cumulative Power
26
IA
17
8c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
26
8
7 : 19
63.2 (8.5, 86.9)
1.7%
77.3%
87.4%
94.7%
98.6%
99.8%
100%
27
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
27
8
7 : 20
65.0 (13.8, 87.5)
1.3%
74.2%
85.3%
93.7%
98.3%
99.8%
100%
28
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.5%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
28
9
8 : 20
60.0 (5.2, 84.8)
2.1%
82.7%
91.3%
96.8%
99.3%
99.9%
100%
29
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
29
9
8 : 21
61.9 (10.5, 85.4)
1.6%
80.1%
89.7%
96.1%
99.1%
99.9%
100%
30
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
30
10
9 : 21
57.1 (2.4, 82.7)
2.4%
87.1%
94.1%
98.1%
99.7%
100%
100%
31
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
31
10
9 : 22
59.1 (7.6, 83.4)
1.8%
84.8%
92.9%
97.6%
99.6%
100%
100%
32
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
32
10
9 : 23
60.9 (12.2, 84.1)
1.4%
82.5%
91.5%
97.1%
99.4%
100%
100%
33
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
33
11
10 : 23
56.5 (5.0, 81.5)
2.1%
88.6%
95.1%
98.6%
99.8%
100%
100%
34
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
34
11
10 : 24
58.3 (9.6, 82.2)
1.6%
86.6%
94.1%
98.2%
99.7%
100%
100%
35
IA
17
― c
3 : 14
78.6 (23.2, 96.1)
0.6%
42.4%
54.9%
68.9%
82.8%
93.8%
99.3%
Final†
35
12
11 : 24
54.2 (2.8, 79.7)
2.4%
91.4%
96.7%
99.2%
99.9%
100%
100%
a Upon completion of the analyses, a multiplicity-adjusted p-value and confidence interval will be calculated using the group-sequential methodology of Jennison and Turnbull 
(2000). 
b Type I error and power shown are cumulative over the IA and final analysis.
c The IA was conducted at 17 total cases based on the plan that the final analysis, if needed, would be conducted at 26 total cases. For this plan, the futility criterion at the IA was 
occurrence of 8 cases in the V160 3-dose group. When the IA was conducted on March 17 2020, the futility criterion was not met. If the final analysis needs to be conducted 
and is conducted at a total of 27 or more cases, the futility criterion at the IA is no longer needed nor relevant since the IA has already been conducted. 
CI = Confidence interval; IA = Interim analysis; VE = Vaccine efficacy.
 
 
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Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
3.9.1.2
Sample Size and Power for Testing the Secondary Efficacy Hypothesis
The power corresponding to the decision rule that will be implemented in testing the secondary 
efficacy hypothesis is based on a fixed-event design. The secondary efficacy hypothesis will 
be tested when a total of 15 CMVi cases in the combined V160 2-dose and placebo arms have 
been observed (see Figure 1 in Section 3.7). When the secondary efficacy hypothesis is tested 
based on a total of 15 CMVi cases, the study has >85% power to demonstrate that the efficacy 
of a V160 2-dose regimen compared to placebo to prevent CMVi is >0% if the underlying 
vaccine efficacy is 85% at an overall one-sided Type 1 error 2.5% (see Table 4). The 
calculations are based on an exact method under a large sample Poisson distribution 
assumption proposed by Chan and Bohidar (1998) [1] and carried out using the R software.
This study will randomize 700 participants into the V160 2-dose regimen and 700 into the 
placebo regimen. Under the same assumptions governing the accumulation of CMVi cases in 
the V160 3-dose and placebo arms, a total of 15 CMVi cases in the combined V160 2-dose 
and placebo arms are projected to be accumulated in ≈18 to 19 months; and a total of 24 in ≈3 
years. 
It is possible that at the time when CMVi cases are counted to determine whether a total of 15 
cases have been accrued in the combined V160 2-dose and placebo arms, the actual CMVi 
cases accumulated will already be greater than 15. In such an eventuality, the testing of the 
secondary efficacy hypothesis will be conducted based on the actual number of cases accrued 
(eg, 16, or 17, or some other count). Table 4 also shows the operating characteristics of decision 
rules that will be used in the eventuality that the testing of the secondary efficacy hypothesis 
is conducted at a total case count greater than 15.
 
 
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PAGE 25
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Table 4
Decision Rule and Corresponding Type 1 Error and Power for Testing the Secondary Efficacy Hypothesis of VE >0% 
Against CMVi (2-Dose Regimen vs. Placebo)
Analysis
Time
Point
Total 
Cases 
Accrued
Case Split
for Success
(Vacc:Pbo)
VE (95% CI)
Type I 
Error
True Vaccine Efficacy
70%
75%
80%
85%
90%
95%
Power
Final
15
3 : 12
75.0 (7.4, 95.5)
1.8%
53.2%
64.8%
76.8%
87.8%
95.9%
99.5%
16
3 : 13
76.9 (16.0, 95.8)
1.1%
47.7%
59.8%
72.9%
85.4%
94.9%
99.4%
17
4 : 13
79.2 (0.4, 92.7)
2.5%
64.7%
75.8%
86.0%
93.9%
98.5%
99.9%
18
4 : 14
71.4 (9.0, 93.2)
1.5%
59.6%
71.6%
83.2%
92.5%
98.1%
99.9%
19
4 : 15
73.3 (16.3, 93.6)
1.0%
54.5%
67.3%
80.1%
90.9%
97.6%
99.8%
20
5 : 15
66.7 (3.5, 90.5)
2.1%
69.3%
80.4%
89.8%
96.2%
99.3%
100%
21
5 : 16
68.8 (10.7, 91.0)
1.3%
64.8%
76.9%
87.7%
95.3%
99.1%
100%
22
5 : 17
70.6 (16.9, 91.5)
0.8%
60.2%
73.3%
85.3%
94.2%
98.8%
100%
23
6 : 17
64.7 (6.2, 88.6)
1.7%
73.2%
84.0%
92.5%
97.7%
99.7%
100%
24
6 : 18
66.7 (12.3, 89.2)
1.1%
69.1%
81.1%
90.9%
97.1%
99.6%
100%
25
7 : 18
61.1 (2.4, 86.3)
2.2%
79.8%
89.1%
95.5%
98.8%
99.9%
100%
26
7 : 19
63.2 (8.5, 86.9)
1.4%
76.4%
86.9%
94.5%
98.5%
99.8%
100%
27
7 : 20
65.0 (13.8, 87.5)
1.0%
72.8%
84.4%
93.2%
98.1%
99.8%
100%
28
8 : 20
60.0 (5.2, 84.8)
1.8%
82.2%
91.0%
96.7%
99.3%
99.9%
100%
29
8 : 21
61.9 (10.5, 85.4)
1.2%
79.1%
89.2%
95.9%
99.1%
99.9%
100%
30
9 : 21
57.1 (2.4, 82.7)
2.1%
86.7%
93.9%
98.0%
99.6%
100%
100%
31
9 : 22
59.1 (7.6, 83.4)
1.5%
84.2%
92.5%
97.5%
99.5%
100%
100%
32
9 : 23
60.9 (12.2, 84.1)
1.0%
81.5%
91.0%
96.9%
99.4%
100%
100%
33
10 : 23
56.5 (5.0, 81.5)
1.8%
88.1%
94.9%
98.5%
99.8%
100%
100%
34
10 : 24
58.3 (9.6, 82.2)
1.2%
85.9%
93.8%
98.1%
99.7%
100%
100%
35
11 : 24
54.2 (2.8, 79.7)
2.0%
91.1%
96.6%
99.1%
99.9%
100%
100%
CI = Confidence interval; VE = Vaccine efficacy.
 
 
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Product:  V160
PAGE 26
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Sample Size and Power for Safety Analyses
Testing of hypotheses relating to safety will not be conducted in this study. Thus, power and Type 
1 error considerations relating to safety analyses are not applicable in this study.
The probability of observing a specific AE in specific treatment arms of this study depends on the 
number of vaccinated participants with safety follow-up and the underlying incidence of that 
specific AE in the study population. In each treatment arm, assuming that all 700 participants 
randomized into the study will have safety follow-up, then there is a >99% chance of observing at 
least one specific AE if the incidence of that AE is at least 1 in every 150 participants; >90% 
chance if the incidence is at least 1 in every 300 participants. If no vaccine-related SAEs are 
observed among the 700 participants randomized in a specific V160 treatment arm, this study will 
provide 97.5% confidence that the underlying percentage of participants with vaccine-related SAE 
is 0.5% (1 in every 200 participants) among V160 vaccinated participants in that V160 arm. This 
is based on calculation of a 1-sided 97.5% upper confidence limit of a binomial proportion using 
the exact binomial method of Clopper and Pearson [3] and was done using PASS 2008. For a 
sample size of 700 per treatment arm, Table 5 provides the critical split of participants with AEs 
between V160 and placebo arms for which it will be possible to state with 95% confidence that 
the risk difference (defined as percent in V160 dose regimen group minus percent in placebo) 
excludes 0%. For a specific AE, given a sample size of 700 per arm, the minimum total number of 
participants with AEs for which it is possible to state with 95% confidence that the risk difference 
excludes 0% is 5. As stated in Section 3.6.3, AEs observed in at least 7 participants (representing 
1% of N=700 participants per group) in any vaccination group will be categorized as a Tier 2 
event. The 95% CIs of the percentage point differences were calculated using the Miettinen and 
Nurminen method [2].
Table 5
Critical Counts of Participants with Adverse Events in V160 and Placebo Arms 
Resulting in the Lower Bound of 95% CI Estimate of Risk Difference to be >0%
Observed Count of Participants
with Adverse Events
Risk Difference† (V160 – Placebo)
Total
V160
(N=700)
Placebo
(N=700)
Percentage Points‡
95% CI§
5
5
0
0.7%
(0.1%, 1.7%)
6
6
0
0.8%
(0.3%, 1.8%)
7
7
0
1.0%
(0.4%, 2.0%)
8
7
1
0.9%
(0.1%, 1.9%)
9
8
1
1.0%
(0.2%, 2.1%)
10
9
1
1.1%
(0.3%, 2.3%)
11
9
2
1.0%
(0.1%, 2.2%)
12
10
2
1.1%
(0.2%, 2.4%)
13
11
2
1.3%
(0.4%, 2.5%)
14
11
3
1.1%
(0.1%, 2.4%)
15
12
3
1.3%
(0.3%, 2.6%)
16
13
3
1.4%
(0.4%, 2.8%)
17
13
4
1.3%
(0.2%, 2.6%)
18
14
4
1.4%
(0.3%, 2.8%)
19
15
4
1.6%
(0.4%, 3.0%)
14
5
1.3%
(0.1%, 2.7%)
 
 
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Product:  V160
PAGE 27
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
Observed Count of Participants
with Adverse Events
Risk Difference† (V160 – Placebo)
Total
V160
(N=700)
Placebo
(N=700)
Percentage Points‡
95% CI§
20
15
5
1.4%
(0.2%, 2.9%)
† Defined as percent of participants in the V160 group with adverse events minus the corresponding percent in 
the placebo group.
‡ Calculated as 100%×(V160 – Placebo)÷700.
§ Calculated based on the Miettinen and Nurminen method.
N = Number of participants randomized.
CI = Confidence interval
3.10
Subgroup Analyses and Effect of Baseline Factors
To determine whether VE is consistent across various subgroups associated with specific baseline 
participant characteristics, the estimate of the VE (with a nominal 95% CI) for the primary efficacy 
endpoint (comparing 3-dose regimen versus placebo) and for the secondary efficacy endpoint 
(comparing 2-dose regimen versus placebo) will be estimated and plotted within each subgroup. 
Subgroup assessments of VE will be conducted for the following baseline participant 
characteristics:

Age category (≤21 vs. >21 years)

Race (white, non-white)

Region (US, Ex-US)
For each of the primary and secondary efficacy endpoints, a Forest plot will be produced, which 
provides the estimated point and 95% CI estimates of VE across the subgroups associated with 
specific baseline participant characteristics listed above. The consistency of VE across subgroups
will be assessed descriptively based on these Forest plots. 
3.11
Compliance
Adherence to Scheduled Vaccinations
Compliance to scheduled vaccinations in this study is defined as receipt of all 3 vaccinations of 
either V160 or placebo. To summarize compliance, the numbers of participants who receive each 
vaccination will be tabulated. For each of vaccination visits 2 and 3, histograms of the time (in 
days) of administration of the vaccine or placebo relative to the target vaccination visit will be 
provided.
Adherence to Scheduled Self-Collection of Urine and Saliva Samples
Accurate and reliable ascertainment of the primary and secondary efficacy endpoints (ie, incidence 
of CMVi) will depend on compliance of study participants to the self-collection of urine and saliva 
samples based on the schedule indicated in Section 2. Thus, compliance of study participants to 
the self-collection of urine and saliva samples will be assessed as follows.
 
 
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Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
On a participant-level (say the i-th participant among the N=2100 study participants), percent 
compliance (denoted by Pi) of a study participant to the self-collection of urine and/or saliva 
samples will be calculated as:
where 
ni = number of visits for which either a urine or saliva sample was received from the study 
participant;
Mi = total number of study visits throughout the study for which a urine or saliva sample is 
expected to be received from the study participant. This number will vary based on the time 
when the study participant enters the study and the time when the study ends.
For each of the V160 3-dose, V160 2-dose, and placebo arms, point and 95% CI estimates of the 
geometric mean of the participant-level percent compliance Pi will be calculated to summarize the 
vaccination group-level compliance of study participants to the self-collection of urine and saliva 
samples.
This assessment of compliance of study participants to the self-collection of urine and saliva 
samples will be conducted in both the PPE and FAS analysis populations.
3.12
Extent of Exposure
As indicated in Section 5.5 of the V160-002-02 protocol, participants who will receive an injection 
of V160 at a vaccination visit will receive a 0.5mL dose containing 100 units of V160. Thus:

Participants randomized to the 3-dose regimen of V160 are expected to be administered a 
cumulative total of 1.5 mL containing 300 units of V160 over a 6-months duration;

Participants randomized to the 2-dose regimen of V160 are expected to be administered a 
cumulative total of 1.0 mL containing 200 units of V160 over a 6-months duration;

Participants randomized to placebo are not expected to receive V160.
 
 
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PAGE 29
Protocol No. 02
Supplemental SAP 2.0
MAY.7, 2019 Amendment 02
References
[1]
Chan ISF, Bohidar NR. Exact power and sample size for 
vaccine efficacy studies. Commun Statist-Theory Meth 
1998;27(6):1305-22.
[2]
Miettinen O, Nurminen M. Comparative analysis of two 
rates. Statist Med 1985;4:213-26.
[3]
Clopper CJ, Pearson ES. The use of confidence or fiducial 
limits illustrated in the case of the binomial. Biometrika 
1934;26(4):404-13.
 
 
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