Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-Seronegative Infants 6 to 24 Months of Age
Phase: PHASE1
Conditions: Respiratory Syncytial Virus Infections
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

IMPAACT 2012 
 
 
Phase I Placebo-Controlled Study of the Infectivity, Safety and 
Immunogenicity of a Single Dose of a Recombinant Live-Attenuated 
Respiratory Syncytial Virus Vaccine, LID cp ΔM2-2, Lot RSV#009B, 
Delivered as Nose Drops to RSV-Seronegative Infants 6 to 24 Months 
of Age 
 
A Study of the International Maternal Pediatric Adolescent  
AIDS Clinical Trials Network  
 
 
 
Sponsored by: 
National Institute of Allergy and Infectious Diseases  
Eunice Kennedy Shriver  
National Institute of Child Health and Human Development  
National Institute of Mental Health 
 
 
 
 
DAIDS ES # 30073 
IND # [TBD] Held by DAIDS 
 
 
 
Protocol Chair: 
Coleen Cunningham, MD 
 
Protocol Vice Chair: 
Ruth Karron, MD 
 
NIAID Medical Officer:  
Devasena Gnanashanmugam, MD 
 
NICHD Medical Officer: 
Jack Moye, Jr., MD 
 
Clinical Trials Specialists: 
Charlotte Perlowski, MSPH 
Jennifer Libous, MS, CCRP 
 
Version 1.0 
08 August 2016 

IMPAACT 2012, Version 1.0 
Page 2 of 94 
08 August 2016 
IMPAACT 2012  
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of 
a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus 
Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-
Seronegative Infants 6 to 24 Months of Age 
 
TABLE OF CONTENTS 
 
TABLE OF CONTENTS ............................................................................................................. 2 
LIST OF TABLES ....................................................................................................................... 5 
LIST OF FIGURES .................................................................................................................... 5 
ABBREVIATIONS AND ACRONYMS ........................................................................................ 6 
PROTOCOL TEAM ROSTER .................................................................................................... 8 
SITE ROSTER ..........................................................................................................................11 
SCHEMA ..................................................................................................................................13 
1 
INTRODUCTION ...........................................................................................................16 
1.1 
Overview ......................................................................................................................................... 16 
1.2 
Background ..................................................................................................................................... 18 
1.3 
Prior Research................................................................................................................................. 21 
1.3.1 Experimental Vaccines against Respiratory Syncytial Virus ....................................................... 21 
1.3.2 Preclinical Studies ....................................................................................................................... 22 
1.3.3 Previous Clinical Experience ....................................................................................................... 24 
1.4 
Rationale ......................................................................................................................................... 25 
1.5 
Hypotheses ..................................................................................................................................... 26 
2 
OBJECTIVES ................................................................................................................27 
2.1 
Primary Objective ............................................................................................................................ 27 
2.2 
Secondary Objectives ...................................................................................................................... 27 
3 
STUDY DESIGN ...........................................................................................................28 
4 
STUDY POPULATION ..................................................................................................29 
4.1 
Inclusion Criteria .............................................................................................................................. 29 
4.2 
Exclusion Criteria............................................................................................................................. 30 
4.3 
Co-Enrollment Considerations ......................................................................................................... 32 
4.4 
Recruitment, Screening, and Enrollment Process ........................................................................... 33 
4.5 
Participant Retention ....................................................................................................................... 33 
4.6 
Participant Withdrawal or Termination from the Study .................................................................... 33 
5 
STUDY PRODUCT CONSIDERATIONS .......................................................................34 
5.1 
Study Products ................................................................................................................................ 34 
5.2 
Study Product Regimens ................................................................................................................. 34 
5.3 
Study Product Formulation .............................................................................................................. 34 
5.3.1 Vaccine ....................................................................................................................................... 34 
5.3.2 Diluent for RSV LID cp ΔM2-2 .................................................................................................... 35 
5.3.3 Placebo for RSV LID cp ΔM2-2 ................................................................................................... 35 
5.4 
Study Product Storage .................................................................................................................... 35 
5.5 
Study Product Preparation .............................................................................................................. 35 
5.5.1 Diluent for RSV LID cp ΔM2-2 .................................................................................................... 35 
5.5.2 Placebo for RSV LID cp ΔM2-2 ................................................................................................... 36 
5.5.3 Live Recombinant Respiratory Syncytial Virus (RSV) LID cp ΔM2-2 .......................................... 36 
5.6 
Study Product Administration/ Inoculation Procedure ..................................................................... 37 
5.7 
Study Product Acquisition ................................................................................................................ 37 
5.8 
Study Product Accountability ........................................................................................................... 37 
5.9 
Disposition of Used/Unused Study Product ..................................................................................... 38 
5.10 
Final Disposition of Study Products ................................................................................................. 38 

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5.11 
Concomitant Medications ................................................................................................................ 38 
5.11.1 Prohibited Concomitant Medications ........................................................................................... 38 
5.11.2 Precautionary Concomitant Medications ..................................................................................... 38 
6 
STUDY VISITS AND PROCEDURES............................................................................38 
6.1 
Screening Visit................................................................................................................................. 39 
6.2 
Enrollment Visit................................................................................................................................ 40 
6.3 
Acute Phase Visits and Contacts .................................................................................................... 42 
6.3.1 Acute Phase Visits: Study Days 3, 5, 7, 10, 12, 14, 17, and 28 (±1 day) .................................... 42 
6.3.2 Acute Phase Contacts: Study Days 1, 2, 4, 6, 8, 9, 11, 13, 15,16, 18, 19, 20, 21, 22, 23, 24, 25, 
26, and 27 (±1 day) ..................................................................................................................... 43 
6.4 
Day 29 Contact (+ 1 day) ................................................................................................................ 43 
6.5 
Post-Acute Phase (Days 30 to 56) .................................................................................................. 44 
6.5.1 Day 56 Visit (+7 Days) ................................................................................................................ 44 
6.6 
Period after Day 56 Visit until October 31st ...................................................................................... 44 
6.7 
Pre-RSV Season Study Visit (October 1st to 31st) ............................................................................ 44 
6.8 
RSV Season Surveillance (November 1st through March 31st following inoculation) ....................... 45 
6.9 
Post-RSV Season Study Visit (April 1st to 30th) ............................................................................... 46 
6.10 
Illness Visit ...................................................................................................................................... 46 
6.11 
Early Discontinuation Study Visit ..................................................................................................... 46 
6.12 
Additional Considerations for Laboratory Procedures ..................................................................... 47 
6.12.1 Specimen Collection ................................................................................................................... 47 
6.12.2 Specimen Preparation, Testing, Storage, and Shipping.............................................................. 47 
6.12.3 Biohazard Containment ............................................................................................................... 49 
7 
SAFETY ASSESSMENT, MONITORING, AND REPORTING .......................................49 
7.1 
Safety-Related Roles and Responsibilities ...................................................................................... 49 
7.1.1 Site Investigators ......................................................................................................................... 49 
7.1.2 Protocol Safety Review Team ..................................................................................................... 49 
7.1.3 Data Safety Monitoring Board ..................................................................................................... 50 
7.1.4 Sponsor Reporting ...................................................................................................................... 50 
7.2 
Safety-Related Recording on Case Report Forms .......................................................................... 50 
7.3 
Expedited Adverse Event (EAE) Reporting ..................................................................................... 52 
7.3.1 Adverse Event Reporting to DAIDS ............................................................................................ 52 
7.3.2 Reporting Requirements for this Study ....................................................................................... 52 
7.3.3 Grading Severity of Events.......................................................................................................... 52 
7.3.4 Expedited AE Reporting Period................................................................................................... 53 
8 
PARTICIPANT MANAGEMENT ....................................................................................54 
8.1 
Management of Adverse Events ..................................................................................................... 54 
8.1.1 Solicited Adverse Events ............................................................................................................ 55 
8.1.2 Serious Adverse Event ................................................................................................................ 56 
8.2 
Pausing and Stopping Rules ........................................................................................................... 56 
9 
STATISTICAL CONSIDERATIONS ...............................................................................57 
9.1 
General Design Issues .................................................................................................................... 57 
9.1.1 General Design ........................................................................................................................... 57 
9.1.2 Description of the Statistical Methods to be Employed ............................................................... 58 
9.2 
Outcome Measures ......................................................................................................................... 58 
9.2.1 Primary Outcome Measures........................................................................................................ 58 
9.2.2 Secondary Outcome Measures ................................................................................................... 58 
9.3 
Sample Size and Accrual ................................................................................................................ 58 
9.3.1 Sample Size and Randomization ................................................................................................ 58 
9.4 
Monitoring ........................................................................................................................................ 61 
9.4.1 Monitoring by the Protocol Team ................................................................................................ 62 
9.4.2 Monitoring by the NIAID Intramural Data and Safety Monitoring Board ...................................... 63 
9.5 
Analyses .......................................................................................................................................... 63 

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9.5.1 Assessment of Primary Objectives .............................................................................................. 63 
9.5.2 Assessment of Secondary Objectives ......................................................................................... 64 
10 
DATA HANDLING AND RECORD KEEPING ................................................................64 
10.1 
Data Management Responsibilities ................................................................................................. 64 
10.2 
Essential and Source Documents and Access to Source Data ....................................................... 65 
10.3 
Clinical Investigator’s Brochure ....................................................................................................... 66 
10.4 
Quality Control and Quality Assurance ............................................................................................ 66 
11 
CLINICAL SITE MONITORING .....................................................................................66 
12 
HUMAN SUBJECTS PROTECTIONS ...........................................................................66 
12.1 
Institutional Review Board/Ethics Committee Review and Approval ............................................... 66 
12.2 
Vulnerable Participants .................................................................................................................... 67 
12.3 
Informed Consent ............................................................................................................................ 67 
12.4 
Potential Benefits............................................................................................................................. 68 
12.5 
Potential Risks ................................................................................................................................. 68 
12.5.1 Venipuncture ............................................................................................................................... 68 
12.5.2 Nasal Wash ................................................................................................................................. 68 
12.5.3 Receipt of Study Product ............................................................................................................ 68 
12.6 
Reimbursement/Compensation ....................................................................................................... 68 
12.7 
Privacy and Confidentiality .............................................................................................................. 69 
12.8 
Management of Incidental Findings ................................................................................................. 69 
13 
ADMINISTRATIVE PROCEDURES ..............................................................................69 
13.1 
Regulatory Oversight ....................................................................................................................... 69 
13.2 
Protocol Registration ....................................................................................................................... 70 
13.3 
Study Implementation ...................................................................................................................... 70 
13.4 
Protocol Deviation Reporting ........................................................................................................... 70 
13.5 
ClinicalTrials.gov ............................................................................................................................. 71 
14 
PUBLICATIONS ............................................................................................................71 
15 
REFERENCES ..............................................................................................................72 
APPENDICES ...........................................................................................................................75 
Appendix I: Tables Referenced in the Background Section .......................................................75 
- 
79 
Appendix II: Schedule of Events: Screening, Acute Phase, and Post-Acute Phase ...................81 
Appendix III: Schedule of Events: RSV Pre-season Sampling, seasonal surveillance, and Post-
season Sampling ...........................................................................................................82 
Appendix IV: Definitions of Solicited Adverse Events ................................................................83 
Appendix V: RSV Seasonality in Baltimore ...............................................................................84 
Appendix VI: Sample Informed Consent Form ..........................................................................85 
 
 
 
 
 
 
 
 
 
 

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LIST OF TABLES 
 
Table 1: Overview of RSV vaccine candidates with M2-2 deletion to be studied in IMPAACT 
2011, 2012, and 2013 ................................................................................................. 17 
Table 2: AE CRF Recording Requirements ............................................................................... 51 
Table 3: Grading Table for Solicited AEs................................................................................... 53 
Table 4: Fever Grading* ............................................................................................................ 53 
Table 5: EAE Reporting ............................................................................................................ 54 
Table 6: The Probability of Observing LRI events in Vaccinees ................................................ 59 
Table 7: Percent of Participants Experiencing LRI or AEs with Exact 90% Confidence 
Intervals ...................................................................................................................... 59 
Table 8: Magnitude of Difference in Responses Detectable with 80% Power ............................ 60 
Table 9: Comparison of Genomic Sequence of LID cp M2-2, LOT RSV#009B, to wt rRSV A2 
D46 ............................................................................................................................. 75 
Table 10: Viral titers of nasopharyngeal swab samples from AGMs inoculated with LID ΔM2-2, 
LID M2-2 1030s, or LID cp M2-2a ........................................................................... 76 
Table 11: Viral titers of tracheal lavage samples from AGMs inoculated with LID ΔM2-2, LID 
M2-2 1030s, or LID cp M2-2 a ................................................................................. 77 
Table 12:  Neutralizing antibody titers of AGMs inoculated with LID ΔM2-2, LID M2-2 1030s, or 
LID cp M2-2a ............................................................................................................ 78 
Table 13:  Viral Titers of Nasopharyngeal Swab Samples from AGMs Inoculated with CTM LID 
cp M2-2, Lot RSV#009B ........................................................................................... 79 
Table 14:  Viral Titers of Tracheal Lavage samples from AGMs Inoculated with CTM LID cp 
M2-2, Lot RSV#009B ................................................................................................ 79 
Table 15:  Viral Titers of Tracheal Lavage samples from AGMs Inoculated with CTM LID cp 
M2-2, Lot RSV#009B ................................................................................................ 80 
 
 
 
LIST OF FIGURES 
Figure 1: Study Overview .......................................................................................................... 15 
Figure 2: Study Phases and Periods of Evaluation .................................................................... 29 
Figure 3: Investigational Product Label (Enlarged Sample) ....................................................... 34 
Figure 4: Power curves for comparisons between vaccine and placebo responses .................. 61 
 
 
 
 
 
 
 

IMPAACT 2012, Version 1.0 
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IMPAACT 2012  
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of 
a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus 
Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-
Seronegative Infants 6 to 24 Months of Age 
 
ABBREVIATIONS AND ACRONYMS 
 
ACIP 
Advisory Committee on Immunization Practices (CDC) 
AE 
adverse event  
AGM 
African green monkey 
AIDS 
Acquired Immunodeficiency Syndrome 
cDNA 
complementary deoxyribonucleic acid 
cGMP 
current good manufacturing practice 
CFR 
Code of Federal Regulations 
CI 
Confidence interval 
CIR 
Center for Immunization Research 
cp 
Cold Passaged  
CRADA 
Cooperative Research and Development Agreement 
CRF  
case report form 
CRL 
Charles River Laboratories 
CRPMC 
Clinical Research Products Management Center 
CSO 
Clinical Safety Office 
CTM 
clinical trial material 
DAERS 
DAIDS Adverse Experience Reporting System 
DAIDS 
Division of AIDS  
DAIDS PRO 
Division of AIDS Protocol Registration Office 
DC 
discontinuation 
DCR 
Division of Clinical Research 
DHHS 
Department of Health and Human Services 
DMC 
Data Management Center 
DMEM 
Dulbecco’s Modified Eagle Medium 
DNA 
deoxyribonucleic acid 
DSMB 
Data and Safety Monitoring Board 
EAE 
Expedited Adverse Event 
ELISA 
enzyme-linked immunosorbent assay 
F protein 
fusion protein (of RSV) 
FDA 
Food and Drug Administration 
FDAAA 
Food and Drug Administration Amendments Act of 2007 
FSTRF 
Frontier Science & Technology Research Foundation, Inc. 
GCP 
good clinical practices 
HEENT 
head, ears, eyes, nose, throat 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HJF 
Henry M. Jackson Foundation for the Advancement of Military Medicine 
HVTN 
HIV Vaccine Trials Network 
IBC 
Institutional Biosafety Committee 
ICF 
informed consent form 
ICH 
International Conference on Harmonisation 
IgA, IgG, IgE immunoglobulin A, G, E 
IMPAACT 
International Maternal Pediatric Adolescent AIDS Clinical Trials Network 
IND 
investigational new drug 
IoR 
Investigator of Record  

IMPAACT 2012, Version 1.0 
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IRB  
institutional review board 
JHSPH 
Johns Hopkins Bloomberg School of Public Health 
L-15 
Leibovitz-15 medium 
LDMS 
Laboratory Data Management System 
LID 
Laboratory of Infectious Diseases 
LPC 
laboratory processing chart 
LRI 
lower respiratory illness 
LRT 
lower respiratory tract 
MA-LRI 
medically attended lower respiratory illness 
MOP 
manual of procedures 
mRNA 
messenger Ribonucleic Acid 
NHP 
nonhuman primate 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIAID CRMS NIAID Clinical Research Management System 
NICHD 
National Institute of Child Health and Human Development 
NIH 
National Institutes of Health 
nt 
nucleotide 
OCRPRO 
Office of Clinical Research Policy and Regulatory Operations 
OHRP 
Office for Human Research Protections 
ORF 
open reading frame 
OTC 
over-the-counter 
PBMC 
Peripheral blood mononuclear cell 
PCR 
polymerase chain reaction 
PE 
physical exam 
PFU 
plaque-forming unit 
PID 
participant identification number 
PMTCT 
prevention of mother-to-child HIV transmission 
PRNT 
plaque reduction neutralization titer 
PSRT 
Protocol Safety Review Team 
r 
recombinant 
RE 
regulatory entity 
RNA 
ribonucleic acid 
rRT-PCR 
Reverse transcription polymerase chain reaction 
RSC 
Regulatory Support Center 
RSV 
respiratory syncytial virus 
SAE 
Serious Adverse Event 
SDAC 
Statistical & Data Analysis Center, Harvard School of Public Health 
SDMC 
Statistical and Data Management Center 
SES 
Subject Enrollment System 
SID 
Study Identification Number 
SOP 
standard operating procedure 
SPG 
sucrose-phosphate-glutamate buffer 
SUSAR 
Serious and unexpected suspected adverse reactions 
TL 
Tracheal lavage  
URI 
upper respiratory illness 
URT 
upper respiratory tract 
US 
United States 
VAR 
vaccine administration record 
wt 
 
wild-type 
 

IMPAACT 2012, Version 1.0 
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IMPAACT 2012 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of 
a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus 
Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-
Seronegative Infants 6 to 24 Months of Age 
 
PROTOCOL TEAM ROSTER 
 
Protocol Chair 
Coleen Cunningham, MD  
Chief, Pediatric Infectious Diseases T915, 
Children’s Health Center Box 3499 DUMC  
Durham, NC 27710  
Phone: 919-668-4851  
Email: coleen.cunningham@duke.edu 
 
Protocol Vice Chair  
Ruth Karron, MD 
Center for Immunization Research (CIR)  
Johns Hopkins Bloomberg School of Public 
Health (JHSPH)  
624 N. Broadway, Room 217  
Baltimore, MD 21205  
Phone: 410-614-0319  
Email: rkarron@jhu.edu 
 
Clinical Trials Specialists 
Charlotte Perlowski, MSPH 
IMPAACT Operations Center 
FHI 360 
359 Blackwell Street, Suite 200 
Durham, NC 27701 
Phone: 919-544-7040 x11480 
Email: cperlowski@fhi360.org 
 
Jennifer Libous, MS, CCRP 
IMPAACT Operations Center 
FHI 360 
359 Blackwell Street, Suite 200 
Durham, NC 27701 
Phone: 919-544-7040 x11393 
Email: jlibous@fhi360.org 
 
NIAID Medical Officer 
Devasena Gnanashanmugam, MD 
Branch Chief 
Maternal, Adolescent and Pediatric Research 
Branch 
PSP/DAIDS/NIAID/NIH 
5601 Fishers Lane, Room 8B21B  
Rockville, MD 20852 
Phone: 301-435-3761 
Email: Devasena.gnanashanmugam@nih.gov 
 
NICHD Medical Officer 
Jack Moye, Jr. MD 
Eunice Kennedy Shriver National Institute of 
Child Health & Human Development 
National Institutes of Health,  
DHHS Bldg. 6100 Rm. 3A01 MSC 7510 
6710B Rockledge Drive, Rm. 2146, MSC 7002  
Bethesda, MD 20817 
Phone: 301-594-8624 
Email: john.moye@nih.hhs.gov 
 
Clinical Research Oversight Managers 
Kelly Cahill, RN, CCRC, RAC (US) 
NIH/NIAID/DCR/OCRPRO 
5601 Fishers Lane, Rm 4B23 
Rockville, MD 20852 
Mail Stop Code: 9820  
Email: cahillke@niaid.nih.gov 
 
Susan Vogel, RN, BSN 
OCRPRO/DCR/NIAID/NIH  
6700-B Rockledge Dr. Room 1113, MSC 7609  
Bethesda, MD 20817-7609  
Phone: 301-451-5138 
Email: svogel@niaid.nih.gov 
 
John Tierney, RN, MPM 
OCRPRO/DCR/NIAID/NIH  
5601 Fishers Lane, Rm. 4B11, MSC 9820 
Bethesda, MD 20892 
Phone: 301-451-5136 
Email: jtierney@niaid.nih.gov 
 

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Protocol Pharmacist 
Lynette Purdue, PharmD  
DAIDS/NIAID/NIH 
5601 Fishers Lane, Room 9E28 
Rockville, MD 20852  
Phone: 240-627-3061  
E-mail: lpurdue@niaid.nih.gov 
 
Site-Based Pharmacist 
Vivian Rexroad, Pharm.D. 
Investigational Drug Service Pharmacy 
Carnegie 180, Johns Hopkins Hospital 
600 North Wolfe St. 
Baltimore, MD 21287-6180 
Phone: 410-955-4505 
Email: vrexroa@jhmi.edu 
 
Clinical Investigators 
Elizabeth (Betsy) J. McFarland, MD  
Children’s Hospital Colorado Pediatric 
Infectious Diseases, Box B055  
13123 East 16th Ave.  
Aurora, CO 80045  
Phone: 303-724-3447  
Email: betsy.mcfarland@ucdenver.edu 
 
Elizabeth Schappell, RN, MSN, CCRP  
CIR, JHSPH  
624 N. Broadway, Room 205  
Baltimore, MD 21205  
Phone: 410-614-9114 
Email: eschapp1@jhu.edu 
 
Scientific Investigators 
Ursula Buchholz, PhD (Project Officer) 
Laboratory of Infectious Diseases (LID), 
National Institute of Allergy and Infectious 
Diseases (NIAID),  
NIH Building 50, Room 6503  
50 South Dr., MSC 8007  
Bethesda, MD 20892  
Phone: 301-594-1533  
Email: ubuchholz@niaid.nih.gov 
 
Peter Collins, PhD  
LID, NIAID, NIH  
Building 50, Room 6517  
50 South Dr., MSC 8007  
Bethesda, MD 20892  
Phone: 240-669-5917  
Email: pcollins@niaid.nih.gov 
 
Protocol Statistician 
Petronella Muresan, MS 
Statistical & Data Analysis Center (SDAC) 
Harvard School of Public Health 
900 Commonwealth Ave., 2nd Floor 
Boston, MA 02215 
Phone: 617-632-2059 
Email: petronm@sdac.harvard.edu 
 
Protocol Data Managers 
Benjamin Johnston 
Frontier Science & Technology Research 
Foundation (FSTRF)  
4033 Maple Rd.  
Amherst, NY 14226-1056  
Phone: 716-834-0900 x7407 
Email: johnston@fstrf.org 
 
Linda Marillo  
Frontier Science & Technology Research 
Foundation (FSTRF)  
4033 Maple Rd.  
Amherst, NY 14226-1056  
Phone: 716-834-0900 x7257  
Email: marillo@fstrf.org 
 
Protocol Laboratory Data Managers 
Oswald Dadson, MS 
Frontier Science & Technology Research 
Foundation (FSTRF)  
4033 Maple Rd.  
Amherst, NY 14226-1056 
Phone: 716-834-0900 x7238 
Email: dadson@fstrf.org 
 
Andee Fox, MPH 
Laboratory Data Manager 
Frontier Science and Technology Research 
Foundation (FSTRF)  
4033 Maple Road 
Amherst, NY 14226 
Phone: 716-834-0900 ext. 7276 
Email: fox@fstrf.org 
 
 

IMPAACT 2012, Version 1.0 
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Laboratory Center Representative 
Dale Dayton, RN, CCRA 
Children’s Hospital of Los Angeles 
4650 Sunset Boulevard 
Los Angeles, CA 90027 
Phone: 301-742-9077 
Email: ddayton@impaactlabcenter.org  
 
Protocol Laboratory Technologist 
Paul Harding, MS 
Pediatric Infectious Diseases  
University of Colorado Denver  
Mail Stop 8604, Building P15  
12700 East 19th Ave., Room 11460-A  
Aurora, CO 80045  
Phone: 303-724-3404 
Email: paul.harding@ucdenver.edu 
 
Westat Clinical Research Associate  
Scott Watson 
Westat, Inc. 
1441 W. Montgomery Ave 
Rockville, MD 20850 
 
Phone: 415-494-5575 
Email: ScottWatson@westat.com 
 
Field Representative 
Emily Barr, CPNP, CNM, MSN 
Children’s Hospital Colorado 
13123 East 16th Ave., Box B055 
Aurora, CO 80045 
Phone: 720-777-6752 
Email: Emily.barr@childrenscolorado.org 
 
 
 
 
 
 
 
 
 
 
 
 

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SITE ROSTER 
 
Site 4001, Lurie Children’s Hospital of Chicago 
Ram Yogev, MD 
Ann & Robert H. Lurie Children's Hospital of 
Chicago 
2300 Children's Plaza, Box #155 
Chicago, IL 60611 
Phone: 773-880-4757 
Email: ryogev@luriechildrens.org  
 
Site 4601, University of California San Diego 
Stephen Spector, MD 
9500 Gilman Drive 
Stein Clinical Research Building 
Room 430, Mail Code 0672 
La Jolla, CA 92093 
Phone: 858-534-7361 
Email: saspector@ucsd.edu  
 
Site 5013, Jacobi Medical Center 
Andrew Wiznia, MD 
Jacobi Medical Center 
1400 Pelham Parkway South 
JACP room 5c15, Building #5 
Bronx, NY 10461 
Phone: 718-918-4664 
Email: andrew.wiznia@einstein.yu.edu  
 
Site 5030, Emory University School of Medicine  
Paul Spearman, MD 
Emory University 
2015 Uppergate Drive 
Atlanta, GA 30322 
Phone: 404-727-5642 
Email: paul.spearman@emory.edu 
 
Site 5040, SUNY Stony Brook 
Sharon Nachman, MD 
SUNY Stony Brook 
Stony Brook, NY 11794-8111 
Phone: 631-444-7692 
Email: Sharon.Nachman@stonybrookmedicine.edu  
 
Site 5048, University of Southern California 
Mikhaela Cielo, MD 
Keck School of Medicine of USC 
HSA 10220, Bldg A10N 
Los Angeles, CA 90089 
Phone: 323-226-4726 
Email: mikhaela.cielo@usc.edu  
 
Site 5052, University of Colorado Denver 
Elizabeth (Betsy) J. McFarland, MD  
Children’s Hospital Colorado Pediatric 
Infectious Diseases, Box B055  
13123 East 16th Ave.  
Aurora, CO 80045  
Phone: 303-724-3447  
Email: betsy.mcfarland@ucdenver.edu 
 
Site 5083, Rush University/Cook County 
Hospital 
Mariam Aziz, MD 
Rush University Medical Center 
600 South Paulina St., Suite 140-142 
Chicago, IL 60612 
Phone: 312-942-4265 
Email: mariam_aziz@rush.edu  
 
Site 5112, David Geffen School of Medicine at 
UCLA 
Jaime Deville, MD 
University of California, Los Angeles 
Department of Pediatrics  
Division of Infectious Diseases 
10833 Le Conte Ave., MDCC 22-442 
Los Angeles, CA 90095 
Phone: 310-825-9660 
Email: jdeville@mednet.ucla.edu  
 
Site 5113, Children’s Hospital of Philadelphia 
Richard Rutstein, MD 
Children’s Hospital of Philadelphia 
Main Bldg, RM 12NW60 
34th and Civic Center Blvd. 
Philadelphia, PA 19104 
Phone: 215-590-1466  
Email: rutstein@email.chop.edu  
 

IMPAACT 2012, Version 1.0 
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Site 6501, St. Jude Children’s Research Hospital 
Nehali Patel, MD 
St. Jude Children’s Research Hospital 
262 Danny Thomas Place 
Memphis, TN 38105 
Phone: 901-595-4646 
Email: Nehali.Patel@stjude.org  
 
Site 31779, Johns Hopkins University Center for 
Immunization Research 
Ruth Karron, MD 
Center for Immunization Research (CIR)  
Johns Hopkins Bloomberg School of Public 
Health (JHSPH)  
624 N. Broadway, Room 217  
Baltimore, MD 21205  
Phone: 410-614-0319  
Email: rkarron@jhu.edu 
 

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IMPAACT 2012 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of 
a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus 
Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-
Seronegative Infants 6 to 24 Months of Age 
 
SCHEMA 
 
Purpose: 
To determine whether LID cp ΔM2-2 vaccine is attenuated and immunogenic 
in this age group. 
 
Design: 
A double-blind, randomized, placebo-controlled study design will be used to 
evaluate the safety and immunogenicity of the vaccine in RSV-seronegative 
infants and children. This protocol, IMPAACT 2012, is a companion 
protocol to the Johns Hopkins University, Center for Immunization Research 
(CIR) protocol CIR 312. 
 
 
Study Population: 
Healthy RSV-seronegative* infants ≥6 months (180 days) to <25 months 
(750 days) of age  
 
 
 
* Throughout the protocol and informed consent documents, seronegativity 
refers to RSV antibody status, which is defined as a serum RSV-neutralizing 
antibody titer <1:40. 
 
Sample Size: 
Approximately 33 (IMPAACT 2012 and CIR 312 combined) 
 
Study Product: 
Eligible RSV-seronegative infants will receive a single dose of LID cp ΔM2-
2 vaccine or placebo intranasally at entry. 
 
 
         
 
 
 
 
**plaque-forming units (PFU) 
 
Study Duration: 
Approximately 10 months total. Accrual is expected to be completed in 
approximately 6 to 8 weeks from first enrollment, and participants will be 
followed through April of the year following enrollment.  Therefore, 
expected duration of follow up for a given participant is between 6 and 10 
months depending on time of enrollment. 
 
Participants will be enrolled in the study between April 1st and October 14th 
(outside of RSV season) and will remain on study until they complete the 
post-RSV season visit between April 1st and April 30th in the calendar year 
following enrollment. For example, a participant enrolled on August 1st, 2016 
will remain on study approximately 8-9 months (completing a final visit in 
April 2017) while a participant enrolled on October 14th, 2016 will remain on 
study approximately 6 months (also completing a final visit in April 2017). 
N 
Product 
Dose 
22 
Vaccine 
105 PFU** 
11 
Placebo 
0 

IMPAACT 2012, Version 1.0 
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Primary Objectives 
1. Safety: To assess the frequency and severity of study product-related solicited and unsolicited 
adverse events (AEs), from Day 0 through midnight of the 28th day following inoculation, in 
vaccinated participants  
2. Safety: To assess study product-related serious adverse events (SAEs) from Day 0 through 
midnight on the 56th day following inoculation for vaccinated participants  
3. Infectivity: To determine the peak titer of vaccine virus shed and duration of virus shedding by 
each participant, where the primary aim is to check if the mean peak titer of shed virus in nasal 
washes is approximately 2.5 log10 
4. Infectivity: To assess the proportion of vaccinated infants infected with study vaccine, where the 
primary aim is to check whether >90% of vaccinees shed vaccine virus detected by infectivity 
assay 
5. Immunogenicity: To characterize antibody responses (Day 56) to the study vaccine 
 
Secondary Objectives 
1. To characterize clinical outcomes (frequency and severity of symptomatic, medically attended 
respiratory and febrile illness) in the vaccine and placebo recipients who experience natural 
infection with wild-type (wt) RSV during the subsequent RSV season  
2. To characterize antibody responses in the vaccine and placebo recipients who experience natural 
infection with wt RSV during the subsequent RSV season 
3. To characterize the B cell response to vaccine and the quality and epitope specificity of RSV F 
specific antibody, and to characterize these responses in the vaccine and placebo recipients who 
experience natural infection to wt RSV during the subsequent RSV season 
 
 
 
 

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IMPAACT 2012 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of 
a Single Dose of a Recombinant Live-Attenuated Respiratory Syncytial Virus 
Vaccine, LID cp ΔM2-2, Lot RSV#009B, Delivered as Nose Drops to RSV-
Seronegative Infants 6 to 24 Months of Age 
 
Figure 1: Study Overview 
 
 
 
 
 
 
 

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1 
INTRODUCTION  
 
1.1 
Overview 
 
Human respiratory syncytial virus (RSV) is the most common viral cause of serious acute lower 
respiratory illness (LRI) in infants and children under 5 years of age worldwide (1). There is 
broad consensus that a vaccine is needed. Attenuated live virus vaccines are a promising strategy 
for RSV since they have not been associated with vaccine enhanced diseases (2) and they have 
the potential of inducing a spectrum of  immune responses that would be similar to immunity 
induced by wild type infection (3). This protocol is part of a multi-year development plan aimed 
at identifying a candidate RSV vaccine that is sufficiently attenuated but still immunogenic 
 
One attenuation strategy that has shown great promise is deletion of a large section of the M2-2 
gene, resulting in an M2-2 deletion mutant. This mutation is associated with increased mRNA 
production linked with reduced RNA replication (4). The increase in mRNA production results in 
increased synthesis of viral antigen, with the potential for increased immunogenicity, and the 
decreased RNA replication results in delayed assembly of new virus particles, resulting in 
attenuation. A large deletion mutation effectively reduces the potential for reversion to wild type, 
an important concern for a live attenuated vaccine. The first candidate RSV vaccine using this 
strategy, MEDI ΔM2-2, was studied sequentially in adults, seropositive children, and then 
seronegative older infants and children (age 6 to 24 months) (5). This study found the vaccine had 
excellent immunogenicity associated with low viral replication. Determination of safety and 
tolerability of this candidate was confounded by a high incidence of adventitious respiratory 
infections during the study period. The MEDI ∆M2-2 phenotype may be suitable for an RSV 
vaccine, pending confirmation of safety. However, because of the overall low titer of shedding, 
and the lack of shedding in a number of vaccinees, it is possible that a virus that replicates 
somewhat more efficiently might be more immunogenic, provided it is suitably attenuated. This 
would be important since immune responses to RSV infection in infants and children are not 
robust. In addition, the MEDI ∆M2-2 virus lacks the temperature sensitive phenotype, a 
phenotype that may increase safety. Therefore, we also are evaluating several other ∆M2-2 
candidates with differences affecting replication and temperature sensitivity. A slightly different 
vaccine, RSV LID ΔM2-2, advanced to clinical trials in RSV-seronegative children in IMPAACT 
2000.  This study demonstrated excellent immunogenicity and increased infectivity.  However, 
the peak viral titers were higher than had been anticipated based on pre-clinical studies, and were 
deemed higher than desired. Therefore, in the interests of caution, IMPAACT 2000 was closed to 
accrual prior to full enrollment of the planned 50 participants [McFarland et al., manuscript in 
preparation], and a decision was made to further attenuate the LID ΔM2-2 vaccine as well as to 
consider several other candidates for evaluation.   
 
In the current protocol and in 2 associated protocols (IMPAACT 2011 and IMPAACT 2013) that 
will open in sequence, 3 further attenuated versions of RSV with M2-2 deletion will be studied, 1 
per protocol. Each of these versions of RSV with ΔM2-2 deletion has a different combination of 
mutations that are known to confer an attenuated phenotype. The 3 vectors are described in Table 
1, and a short description of each candidate follows.   
 

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Table 1: Overview of RSV vaccine candidates with M2-2 deletion to be studied in 
IMPAACT 2011, 2012, and 2013 
 
Protocol 
Vaccine name 
Mutations in addition to M2-2 deletion 
Expected effect 
2011 
LID RSV ∆M2-2 1030s 
genetically stable "1030" attenuating point 
mutation 
Attenuation 
Temperature 
sensitivity 
2012 
LID RSVcp ∆M2-2 
five point mutations in the RSV 
nucleoprotein, fusion protein, and 
polymerase protein 
Attenuation 
2013 
RSV/NS2/N/∆M2-2-
HindIII 
Contains the SH noncoding region that is 
deleted in the other two candidates and in the 
previous LID ∆M2-2, one point mutation 
each in the NS2 and N proteins, and a 
modified version of the M2-2 deletion, based 
on RSV MEDI ∆M2-2.  
Lower level of 
replication than 
LID ∆M2-2 due 
to the presence of 
the SH noncoding 
region 
 
The IMPAACT 2011 virus (LID ∆M2-2 1030s) is LID ∆M2-2 with the addition of a temperature-
sensitivity mutation (1030s) in the L polymerase protein. The 1030s mutation is a genetically 
stabilized version of a previous mutation called 1030. The 1030 and 1030s mutations (which 
confer the same level of temperature sensitivity and attenuation) have been characterized in a 
number of experimental animals including chimpanzees, and have been combined with other 
mutations in vaccine candidates including ones evaluated in infants and children. The LID ∆M2-2 
1030s virus was highly attenuated in African green monkeys.   
 
The IMPAACT 2012 virus (LID cp ∆M2-2) is LID ∆M2-2 with the addition of 5 amino acid 
substitutions in 3 proteins that in aggregate are called the “cp” mutations, and were originally 
identified in a cold-passaged RSV vaccine candidate cpRSV. The cp mutations have been shown 
to confer a relatively small amount of attenuation in adults and chimpanzees. They also have been 
combined with other mutations in vaccine candidates, including those evaluated in infants and 
children (6-9). We expect the cp mutations to confer a smaller increment of attenuation than 
1030s. This virus also was highly attenuated in African green monkeys.   
 
The IMPAACT 2013 virus (RSV D46/NS2/N/∆M2-2-HindIII) is designed to incorporate the 
most significant differences between MEDI ∆M2-2 and LID ∆M2-2. In particular, it uses the LID 
backbone, but without a deletion and nucleotide substitutions in the SH gene. The 
D46/NS2/N/∆M2-2-HindIII virus also has the MEDI assignments at the only two amino acid 
positions (in the NS2 and N ORFs) that differ between MEDI ∆M2-2 and LID ∆M2-2. In 
addition, the deletion of M2-2 in D46/NS2/N/∆M2-2-HindIII is comparable to that of MEDI 
∆M2-2. The level of attenuation of this virus in African Green Monkeys (AGMs) was slightly 
lower than that of LID cp ∆M2-2. 
 
Between these three planned studies, we hope to identify a candidate with the following 
characteristics:  
 
 
>90% of vaccinees should shed vaccine virus detected by infectivity assay.  
 
The mean peak titer of shed virus in nasal washes should be approximately 2.5 log10 PFU.  
 
RSV-neutralizing serum antibody titers (measured 56 days post inoculation) should be similar 
to or better than MEDI ∆M2-2 (geometric mean titer of >1:97).   
 
Post-vaccination surveillance during the RSV season (Nov 1 to Mar 31) following 
vaccination should reveal substantial rises in RSV-neutralizing serum antibodies in a subset 

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of vaccine recipients in the absence of reported RSV disease, comparable to that observed 
with MEDI and LID ∆M2-2. 
 
If a promising candidate can be identified in the planned studies, this candidate will be selected 
for further evaluation in expanded Phase I studies or Phase II studies. 
 
1.2 
Background  
 
Epidemiology, Disease Burden, and the Need for a Vaccine 
 
In the United States alone, RSV is responsible for 75,000 to 125,000 hospitalizations of infants 
yearly (10), and worldwide, RSV infects at least 34 million children under 5 years, with an 
estimated 3.4 million RSV LRI hospitalizations and 66,000 to 199,000 RSV-attributable deaths 
each year (1). In temperate climates, annual RSV epidemics occur in late winter and early spring, 
and nearly all children are infected within the first 2 years of life.  RSV illness can range from 
mild upper respiratory tract illness (URI) including rhinitis, pharyngitis, and coryza to severe LRI 
including bronchiolitis and pneumonia. Beyond the acute burden of disease caused by RSV, 
severe RSV disease in infancy may predispose to reactive airways disease during childhood (11, 
12). 
 
 
RSV is an enveloped RNA virus that is a member of the Paramyxoviridae family, genus 
Pneumovirus (13). RSV has a single negative-sense strand RNA genome of 15.2 kilobases 
encoding 10 mRNAs. Each mRNA encodes a single protein, with the exception of the M2 
mRNA, which contains 2 overlapping open reading frames (ORFs). The 11 RSV proteins are: the 
viral RNA-binding nucleocapsid protein N, the phosphoprotein P, the large polymerase protein L, 
the attachment glycoprotein G, the fusion glycoprotein F, the small hydrophobic surface 
glycoprotein SH, the internal matrix protein M, the 2 nonstructural proteins NS1 and NS2, and 
the M2-1 and M2-2 proteins encoded by the M2 mRNA. The gene order is: 3'-NS1-NS2-N-P-M-
SH-G-F-M2-L-5'. RSV transcription and genome replication take place exclusively in the 
cytoplasm, and virions form by budding from the apical plasma membrane of respiratory 
epithelial cells.  
 
Currently, no licensed vaccine against RSV is available, although there is broad consensus that 
such a vaccine is urgently needed and should be a global health priority. Although passive 
immunoprophylaxis with the monoclonal RSV-neutralizing antibody palivizumab (Synagis®; 
MedImmune) is available for high-risk infants, this approach is not feasible for general use. A 
formalin-inactivated vaccine against RSV was evaluated clinically in the 1960s and did not 
confer protection; instead, disease enhancement following infection of vaccinees with wt RSV 
was observed (14). Studies in experimental animals established that disease enhancement was 
specific to non-replicating RSV vaccines and not seen with infectious RSV or replicating vaccine 
vectors (15, 16).   
 
Following the failure of the formalin-inactivated RSV vaccine, attempts at developing RSV 
vaccines at National Institute of Allergy and Infectious Diseases (NIAID) have focused largely on 
live-attenuated approaches (3). Importantly, over a period of over 20 years, a number of live-
attenuated investigational RSV vaccines have been evaluated in RSV-naïve infants and children, 
and enhanced disease following wt RSV infection of vaccinees has not been observed (2). Apart 
from the absence of enhanced disease, live-attenuated RSV vaccines have a number of known 
advantages over non-replicating RSV vaccines: they can be administered intranasally, induce 
protective mucosal immunity in the respiratory tract (as well as systemic immunity), infect in the 

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presence of maternally-derived RSV serum antibody, and have been well-tolerated and 
immunogenic when administered to infants as young as 4 weeks of age (8).  
 
Human RSV has a single serotype with 2 antigenic subgroups, A and B. The 2 subgroups exhibit 
a 3- to 4-fold reciprocal difference in neutralization by polyclonal convalescent serum. Analysis 
of glycoprotein-specific responses in infants by enzyme-linked immunosorbent assay (ELISA) 
with purified F and G glycoproteins showed that the fusion proteins (F proteins) were 50% 
related antigenically, and the G proteins were 7% related (17). Consistent with this level of 
antigenic relatedness, F protein expressed by a recombinant vaccinia virus was equally protective 
in cotton rats against challenge with either subgroup A or B, whereas the G protein was 13-fold 
less effective against the heterologous subgroup (18). Thus, the F protein is responsible for most 
of the observed cross-subgroup neutralization and protection, and a subgroup A vaccine virus is 
likely to induce a broad immune response against wt RSV of either subgroup. Antibodies to the F 
protein are one of the endpoints evaluated in this study. 
 
The RSV vaccine to be evaluated in this study was derived using a recombinant deoxyribonucleic 
acid (DNA)-based technique called reverse genetics (19).  The technique of reverse genetics has 
been used to produce a number of licensed vaccines; among them is FluMist® (MedImmune), the 
live-attenuated influenza vaccine currently licensed for adults and children. This technique allows 
de novo recovery of infectious virus entirely from complementary DNA (cDNA) in a qualified 
cell substrate under defined conditions. Reverse genetics provides a means to introduce 
predetermined mutations into the RSV genome via the cDNA intermediate. Specific attenuating 
mutations were characterized in preclinical studies and combined to achieve the desired level of 
attenuation of this investigational vaccine. Derivation of vaccine virus from cDNA minimizes the 
risk of contamination with adventitious agents and helps to keep the passage history brief and 
well documented. Once recovered, the vaccine virus is propagated in the same manner as a 
biologically derived virus. As a result of repeated passage and amplification, the drug substance 
(clinical trials material) does not contain any recombinant DNA.  
 
This vaccine is a derivative of strain A2, subgroup A, with a deletion of the M2-2 ORF. The RSV 
M2-2 protein is a small protein (90 amino acids in RSV strain A2) encoded by the second, 
downstream ORF in the M2 mRNA, which slightly overlaps the 5'-proximal, upstream M2-1 
ORF (20). M2-2 is expressed intracellularly at a low level (21), and it is not known whether it is 
packaged into the virion. cDNA-derived RSV mutants, in which the M2-2 ORF has been silenced 
or deleted, grow more slowly in vitro than wt RSV (4, 22). Deletion of M2-2 results in increased 
accumulation of intracellular viral mRNA and decreased accumulation of genome and 
antigenome. This finding suggests that, during infection by wt RSV, M2-2 plays a role in shifting 
the balance of RNA synthesis from transcription to RNA replication (4). The increase in mRNA 
accumulation in cells infected with an M2-2 deleted RSV (∆M2-2) was accompanied by an 
increase in the expression of RSV proteins, including expression of the F and G glycoproteins, 
suggesting that M2-2 deletion mutants might be more immunogenic than wt RSV. M2-2 deletion 
mutants are highly attenuated in non-human primates but do replicate to detectable titers (22, 23). 
In addition, 2 previous clinical studies have shown that M2-2 deletion mutants of RSV seem to be 
refractory to genetic or phenotypic reversion to wt RSV. In order to further attenuate the vaccine 
virus, additional mutations have been added as described below. 
 

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Vaccine Description 
 
The Drug Substance, LID cp M2-2, is a recombinant RSV that contains a 241 nt deletion of the 
M2-2 ORF. To generate the vaccine candidate LID cp M2-2, the M2-2 deletion was combined 
with the cp mutations, 5 amino acid substitutions in 3 proteins that in aggregate are called the 
“cp” mutations. These mutations were originally identified in a cold-passaged RSV vaccine 
candidate cpRSV. The cp mutations have been shown to confer a relatively small amount of 
attenuation in adults and chimpanzees (24-26). The cp mutations also have been combined with 
other mutations in vaccine candidates, including those evaluated in infants and children (6-9). 
Genetic stability of live-attenuated RNA viruses is a concern in clinical vaccine development. 
The deletion of an entire ORF, here M2-2, should be refractory to genetic or phenotypic 
reversion. In a previous clinical study where the cp mutations were present in combination with 
other attenuating mutations, no further mutations of any of the cp mutations were detected, 
indicating that the cp mutations are genetically stable (8).  
 
To generate an RSV antigenomic cDNA with an M2-2 deletion, a cDNA fragment of 241 nt 
length was removed from a wt RSV antigenomic cDNA, and 3 potential translation initiation 
codons were silenced by site directed mutagenesis. In addition, the LID cp ΔM2-2 full length 
plasmid contains a deletion of 112 nucleotides from the downstream nontranslated region of the 
SH gene, and 5 translationally silent nucleotide changes in the downstream end of the SH open 
reading frame (Appendix I, Table 9). These changes in the SH noncoding region stabilize RSV 
full-length cDNA plasmids during propagation in bacteria, and were deemed to be phenotypically 
inconsequential based on in-vitro and in-vivo studies (27). The cp mutations (V267I in N, E218A 
and T523I in F, and C319Y and H1690Y in L) (Appendix I, Table 9)  were subsequently 
introduced by site directed mutagenesis of the full length plasmid. The resulting plasmid, 
designated pSK LID cp ΔM2.2, was used for the recovery of the drug substance. Sequence 
analysis of the seed virus genome indicated that its consensus sequence perfectly matched that of 
the pSK LID cp ΔM2.2 cDNA, except for a single point mutation at nucleotide 9972 
(corresponding to nt 10325 of wt recombinant RSV strain A2 D46 (19); Genbank accession 
number KT992094, Appendix I, Table 9). This mutation is silent on the amino acid level and 
considered inconsequential. 
 
The seed virus was generated at the Laboratory of Infectious Diseases (LID), NIAID (non-GMP), 
and transferred to Charles River Laboratories [CRL; Malvern, PA; operated under cGMP (current 
Good Manufacturing Practice)]. The seed virus passed pre-production testing (Sterility, 
Mycoplasma, Bacteriostasis/Fungistasis, and testing for porcine circovirus types 1 and 2; testing 
performed at CRL under cGMP), and was accepted for manufacturing of the Drug Product under 
cGMP. For the production of the Drug Product at CRL, Vero cells (MF 11702) were grown in 
OptiPRO™ serum-free medium. On day 3 post-infection, OptiPro™ medium was removed, and 
replaced by Dulbecco’s modified Eagle medium (DMEM). Antibiotics were not used in any stage 
of cell passage, virus growth, or vaccine development. The virus-containing supernatant was 
harvested on Day 6 post-infection and clarified by centrifugation. Intact cells were removed by 
filtration. 
 
Clarified supernatant in 1X SPG (sucrose, 0.218 M; KH2PO4, 0.0038 M; K2HPO4, 0.0072 M; L-
Glutamic Acid, 0.0054 M) was dispensed in 0.6 mL aliquots into labeled 2.0 mL cryogenic vials. 
Vials are snap-frozen and stored at -80ºC ± 15ºC. The Drug Product is a concentrated suspension 
of live recombinant LID cp ΔM2-2 Vero Grown Virus Vaccine (Lot RSV#009B) in DMEM 
without phenol red with 1X SPG (sucrose, 0.218 M; KH2PO4, 0.0038 M; K2HPO4, 0.0072 M; L-
Glutamic Acid, 0.0054 M). The Drug Product has a potency of about 5.6 log10 PFU/mL and is 
diluted to dose on site. 

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The Final Drug Product, LID cp ΔM2-2, Lot RSV # 009B, passed all in-vitro and in-vivo testing 
required for viral vaccines (Detection of Inapparent Viruses in a Viral Vaccine Product, in vitro 
Tuberculosis Testing, PCR-based Reverse Transcriptase Testing, Porcine Circovirus Testing, 
Sterility, Mycoplasma, Bacteriostasis/Fungistasis, Residual DNA testing, DNA Sizing, 
Endotoxin, General Safety, Determination of the Sucrose Level, pH Determination, Intact Cell 
Assay, Potency/Infectivity, Identity, Purity, Toxicology and Pharmacology testing).  Sequence 
analysis confirmed that the seed virus and Drug Product, RSV Lot #009B, were of identical 
sequence. LID cp ΔM2-2, Lot RSV #009B, was released by CRL for use as an investigational 
vaccine. 
 
1.3 
Prior Research 
 
1.3.1 
Experimental Vaccines against Respiratory Syncytial Virus 
 
Efforts have been directed toward the development of a live-attenuated RSV vaccine because of 
the advantages of live-attenuated vaccines over inactivated or subunit vaccines. These advantages 
include the ability to (i) induce the full spectrum of protective immune responses including serum 
and local antibodies as well as CD4+ and CD8+ T cells and innate immunity, (ii) infect and 
replicate in the presence of maternal antibody permitting immunization of young infants, and (iii) 
produce an acute, self-limited, attenuated infection that is well tolerated and readily eliminated 
from the respiratory tract. Another important advantage is the absence of vaccine-related 
enhanced disease, as has been confirmed in clinical studies (2). 
 
Several live-attenuated RSV vaccines have been evaluated in clinical trials in adult and pediatric 
populations as part of NIAID's ongoing RSV vaccine development program (6-9).  
 
Two versions of RSV vaccine candidates with M2-2 deletion have recently been evaluated. The 
first vaccine candidate, designated RSV MEDI ΔM2-2, was sequentially evaluated in adults, 
RSV-seropositive children, and RSV-seronegative infants and children (5). RSV MEDI ΔM2-2 
was highly restricted in replication; no shedding was observed in RSV-seropositive children, and 
minimal shedding was detected in RSV-seronegative children, in whom the mean peak titer of 
virus shed was 101.5 PFU/mL. Rhinorrhea occurred more frequently in recipients of RSV MEDI 
ΔM2-2 (85%) than placebo (44%), although this difference was not significant. Despite its 
restriction in replication, RSV MEDI ΔM2-2 was highly immunogenic. When we compared 
vaccine virus replication and antibody responses in RSV-seronegative children who received 
RSV MEDI ΔM2-2 to those achieved with rA2cp248/404/1030ΔSH, a live-attenuated RSV 
vaccine candidate that was well tolerated and immunogenic in pediatric Phase I studies (8), we 
found that RSV MEDI ΔM2-2 vaccine virus shedding was significantly more restricted, yet the 
post-vaccination RSV-neutralizing serum antibody titers achieved (geometric mean titer [GMT] = 
1:97) were significantly greater.  
 
Surveillance during the subsequent RSV season showed that several RSV seronegative RSV 
MEDI ∆M2-2 recipients had substantial antibody rises without reported illness, suggesting that 
the vaccine was protective yet primed for anamnestic responses to RSV. The M2-2 deletion was 
stable in all shed vaccine virus samples that were tested (5).  The MEDI ∆M2-2 phenotype may 
be suitable for an RSV vaccine, pending confirmation of safety. However, because of the overall 
low titer of shedding, and the lack of shedding in a number of vaccinees, it is possible that a virus 
that replicates somewhat more efficiently might be more immunogenic, provided it is suitably 
attenuated. Therefore, we also are evaluating several other ∆M2-2 candidates with differences 
affecting replication and temperature sensitivity.  

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The closely related RSV LID ΔM2-2 candidate vaccine virus was evaluated in RSV-seronegative 
children (IMPAACT 2000/CIR 291, ClinicalTrials.gov identifiers NCT02237209, 
NCT02040831), 20 of whom received vaccine and 9 of whom received placebo. RSV LID ΔM2-
2 was found to be highly infectious; 95% of vaccinees shed vaccine virus, with a mean peak titer 
of 103.4 PFU/mL by viral culture and a mean peak titer of 105.9 log10 copies/mL by quantitative 
real-time polymerase chain reaction (qRT-PCR). The level of replication of RSV LID ΔM2-2 in 
seronegative children was greater than expected based on previous study of RSV MEDI ∆M2-2. 
Respiratory or febrile illnesses occurred frequently in both recipients of RSV LID ΔM2-2 (95%) 
and placebo (78%). One vaccinee experienced a mild LRI (rhonchi) accompanied by shedding of 
vaccine virus and rhinovirus. It was not possible to determine whether the vaccine virus played a 
causal role in this participant's LRI, since an additional respiratory pathogen was also present.  
However, based upon the overall high level of vaccine virus replication and the concern that this 
might be a marker for under-attenuation, a decision was made to stop accrual to the study at 29 
rather the targeted 51 participants.    
 
Because of the unique properties of the M2-2 deletion mutation, which increases antigen 
production and seems to increase the inherent immunogenicity per infectious unit of virus, a 
decision was made to introduce additional attenuating mutations into the LID ΔM2-2 backbone to 
create vaccine candidates that would be more restricted in replication than LID ΔM2-2. The 
vaccine for the current protocol, LID cp ΔM2-2, is closely related to LID ΔM2-2.  It was 
generated by reverse genetics and contains two attenuating elements, namely the M2-2 deletion, 
and the further attenuating set of "cp" mutations (V267I in N, E218A and T523I in F, and C319Y 
and H1690Y in L). The "cp" mutations are expected to confer further attenuation to LID M2-2.  
Biologically derived cpRSV, which carries the cp mutations alone, was attenuated in early studies 
in adults (26) and in RSV seropositve children (28), but was underattenuated for RSV 
seronegative children (28). The combination of the “cp” mutations with the M2-2 deletion 
mutation is expected to result in further attenuation of the vaccine without decreasing 
immunogenicity.  
 
1.3.2 
Preclinical Studies 
 
RSV with M2-2 deletion has been extensively studied in vitro and in vivo (4, 21-23).  The results 
indicate that RSV ΔM2-2 is attenuated in 2 small animal models and in non-human primates. 
RSV ΔM2-2 was also found to be immunogenic in all 3 animal models. Attenuation of the Seed 
Lot and the clinical trial material (CTM) of LID cp ΔM2-2 was confirmed in a nonhuman primate 
model in African green monkeys (AGMs). Additional information about the preclinical 
evaluation of LID cp ΔM2-2 can be found in the Investigator’s Brochure (IB).  
 
Evaluation of the Attenuation Phenotype of LID cp ΔM2-2 in Nonhuman Primates 
 
LID cp ΔM2-2 was evaluated for its ability to replicate in the upper and lower respiratory tract 
(URT and LRT, respectively) of nonhuman primates (NHPs) [AGMs] in 2 independent non-GLP 
studies. AGMs are semi-permissive for RSV. The first NHP study was done to evaluate an 
Experimental Lot of LID cp ΔM2-2. Data from studies of related RSV vaccine candidates with 
M2-2 deletion (LID ΔM2-2 and LID ΔM2-2 1030s), performed in AGMs from the same group 
and origin, is included for comparison (Appendix I, Table 10, d A value of 0.35 was used for 
samples with no detectable virus. 
e The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve).   
 
 
 

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Table 11, and Table 12). The total doses administered were 2 x 106 PFU per animal.  The second 
study (total dose administered: 1 x 106 PFU per animal, n=4 animals, Appendix I, Table 13, Table 
14, and Table 15) was done to test the non-clinical safety and immunogenicity of the CTM LID 
cp ΔM2-2 in AGMs. 
 
Replication and Immunogenicity of an Experimental Lot of LID cp ΔM2-2 in AGMs 
 
AGMs (n=3 per group) seronegative for RSV were inoculated intranasal (i.n.) and intratracheal 
(i.t.) with LID cp ΔM2-2. Results from studies performed in animals from the same group and 
origin, inoculated with LID ΔM2-2 and LID ΔM2-2 1030s at the same dose (n=4 per group), 
were included for comparison. A dose of 1 x 106 PFU per site was administered to sedated 
juvenile male and female AGMs in a 1 mL volume per site (total dose: 2 x106 PFU/AGM). 
Nasopharyngeal (NP) swabs were collected daily on Days 0 through 10 and Day 12, and tracheal 
lavage (TL) samples were collected every other day from Day 2 through Day 12 from all animals 
included in the study, and virus shedding was analyzed by plaque assay. Serum RSV neutralizing 
antibody titers were determined by a complement-enhanced 60% plaque reduction assay with 
GFP-expressing RSV A2 on Vero cell monolayer cultures incubated at 37°C. Studies were 
approved by the Animal Care and Use Committee of NIAID, NIH. 
 
Substantial shedding of the RSV LID M2-2 control virus was detectable by plaque assay from 
the upper and lower respiratory tract over several days, with mean peak titers of 2.9 log10 PFU per 
mL in the URT, and 4.2 log10 PFU per mL in the LRT (Appendix I, Table 10 and d 
A value 
of 0.35 was used for samples with no detectable virus. 
e The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve).   
 
 
 
 
 
 
Table 11). In contrast, shedding of LID cp ΔM2-2 was only detectable in a single animal on a 
single day at very low titer in the upper respiratory tract, and in all animals on a single day in the 
lower respiratory tract. Remarkably, despite a very low level of virus replication, LID cp ΔM2-2 
induced serum neutralizing antibody titers comparable to LID ΔM2-2 in AGMs (Appendix I, 
Table 12). These results show that at a total dose of 2 x 106 PFU, administered i.n. and i.t., LID 
cp ΔM2-2 is highly attenuated, yet highly immunogenic in AGMs. 
 
Replication and Immunogenicity of LID cp ΔM2-2 (CTM) in AGMs 
 

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In a second study, the CTM LID cp M2-2 was evaluated at a total dose of 1 x 106 PFU per 
animal for its ability to replicate and induce an immune response in AGMs inoculated intranasally 
and intratracheally, following the same protocol described above. A dose of 5 x 105 PFU per site 
was administered to sedated juvenile male and female AGMs in a 1.25 mL volume per site (total 
dose: 1 x 106 PFU/AGM). At this dose, LID cp M2-2 was highly restricted for replication in 
AGMs (Table 13, Table 14). Shedding was undetectable, except for a single animal that shed a 
low level of LID cp ΔM2-2 virus from the lower respiratory tract on a single day (Day 8) post-
immunization. Despite the low or undetectable shedding, a robust serum neutralizing antibody 
response was induced by the CTM LID cp M2-2 (Table 15), confirming that the CTM LID cp 
ΔM2-2 is highly attenuated yet immunogenic for AGMs. 
 
In summary, compared to the previously characterized RSV vaccine candidate LID ΔM2-2, 
replication of LID cp ΔM2-2 was substantially reduced in AGMs, showing that the "cp" 
mutations resulted in further attenuation. Despite the low level of replication, intranasal and 
intratracheal inoculation with LID cp ΔM2-2 induced a strong neutralizing serum antibody 
response.  
  
The Drug Product Live Recombinant LID cp ΔM2-2 Vero Grown Virus Vaccine is anticipated to 
be more attenuated than the previous RSV vaccine candidate LID ΔM2-2. Intranasal 
administration of LID cp ΔM2-2 to RSV seronegative infants and children is anticipated to result 
in infection, limited vaccine replication, and the induction of a robust neutralizing antibody 
response to RSV.  
 
1.3.3 
Previous Clinical Experience 
 
The live attenuated recombinant LID cp ∆M2-2 vaccine virus is being evaluated for the first time 
in humans. This vaccine is a further attenuated derivative of the live attenuated recombinant RSV 
LID M2-2 vaccine virus, which is genetically similar to RSV MEDI ΔM2-2, which each have 
been studied in RSV-seronegative children. 
 
RSV MEDI ΔM2-2 
 
RSV MEDI ΔM2-2, Lot RSV #002A, was evaluated in adults, RSV-seropositive children, and 
RSV-seronegative children. Fifteen healthy adults received a 106 PFU dose of this vaccine in an 
open-label study. The vaccine was generally well tolerated, and vaccine virus was not detected in 
nasal washes collected from any of the vaccines. Serum antibody responses were not detected in 
any of these adult vaccinees. Thus, there was no evidence of replication of RSV MEDI ΔM2-2 in 
adult vaccinees. A 106 PFU dose of RSV MEDI ΔM2-2 was subsequently evaluated in 
RSV-seropositive children ages 12-59 months. Ten children in this RSV-seropositive cohort 
received a 106 PFU dose of vaccine, and 5 received placebo. Among the vaccinees, 5 children had 
rhinorrhea or nasal congestion, which was associated in all cases with shedding of rhinovirus and 
with shedding of adenovirus (1 child) or enterovirus (1 child). All illnesses were mild in severity. 
None of the vaccinees shed vaccine virus, nor did they have antibody responses to RSV, 
indicating that there was also no evidence of replication of RSV MEDI ΔM2-2 in RSV-
seropositive children 
 
RSV MEDI ∆M2-2 was subsequently evaluated at a 105 PFU dose in RSV-seronegative children. 
RSV MEDI ∆M2-2 replicated at low titers yet induced substantial RSV neutralizing antibody 
responses in RSV-seronegative children. Vaccine virus was detected by culture in 12 of 20 RSV-
seronegative vaccinees with a mean peak titer of 101.5 PFU/mL; 17 of 20 had vaccine virus 

IMPAACT 2012, Version 1.0 
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detected by qRT-PCR. Four-fold or greater increases in RSV neutralizing antibody occurred in 19 
of 20 children, with mean log2 titers of 2.7 ± 0.9 before vaccination and 6.6 ± 1.1 following 
vaccination. Respiratory illnesses were observed in 85% of vaccinees and 70% of placebo 
recipients, including fever (20% vs. 30%), rhinorrhea (85% vs. 50%), cough (35% vs. 30%), and 
otitis media (5% vs. 0%). LRI was not detected in any participant. Transmission of vaccine virus 
occurred in a set of 13-month-old twin study participants; both were minimally symptomatic and 
vaccine virus shed retained the M2-2 deletion. When we compared data on vaccine virus 
infectivity and immunogenicity in RSV-seronegative children to those achieved with rA2 
cp248/404/1030/ΔSH, a previous live-attenuated RSV vaccine candidate that was well tolerated 
and immunogenic in pediatric Phase I studies (8), we found that vaccine virus shedding was 
significantly more restricted. However, the post-vaccination RSV-neutralizing serum antibody 
achieved (GMT = 1:97) was significantly greater with RSV MEDI ∆M2-2 than with rA2 
cp248/404/1030/ΔSH. Surveillance during the subsequent RSV season showed that several RSV-
seronegative RSV MEDI ∆M2-2 recipients had substantial antibody rises without reported illness, 
suggesting that the vaccine was protective yet primed for anamnestic responses to RSV (5). 
However, conclusions about the safety and tolerability of this candidate were confounded by a 
high incidence of adventitious respiratory infections among participants during the trial. 
RSV LID M2-2  
 
Based upon these data, with the intent to gain additional safety information about the vaccine 
candidates with M2-2 mutation, we evaluated the closely genetically related vaccine RSV LID 
M2-2 in RSV-seronegative children at a dose of 105 PFU in 0.5 mL. In pre-clinical studies, RSV 
LID M2-2 behaved similarly to MEDI M2-2. As noted in Section 1.1, clinical data show that 
LID M2-2 replicated to higher viral titers than RSV MEDI ∆M2-2, as measured in nasal washes 
from RSV-seronegative vaccinees. MEDI M2-2 and LID M2-2 differ by 2 amino acids, which 
may confer a small amount of additional attenuation to MEDI M2-2. The modifications in the SH 
gene in the LID backbone, especially the 112 nt deletion of the 3' noncoding region in the LID 
M2-2 SH gene, may confer a small increase in replication in vivo. The available pre-clinical 
assays and experimental animals have been too insensitive to reliably detect these effects.  
 
1.4 
Rationale 
 
The IMPAACT Network is testing three further-attenuated versions of RSV ∆M2-2, each bearing 
additional attenuating elements for increased safety, in RSV-seronegative infants 6-24 months of 
age in three studies with a design similar to IMPAACT 2000.  
 
The candidates for each protocol described in Table 1, Section 1.1 include: 
 
IMPAACT Protocol 2011: LID ∆M2-2 1030s  
 
IMPAACT Protocol 2012: LID cp ∆M2-2  
 
IMPAACT Protocol 2013: RSV/NS2/N/∆M2-2-HindIII 
 
Each candidate is more attenuated in non-human primates than the candidate studied in 
IMPAACT 2000: LID ∆M2-2.  
 

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Each of these protocols monitors safety, infectivity, and immunogenicity, with particular attention 
to vaccine virus infectivity and replication (i.e., peak vaccine virus titer in nasal washes, as well 
as duration of shedding), which are the most quantifiable metrics for the level of attenuation. 
Each trial stands alone with the purpose to assess safety and immunogenicity in RSV-
seronegative infants 6-24 months of age. However, since the studies will have the same study 
design with centralized laboratory testing, comparison across the studies will be possible to 
evaluate which candidate vaccine(s) is most promising. It is anticipated that the most promising 
one or two of the candidates will move forward to an expanded study enrolling additional 
participants to further evaluate safety and immunogenicity.   
 
In previous studies (summarized in Section 1.3.3), 2 RSV vaccine candidates with M2-2 deletions 
have been shown to be safe and immunogenic in seronegative infants and children at a dose of 
105 PFU. Specifically, RSV MEDI M2-2 was the first RSV vaccine candidate with an M2-2 
deletion that was tested clinically, and was therefore sequentially evaluated (at the higher dose of 
106 PFU) in adults and RSV-seropositive infants and children, followed by evaluation of a dose 
of 105 PFU in RSV-seronegative infants and children. As mentioned in Section 1.3.3, in RSV-
seropositive cohorts, virus replication and RSV-specific antibody responses were undetectable 
after intranasal administration at a dose of 106 PFU. These results showed that RSV MEDI M2-
2 is highly attenuated and does not infect RSV-experienced individuals. Each of the vaccine 
candidates in IMPAACT 2011, 2012, and 2013 contains the M2-2 deletion, together with an 
additional attenuating element, and pre-clinical testing showed that this yielded further 
attenuation in nonhuman primates. Since it was previously shown that RSV MEDI M2-2 did not 
infect RSV-seropositive individuals, we would not expect to gain any relevant safety information 
by testing these further attenuated candidates in RSV-seropositive individuals.  
 
The primary immunogenicity endpoints to be evaluated are RSV neutralizing antibody titer, and 
RSV F protein antibody (by ELISA). Neutralizing antibody is a well-established and important 
surrogate marker of effective immunity to RSV disease. Antibodies to the F protein are also 
associated with cross-subgroup neutralization and protection (18). These assays will be performed 
at a central laboratory at the CIR at JHU that has performed the assays for the preceding clinical 
trials.      
 
As a secondary objective, we will study additional details of the B cell response to RSV. RSV 
neutralizing serum antibody levels represent the most reliable correlate of protection from RSV 
LRI; the protective role of antibody has been established infants over years of preventive use of 
palivizumab (29). Recent findings suggest that antibodies to the post-fusion form of the F protein 
may be most effective in neutralizing RSV. However, new experimental approaches to discern 
antibody specificities to epitopes present on the pre- and post-fusion forms of the fusion protein 
have become available. We are planning to study the epitope specificity and the quality (affinity 
and avidity) of the primary immune response to RSV vaccines. The induction of memory B cells 
is essential for long-term protection from severe RSV disease. RSV F protein specific B cells will 
be isolated, and studies on class switching, antibody maturation, and induction of B cell memory 
will be performed. 
 
1.5 
Hypotheses 
 
LID cp ∆M2-2 will be safe and immunogenic in RSV-naive infants.  
 

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2 
OBJECTIVES 
 
2.1 
Primary Objective 
 
The primary objectives of this study are the following:  
 
2.1.1 
Safety: To assess the frequency and severity of study product-related solicited and 
unsolicited adverse events (AEs) from Study Day 0 through Study Day 28 visit in 
vaccinated participants 
 
2.1.2 
Safety: To assess study product-related SAE from Study Day 0 through midnight on the 
56th day following inoculation for vaccinated participants 
 
2.1.3 
Infectivity: To determine the peak titer of vaccine virus shed and duration of virus 
shedding by each participant, where the primary aim is to check if the mean peak titer of 
shed virus in nasal washes is approximately 2.5 log10 
 
2.1.4 
Infectivity: To assess the proportion of vaccinated infants infected with study vaccine, 
where the primary aim is to check whether >90% of vaccinees shed vaccine virus 
detected by infectivity assay 
 
2.1.5 
Immunogenicity: To characterize antibody responses (Day 56) to the study vaccine 
 
2.2 
Secondary Objectives 
 
The secondary objectives of this study are to:  
 
2.2.1 
To characterize clinical outcomes (frequency and severity of symptomatic, medically 
attended respiratory and febrile illness) in the vaccine and placebo recipients who 
experience natural infection with wt RSV during the subsequent RSV season  
 
2.2.2 
To characterize antibody responses in the vaccine and placebo recipients who experience 
natural infection to wt RSV during the subsequent RSV season 
 
2.2.3 
To characterize the B cell response to vaccine and the quality and epitope specificity of 
RSV F specific antibody, and to characterize these responses in the vaccine and placebo 
recipients who experience natural infection to wt RSV during the subsequent RSV season 
 

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3 
STUDY DESIGN 
 
IMPAACT 2012 is a companion study to the Johns Hopkins University (JHU) Center for 
Immunization Research (CIR) protocol 312. The CIR 312 and IMPAACT 2012 protocols have 
identical primary and secondary objectives, investigational agents, inoculation schedules, 
evaluation assays and schedules, and safety monitoring and reporting. Because the CIR site does 
not enroll HIV-exposed infants, the eligibility criteria pertaining to that population are not 
included in CIR 312. 
 
The study will be conducted in infants at the JHU CIR and selected IMPAACT sites in the United 
States. The vaccine will be evaluated in RSV-seronegative (i.e., RSV-naïve) infants ≥6 months 
(180 days) to <25 months (750 days) of age. For the purpose of this study, RSV-seronegative is 
defined as having a serum neutralizing antibody titer of <1:40. This definition has been used in 
previous evaluations of live attenuated RSV vaccines (6, 8, 30). In these previous studies, live-
attenuated RSV vaccines were highly restricted in replication and poorly immunogenic in 
children with titers ≥1:40 but were far less restricted in replication and highly immunogenic in 
children with titers <1:40. These data suggest that this neutralizing antibody cutoff can 
distinguish effectively between RSV-experienced and RSV-naïve children. 
 
The study will be double-blind, randomized, and placebo-controlled. Thirty-three RSV-
seronegative participants will be randomized at a ratio of 2:1 to receive vaccine or placebo, 
respectively. Placebo recipients are needed in pediatric studies to distinguish the background 
respiratory and febrile illnesses that occur in infants and children from those attributable to study 
vaccine. These numbers were chosen based upon experience with Phase I evaluation of other 
live-attenuated respiratory virus candidate vaccines (6, 7, 9) and statistical considerations (see 
Section 9).   
 
Enrollment will occur between April 1st and October 14th to avoid the time during which wt RSV 
typically circulates in the community. Specific study phases are described in the paragraphs that 
follow. 
 
Duration of participation in the initial phase of the study is 56 days, which consists of an Acute 
and a Post-Acute Phase. During the Acute Phase (Study Day 0 to midnight on the 28th day 
following inoculation), participants will be contacted daily. These contacts will consist either of: 
1) in-person evaluation of interim medical history, clinical assessment, and nasal wash or 2) 
interim medical history conducted by phone, text, or email. During the Post-Acute Phase (Study 
Day 29 to midnight on the 56th day following inoculation), study participants will have a 
scheduled visit on Day 56. The schedule of evaluations during the Acute Phase and Post-Acute 
Phase is shown in Appendix II. 
 
The study has a third phase that assesses the incidence and severity of illness suggestive of RSV 
occurring during the RSV season following inoculation. During the RSV Season Surveillance 
Period, encompassing November 1st to March 31st, site study staff will make weekly contact with 
the participants to identify medically attended episodes of fever, URI or LRI, or otitis media. 
Participants who have such an episode will have a study visit to perform a nasal wash to evaluate 
for RSV and other respiratory pathogens (adventitious agents) (see Appendix III).  
 
Participants will also have a study visit during the pre-RSV season (between October 1st and 31st) 
to collect a blood sample for immunological assays, which will be used to assess the durability of 
the vaccine response and to serve as a pre-RSV season specimen. Participants will have a post-
RSV season visit (April 1st to April 30th) to collect blood for measurement of RSV immune 

IMPAACT 2012, Version 1.0 
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response to further assess the durability of the vaccine response and to assess the immune 
response to naturally occurring wt RSV infection. Thus, the maximum duration of participation 
will be up to 395 days, depending upon the time of enrollment relative to the RSV season. 
 
Figure 2 summarizes the study phases and periods of evaluation. There may be overlap in these 
various phases and periods. Accrual will stop effective on October 14th. 
 
Figure 2: Study Phases and Periods of Evaluation 
 
4 
STUDY POPULATION 
 
The vaccine will be evaluated in RSV-seronegative infants ≥6 months (180 days) to <25 months 
(750 days) of age. Approximately 33 evaluable participants will be enrolled (22 receiving the 
vaccine and 11 receiving a placebo) from US sites only. Infants will be selected for participation 
according to the criteria in Section 4.1 and 4.2. The sections that follow describe study-specific 
co-enrollment considerations; the recruitment, screening, and enrollment process; and participant 
retention and withdrawal or termination Sites are expected to obtain the infant’s medical records 
from the infant’s primary care provider to review for eligibility. The criteria related to the health 
status and age of household members, day care attendance, scheduled vaccine administration in 
relation to inoculation of study product, and use of salicylates (except as noted in the medical 
record) may rely on parent report.  
 
Any questions regarding interpretation of the inclusion/exclusion criteria or other considerations 
described in this section should be forwarded to the Protocol Team. 
 
4.1 
Inclusion Criteria 
 
Potential participants must meet all of the following criteria in order to be included in this study: 
 
 
4.1.1 
≥6 months (defined as ≥180 days) of age at the time of screening and <25 months 
(defined as < 750 days) of age 
 

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4.1.2 
The participant is in good health based on review of the medical record, history, and 
physical examination, without evidence of chronic disease.  
 
4.1.3 
Parents/guardians are willing and able to provide written informed consent as described 
in protocol Section 12.3.  
 
4.1.4 
Seronegative for RSV antibody, defined as a serum RSV-neutralizing antibody titer 
<1:40 at screening from a sample collected no more than 42 days prior to inoculation. 
Note: results from specimens collected during screening for IMPAACT 2011 are 
acceptable as long as within the 42-day window. 
 
4.1.5 
Is growing at a normal velocity for age (as demonstrated on a standard growth chart) 
AND 
 
If less than 1 year of age: has a current height and weight above the 5th percentile  
 
If 1 year of age or older: has a current height and weight above the 3rd percentile 
for age. 
 
4.1.6 
Participant has received routine immunizations appropriate for age (as per national 
Center for Disease Control Advisory Committee on Immunization Practices [ACIP]).  
 
4.1.7 
Participant is expected to be available for the duration of the study. 
 
4.1.8 
If born to an HIV-infected woman, participant must not have been breastfed and must 
have documentation of 2 negative HIV nucleic acid (RNA or DNA) test results from 
samples collected on different dates with both collected when ≥1 month of age and at 
least one collected when ≥4 months of age, and no positive HIV nucleic acid (RNA or 
DNA) test; or 2 negative HIV antibody tests, both from samples collected at ≥ 6 months 
of age.    
 
4.2 
Exclusion Criteria 
 
Potential participants who meet any of the following criteria will be excluded from this study: 
 
4.2.1 
Known or suspected HIV infection or impairment of immunological functions. 
 
4.2.2 
Receipt of immunosuppressive therapy, including any systemic, including either nasal or 
inhaled, corticosteroids within 28 days of enrollment.  Note: Cutaneous (topical) steroid 
treatment is not an exclusion. 
 
4.2.3 
Bone marrow/solid organ transplant recipient. 
 
4.2.4 
Major congenital malformations (such as congenital cleft palate) or cytogenetic 
abnormalities. 
 
4.2.5 
Previous receipt of a licensed or investigational RSV vaccine (or placebo in any 
IMPAACT RSV study) or previous receipt of or planned administration of any anti-RSV 
product (such as ribavirin or RSV IG or RSV mAb). 
 
4.2.6 
Previous anaphylactic reaction.  
 

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4.2.7 
Previous vaccine-associated adverse reaction that was Grade 3 or above. 
 
4.2.8 
Known hypersensitivity to any study product component. 
 
4.2.9 
Heart disease. Note: Participants with cardiac abnormalities documented to be clinically 
insignificant and requiring no treatment may be enrolled. 
 
4.2.10 Lung disease, including any history of reactive airway disease or medically documented 
wheezing.   
 
4.2.11 Member of a household that contains, or will contain, an infant who is less than 6 months 
of age at the enrollment date through Day 28. 
 
4.2.12 Member of a household that contains another child who is, or is scheduled to be, enrolled 
in IMPAACT 2011, 2012 or 2013 AND there has been or will be an overlap in residency 
during that other child’s participation in the study’s Acute Phase (Days 0 to 28). 
 
4.2.13 Member of a household that contains an immunocompromised individual, including, but 
not limited to: 
 
a person who is greater than or equal to 6 years of age with HIV-related 
immunodeficiency, defined as having a most recent CD4 T lymphocyte cell count 
<300 cells/mm3. CD4 T lymphocyte count must have been measured within 6 
months prior to enrollment, or 
 
a person age 1 year up to less than 6 years with HIV-related immunodeficiency, 
defined as having a most recent CD4 T lymphocyte cell percentage <25 or CD4 T 
lymphocyte count <750 cells/mm3 (if both values are available, use the lower of the 
two). CD4 T lymphocyte parameter must have been measured within the 6 months 
prior to enrollment; or 
 
a person age less than 1 year with HIV-related immunodeficiency, defined as 
having a most recent CD4 T lymphocyte cell percentage <30 or CD4 T lymphocyte 
count <1000 cells/mm3 (if both values are available, use the lower of the two). CD4 
T lymphocyte parameter must have been measured within the 6 months prior to 
enrollment; or  
 
a person who has received chemotherapy within the 12 months prior to enrollment; 
or 
 
a person receiving immunosuppressant agents; or 
 
a person living with a solid organ or bone marrow transplant. 
 
Verbal report of CD4 T cell lymphocyte is sufficient documentation if the 
parent/guardian is confident of history. 
 
4.2.14 Attends a daycare facility and shares a room with infants less than 6 months of age, and 
parent/guardian is unable or unwilling to suspend daycare for 28 days following 
inoculation.  
 
4.2.15 Any of the following events at the time of enrollment: 
 
fever (rectal temperature of ≥100.4°F (38°C)), or  
 
upper respiratory signs or symptoms (rhinorrhea, cough, or pharyngitis) or 
 
nasal congestion significant enough to interfere with successful inoculation, or  

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 
otitis media. 
 
4.2.16 Receipt of the following prior to enrollment: 
 
any killed vaccine or live-attenuated rotavirus vaccine within the 14 days prior, or  
 
any live vaccine, other than rotavirus vaccine, within the 28 days prior, or 
 
another investigational vaccine or investigational drug within 28 days prior 
 
4.2.17 Scheduled administration of the following after planned inoculation: 
 
killed vaccine or live-attenuated rotavirus vaccine within the 14 days after, or 
 
any live vaccine other than rotavirus in the 28 days after, or 
 
another investigational vaccine or investigational drug in the 56 days after 
 
4.2.18 Receipt of immunoglobulin, any antibody products, or any blood products within the past 
6 months 
 
4.2.19 Receipt of any of the following medications within 3 days of study enrollment: 
 
systemic antibacterial, antiviral, antifungal, anti-parasitic, or antituberculous 
agents, whether for treatment or prophylaxis, or 
 
intranasal medications, or 
 
other prescription medication except as listed below 
 
Permitted concomitant medications (prescription or non-prescription) include nutritional 
supplements, medications for gastroesophageal reflux, eye drops, and topical 
medications, including (but not limited to) cutaneous (topical) steroids, topical 
antibiotics, and topical antifungal agents. 
 
4.2.20 Receipt of salicylate (aspirin) or salicylate-containing products within the 28 days prior to 
enrollment. 
4.2.21 Born at less than 34 weeks gestation. 
 
4.2.22 Born at less than 37 weeks gestation and less than 1 year of age at the time of enrollment. 
 
4.2.23 Suspected or documented developmental disorder, delay, or other developmental 
problem. 
 
4.2.24 Previous receipt of supplemental oxygen therapy in a home setting. 
 
 
4.3 
Co-Enrollment Considerations 
 
Co-enrollment to an interventional study is not allowed during the Acute Phase or Post-Acute 
Phase.  After the Post-Acute Phase, co-enrollment may be considered if both protocol teams 
agree. 
 
Note: co-enrollment into IMPAACT 2012 is allowable for participants already enrolled in 
IMPAACT P1112, Dose Group 1 or 2, provided all eligibility criteria are met. The P1112 team 
should be queried in each case to confirm. 
 

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4.4 
Recruitment, Screening, and Enrollment Process 
 
Recruitment will take place at IMPAACT sites selected on the ability to recruit and enroll both 
HIV-exposed, uninfected and HIV-unexposed infants in RSV vaccine studies. Each site will 
identify the specific clinics where recruitment will occur as part of the site selection process, 
which will be reviewed and approved by the Protocol Team. All recruitment materials must be 
reviewed and approved by site IRBs. 
 
The IMPAACT Operations Center will monitor screening and enrollment through close contact 
with sites. These data will be provided to the team during regular team calls. 
 
The IMPAACT Data Management Center (DMC) Subject Enrollment System (SES) will be used 
to track enrollment. When informed consent is obtained, participant identification numbers 
(PIDs) will be assigned to the infant through the SES. For infants found to be eligible, 
randomization will occur upon successful entry of required eligibility data into the SES.  
Successful entry into the SES will generate a study identification number (SID) and blinded 
prescribing information for the study vaccine regimen to which the infant has been randomly 
assigned.  For infants who are found to be ineligible for the study, or who do not enroll in the 
study for any reason, a case report form (CRF) will be completed to record the screening 
outcome.  Refer to Section 9.4 for more information on monitoring participant accrual in this 
study.  
 
4.5 
Participant Retention 
 
Once an infant is enrolled in this study, study staff will make every effort to retain him or her in 
follow-up for the protocol-specified duration of follow-up, i.e., through the Post-RSV Season 
Study Visit, thereby minimizing potential biases associated with loss to follow-up.   
 
4.6 
Participant Withdrawal or Termination from the Study 
 
Regardless of the participant retention procedures referenced above, infants participating in this 
study may voluntarily withdraw from the study at any time. Any participant who has received 
study product will be encouraged to remain in the safety evaluation for the duration of the study 
even if sample collection is refused.  
 
A participant may withdraw or be terminated from the study early for any of the following 
reasons: 
 
 
Withdrawal of consent – applies to a parent/guardian who verbally or in writing 
withdraws consent to participate in the study for any reason. 
 
Noncompliant with protocol – applies to a parent/guardian who does not comply with 
protocol-specific visits or evaluations on a consistent basis, such that adequate follow-up 
is not possible and the participant’s safety would be compromised by continuing in the 
study. 
 
Participant withdrawal may occur if the investigator believes that it is in the best interest 
of the participant. 
 
Other – a category used when previous categories do not apply; requires an explanation. 
 
The study may be terminated for the following reasons: 
 

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 
Research terminated by sponsor or investigator – applies to the situation where the entire 
study is terminated by the sponsor or investigator for any reason. 
 
The study sponsor, IMPAACT, the site IRB, the Office for Human Research Protections 
(OHRP), NIAID, or the US FDA may decide to end the study. 
 
For any participant who withdraws or is terminated from the study prior to scheduled completion 
of follow-up, study staff will document the reason for the withdrawal or termination in detail and 
will make every effort to complete final evaluations as described in Section 6.11.  In the event 
that the circumstances that led to a participant’s withdrawal or termination change (e.g., he or she 
returns to the study site area after having re-located previously), the site investigator or designee 
should contact the protocol team to discuss options for resumption of follow-up. 
 
 
5 
STUDY PRODUCT CONSIDERATIONS 
 
Site pharmacists should consult the Pharmacy Guidelines and Instructions for DAIDS Clinical 
Trials Networks for standard pharmacy operations.  Refer to Figure 1 for an overview of the 
study design and to the Investigator’s Brochures (IBs) for further information about the study 
product. 
 
 
5.1 
Study Products 
 
The products that will be administered in this study are: 
 
Live Recombinant Respiratory Syncytial Virus (RSV) LID cp ΔM2-2 105 PFU per 0.5ml 
vaccine 
 
Placebo for RSV vaccine will be 1X L-15 Leibovitz medium (1X L-15) 0.5ml 
 
5.2 
Study Product Regimens 
 
Enrolled study participants will receive a single dose of RSV LID cp ΔM2-2 vaccine or placebo, 
administered as nose drops within 3 days of randomization. Ideally, the date of randomization 
will be the same as the date of inoculation (see Section 6.2).  
 
5.3 
Study Product Formulation  
 
5.3.1 
Vaccine 
 
The RSV LID cp ΔM2-2 is provided in a sterile 2.0 mL cryovial, each containing 0.6 mL of 
Vaccine (Lot RSV #009B), with a titer of approximately 105.6 PFU/mL The vaccine virus 
concentrate is diluted to an appropriate dose by designated licensed pharmacy personnel to 
prepare a dose of 105 PFU in a 0.5 mL volume. The vaccine vial is labeled as shown below in 
Figure 3. 
  
Figure 3: Investigational Product Label (Enlarged Sample) 
 

IMPAACT 2012, Version 1.0 
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Live Recombinant Respiratory Syncytial Virus 
LID cp ∆M2-2 
VERO GROWN VIRUS VACCINE 
Date: 10MAY2016 
Vial#:XXXX 
Lot: RSV #009B  
 
CAUTION:NEW DRUG LIMITED 
 BY FEDERAL (USA) LAW 
TO INVESTIGATIONAL USE 
Store at –80C ± 15C 
Charles River Laboratories, Malvern, PA 
 
5.3.2 
Diluent for RSV LID cp ΔM2-2 
 
The diluent for RSV LID cp ΔM2-2 is 1X Leibovitz L-15 medium.  
 
Sterile Water For Injection, USP (SWFI) and 2X Leibovitz L-15 medium are required to prepare 
the diluent for the vaccine. 2X Leibovitz L-15 medium is a specific lot of Leibovitz L-15 
medium. It is a solution of amino acids, sugar, and salt that has been safety tested as described in 
a Master File (MF 12959), which has been submitted to the FDA.  
 
5.3.3 
Placebo for RSV LID cp ΔM2-2 
 
The placebo for RSV LID cp ΔM2-2 is 1X Leibovitz L-15 medium.  
 
Sterile Water For Injection, USP (SWFI) and 2X Leibovitz L-15 medium are required to prepare 
the Placebo for the vaccine.  
 
5.4 
Study Product Storage 
 
Vaccine will be stored in a secure freezer -80°C ±15°C. It must remain frozen until time of use. 
Once the vaccine is thawed, it should never be refrozen for reuse. Vaccine will be prepared from 
new, unopened containers for each use.  
 
Leibovitz L-15 medium will be stored in a secure refrigerator at 2°C to 8°C. Vaccine 
diluent/placebo will be prepared from new, unopened containers for each use. 
Sterile Water for Injection, USP (SWFI) must also be stored in the refrigerator at 2°C to 8°C.  
 
Procedures for managing the vaccine and diluent/placebo shipment are in the Manual of 
Procedures (MOP). 
 
5.5 
Study Product Preparation 
 
The diluent for the vaccine, the placebo for the vaccine and the RSV vaccine must be prepared by 
following the detailed instruction in the MOP. 
 
Prior to inoculation, an authorized prescriber will supply a prescription to the pharmacy. The 
prescription must include the information outlined in the MOP. Designated licensed pharmacy 
personnel will prepare the correct dose of vaccine for each participant in a Biological Safety 
Cabinet (BSC) or Compounding Aseptic Containment Isolator (CACI) using aseptic technique.  
To preserve blinding, yellow overlays will be applied to all prepared syringes. 
 
5.5.1 
Diluent for RSV LID cp ΔM2-2 
 

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The diluent is prepared by mixing concentrated 2X Leibovitz L-15 medium with sterile water for 
injection in 1:1 ratio. The prepared vaccine diluent will be 1X Leibovitz L-15 medium.  
 
Please follow the MOP for detailed instructions on diluent preparation. 
 
5.5.2 
Placebo for RSV LID cp ΔM2-2 
 
Placebo for RSV LID cp ΔM2-2 will be prepared by mixing the concentrated 2X Leibovitz L-15 
medium with sterile water for injection in 1:1 ratio. The prepared product will be 1X Leibovitz L-
15 medium.  
 
Placebo will be drawn up to a volume of 0.5 mL in a sterile 1 mL oral syringe and labeled per 
instructions in MOP. An auxiliary label stating “FOR INTRANASAL ADMINISTRATION 
ONLY” will be affixed to the syringe or outside bag. The labeled, filled syringes will be 
transported in a cooler monitored and maintained at 2ºC to 8ºC with ice or cold packs to the 
clinical site for administration. Placebo must be administered within 4 hours of concentrated 2X 
Leibovitz L-15 being removed from the refrigerator.  
 
Please follow the MOP for detailed instructions on preparation of placebo. 
 
5.5.3 
Live Recombinant Respiratory Syncytial Virus (RSV) LID cp ΔM2-2 
 
Diluent must be prepared prior to removal of the RSV vaccine from the freezer.  
 
If the -80°C freezer where the RSV vaccine is stored is not in close proximity to where the 
preparation is being done, the vaccine vials should be transported on dry ice from the freezer 
to the biologic safety cabinet/isolator. Do not thaw this product on the bench top or allow the 
vial to thaw completely before putting onto wet ice. RSV is extremely sensitive to 
temperature fluctuations. Please follow the MOP for proper handling of the study product.  
 
Three vials per dose of undiluted vaccine are always used to prepare the vaccine dose to account 
for potential differences in titers of the concentrated vaccine. When manipulating the undiluted 
vaccine, use as small a gauge needle as possible to avoid loss of vaccine in the needle and 
syringe hub. Concentration of the undiluted vaccine is about 105.6 PFU per mL. The frozen 
vaccine will be thawed and diluted with 1X L-15 to a dose of 105 PFU in 0.5 mL prior to 
administration.  
 
The diluted vaccine will be drawn up to a volume of 0.5 mL in a sterile 1 mL oral syringe and 
labeled per instructions in MOP. An auxiliary label stating “FOR INTRANASAL 
ADMINISTRATION ONLY” will be affixed to the syringe or outside bag. The labeled, filled 
syringes will be transported in a cooler monitored and maintained at 2ºC to 8ºC with ice or 
cold packs to the clinical site for administration. Vaccine must be prepared and administered 
within 4 hours of being removed from the freezer and thawing. However, the expiration time 
is assigned based on the time the concentrated 2X Leibovitz L-15 is removed from the 
refrigerator in order to maintain the blind. 
 
Samples of undiluted (if available) and diluted vaccine will be aliquoted from the vaccine 
remaining after vaccine has been prepared and delivered to the clinical staff. The samples will 
be snap frozen as per the MOP and stored at -80°C ±15°C separate from the study product in 
the investigational pharmacy until they are shipped to the central laboratory for titration.   
Titration of vaccine is done to confirm the titer of the vaccine administered to the participants. 

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To “snap freeze” diluted and undiluted vaccine aliquots, follow BioCision CoolBox procedures 
described in the MOP. 
 
Please follow the MOP for detailed instructions on vaccine storage, handling, preparation, 
labelling and transport to the clinic. 
 
5.6 
Study Product Administration/ Inoculation Procedure 
 
All study participants will receive a single dose of study product, administered as a nose drops. 
There is no nasal preparation prior to administration. While the participant is supine, a volume of 
0.5 mL of study product will be delivered as nose drops (approximately 0.25 mL per nostril) 
using a sterile, needle-less 1 mL oral syringe. Participant will remain supine for approximately 60 
seconds following inoculation. 
 
5.7 
Study Product Acquisition 
 
The clinical lot preparation of RSV LID cp ΔM2-2 was generated by Charles River Laboratories 
using the seed virus provided by the National Institutes of Health (NIH).  
 
Specific lot concentrated 2X L-15 Leibovitz Medium that will be used to prepare the Diluent and 
Placebo is provided by the National Institutes of Health (NIH). 
 
Sterile Water for Injection, USP (SWFI) must be obtained by each site.   
Upon successful completion of protocol registration procedures, the Clinical research site (CRS) 
pharmacists can order the vaccine, the 2X L-15 Leibovitz Medium (which is used to make the 
diluent and placebo), sterile oral 1ml syringes (commercially available, individually packaged), 
sterile syringe caps and yellow overlays from the CRPMC by following the instructions in 
Pharmacy Guidelines and Instructions for DAIDS Clinical Trials Networks.  
 
Please refer to the MOP for details on the vaccine and diluent/placebo shipment. 
 
5.8 
Study Product Accountability 
 
The site pharmacist is responsible for maintaining an accurate inventory and accountability record 
of vaccine and Leibovitz L-15 medium supplies for this study. The vaccine will be prepared as 
instructed with the site pharmacist serving as the unblinded dispenser. A copy of the 
randomization code will be retained by the site pharmacist. Without the Protocol Chair’s written 
request to unblind, the randomization code may not be released. The site pharmacist will be 
responsible for maintaining the blind, and pharmacy records will be maintained in the pharmacy 
only. Partially used vials of vaccine and Leibovitz L-15 medium components may not be saved 
and reused at a later time. 
 

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5.9 
Disposition of Used/Unused Study Product 
 
After the designated licensed pharmacy personnel dilutes the vaccine and draws up the vaccine 
into a syringe for administration, he/she will remove the label from all the three vaccine vials and 
place it in the accountability log. Please see the IMPAACT 2012 MOP Appendix VI for the study 
preparation and accountability documents. In this manner, monitoring personnel will be able to 
verify the accountability of all vaccine vials used for the study. If there is any vaccine left after 
the syringes have been drawn up and aliquots have been removed for titering, it will be destroyed 
by pharmacy personnel as per the MOP. Any unused vaccine will be destroyed as per the MOP. 
 
5.10 
Final Disposition of Study Products 
 
All unused study products must be returned to the CRPMC after the study is completed or 
terminated, unless otherwise instructed by the CRPMC. Procedures and relevant forms are 
provided in the Pharmacy Guidelines and Instructions for DAIDS Clinical Trials Networks. 
 
5.11 
Concomitant Medications 
 
5.11.1 Prohibited Concomitant Medications 
 
The use of prophylactic antipyretics, decongestants, or antihistamines is not permitted during the 
Acute Phase (28 days following inoculation); however, use of these medications for treatment of 
symptoms is allowed.  
 
5.11.2 Precautionary Concomitant Medications  
 
 
Due to their potentially confounding effect on study immunogenicity results, the following 
treatments should be avoided after inoculation unless determined to be clinically indicated after 
consultation with the PSRT:  
 
Use of an investigational drug or investigational vaccine other than the study product 
within 56 days after receiving study product 
 
Systemic corticosteroids for more than 14 days at a dosage equivalent to prednisone at >2 
mg/kg or 20 mg daily or other immune-modifying drugs  
 
Licensed killed vaccine or live-attenuated rotavirus vaccine within 14 days after 
receiving study product  
 
Immunoglobulins and/or any blood products  
 
Licensed live virus vaccine, other than rotavirus vaccine, within 28 days after receiving 
study product 
 
6 
STUDY VISITS AND PROCEDURES 
 
Study visits, except inoculation, may be conducted at one of the clinical sites or as home visits. 
Inoculation must be conducted at one of the clinical sites. 
 
An overview of the study visits, evaluation schedule, and specimen collection and volumes are 
provided in Appendices II and III. Presented in this section is additional information on visit-
specific study procedures.  Study phases and periods of evaluation are summarized in Figure 2 of 
Section 3. 
 

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Unless otherwise specified, visits may be split, with required procedures conducted on more than 
one day within the allowable visit window, if necessary. All visits and procedures must be 
documented in accordance with the NIAID Division of AIDS (DAIDS) policies for source 
documentation; refer to Section 10 for more information on documentation requirements and 
completion of CRFs. Refer to Section 7 for information on expedited adverse event (EAE) 
reporting, which is required at specified times during follow-up.   
 
In addition to the protocol-specified procedures listed in this section, study staff may complete 
other tasks consistent with site SOPs, including but not limited to collecting, reviewing, and 
updating demographic and locator information; reviewing elements of informed consent; 
scheduling telephone contacts and visits; providing instructions for contacting study staff between 
visits; providing visit reminders; and following up on missed visits. All such tasks should be 
documented consistent with site SOPs. Clinical evaluations must be performed by a medical 
professional. Study staff should inform participant parents/guardians of clinically meaningful 
physical exam findings and laboratory test results when available. 
 
6.1 
Screening Visit 
 
As sites consider scheduling for the screening visit, it is important to also consider potential dates 
of randomization, given that inoculation cannot occur 42 days or more after the screening 
evaluation.  Sites should schedule randomization and inoculation to be as soon as feasible after 
screening, allowing for the turnaround time for the RSV serology testing. 
 
The parent/guardian must complete the informed consent process and sign the informed consent 
form before any study-related procedures are performed. Parents/guardians will also be offered a 
signed copy of the informed consent form (see Section 12.3). 
 
As in previous Phase I trials of other live-attenuated RSV vaccines (6-9), screening laboratory 
tests other than the RSV antibody will not be performed on participants. Such tests are not 
routinely performed as part of well-child care, given that the risk of undiagnosed hepatic, 
metabolic, and renal diseases is much lower in children than in adults (31).  
 

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Screening Visit Procedures (no more than 42 days prior to enrollment) 
Administrative 
and Regulatory 
 Obtain written informed consent 
 Confirm parent/guardian’s informed consent comprehension using 
the template quiz provided in the MOP or other, site-specific 
method.  
 Assign participant identification number (PID) 
 Assess eligibility 
Clinical 
 Obtain available medical records, including immunization record.  
If necessary, obtain release of medical records from 
parent/guardian to review the participant’s medical record as 
required per Health Insurance Portability and Accountability Act 
(HIPAA). 
 Obtain medical history, which should include demographics, prior 
diagnoses, current medications, signs and symptoms, and 
developmental status. 
 Perform complete infant physical examination including 
temperature, heart rate, respiratory rate, weight, length and 
assessment of HEENT [Head, Ears, Eyes, Nose, Throat], lungs, 
heart, abdomen, musculoskeletal, age-appropriate neurological and 
skin exam. 
Laboratory 
Blood  
Collect blood for: 
 Screening and pre-study inoculation for RSV serum antibody 
testing.  Note: if the potential participant had been screened but not 
enrolled into IMPAACT 2011, the results from the sample 
collected for that study may be used if the sample was collected 
within the 42-day enrollment window for IMPAACT 2012; it is not 
necessary to collect another sample. 
 Only from participants who are at sites with capacity for 
processing viable PBMCs: Cryopreservation of PBMCs to 
characterize the B cell response.  Remaining plasma to be stored. 
Study Product 
Not applicable 
Schedule Enrollment Visit 
Note that Enrollment must be no more than 42 days after screening 
and within 3 days of randomization.  
 
6.2 
Enrollment Visit 
 
Study enrollment and inoculation must occur between April 1st and October 14th inclusive, 
(outside of the RSV season) and after: 
 
the screening sample confirms that the infant is RSV-seronegative, and 
 
he/she meets all other inclusion/exclusion criteria.  
 
Day 0 will correspond to the day of inoculation with study product. Whenever possible, 
inoculation is to occur on the same day as randomization.  Although sites are allowed up to 3 
days after randomization to conduct the enrollment visit and administer study product, sites 
should not proceed to randomization until a) final eligibility determination has been confirmed 
and b) it has been confirmed that study product can be administered within this window. If these 
2 conditions cannot be met, randomization should be postponed. 
 
If participant is randomized and subsequently noted to have any of the following, inoculation 
must be deferred: 
 
fever (rectal temperature of ≥100.4°F (38°C)), or  
 
upper or lower respiratory symptoms or signs (including but not limited to rhinorrhea, 

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cough, or pharyngitis), or 
 
nasal congestion significant enough to interfere with successful inoculation, or  
 
otitis media 
Note: if the inoculation will not occur within 3 days of randomization, the site MUST contact the 
Protocol Safety Review Team (PSRT) via email at impaact.psrt2012@fstrf.org to explain the 
circumstances and obtain permission to inoculate after the 3-day window period.   
 
If the 42-day window from screening to inoculation is exceeded during the deferral and the infant 
has already been randomized, the team may grant permission for the infant to be rescreened.  If 
found eligible, the infant will be taken off study, re-enrolled and randomized again (possibly to a 
different arm than the original randomization). Sites are advised to avoid this situation by 
scheduling the enrollment visit early in the screening window. 
 
Enrollment Visit Procedures   
Must be no more than 42 days from screening and  within 3 days of randomization  
Administrative 
and Regulatory 
 Complete eligibility determination and confirmation*  
 Complete paper-based eligibility checklist*, enter checklist data 
into SES to enroll/randomize the infant, print and file a copy of the 
confirmation file 
Clinical 
 Obtain interim history  
 Clinical examination: 
o Perform focused clinical examination including temperature, 
heart rate, respiratory rate, HEENT, lung, heart, and lymph 
nodes. 
o Record temperature, pulse, and respirations 
 
Note: Confirm eligibility including clinical examination prior to 
administering study product. 
Laboratory 
 
Blood  
If insufficient volume collected at screening, collect blood: 
 To ensure that back-up aliquot is available for comparing pre- and 
post sera.  
Nasal wash 
Collect nasal wash for: 
 RSV antibody assays 
 RSV viral detection and quantification  
 
Note: The nasal wash must be performed prior to administering the 
study product. 
Study Product 
 Prescribe and prepare/dispense study product 
 Administer study product and maintain participant in a supine 
position for 1 minute. 
 Observe for at least 30 minutes after inoculation to evaluate for 
immediate hypersensitivity reactions. 
Prepare for follow-up 
 Provide the following: temperature card with explanation, temporal 
and rectal thermometers with instructions for use, illness criteria 
explanation, and study personnel contact information.  Schedule 
non-visit day contact for Days 1 and 2 and schedule an in-person 
visit for Day 3. 
*Perform prior to enrollment/randomization 
 
Following the Entry Visit, the parent/guardian will record the infant’s temperatures and signs of 
illness on the temperature card and provide these to study personnel during an in-person visit or 
non-visit day phone, email, or text contact. New rectal thermometers will be given, and temporal 

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artery thermometers will be provided, to parents/guardians for use during the study. For 
temperature measurements, parents/guardians will be instructed to use the study-provided 
temporal artery thermometer to screen for elevated temporal artery temperatures. This device is 
used to minimize the number of rectal temperature measurements and has been shown to be an 
effective screening tool for rectal fever (32). The parent/guardian will measure temporal artery 
temperatures following the manufacturer’s directions.  If any temporal artery temperature is 
≥100.0°F, parents/guardians will be asked to measure a rectal temperature within 20 minutes (32). 
For study-specific management and grading of temperatures, see Section 8.1.1 and Table 4. 
 
6.3 
Acute Phase Visits and Contacts  
 
Refer to Figure 1 for a timeline of study visits. The Acute Phase begins with inoculation and ends 
at midnight on the 28th day after inoculation. During the Acute Phase of the study, a study 
physician, physician assistant, nurse practitioner, or study nurse will be available by telephone 24 
hours a day for consultation with parents/guardians regarding any illnesses that may occur during 
this period. 
 
Study personnel will have daily contact with participant’ parents/guardians for the first 28 days 
after inoculation. This 28-day period is consistent with the duration of shedding of live-attenuated 
respiratory virus vaccines in RSV-seronegative infants and children (24, 25, 32, 33). A clinical 
assessment will be completed during visits on Days 3, 5, 7, 10, 12, 14, 17, and 28 (each visit ±1 
day) after inoculation. This clinical assessment must be performed by a medical professional. 
 
On non-visit days, study staff will contact the parent/guardian and will record the 
parent/guardian-provided temperatures and signs of illness. Participants with illness may have 
additional visits to assess the severity of the illness (see Section 6.10 for Illness Visits).   
 
There will be a final non-visit contact on Day 29 to obtain interim history through midnight on 
the 28th day following inoculation. 
 
Certain events may trigger study pause or stop during the first 56 days following inoculation, 
review Section 8.2 for details associated with pausing and stopping rules. 
 
6.3.1 
Acute Phase Visits: Study Days 3, 5, 7, 10, 12, 14, 17, and 28 (±1 day) 
 
Study visits during the Acute Phase are scheduled to be conducted at the frequency noted above 
with a visit window of ± 1 day.  If an in-person visit is moved by ± 1 day, then telephone contact 
is conducted in place of the original interim visit date.   
 
Events that took place through midnight of Day 28 are considered to have occurred during the 
Acute Phase and will be reported on the non-visit contact conducted on Day 29.  Note that it is 
not necessary to also have a “Day 29” contact (Section 6.4) if the Day 28 Visit is conducted on 
this day. 

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Days 3, 5, 7, 10, 12, 14, 17, and 28 Visit Procedures (each visit from day of inoculation ± 1 day) 
Clinical 
 Obtain interim history 
 Perform focused clinical examination including temperature, heart 
rate, respiratory rate, HEENT, lung, heart, and lymph nodes. 
 Record temperature, pulse, and respirations.   
Laboratory 
 
Nasal wash 
Collect nasal wash for: 
 RSV viral detection and quantification 
Prepare for follow-up 
 Schedule non-visit day contact and follow-up in-person visits.  
 Day 28 only: review SAE criteria with participants and how to 
contact study personnel during Post-Acute Phase (Study Day 30 to 
the Day 56 Visit). 
 
If the infant is diagnosed as having an LRI or otitis media, fever or URI (per Appendix IV) during 
the Acute Phase, evaluations required for the Illness Visit need to be performed and processed. If 
illness criteria are met or suspected (see Section 6.10 and Appendix IV), also request that an 
Adventitious Agent Assay be performed on the nasal wash (see MOP and LPC).    
 
6.3.2 
Acute Phase Contacts: Study Days 1, 2, 4, 6, 8, 9, 11, 13, 15,16, 18, 19, 20, 21, 22, 23, 24, 25, 
26, and 27 (±1 day)  
 
The non-visit contacts during the Acute Phase will be conducted via phone or email.   
 
If the parent reports symptoms suspicious for an LRI, otitis media, fever or URI (per Appendix 
IV), then an Illness Visit should be scheduled (see Section 6.10). 
 
Days 1, 2, 4, 6, 8, 9, 11, 13, 15, 16, 18,19, 20, 21, 22, 23, 24, 25, 26 and  27 Contact Procedures (each 
visit from day of inoculation ± 1 day) 
Interim History 
 Obtain and document from parent/guardian the participant’s 
previous days’ interim history, including any changes in 
medications and/or immunizations.   
 Document highest temperature (temporal or rectal) 
Prepare for follow-up 
 Address any concerns and schedule appointment if necessary. 
 
 
6.4 
Day 29 Contact (+ 1 day) 
 
There will be a non-visit contact on Day 29 to obtain interim history through midnight on the 28th 
day following inoculation. If the Day 28 Visit is conducted on Day 29, it is not necessary to have 
an additional telephone or email contact with the family on Day 29. 
 
Day 29 Non-Visit Procedures (+ 1 day) 
Interim History 
 Obtain interim history through midnight of the 28th day following 
inoculation 
 Document highest temperature (temporal or rectal) 
Prepare for follow-up 
 Address any concerns. 
 Review SAE criteria with participants and how to contact study 
personnel during Post-Acute Phase (Day 30 to the Day 56 Visit). 
 

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6.5 
Post-Acute Phase (Days 30 to 56) 
 
The Post-Acute Phase begins at 12:01 am on the 29th day after inoculation and ends at midnight 
on the 56th day after inoculation. During the Post-Acute Phase, parents/guardians will be 
instructed to monitor for and contact the study site if their infant has symptoms that are 
suggestive a Serious Adverse Event (SAE). If the parent reports an SAE that may meet the study 
pause or stop criteria (Section 8.2) then an Illness Visit should be scheduled (see Section 6.10). 
 
Certain events may trigger study pause or stop during the first 56 days following inoculation; 
review Section 8.2 for details associated with pausing and stopping rules. 
 
6.5.1 
Day 56 Visit (+7 Days) 
 
The Day 56 Visit should be conducted between 56 and 63 days following inoculation. Because 
the Post-Acute Phase ends as of midnight on the 56h day following inoculation, only events 
through that time should be evaluated as having occurred during the Post-Acute Phase. If the Day 
56 visit is conducted on the 56th day after inoculation, sites should arrange a non-visit contact the 
next day to confirm that there were no events between the time of the study visit and midnight of 
the 56th day after inoculation. The evaluations expected at this visit are outlined below.  
 
Day 56 Visit (from day of inoculation +7 Days) 
Clinical 
 Obtain interim history since last contact  
 Record temperature, pulse, and respirations.  
Laboratory 
 
Blood 
Collect blood for: 
 Serum antibodies to RSV 
 Only from participants who are at sites with capacity for 
processing viable PBMCs: Cryopreservation of PBMCs to 
characterize the B cell response. Remaining plasma to be stored. 
Nasal Wash 
Collect nasal wash for: 
 RSV antibody assays 
Prepare for follow-up 
Follow-up depends on when, during the calendar year, the Day 56 
visit is conducted. If the Day 56 Visit is conducted: 
 Prior to October 1st, schedule for Pre-RSV Season Visit (Section 
6.7) 
 On or after October 1st the Day 56 Visit will also be the Pre-RSV 
Season Visit. Therefore, review plans for weekly contact during 
the RSV Season Surveillance Period (see Section 0). 
 
6.6 
Period after Day 56 Visit until October 31st 
 
During this period, contact with the participant is not required except for the Pre-RSV Season 
Study Visit described in Section 6.7. No clinical data will be recorded on CRFs or reported under 
this protocol except for data as outlined in Table 2 in Section 7.2. 
 
6.7 
Pre-RSV Season Study Visit (October 1st to 31st) 
 
The Pre-RSV Season Study Visit is not required if the Day 56 Visit is conducted on or after 
October 1st (i.e., the sample collected at the Day 56 Visit is sufficient for the Pre-RSV Season 
Study Visit). Otherwise, an in-person visit is expected of participants during the Pre-RSV Season 
for collection of a blood sample.  
 

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Pre-RSV Season Study Visit (October 1st to 31st) 
Laboratory 
 
Blood 
Collect blood for: 
 Serum antibodies to RSV  
 Only from participants who are at sites with capacity for 
processing viable PBMCs: Cryopreservation of PBMCs to 
characterize the B cell response. Remaining plasma to be stored. 
Prepare for follow-up 
 Review plans for weekly contact during the RSV Season 
Surveillance Period (see Section 0). 
 
6.8 
RSV Season Surveillance (November 1st through March 31st following inoculation) 
 
Based on previous data regarding the seasonality of RSV in the Baltimore, MD area (Appendix 
V), surveillance for RSV-associated disease will be conducted between November 1st and March 
31st. Note, RSV Season Surveillance may overlap with the Study Acute and Post-Acute Phase. In 
this case, evaluations required for each of the relevant phases of the study will be conducted. 
During the RSV season following receipt of study product, participants enrolled in this study will 
be monitored for symptomatic, medically attended, RSV-like illnesses listed below. These 
contacts may be by weekly telephone, email, or an in-person visit. Information about these 
illnesses will be obtained during the RSV Season Surveillance Period by weekly 
communication between study personnel and the participant’s parent/guardian. For this period, 
determine if any of the following medically attended events occurred.  Please note that the 
symptoms below do not need to meet the Appendix IV criteria. 
o Medically attended fever 
o Medically attended upper respiratory illness  
o Medically attended otitis media 
o Medically attended lower respiratory illness  
 
An Illness Visit should be scheduled within 3 days of a site’s study staff notification of any of 
these events (see Section 6.10).   
 
RSV Season Surveillance (November 1st through March 31st following inoculation) 
Clinical 
 Obtain interim history 
Prepare for follow-up 
 Continue with weekly contacts through March 31st  
 Schedule an Illness Visit if warranted  
 Schedule Post-RSV Season Study Visit to take place between April 
1st and 30th  
 
 
 

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6.9 
Post-RSV Season Study Visit (April 1st to 30th) 
 
There will be a single in-person visit between April 1st and 30th.   
 
Post-RSV Season Study Visit (April 1st to 30th) 
Administrative 
and Regulatory 
 Inform participant’s parent/guardian of study randomization, if 
known. 
Laboratory 
 
Blood 
Collect blood for: 
 Serum antibodies to RSV. 
 Only from participants who are at sites with capacity for 
processing viable PBMCs: Cryopreservation of PBMCs to 
characterize the B cell response. Remaining plasma to be stored. 
 
6.10  Illness Visit 
 
The timeframe after site notification in which the Illness Visit must occur depends on grading of 
the fever and respiratory symptoms per Section 7.3.3 and Appendix IV and the phase of the 
study. If the Illness Visit occurs on a day concurrent with a routine Study Visit, a single nasal 
wash collection is required, however, an Adventitious Agent request form should be completed. 
Following an Illness Visit, sites should continue to follow participants until resolution of 
symptoms. Illness visits may occur during any of the following phases of the study.  
  
 
Acute Phase: For fever, otitis media, or URI, the Illness Visit must be conducted within 3 
days if Grade 1 and within 2 days if Grade ≥ 2. 
 
Acute Phase: For a possible LRI, with any grade, the assessment will occur within 1 day. 
 
Post-Acute Phase: For an SAE that may meet the study pause or stop criteria (Section 
8.2), an Illness Visit must be conducted within 3 days of site notification. 
 
RSV Season Surveillance Period (between November 1st and March 31st): The Illness 
Visit should be scheduled within 3 days of site notification of a medically-attended 
illness of the following types: fever, URI, LRI or otitis media. However, if this phase 
overlaps with the Acute or Post-Acute Phases, the time frames specified for the relevant 
Acute or Post-Acute phase should be used. 
 
Illness Visit Procedures 
Clinical 
 Obtain interim history 
 Perform focused clinical examination including temperature, heart 
rate, respiratory rate, HEENT, lung, heart, and lymph nodes. 
 Record temperature, pulse, and respirations.   
Laboratory 
 
Nasal Wash 
Collect nasal wash for: 
 Viral detection and quantification 
 Complete Adventitious Agent Assay Request for rRT/PCR on 
nasal wash for adventitious agents (see MOP and LPC). 
Prepare for follow-up 
 Schedule follow-up as appropriate 
 
 
6.11 Early Discontinuation Study Visit 
 
In the event that a participant is unable to continue participation in the study, every effort should 
be made to schedule a final Early Discontinuation Visit. 
 

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Early Discontinuation 
Clinical 
 Obtain interim history   
Laboratory 
 
Blood 
Collect blood for: 
 Serum antibodies to RSV 
 Only from participants who are at sites with capacity for 
processing viable PBMCs: Cryopreservation of PBMCs to 
characterize the B cell response. Remaining plasma to be stored. 
Nasal Wash 
If Early Discontinuation Visit is conducted within 56 Days of 
inoculation (Appendix II), collect nasal wash for: 
 Viral detection and quantification 
 RSV antibody assays 
 
6.12 Additional Considerations for Laboratory Procedures 
 
Each study site and laboratory involved in this study will comply with the DAIDS policy on 
Requirements for DAIDS Funded and/or Sponsored Laboratories in Clinical Trials Policy, which 
is available at: 
http://www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/Laboratories.aspx  
 
6.12.1 Specimen Collection 
 
Specimens will be collected for this study as indicated in the Schedule of Evaluations and per 
detailed guidance provided in the Laboratory Processing Chart (LPC), which will be available on 
the IMPAACT web site: www.impaactnetwork.org. Further information on collection of blood 
and nasal wash specimens will also be provided in the MOP. 
 
In accordance with US National Institutes of Health (NIH) recommendations, pediatric blood 
collection will not exceed 5 mL/kg in a single day or 9.5 mL/kg in any eight-week period.   
 
Virus Detection 
 
Specimens for viral culture and quantification of vaccine virus shedding will be collected by nasal 
wash with approximately 20 mL of Ringer’s lactate solution once before and approximately 8 
times after inoculation as shown in Appendix II. These specimens may also be tested for 
adventitious agents if the participant is ill. Additional nasal wash specimens for detection of RSV 
and adventitious respiratory viruses by culture and rRT-PCR will also be collected from 
participants who meet illness criteria during the initial phase (Day 0 to the Day 56 Visit) and RSV 
Season Surveillance Period (November 1st – March 31st). Laboratory testing will be performed by 
personnel that are not involved with clinical assessment to maintain the blinding of the study. 
 
6.12.2 Specimen Preparation, Testing, Storage, and Shipping 
 
All specimens collected for this study will be labeled, transported, processed, tested, stored and/or 
shipped in accordance with the DAIDS policy referenced in Section 6.12, site and local 
laboratory SOPs, and the LPC. The frequency of specimen collection and testing will be directed 
by the Schedule of Evaluations (Appendices II and III). The Laboratory Data Management 
System (LDMS) will be used to document specimen collection, testing, storage, and shipping as 
specified in LPC.  
 

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Virologic and Immunologic Assays 
 
Serum specimens will be collected up to 42 days prior to inoculation for the screening and the 
study pre-inoculation, and at the Day 56 Visit for measurement of post-inoculation serum 
antibodies to RSV. In addition, pre-RSV season and post-RSV season serum specimens will be 
collected. These samples will be used to determine whether a fourfold or greater rise in antibody 
titer has occurred during the RSV season, which would signify infection with wt RSV. This 
will allow comparison of the rate and severity of significant RSV illness following infection 
with wt virus, as well as comparison of the antibody responses, in vaccine and placebo 
recipients.  
 
Blood specimens for cryo-preservation of PBMCs will be collected up to 42 days prior to 
inoculation and at the Day 56 Visit to analyze the B cell response to the vaccine. In addition, pre-
RSV season and post-RSV season PBMCs will be collected. These samples will be used to 
analyze the B cell response to vaccination, and to wt RSV infection during the surveillance 
season. Plasma leftover from the processing of the PBMCs will be saved. Analysis of the B cell 
response may include sequencing of participant genes related to their immune response to the 
vaccine or RSV if the parent/guardian consents to limited genetic testing. 
 
Nasal wash specimens for measurement of secretory immunity will be collected from participants 
before inoculation and at the Day 56 Visit after inoculation. These specimens may be generated 
from the same nasal wash used for viral culture, except that the aliquot for measurement of 
secretory immunity will not contain viral transport medium. 
 
Quantitation of the amount of vaccine virus shed, assays to measure immune responses before 
and after inoculation, and assessment of nasal washes for the presence of adventitious viral agents 
will be performed at the JHU CIR. Cytokine/chemokine assays may also be performed on nasal 
washes from participants infected with vaccine virus if sufficient material is available. Selected 
specimens may be sent to LID, NIAID for confirmatory testing.  PBMC analyses will be 
performed at LID, NIAID. 
 
Plan for Future Use and Storage of Biological Sample 
 
All specimens collected as part of this study may, with the parent/guardian’s permission, be 
stored for future research as part of JHU CIR’s approved biospecimen repository for vaccine 
research or at the LID, NIAID. These samples may be used for future screening for respiratory 
virus vaccine studies and to learn more about RSV infection and other diseases. These samples 
will not be sold or used to make commercial products. With permission, limited human genetic 
tests may be performed on these samples. Samples will be stored only with the parent/guardian’s 
permission.  
 
All samples stored in the repository will be labeled with the participant identification (PID) 
numbers of the participants that, by themselves, cannot identify study participants but are linkable 
to the study databases generated by the main study. The repository database will contain only the 
study participants’ PID numbers. A master log linking the study participants’ PID numbers is 
maintained at the individual enrolling site and will not be shared with the Protocol Team or the 
laboratory at CIR. Study participants, or their parents/guardians, may withdraw consent for future 
testing of their specimens at any time. 
 
In the future, other investigators (both at NIH and outside) may wish to study these samples 
and/or data. In that case, IRB approval must be sought prior to any sharing of samples and/or 

IMPAACT 2012, Version 1.0 
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data. Any clinical information shared about the sample would similarly require prior IRB 
approval. 
 
6.12.3 Biohazard Containment 
 
As the transmission of blood-borne pathogens can occur through contact with contaminated 
needles, blood, and blood products, appropriate blood and secretion precautions will be employed 
by all personnel in the drawing of blood and shipping and handling of all specimens for this 
study, as currently recommended by the Centers for Disease Control and Prevention and the 
National Institutes of Health. Additional specimen collection and processing precautions are 
included in the MOP. All infectious specimens will be transported using packaging mandated in 
Title 42 of the Code of Federal Regulations, Part 72 (42 CFR 72) and in accordance with 
individual carrier guidelines (e.g., Federal Express, Airborne Express). 
 
7 
SAFETY ASSESSMENT, MONITORING, AND REPORTING 
 
Participant safety will be carefully assessed, monitored, and reported at multiple levels 
throughout this study. Sections 7.1-7.3 describe safety-related roles, responsibilities, and 
procedures for site investigators. The safety monitoring roles of the Protocol Safety Review Team 
(PSRT) and the Data Safety and Monitoring Board (DSMB) are briefly referenced in Section 7.1 
and described in detail in Sections 9.4.1 and 9.4.2. 
 
7.1 
Safety-Related Roles and Responsibilities 
 
7.1.1 
Site Investigators 
 
Site investigators are responsible for continuous monitoring of all study participants and for 
alerting the Protocol Team if unexpected concerns arise. Site investigators will record safety-
related data on CRFs as indicated in Section 7.2 and complete expedited adverse event (EAE) 
reporting as indicated in Section 7.3.  Site investigators are also responsible for prompt reporting 
to their IRBs/IBCs and other applicable review bodies of any unanticipated problems involving 
risks to participants or others. 
 
7.1.2 
Protocol Safety Review Team 
 
A Protocol Safety Review Team (PSRT) will routinely review clinical and laboratory safety data 
reports prepared by the SDMC. Designees for PSRT members will be allowed in the event of 
their non-availability for a review. To meet minimum quorum for a safety data review the PSRT 
must include (but is not limited to): 
 
the Protocol Chair or Vice Chair 
 
Data Manager 
 
DAIDS or NICHD Medical Officer 
 
the Protocol Statistician 
 
The content, format and frequency of safety monitoring will be agreed upon in advance by the 
PSRT.  Representatives of the product developer may also be included on PSRT discussions, but 
not in decisions. 
 
The PSRT will convene via teleconference or other means routinely throughout the study to 
review data relevant to safety monitoring and discuss any safety concerns – at least twice per 

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month during the active immunization phases -- and at least once a month thereafter, as well as on 
an ad hoc (as needed) basis outside of regularly scheduled calls.  The PSRT will also provide 
rapid consultation to site clinicians regarding toxicity management as needed. 
 
On behalf of the full Protocol Team, the PSRT will monitor participant safety through routine 
review of study data reports as described in Section 9.4.1.  
 
7.1.3 
Data Safety Monitoring Board 
 
An independent DSMB will monitor participant safety through routine and as needed reviews of 
study data.  Refer to Section 9.4.2 for more information on the composition and role of the 
DSMB in monitoring of this study.   
 
7.1.4 
Sponsor Reporting  
 
Serious and unexpected suspected adverse reactions (SUSAR) as defined in 21 CFR 312.32 will 
be determined by the IND Sponsor and reported to the FDA and all participating investigators as 
IND Safety Reports. The sponsor will also submit a brief report of the progress of the 
investigation to the FDA on an annual basis as defined in 21 CFR 312.33  
 
7.2 
Safety-Related Recording on Case Report Forms 
 
Any event that occurs during protocol-specified AE reporting periods (see Table 2), following 
inoculation with study product, is to be considered an AE and should be evaluated as a potential 
Expedited AE (per DAIDS). The current section outlines which events should be collected on 
source documents and which should be recorded on CRFs for inclusion in the database. 
 
AEs may be observed by the site investigator, elicited from the parent/guardian or participant, 
captured on participant’s temperature cards, or volunteered by the parent/guardian or participant. 
Assessment of safety will include clinical observation and monitoring of hematological, 
chemical, and immunologic parameters as necessary. Follow-up such as history, physical 
examination, and laboratory testing and/or treatment may be necessary if a participant 
experiences an AE. Details of AEs will be properly collected on the source documents, recorded 
on CRFs, and provided to PSRT and the DSMB in separate semi-annual and annual reports. AEs 
will be provided to the IRB as defined by the individual IRB policy. 
 
This study has several periods of AE surveillance that have different AE CRF recording 
requirements. In addition, there may be a period when no AEs are recorded on CRFs if the Day 
56 Visit is conducted in advance of the start of the RSV Season Surveillance Period (November 
1st). The adverse events (solicited and unsolicited; and SAEs) to be recorded on CRFs and the 
study phase and the calendar dates during which they are to be reported are defined in Table 3. 
 
Adverse events identified in this study will be recorded on CRFs as signs, symptoms, laboratory 
test results, and diagnoses, as described in Table 2.  Expectations regarding the recording of 
Concomitant Medications are also detailed in this table. 

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Table 2: AE CRF Recording Requirements 
Study Phase at the 
time of event onset 
Calendar Date 
AEs to record on CRFs 
Concomitant Medications 
to record on CRFs 
Days 0 through 
midnight of 28th 
day following 
inoculation 
(Acute Phase) 
ANY 
 All SAEs 
 All solicited AEs that meet Appendix 
IV criteria  
 All unsolicited AEs (Grades 1 to 4), 
with the exception of the following 
conditions if not treated with 
prescription medication or OTC 
medications with antipyretic 
properties: diaper rashes, teething 
pain, and spitting up. 
 
Note: SAEs and LRIs must be reported 
via DAIDS Adverse Experience 
Reporting System (DAERS; see Section 
7.3.4). 
 All cough and cold 
remedies including 
decongestants, cough 
suppressants, 
expectorants 
 All nasal sprays (except 
saline spray) 
 All antihistamines 
 All antipyretics 
 All prescription 
medications 
 
For SAE and LRIs: 
 All concomitant 
medications related to 
the recorded event 
From 12:01 am on 
29th day after 
inoculation to 
midnight of the 
56th day following 
inoculation 
(Post-Acute Phase) 
ANY 
 All SAEs 
 
Note: SAEs must be reported via 
DAERS (see Section 7.3.4). 
 All concomitant 
medications related to 
the recorded event 
After Day 56 Visit 
and until RSV 
Season 
Surveillance Period 
Up to October 
31st in year of 
inoculation 
 Grade ≥3 AE or SAE that is deemed 
related to Pre-RSV Season Study Visit 
procedures. 
 All concomitant 
medications related to 
the recorded event 
RSV Season 
Surveillance Period 
November 1st 
to March 31st 
following 
inoculation 
 Fever, LRI, URI, and/or otitis media 
that are medically attended  
 All SAEs  
 
Note: these events do not need to meet 
the Appendix IV criteria. SAEs and 
medically attended LRIs must be 
reported via DAERS (see Section 
7.3.4). 
For SAE and LRIs: 
 All concomitant 
medications related to 
the recorded event 
 
Medications related to 
recorded medically 
attended illness should be 
documented in source 
notes. 
Post-RSV Season 
April 1st-April 
30th in the year 
after the 
inoculation 
 Grade ≥3 AE or SAE that is deemed 
related to Post-RSV Season Study 
Visit procedures. 
 All concomitant 
medications related to 
the recorded event 
Throughout study 
ANY 
 Unresolved AE or SAE with onset 
date during Day 0 to midnight on the 
28th day after inoculation 
 Unresolved SAE with onset date prior 
to midnight on the 56th day following 
inoculation 
 Unresolved SAE with onset date 
during RSV Surveillance Period or 
related to the Pre- or Post-RSV Season 
Study Visit 
 All concomitant 
medications related to 
the recorded event 
 

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7.3 
Expedited Adverse Event (EAE) Reporting 
 
7.3.1 
Adverse Event Reporting to DAIDS 
 
Requirements, definitions, and methods for expedited reporting of adverse events (AEs) are 
outlined in Version 2.0 of the DAIDS EAE Manual, which is available on the RSC website at 
http://rsc.tech-res.com/safetyandpharmacovigilance. 
 
The DAIDS Adverse Experience Reporting System (DAERS), an internet-based reporting 
system, must be used for expedited AE reporting to DAIDS. In the event of system outages or 
technical difficulties, expedited AEs may be submitted via the DAIDS EAE Form. For questions 
about DAERS, please contact NIAID CRMS at CRMSsupport@niaid.nih.gov. Site queries may 
also be sent from within the DAERS application itself. 
 
Where DAERS has not been implemented, sites will submit expedited AEs by documenting the 
information on the current DAIDS EAE Form. This form is available on the RSC website: 
http://rsc.tech-res.com/safetyandpharmacovigilance/. For questions about expedited reporting, 
please contact DAIDS RSC (DAIDSRSCSafetyOffice@tech-res.com). 
 
7.3.2 
Reporting Requirements for this Study 
 
 
The SAE Reporting Category, as defined in Version 2.0 of the DAIDS EAE Manual, will 
be used by this study. 
 
In addition to the EAE Reporting Category identified above, other AEs that must be 
reported in an expedited manner are LRIs as defined in Appendix IV if occurring from 
Day 0 to midnight on the 28th day following inoculation. 
 
The study agent for which expedited reporting is required is recombinant live-attenuated 
respiratory syncytial virus vaccine LID cp ΔM2-2, Lot RSV#009B/placebo. 
 
After the protocol-defined AE reporting period, unless otherwise noted, only SUSARs as 
defined in Version 2.0 of the EAE Manual will be reported to DAIDS if the study staff 
become aware of the events on a passive basis (from publicly available information). 
 
With respect to the relationship categories specified for purposes of participant and 
toxicity management in Section 8.1, the categories of definitely related, probably related, 
and possibly related will correspond to an assessment of “related” for EAE reporting; the 
categories of probably not related and not related will correspond to an assessment of 
“not related” for EAE reporting. 
 
7.3.3 
Grading Severity of Events 
 
All solicited AEs and fever will be graded following protocol-defined grading system outlined in 
Table 3 and Table 4. Other AEs (i.e., excluding solicited AEs and fever) will be assessed for 
severity by the site investigator using the Division of AIDS Table for Grading the Severity of 
Adult and Pediatric Adverse Events (DAIDS AE Grading Table): Version 2.0, dated November 
2014. In the event that this table is updated following protocol implementation, events will 
continue to be evaluated per this version of the DAIDS AE Grading Table.  The DAIDS AE 
Grading Table is available on the RSC website at http://rsc.tech-
res.com/safetyandpharmacovigilance. 
 
 
 

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Solicited AE Grading 
 
Table 3: Grading Table for Solicited AEs 
 
Severity 
Defined 
Grade (0) None 
Not applicable 
Grade (1) Mild 
No medical intervention required; may include use of over-the-counter 
medications managed by the caregiver for treatment of symptoms 
Grade (2) Moderate 
Outpatient medical intervention by a health care provider required; may 
include use of over-the-counter and/or prescription medications 
Grade (3) Severe 
Prolonged medical intervention and/or hospitalization required 
Grade (4) Life 
threatening 
Illness requiring hospitalization with intensive care  
Grade (5) Death 
Event resulting in fatal outcome to the participant 
 
Fever Grading: Temperature Measurement 
Table 4: Fever Grading* 
 
 
 
 
 
Severity 
Defined 
Grade (0) 
≥100.0ºF but <100.4ºF (≥37.8ºC but <38ºC) 
Grade (1) 
≥100.4ºF but ≤101.4ºF (≥38ºC but <38.6ºC) 
Grade (2) 
≥101.5ºF but ≤102.4ºF (≥38.6ºC but ≤39.1ºC) 
Grade (3) 
≥102.5ºF but ≤104.8ºF (≥39.2ºC but ≤40.4ºC) 
Grade (4) 
≥104.9ºF (≥40.5ºC)  
*Applies to any modality of temperature measurement Expedited AE Reporting Period 
 
The expedited AE reporting period for this study is defined in Section 7.3.4. 
 
7.3.4 
Expedited AE Reporting Period 
 
Table 5 details the events that must be reported in an expedited fashion via DAERS during 
specific periods of the study. 
 
 

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Table 5: EAE Reporting 
 
Study Phase at the time of 
event onset 
Calendar Date 
Events to Report via 
DAERS 
Days 0 through midnight of 28th day 
following inoculation 
(Acute Phase) 
ANY 
 SAEs  
 LRIs 
From 12:01 am on 29th day after 
inoculation to midnight of the 56th day 
following inoculation 
(Post-Acute Phase) 
ANY 
 SAEs 
 
After Day 56 Visit and until RSV 
Season Surveillance Period 
 
Up to October 31st in year of 
inoculation 
 Follow-up related to 
SAEs/LRIs already 
reported 
Pre-RSV Season 
October 1st-October 31st in the year 
of inoculation 
 SAEs related to Pre-
RSV Season Study Visit 
procedures 
 
RSV Season Surveillance Period 
November 1st to March 31st 
following inoculation 
 
 SAEs 
 Medically attended 
LRIs 
 
Post-RSV Season 
April 1st-April 30th in the year after 
the inoculation 
 SAEs related to Post-
RSV Season Study Visit 
procedures  
 
 
 
 
8 
PARTICIPANT MANAGEMENT 
 
8.1 
Management of Adverse Events 
 
An adverse event (AE) is any untoward medical occurrence in a patient or clinical investigation 
participant administered a pharmaceutical product and which does not necessarily have a causal 
relationship with this treatment. An AE can therefore be any unfavorable and unintended sign 
(including an abnormal laboratory finding), symptom, or disease temporally associated with the 
use of a medicinal (investigational) product, whether or not related to the medicinal 
(investigational) product. This includes exacerbation of pre-existing conditions and intercurrent 
illnesses.   
 
All adverse events identified in this study will be source documented in infant research records, 
consistent with the policies and procedures referenced in Section 10.  Among other details, source 
documentation will include the severity of each event (graded as described in Section 7.3.3) and 
its relationship to study product, assessed by the site clinician (see below).   Further standardized 
guidance on determining whether there is a reasonable possibility of a relationship is available in 
the DAIDS EAE Manual, referenced in Section 7.3.1. 
 
Relationship categories for Adverse Events are as follows: 
 
 
Definitely related 
The event and administration of study drug are related in time, and a 
direct association can be demonstrated. 

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Probably related 
The event and administration of study drug are reasonably related in time 
and the event is more likely explained by study drug than other causes. 
 
Possibly related 
The event and administration of study drug are reasonably related in time 
and the event can be explained equally well by causes other than study 
drug. 
 
Probably not related A potential relationship between the event and study drug could exist 
(i.e., the possibility cannot be excluded), but the event is most likely 
explained by causes other than study drug. 
 
Not related 
The event is clearly explained by another cause not related to study drug.  
 
There are two categories of AEs specific to IMPAACT 2012: solicited and unsolicited.  Solicited 
AEs are described in Section 8.1.1. Unsolicited AEs are all other AEs. 
 
Serious Adverse Events (SAEs) are described in Section 8.1.2. 
 
8.1.1 
Solicited Adverse Events 
 
Solicited AEs are predefined AEs that can occur after inoculation with study product, may be 
expected to occur if the study product is insufficiently attenuated, and have protocol-specific 
criteria for reporting.  
 
Solicited AEs, whether identified by a parent/guardian or clinician, are only recorded on CRFs if 
they met the definitions per Appendix IV. Individual symptoms listed in the “events” column that 
fail to meet the criteria in the “definition” column in Appendix IV are recorded in source 
documents but are not recorded on the CRFs.  During the Acute Phase of this study, Days 0 to 28, 
solicited AEs meeting the criteria for reporting will be recorded on CRFs, assigned a severity 
grade (Section 7.3.3 and Table 3 and Table 4), and assessed for relationship to study product (see 
Section 8.1).  For this study, the solicited AEs are defined in Appendix IV and include the 
following:  
 
1. Fever 
2. Upper respiratory illness (URI) 
a. Rhinorrhea, 
b. Pharyngitis,  
c. Cough without LRI, or 
d. Hoarseness. 
3. Otitis Media 
4. Lower respiratory illness (LRI) 
a. Wheezing, 
b. Pneumonia, 
c. Laryngotracheobronchitis (croup), 
d. Rhonchi, or 
e. Rales. 
 

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Solicited Adverse Events Elicited by History Unconfirmed by Clinical Assessment 
 
Solicited AEs and symptoms reported by parents/guardians are NOT recorded on CRFs if a 
clinical assessment done on the day of the event(s) does/did not confirm their presence. For 
example, if a parent/guardian reports rhinorrhea on the day of visit, and there is/was no rhinorrhea 
upon exam, then the participant is considered to not have rhinorrhea that day. 
 
Events elicited by parent/guardian history for days on which there was no clinical exam will be: 
 
 
Recorded on the CRFs as AEs if the parent/guardian description meets the “definition” 
column criteria in Appendix IV. 
 
Recorded only on the source document, and NOT on the CRF, if the parent/guardian 
description fails to meet the “definition” column criteria in Appendix IV. For example, both 
rhinorrhea and cough must each occur on 2 consecutive days to meet the definition required 
for reporting per Appendix IV. 
 
8.1.2 
Serious Adverse Event 
 
A Serious Adverse Event (SAE) is an AE, whether considered related to the study product or not, 
that: 
1. Results in death during the period of protocol-defined surveillance 
2. Is life threatening: defined as an event in which the participant was at immediate risk 
of death at the time of the event; it does not refer to an event that hypothetically 
might have caused death were it more severe 
3. Requires inpatient hospitalization (or prolongation of existing hospitalization): 
defined as at least an overnight stay in the hospital or emergency ward for treatment 
that would have been inappropriate if administered in the outpatient setting 
4. Results in a persistent or significant disability/incapacity 
5. Is a congenital anomaly or birth defect 
6. Is an important medical event that may not be immediately life threatening or result 
in death or hospitalization but may jeopardize the patient or may require intervention 
to prevent one of the outcomes listed above. 
 
8.2 
Pausing and Stopping Rules 
 
If any of the following occur in a participant during the specified period after he/she receives 
study product), additional inoculations and continued enrollment will be temporarily suspended at 
all sites: 
 
 
Through midnight of the 56th day following inoculation: 
o An SAE that cannot be attributed to an etiology or cannot be attributed to a cause 
unrelated to the study product, OR 
 
Through midnight of the 28th day following inoculation: 
o An LRI per Appendix IV, OR 
o A fever of Grade 4 OR  
o Any Grade 3 or above solicited AE (other than fever)  
 
If any of these events occur, the site will report the event (as outlined in Sections 7.3) AND will 
notify the PSRT of the event (including a description of the event) via email at 
impaact.psrt2012@fstrf.org within 24 hours of site notification. The Protocol Team will notify all 

IMPAACT 2012, Version 1.0 
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sites to suspend enrollment and inoculation. The site reporting the event will complete the event 
assessment including the collection of viral samples. Respiratory viral samples collected from the 
participant up to that point will be shipped to the Johns Hopkins University laboratory as soon as 
possible (see MOP). Study accrual will remain suspended while the SDMC checks study product 
assignment and virology studies are started. The DSMB will be informed and receive pertinent 
data regarding the event from the Protocol Chair. The study sponsor will also be notified. Follow-
up visits for participants already inoculated will continue as outlined in Appendix II. 
 
If the event is determined to have occurred in a participant who received active agent (vaccine), 
and the event meets one of the following stopping rule criteria, then the event will be reviewed by 
the DSMB prior to resuming enrollment. (Note: If the event is determined to have occurred in a 
participant who received placebo or in a participant who received active agent (vaccine) but had 
no viral shedding associated with the event, the DSMB may review the event by phone without 
calling a formal meeting.) 
 
1. One or more participants experiences an SAE that cannot be attributed to an etiology 
or cannot be attributed to a cause unrelated to the study vaccine OR   
2. One or more participants develops LRI associated with shedding of vaccine virus at 
the time of the LRI (even if another pathogen is identified, unless the RSV is 
confirmed to be wt RSV), OR 
3. One or more participants develops LRI that is not explained by a diagnosis unrelated 
to the vaccine virus, OR  
4. One or more participants experiences a Grade 4 fever or any Grade 3 or Grade 4 
solicited AE other than fever associated with shedding of vaccine virus, OR 
5. Any pattern of research laboratory values or clinical symptoms is observed that the 
Protocol Team considers a significant safety issue for participants. 
 
The DSMB will notify the Protocol Chair (via the study sponsor) of their recommendations, and 
the sponsor and PSRT will determine if enrollment can resume, or if the study needs to be 
stopped. In the event of an SAE, the study may be resumed if it can be demonstrated to the 
DSMB that there is no proven causal relationship with the vaccine. In all cases, once a pause 
occurs, the sites cannot resume enrollment or inoculation until notified to do so by the Protocol 
Team. 
 
9 
STATISTICAL CONSIDERATIONS 
 
9.1 
General Design Issues 
 
9.1.1 
General Design 
 
The goal of this Phase I, blinded, randomized, placebo-controlled vaccine trial is to assess the 
safety, infectivity, and immunogenicity of the LID cp ∆M2-2 vaccine candidate in RSV-
seronegative pediatric participants. Thirty-three participants will be randomized in a 2:1 ratio to 
receive either the candidate vaccine or placebo. Data from this study and from a companion study 
of this vaccine conducted at the Center for Immunization Research (CIR), Johns Hopkins 
Bloomberg School of Public Health, will be maintained in the same database and will be analyzed 
together. A subset of the study team will be unblinded to study results prior to completion of the 
study follow-up (see Section 9.4.1 for details).  
  

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9.1.2 
Description of the Statistical Methods to be Employed 
 
This study, like other Phase I studies, is exploratory, rather than confirmatory; its purpose is to 
assess frequencies of adverse events and patterns of immune responses. Descriptive approaches 
will be used to meet the protocol objectives as stated in Section 2 of this protocol, as well as 
formal statistical tests as outlined below.  
 
9.2 
Outcome Measures 
 
9.2.1 
Primary Outcome Measures 
 
o Safety: types and grades of study product-related: 
 
solicited AEs as defined in Appendix IV from Study Day 0-28 
 
unsolicited AEs from Study Day 0-28 
 
SAE (as defined in Section 8.1.2) from Study Day 0-56 
o Infectivity: 
 
infection with RSV defined as 1) vaccine virus identified in a nasal wash from Study 
Day 0-28 (a binary outcome based on nasal washes done throughout the study period; 
Day 0 nasal wash will be counted as baseline) or 2) ≥4-fold rise in RSV neutralizing 
antibody titer from Study Day 0-56 
 
peak titer of vaccine virus shed from Study Day 0-28 
 
duration of virus shedding in nasal washes as determined by a) culture and b) RT-
PCR from Study Day 0-28 
 
o Immunogenicity: 
 
≥4-fold rise in RSV-neutralizing antibody titer from study entry to Study Day 56 
 
antibody responses to RSV F glycoprotein as assessed by ELISA at study entry and 
Study Day 56 
 
9.2.2 
Secondary Outcome Measures 
 
 
The frequency and severity of symptomatic, medically attended respiratory and febrile illness in 
the vaccine and placebo recipients who experience natural infection with wt RSV during the 
subsequent RSV season. 
 
The antibody responses in the vaccine and placebo recipients who experience natural infection 
with wt RSV during the subsequent RSV season. 
 
The B cell responses to vaccine and the quality and epitope specificity of RSV F specific 
antibody 
 
9.3 
Sample Size and Accrual 
 
 
 
9.3.1 
Sample Size and Randomization 
 
Approximately 33 RSV-seronegative infants will be enrolled in the study (IMPAACT 2012 and 
CIR 312 combined) and will receive either vaccine or placebo in a 2:1 ratio. Assuming an 
attrition rate of about 10%, approximately 20 vaccine and 10 placebo recipients will provide data 
for the primary objectives. The sample size was chosen based upon past experience with Phase I 
evaluation of other live-attenuated respiratory virus candidate vaccines (5-7). The 2:1 
randomization ratio will be used to maximize the information obtained regarding the response of 

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infants to the LID cp ∆M2-2 vaccine. Permuted block randomization will be used to ensure that 
the 2:1 ratio of treated to control participants will be maintained across time. 
 
Given the small sample size, the study will have limitations with respect to detecting AEs and in 
estimating the rates of such events in the population represented by the study sample. 
 
The following calculations focus on the assessment of the tolerability of the vaccine (Section 2.1) 
and in particular, occurrence of LRI, which occurs very infrequently in children who participate 
in these types of studies but would be considered a sentinel safety event if observed in 
participants infected with vaccine virus.   
 
Table 6 shows the probability of observing 0 events of LRI within the sample of 20 vaccinees, as 
well as the probability of observing 1 or more events, or 2 or more events, under a range of 
assumptions concerning the true rate of such events in the participant population represented by 
this sample. From this table, it can be seen that if the true proportion of LRI (or other AE) is at 
least 10%, there is a 61% chance of observing 2 or more events in a group of size 20, and an 88% 
chance of observing at least a single event. 
 
 
 
Table 6: The Probability of Observing LRI events in Vaccinees 
 
N=20 
True underlying probability of LRI 
or AEs 
Pr  
(0 events) 
Pr  
(1+ events) 
Pr 
(2+ events) 
.01 
.82 
.18 
.02 
.03 
.54 
.46 
.12 
.05 
.36 
.64 
.26 
.1 
.12 
.88 
.61 
.15 
.04 
.96 
.82 
 
Table 7 presents 90% confidence intervals (CIs) around potential rates of LRI or AEs that might 
be observed in the sample of 20 vaccinees.  The CIs around similar rates in a sample of 10 
placebo recipients are also presented.  Note that if no LRI or AEs are detected among the 20 
vaccinees, we are 90% confident that the true probability of AEs in the population from which the 
sample is drawn is between 0 and 14%. 
 
Table 7: Percent of Participants Experiencing LRI or AEs with Exact 90% 
Confidence Intervals 
 
N 
% LRI or AEs 
90% CI 
10 
0% 
0% -- 26% 
20 
0% 
0% -- 14% 
10 
10% 
1% -- 39% 
20 
10% 
2% -- 28% 
10 
20% 
4% -- 51% 
20 
20% 
7% -- 40% 
10 
30% 
9% -- 61% 
20 
30% 
14% -- 51% 
 

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Group sample sizes of 20 in the vaccinated group and 10 in the placebo group would achieve 
80% power to detect a difference between the group proportions of about 0.45. The test statistic 
used is the one-sided Fisher's Exact test. The alpha level of the test was targeted at 0.05. Table 8 
presents examples of true group differences that can be detected with 80% power, and Figure 4 
displays graphically power curves for 90% power, 80% power, and 70% power given the sample 
sizes. 
 
Table 8: Magnitude of Difference in Responses Detectable with 80% Power 
 
Response 
Proportion in the 
Placebo Group 
Response Proportion in 
the Vaccinated Group 
Difference 
0.05 
0.52 
0.47 
0.1 
0.59 
0.49 
0.15 
0.65 
0.50 
0.2 
0.71 
0.51 
0.5 
0.95 
0.45 
 

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Response proportion in the Vaccine Group 
Response proportion in the Placebo Group 
Figure 4: Power curves for comparisons between vaccine and placebo responses 
 
 
 
(Alpha=.05; N Vaccine=20; N Placebo=10; 1-Sided Exact Test) 
 
With a sample size of 20 vaccinees, the 90% CI around a sample mean peak titer of 2.5 log10, 
with a SD of 1.5 (from IMPAACT 2000) is (1.92, 3.08). This ensures with 90% confidence that 
the true population mean peak titer is between 1.92 and 3.08 log10, and with 95% confidence that 
the true population mean is not lower than 1.92 log10. 
 
With the same sample size of 20, the 90% CI around a proportion of 18/20 (90%) vaccinees who 
shed vaccine virus is (72%-98%). For a proportion of 19/20 (95%) the 90% CI is (78%-99.7%), 
and for 20/20 (100%) the 90% CI is (86%-100%). For the target proportion of 95%, this ensures 
with 95% confidence that the true proportion of vaccinees who shed vaccine virus is not lower 
than 78%. 
 
9.4 
Monitoring 
 
Accrual to this study will be monitored by the IMPAACT leadership in accordance with standard 
operating procedures. The team will monitor feasibility monthly, first based on site activation and 
then on accrual. Initially, the team will monitor site activation weekly to ensure that an adequate 
number of sites have been activated to participate in the study.  If relatively few of the eligible 
sites have been activated after the study has been approved for 1 month, the team will re-assess 

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the feasibility of the study and the reasons why sites have not been activated, and may amend the 
protocol accordingly.   
 
9.4.1 
Monitoring by the Protocol Team 
 
Study Progress and Quality of Study Conduct 
 
The Protocol Team is responsible for continuous monitoring of study progress, including timely 
achievement of key milestones, and quality of study conduct.   
 
The team will closely monitor participant accrual and retention based on reports that will be 
generated at least monthly by the Statistical and Data Management Center (SDMC). The team has 
developed a study accrual plan that includes site-specific and total enrollment projections over the 
course of the accrual period, and actual accrual will be monitored relative to these projections. 
The team will monitor the timing of site-specific study activation, which will determine when 
each site will begin accruing participants, and actual accrual following activation. Accrual 
performance will be reported by the DMC, by site and across sites, and the team will review and 
discuss study progress at least monthly.  For any site that is delayed in completing the study 
activation process, or that falls short of its accrual projections, the team will communicate with 
the site to identify the barriers the site has encountered and the operational strategies and action 
plans to address these. 
 
The Protocol Team will similarly review participant retention and other key indicators of the 
quality of study conduct (e.g., data quality, and data and specimen completeness) based on reports 
generated by the SDMC and take action with study sites as needed to ensure high quality study 
conduct throughout the period of study implementation.  
 
Participant Safety 
 
The Protocol Team and the PSRT will closely monitor participant safety through routine review 
of safety data reports generated by the SDMC.  These reports will provide tabulations of adverse 
events (defined in Section 8.1) identified in enrolled infants, including abnormal laboratory test 
results, signs, symptoms, and diagnoses, pooled across arms (with the vaccine and placebo arms 
presented together). The PSRT will review these reports via conference call or other meeting at 
least twice a month during the first 56 days post inoculation and at least monthly thereafter. At 
the time of each call, the DAIDS Medical Officer will also review any EAEs (defined in Section 
7.3) reported to the DAIDS Safety Office that are not yet reflected in the data reports.  The PSRT 
will continually evaluate the pattern and frequency of reported events and assess for any 
individual occurrences or trends of concern.    
 
If there is a report of any event that meets the pause/stop criteria, procedures as per 8.2 will be 
followed. 
Blinding/Unblinding 
 
At the end of the RSV Season Surveillance Period, each site will receive a list of the assignments 
so that each participant’s parent/guardian may be informed of their infant’s assignment at the 
final visit (between April 1st and 30th following inoculation). 
 
If the need arises to unblind a specific participant’s assignment in the event of a serious illness 
prior to completion of the RSV Season Surveillance Period, IMPAACT procedures for 

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unblinding will be followed. In the event that unblinding is required, only that specific 
participant’s assignment will be unblinded. Whenever possible, the Protocol Chair will make a 
decision regarding early unblinding in collaboration with the Data and Safety Monitoring Board 
(DSMB). The sponsor and the DSMB Executive Secretary will also be notified of the event in 
real time. 
 
A subset of protocol team members limited to the Protocol Statistician, Protocol Vice Chair, and 
the two Scientific Investigators of the Laboratory of Infectious Diseases will be unblinded to all 
data at the completion of the Post-Acute Phase of follow-up (Day 56) for the last participant 
enrolled in each calendar year to enable more efficient and timely study evaluation and planning 
for appropriate next steps with respect to RSV candidate vaccine development. Unblinding will 
occur after all the relevant data that are needed to make decisions have been entered into the 
database and analyzed. 
 
9.4.2 
Monitoring by the NIAID Intramural Data and Safety Monitoring Board 
 
The NIAID Intramural DSMB is constituted to review the safety data of Intramural NIAID 
clinical studies that require DSMB oversight. The NIAID Intramural DSMB includes independent 
experts in infectious diseases, biostatistics, and clinical research that do not have direct 
involvement in the conduct of the study and have no significant conflicts of interests as defined 
by NIAID policy. The DSMB will review the protocol prior to opening the study to enrollment. 
The DSMB will meet at least twice a year or on a schedule specified by the DSMB to review the 
completeness of the study data, the adherence to the protocol, and AE data. Cumulative safety 
data (pooled across arms, with the vaccine and placebo arms presented together) will be 
submitted to the DSMB Executive Secretary for DSMB review. The DSMB will be notified in the 
event of unintentional or intentional unblinding and consulted in the event that pausing or halting 
criteria are met. The DSMB Executive Secretary will provide the Protocol Chair with DSMB 
recommendations promptly, and the official DSMB Report will then be provided in a timely 
fashion through the office of the NIAID Clinical Director. The Protocol Chair will submit the 
written DSMB recommendations to the sites’ IRBs upon receipt. All SAEs, LRIs, and all IND 
Safety Reports as specified in Section 7.2 will be reported by the Protocol Chair to the DSMB at 
the same time they are submitted to the IND sponsor or FDA. The PI will notify the DSMB of 
any cases of intentional or unintentional unblinding as soon as possible. The PI will notify the 
Board at the time pausing or halting criteria are met and obtain a recommendation concerning 
continuation, modification, or termination of the study. 
 
 
9.5 
Analyses 
 
9.5.1 
Assessment of Primary Objectives  
 
Safety data from all study participants who have been inoculated will be summarized, including 
data from participants who discontinue study early or have some missed visits. In the 
immunogenicity analyses, those who do not provide data for the Day 56 Visit follow-up (due to 
early discontinuation or missed visit) will be treated as “failures”. Sensitivity analyses will be 
performed to check if the results are consistent with those when these participants are excluded. 
Participants who receive any of the disallowed treatments listed in Section 5.11 after inoculation 
may be excluded from the immunogenicity evaluations after the time of the treatment. These 
participants will, however, be included in the safety evaluations for the duration of the study.  
These participants will not be replaced. Details of the analyses listed below will be included in 
the Statistical Analysis Plan.  

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The frequency of solicited AEs and unsolicited AEs, along with 90% confidence intervals, during 
Study Days 0 to 28 and of vaccine-related SAE during Study Day 0 to the Day 56 Visit will be 
summarized. In addition, line listing of individual clinical solicited AEs and unsolicited AEs 
during Study Days 0 to 28 and vaccine-related SAE during Study Day 0 to the Day 56 Visit, 
graded by severity, will be prepared. 
 
The proportion of participants with infection defined as recovery of vaccine virus from a nasal 
wash and/or a > fourfold rise in neutralizing antibody titer, as determined by culture and RT-
PCR, will be summarized. A line listing of the individual peak titer of vaccine virus shed and 
duration of virus shedding in nasal washes by each individual will be prepared. In addition, the 
geometric mean peak titer and mean duration of virus shed will be provided, for each treatment 
group.  
 
The proportion of participants that develop fourfold or greater rises in RSV-neutralizing antibody 
titer following vaccination will be summarized. A line listing of the individual RSV antibody titer 
pre- and post-vaccination will be prepared. In addition, the geometric mean and median antibody 
titers will be provided, for each treatment group. Line listings of individual RSV-neutralizing 
antibody responses as well as of antibody responses to the RSV F glycoprotein will be prepared 
as well.  
 
Where appropriate, the Wilcoxon rank sum test will be used to determine the statistical 
significance of differences between vaccine and placebo recipients. A 1-tailed Fisher’s exact test 
will be used to test the hypothesis that the vaccinated group will exhibit a greater proportion of 
participants who develop fourfold or greater rises in RSV-neutralizing antibody titer following 
vaccination compared to the placebo group.  
 
These will be the only formal statistical comparisons between the vaccinated and placebo groups. 
These tests will be carried out at a 5% significance level. 
 
The study results will be compared with the criteria listed in Section 1.1 to determine if this 
vaccine is a promising candidate for further evaluation in expanded Phase I studies or Phase II 
studies. 
 
9.5.2 
Assessment of Secondary Objectives  
 
The summary of the frequency and severity of symptomatic, medically attended respiratory and 
febrile illness in the vaccine and placebo recipients who experience natural infection with wt RSV 
during the subsequent RSV season will be presented. A line listing of the individual RSV 
antibody titer pre- and post- RSV Season Surveillance Period will be prepared. In addition, the 
geometric mean and median antibody titers will be provided for each treatment group. The B cell 
responses to vaccine as well as the quality and epitope specificity of RSV F specific antibody will 
be summarized for each treatment group.  
 
10 
DATA HANDLING AND RECORD KEEPING 
 
10.1 
Data Management Responsibilities 
 
As described in Section 4.4, data on screening and enrollment in this study will be collected using 
the DMC Subject Enrollment System.   

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Study sites must maintain adequate and accurate research records containing all information 
pertinent to the study for all screened and enrolled participants, including CRFs and supporting 
source data. In maintaining these records, sites must comply with the standards of source 
documentation specified in the DAIDS policy on Requirements for Source Documentation in 
DAIDS Funded and/or Sponsored Clinical Trials.  
All DAIDS policies referenced in this section are available at: 
www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/ClinicalSite.aspx  
 
CRFs are completed by study site staff and, following quality control and quality assurance 
reviews, are entered using a remote data entry system and transferred electronically to the DMC. 
Selected laboratory data are transferred electronically to the DMC through the LDMS.   
 
At the DMC, computerized checks are applied to the transferred data and, when required, data 
queries are issued for resolution by study site staff. All data must be transferred to the DMC 
within timeframes specified in the forms instructions; queries must also be resolved in a timely 
manner.  
 
Further information on the study CRFs and IMPAACT data management procedures, including a 
Forms Manual: Policies and Procedures for Forms Completion for DAIDS-Sponsored Clinical 
Trials, a comprehensive Computing Manual, and a User Manual for the Subject Enrollment 
System, are available on the DMC portal at www.fstrf.org.   
 
10.2 
Essential and Source Documents and Access to Source Data 
 
All DAIDS policies referenced in this section are available at: 
www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/ClinicalSite.aspx  
 
Study-related documentation will be completed as required by the IRB, the sponsor, and 
regulatory authorities. Continuing review documentation will be submitted by the site investigator 
to the IRB as specified by the IRB. An annual report will be submitted by the sponsor to the FDA 
based on the anniversary date that the IND for the LID cp M2-2 vaccine went into effect. These 
reports will provide a brief description of the progress of the investigation as outlined in 21 CFR 
312.33 and will include any revisions of the protocol not previously submitted to the FDA. 
 
Study-related documents will be maintained by the site investigator for a period of at least 2 years 
after final marketing approval of the vaccine, or at least 2 years after the formal discontinuation 
of clinical development of the product (or longer based upon local laws). The sponsor is required 
to inform the site investigator as to when such documents need no longer be retained. No study 
document should be destroyed without prior written agreement between the sponsor and the 
Protocol Chair. Storage of all study-related documents will be such that confidentiality will be 
strictly maintained. These records are also to be maintained in compliance with IRB, state, and 
federal medical records retention requirements, whichever are longest. Should the site 
investigator wish to assign the study records to another party and/or move them to another 
location, the site investigator must provide written notification of such intent to the sponsor with 
the name of the person who will accept responsibility for the transferred records and/or their new 
location. The sponsor must be notified in writing, and written permission must be received by the 
site from the sponsor prior to destruction or relocation of research records. 
 

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All study records must be accessible for inspection, monitoring, and/or auditing during and after 
the conduct of the study by authorized representatives of the study sponsors and their contracted 
monitors, IMPAACT, the US Food and Drug Administration, site drug regulatory authorities, site 
IRBs/IBCs, OHRP, and other applicable regulatory entities.  Records must be kept on-site 
throughout the period of study implementation; thereafter, instructions for off-site storage may be 
provided by NIH.  No study records may be removed to an off-site location or destroyed prior to 
receiving approval from NIH.  
 
10.3 
Clinical Investigator’s Brochure 
 
Investigators will receive the current version of the Clinical Investigator’s Brochure (IB) that 
comprehensively describes all the available preclinical experience with the experimental vaccine. 
If relevant new information becomes available during the course of the trial, the investigators will 
receive a revised IB or an amendment to the current version. 
 
10.4 
Quality Control and Quality Assurance 
 
Study sites must ensure that essential documents and participant research records are subject to 
continuous quality control and quality assurance procedures consistent with the DAIDS policy on 
Requirements for Clinical Quality Management Plans, which is available at: 
www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/ClinicalSite.aspx  
 
11 
CLINICAL SITE MONITORING 
 
Site monitors under contract to NIAID or NICHD will visit study sites to inspect study facilities 
and review participant study records including consent forms, CRFs, medical records, laboratory 
records, and pharmacy records, to ensure protection of study participants, compliance with the 
IRB/IBC-approved protocol, and accuracy and completeness of records. The monitors also will 
review essential document files to ensure compliance with all applicable regulatory requirements.  
Site investigators will make study facilities and documents available for inspection by the 
monitors.   
 
The trial will be conducted in compliance with this protocol, ICH GCP guidelines, and any 
applicable regulatory requirement(s). The study site monitoring will be conducted according to 
the “NIAID/DAIDS and NICHD Clinical Research Site Monitoring Guidelines”.   
 
The site investigator or designee will make study documents (e.g., consent forms, CRFs) and 
pertinent medical or clinical records readily available for inspection by the local IRB, the FDA, 
the site monitors, and the NIAID staff for confirmation of study data. The sponsor will retain 
originals of the Form FDA 1572 and copies of other study documents as deemed necessary. 
 
12 
HUMAN SUBJECTS PROTECTIONS 
 
12.1 
Institutional Review Board/Ethics Committee Review and Approval  
 
Prior to study initiation, site investigators must obtain IRB/IBC review and approval of this 
protocol and site-specific ICFs in accordance with 45 CRF 46; subsequent to initial review and 
approval, IRBs/IBCs must review the study at least annually. Site investigators must also 
promptly report to the IRB/IBC any changes in the study and any unanticipated problems 
involving risks to participants or others. 

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All IRB/IBC policies and procedures must be followed and complete documentation of all 
correspondence to and from the IRBs/IBCs must be maintained in site essential document files.  
Sites must submit documentation of both initial review and approval and continuing review to the 
DAIDS Protocol Registration Office (PRO) in accordance with the DAIDS Protocol Registration 
Manual (Section 13.2). 
 
A copy of the study approval (including approval of the informed consent form) is to be 
maintained in the site investigator’s study document binder, and a copy will be supplied to the 
sponsor.  
 
During the study, the site investigator is responsible for providing the IRB with all documents 
subject to review (i.e., protocol amendments, informed consent form updates, advertisements, and 
any written information that may be provided to the participant’s parents/guardians). Study 
progress reports will be made to the IRB by the investigator in accordance with IRB guidelines 
and government regulations. 
 
12.2 
Vulnerable Participants 
 
The NIH is mandated by law to ensure that children be included in clinical research when 
appropriate (34, 35). This study responds to that mandate and will provide clinical research data 
to inform RSV vaccine infectivity, safety and immunogenicity in children. Nonetheless, the 
infants who take part in this study are considered vulnerable participants per the US Code of 
Federal Regulations, and site IRBs/IBCs must consider the potential risks and benefits to child 
participants as described in 45 CFR 46 Subpart D (for children). 
 
With respect to 45 CFR 46 Subpart D, IRBs/IBCs must determine the level of risk to children in 
the categories specified in 45 CFR 46.404-407. Documentation of this determination is required 
to complete the DAIDS protocol registration process described in Section 13.2, and the risk 
category assigned by the IRB/IBC further determines the parental informed consent requirements 
for the study at each site.  Per 45 CFR 46.408 (b), the IRB/IBC may find that the consent of one 
parent is sufficient for research to be conducted under 46.404 or 46.405. If the IRB/IBC finds that 
the research is covered by 46.406 or 46.407, both parents must give their consent, unless one 
parent is deceased, unknown, incompetent, or not reasonably available or when only one parent 
has legal responsibility for the care and custody of the child (as determined locally). IRBs/IBCs 
must document their risk determination, and study sites should adapt the signature pages of their 
site-specific ICFs as needed to accommodate the parental consent requirements associated with 
the IRB/IBC determination. 
 
Study sites must comply with the requirements of the DAIDS policy on Enrolling Children 
(including Adolescents) in Clinical Research, which is available at: 
www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/ClinicalSite.aspx 
 
12.3 
Informed Consent 
 
In obtaining and documenting informed consent, the site investigator must comply with the 
applicable regulatory requirements, ICH GCP guidelines, and ethical principles. The written 
informed consent form must be approved by the IRB prior to its use. 
 

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Written informed consent for infant study participation will be obtained before any study-specific 
procedures are performed. The informed consent process will include information exchange, 
detailed discussion, and assessment of understanding of all required elements of informed 
consent, including the potential risks, benefits, and alternatives to study participation.  The 
process will emphasize the unproven efficacy of the study vaccine products. 
  
As part of the informed consent process, parents/guardians will also be asked whether they agree 
to storage and future research testing of biological specimens remaining after all protocol-
specified testing has been completed.  Future research testing of residual specimens may be 
declined with no impact on other aspects of infant study participation.   
 
Parental consenting requirements at each site will depend on the IRB/IBC risk determination 
described in Section 12.2; all IRB/IBC requirements will be followed.   
 
12.4 
Potential Benefits 
 
Participants may not receive any direct study product-related benefit from enrollment in this 
study. It is hoped that information gained in this study will contribute to the development of a 
safe and effective vaccine for the prevention of illness associated with RSV infection. 
 
12.5 
Potential Risks  
 
12.5.1 Venipuncture 
 
Risks occasionally associated with venipuncture include pain and bruising at the site of 
venipuncture, lightheadedness, infection, and syncope (rarely). 
 
12.5.2 Nasal Wash 
 
Risks occasionally associated with nasal wash include pain or discomfort and occasionally 
epistaxis. Nasal washes are not standard care in well children and are not usually performed on ill 
children, although many parents/guardians are advised to use saline nose drops and nasal bulb 
suction (the 2 components of the nasal wash procedure used in this study) to clear a young child’s 
congested nostrils during a URI. 
 
12.5.3 Receipt of Study Product 
 
If the LID cp M2-2 vaccine is insufficiently attenuated, participants could experience 
rhinorrhea, cough, fever, hoarseness, otitis media, or LRI. Immediate hypersensitivity reactions—
which could be life threatening—including urticaria, anaphylaxis, or other Immunoglobulin E 
(IgE)-mediated responses are possible, as with any vaccine. There is a theoretical possibility, as 
with any investigational vaccine, of risks about which we have no present knowledge. 
Parents/guardians will be informed of any such risks should further data become available. 
 
12.6 
Reimbursement/Compensation 
 
Compensation will be provided to the participant’s parent/guardian based on each site’s standard. 
The amount must be reviewed and approved by each sites’ IRB. Compensation will be in 
accordance with each institution’s IRB policies and will be specified in site-specific ICFs or other 
materials if applicable per IRC/EC policies and procedures. 

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12.7 
Privacy and Confidentiality 
 
All study procedures will be conducted in private and every effort will be made to protect 
participant privacy and confidentiality to the extent possible. Participant information will not be 
released without written permission to do so except as necessary for review, monitoring, and/or 
auditing as described in Section 10.2. 
 
All study-related information will be stored securely. Participant research records will be stored 
in locked areas with access limited to study staff. All laboratory specimens, CRFs, and other 
documents that may be transmitted off-site (e.g., EAE report forms, photographs of observed 
reactions) will be identified by PID only. Likewise, communications between study staff and 
Protocol Team members regarding individual participants will identify participants by PID only.   
 
Study sites are encouraged but not required by DAIDS policies to store study records that bear 
participant names or other personal identifiers separately from records identified by PID. All local 
databases must be secured with password protected access systems. Lists, logbooks, appointment 
books, and any other documents that link PID numbers to personal identifying information should 
be stored in a separate, locked location in an area with limited access.   
 
In addition to the above, a Certificate of Confidentiality has been obtained for this study from the 
US Department of Health and Human Services. This certificate protects study staff from being 
compelled to disclose study-related information by any US Federal, state, or local civil, criminal, 
administrative, legislative, or other proceedings.  It thus serves to protect the identity and privacy 
of study participants. Because the certificate cannot be enforced outside of the US, however, it 
applies only to US sites and participants. 
 
12.8 
Management of Incidental Findings 
 
Site clinicians will inform parents (or other authorized guardians if applicable) of all clinically 
meaningful physical exam findings and laboratory tests. When applicable, site clinicians will 
provide referrals to non-study sources of medical care for further evaluation and/or treatment of 
these findings.  
 
13 
ADMINISTRATIVE PROCEDURES 
 
13.1 
Regulatory Oversight 
 
This study is sponsored by the National Institute of Allergy and Infectious Diseases (NIAID), 
Eunice Kennedy Shriver National Institute of Child Health and Development (NICHD), and 
National Institute of Mental Health (NIMH), which are part of the United States National 
Institutes of Health (NIH).   
 
The Division of AIDS (DAIDS) within the NIAID is responsible for regulatory oversight of this 
study and holds the Investigational New Drug (IND) application under which the study will be 
conducted. DAIDS will distribute safety-related information pertaining to the study product prior 
to and during the conduct of the study, in accordance with its sponsor obligations.  
 

IMPAACT 2012, Version 1.0 
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08 August 2016 
NIAID and NICHD provide funding to the clinical research sites at which this study will be 
conducted.  Each institute contracts with independent clinical site monitors who will perform 
monitoring visits as described in Section 11.  As part of these visits, monitors will inspect study-
related documentation to ensure compliance with all applicable US and local regulatory 
requirements. 
 
13.2 
Protocol Registration 
 
Prior to implementation of this protocol, and any subsequent full version amendments, each site 
must have the protocol and the protocol ICFs approved, as appropriate, by their local IRB/IBC, 
local IBC, and any other applicable regulatory entity.  Upon receiving final approval, sites will 
submit all required protocol registration documents to the DAIDS PRO at the RSC.  The DAIDS 
PRO will review the submitted protocol registration packet to ensure that all of the required 
documents have been received.   
 
Site-specific ICFs will be reviewed and approved by the DAIDS PRO and sites will receive an 
Initial Registration Notification from the DAIDS PRO that indicates successful completion of the 
protocol registration process. A copy of the Initial Registration Notification should be retained in 
the site's regulatory files. 
 
For any future protocol amendments, upon receiving final IRB/IBC and any other applicable 
regulatory entity approvals, sites should implement the amendment immediately. Sites are 
required to submit an amendment registration packet to the DAIDS PRO at the RSC. The DAIDS 
PRO will review the submitted protocol registration packet to ensure that all the required 
documents have been received. Site-specific ICFs will not be reviewed and approved by the 
DAIDS PRO and sites will receive an Amendment Registration Notification when the DAIDS 
PRO receives a complete registration packet. A copy of the Amendment Registration Notification 
should be retained in the site's regulatory files. 
 
For additional information on the protocol registration process and specific documents required 
for initial and amendment registrations, refer to the current version of the DAIDS Protocol 
Registration Manual, which is available on the RSC website:  
http://rsc.tech-res.com/protocolregistration/ 
 
 
13.3 
Study Implementation 
 
This study will be conducted in accordance with the protocol, international good clinical practice 
guidelines, and all applicable US and local regulations. Study implementation will also be guided 
by the IMPAACT Network MOP, study-specific MOP, LPC, and other study implementation 
materials, which will be available on the IMPAACT web site: www.impaactnetwork.org. 
 
13.4 
Protocol Deviation Reporting 
 
Per the policy for Requirements for Source Documentation in DAIDS Funded and/or Sponsored 
Clinical Trials, all protocol deviations must be documented in participant research records. 
Reasons for the deviations and corrective and preventive actions taken in response to the 
deviations should also be documented. 
 
All DAIDS policies referenced in this section are available at: 

IMPAACT 2012, Version 1.0 
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08 August 2016 
www.niaid.nih.gov/labsandresources/resources/daidsclinrsrch/Pages/ClinicalSite.aspx  
 
Deviations should be reported to site IRBs/IBCs and other applicable review bodies in 
accordance with the policies and procedures of these review bodies.  Serious deviations that are 
associated with increased risk to 1 or more study participants and/or significant impacts on the 
integrity of study data must also be reported within IMPAACT, following procedures specified in 
the IMPAACT Network MOP. 
 
13.5 
ClinicalTrials.gov 
 
This protocol is not subject to the United States Food and Drug Administration Amendments Act 
of 2007 (FDAAA). However, it will be registered in ClinicalTrials.gov to meet International 
Committee of Medical Journal Editors requirements. 
 
 
14 
PUBLICATIONS 
 
All presentations and publications of data collected in this study are governed by IMPAACT 
policies, which are available in the IMPAACT Network MOP, and NIAID policies. Any 
presentation, abstract, or manuscript will be made available for review by the pharmaceutical and 
NIAID sponsors prior to submission. Publication or presentation approval will conform to any 
CRADA or other collaborative agreement in place. 
 
 

IMPAACT 2012, Version 1.0 
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08 August 2016 
15 
REFERENCES 
 
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burden of acute lower respiratory infections due to respiratory syncytial virus in young 
children: a systematic review and meta-analysis. Lancet. 2010;375(9725):1545-55. 
2. 
Wright PF, Karron RA, Belshe RB, Shi JR, Randolph VB, Collins PL, et al. The absence 
of enhanced disease with wild type respiratory syncytial virus infection occurring after 
receipt of live, attenuated, respiratory syncytial virus vaccines. Vaccine. 
2007;25(42):7372-8. 
3. 
Collins PL, Murphy BR. New generation live vaccines against human respiratory 
syncytial virus designed by reverse genetics. ProcAm ThoracSoc. 2005;2(2):166-73. 
4. 
Bermingham A, Collins PL. The M2-2 protein of human respiratory syncytial virus is a 
regulatory factor involved in the balance between RNA replication and transcription. 
ProcNatlAcadSciUSA. 1999;96(20):11259-64. 
5. 
Karron RA, Luongo C, Thumar B, Loehr KM, Englund JA, Collins PL, et al. A gene 
deletion that up-regulates viral gene expression yields an attenuated RSV vaccine with 
improved antibody responses in children. Science translational medicine. 
2015;7(312):312ra175. 
6. 
Wright PF, Karron RA, Belshe RB, Thompson J, Crowe JE, Jr., Boyce TG, et al. 
Evaluation of a live, cold-passaged, temperature-sensitive, respiratory syncytial virus 
vaccine candidate in infancy. J Infect Dis. 2000;182(5):1331-42. 
7. 
Wright PF, Karron RA, Madhi SA, Treanor JJ, King JC, O'Shea A, et al. The interferon 
antagonist NS2 protein of respiratory syncytial virus is an important virulence 
determinant for humans. J Infect Dis. 2006;193(4):573-81. 
8. 
Karron RA, Wright PF, Belshe RB, Thumar B, Casey R, Newman F, et al. Identification 
of a recombinant live attenuated respiratory syncytial virus vaccine candidate that is 
highly attenuated in infants. J Infect Dis. 2005;191(7):1093-104. 
9. 
Karron RA, Wright PF, Crowe JE, Jr., Clements-Mann ML, Thompson J, Makhene M, et 
al. Evaluation of two live, cold-passaged, temperature-sensitive respiratory syncytial 
virus vaccines in chimpanzees and in human adults, infants, and children. J Infect Dis. 
1997;176(6):1428-36. 
10. 
Hall CB, Weinberg GA, Iwane MK, Blumkin AK, Edwards KM, Staat MA, et al. The 
burden of respiratory syncytial virus infection in young children. The New England 
journal of medicine. 2009;360(6):588-98. 
11. 
Hall CB. Respiratory syncytial virus and parainfluenza virus. The New England journal 
of medicine. 2001;344(25):1917-28. 
12. 
Martinez FD. Respiratory syncytial virus bronchiolitis and the pathogenesis of childhood 
asthma. PediatrInfect DisJ. 2003;22(2 Suppl):S76-S82. 
13. 
Collins PL, Karron RA. Respiratory syncytial virus and metapneumovirus. In: Knipe 
DM, Howley PM, Cohen JI, Griffin DE, Lamb RA, Martin MA, et al., editors. Fields 
Virology, Sixth Edition. 1. Sixth ed. Philadelphia: Lippincott Williams & Wilkins; 2013. 
p. 1086-123. 
14. 
Kim HW, Canchola JG, Brandt CD, Pyles G, Chanock RM, Jensen K, et al. Respiratory 
syncytial virus disease in infants despite prior administration of antigenic inactivated 
vaccine. Am J Epidemiol. 1969;89(4):422-34. 
15. 
Collins PL, Murphy BR. Vaccines against Human Respiratory Syncytial Virus. 
Respiratory Syncytial Virus. 2007;14:233-78. 

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16. 
Acosta PL, Caballero MT, Polack FP. Brief history and characterization of enhanced 
respiratory syncytial virus disease. Clinical and vaccine immunology : CVI. 2015. 
17. 
Hendry RM, Burns JC, Walsh EE, Graham BS, Wright PF, Hemming VG, et al. Strain-
specific serum antibody responses in infants undergoing primary infection with 
respiratory syncytial virus. J Infect Dis. 1988;157(4):640-7. 
18. 
Johnson PR, Jr., Olmsted RA, Prince GA, Murphy BR, Alling DW, Walsh EE, et al. 
Antigenic relatedness between glycoproteins of human respiratory syncytial virus 
subgroups A and B: evaluation of the contributions of F and G glycoproteins to 
immunity. J Virol. 1987;61(10):3163-6. 
19. 
Collins PL, Hill MG, Camargo E, Grosfeld H, Chanock RM, Murphy BR. Production of 
infectious human respiratory syncytial virus from cloned cDNA confirms an essential 
role for the transcription elongation factor from the 5' proximal open reading frame of the 
M2 mRNA in gene expression and provides a capability for vaccine development. 
ProcNatlAcadSciUSA. 1995;92(25):11563-7. 
20. 
Collins PL, Hill MG, Johnson PR. The two open reading frames of the 22K mRNA of 
human respiratory syncytial virus: sequence comparison of antigenic subgroups A and B 
and expression in vitro. J GenVirol. 1990;71 ( Pt 12):3015-20. 
21. 
Ahmadian G, Randhawa JS, Easton AJ. Expression of the ORF-2 protein of the human 
respiratory syncytial virus M2 gene is initiated by a ribosomal termination-dependent 
reinitiation mechanism. EMBO J. 2000;19(11):2681-9. 
22. 
Jin H, Cheng X, Zhou HZ, Li S, Seddiqui A. Respiratory syncytial virus that lacks open 
reading frame 2 of the M2 gene (M2-2) has altered growth characteristics and is 
attenuated in rodents. J Virol. 2000;74(1):74-82. 
23. 
Teng MN, Whitehead SS, Bermingham A, St CM, Elkins WR, Murphy BR, et al. 
Recombinant respiratory syncytial virus that does not express the NS1 or M2-2 protein is 
highly attenuated and immunogenic in chimpanzees. J Virol. 2000;74(19):9317-21. 
24. 
Crowe JE, Jr., Bui PT, London WT, Davis AR, Hung PP, Chanock RM, et al. 
Satisfactorily attenuated and protective mutants derived from a partially attenuated cold-
passaged respiratory syncytial virus mutant by introduction of additional attenuating 
mutations during chemical mutagenesis. Vaccine. 1994;12(8):691-9. 
25. 
Whitehead SS, Juhasz K, Firestone CY, Collins PL, Murphy BR. Recombinant 
respiratory syncytial virus (RSV) bearing a set of mutations from cold-passaged RSV is 
attenuated in chimpanzees. J Virol. 1998;72(5):4467-71. 
26. 
Friedewald WT, Forsyth BR, Smith CB, Gharpure MA, Chanock RM. Low-temperature-
grown RS virus in adult volunteers. Jama. 1968;204(8):690-4. 
27. 
Bukreyev A, Belyakov IM, Berzofsky JA, Murphy BR, Collins PL. Granulocyte-
macrophage colony-stimulating factor expressed by recombinant respiratory syncytial 
virus attenuates viral replication and increases the level of pulmonary antigen-presenting 
cells. Journal of virology. 2001;75(24):12128-40. 
28. 
Kim HW, Arrobio JO, Pyles G, Brandt CD, Camargo E, Chanock RM, et al. Clinical and 
immunological response of infants and children to administration of low-temperature 
adapted respiratory syncytial virus. Pediatrics. 1971;48(5):745-55. 
29. 
Palivizumab, a Humanized Respiratory Syncytial Virus Monoclonal Antibody, Reduces 
Hospitalization From Respiratory Syncytial Virus Infection in High-risk Infants. 
Pediatrics. 1998;102(3):531-7. 

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30. 
McFarland EJ, Buchholz UJ, Muresan P, Cunningham CK, Luongo C, Thumar B, et al., 
editors. High Infectivity of recombinant live-attenuated respiratory syncytial virus 
vaccine RSV LID ΔM2-2 in RSV-seronegativeinfants and children. ID Week Advancing 
Science, Improving Care; 2015; San Diego, CA. 
31. 
American Academy of Pediatrics, Committee on Practice and Ambulatory Medicine. 
Recommendations for Preventive Pediatric Health Care. Pediatrics. 2007;120(6):1376. 
32. 
Siberry GK, ener-West M, Schappell E, Karron RA. Comparison of temple temperatures 
with rectal temperatures in children under two years of age. ClinPediatr(Phila). 
2002;41(6):405-14. 
33. 
Gharpure MA, Wright PF, Chanock RM. Temperature-sensitive mutants of respiratory 
syncytial virus. J Virol. 1969;3(4):414-21. 
34. 
National Institutes of Health. Guidelines for the Inclusion of Women and Minorities as 
Subjects in Clinical Research. Federal Register. 1994 March 18. Report No. 
35. 
National Institutes of Health. NIH Policy and Guidelines on the Inclusion of Children as 
Participants in Research Involving Human Subjects. 1998 March 6. Report No. 
 

 
  
 
IMPAACT 2012, Version 1.0 
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08 August 2016 
APPENDICES 
 
Appendix I: Tables Referenced in the Background Section 
 
Table 9: Comparison of Genomic Sequence of LID cp M2-2, LOT RSV#009B, to wt rRSV A2 D46 
 
Gene 
Region 
RSV Nucleotide1 
 
Encoded Amino 
Acid, Position in 
ORF 
Notes 
Motif; rationale for mutation 
nt position1 
rRSV A2 D46 
(KT992094) 
 
LID cp ∆M2-2  
cDNA 
LID cp ∆M2-2  
Seed Lot and 
Lot RSV#009B 
N 
1938 
A 
 
G 
G 
S266 
ClaI restriction site at "cp" mutation 
N 
1939 
G 
 
A 
A 
V267I 
"cp" mutation 
SH 
4489 
C 
 
T 
T 
V62 
Silent change stabilizes RSV cDNA n E. coli 
SH 
4492 
C 
 
T 
T 
N63 
Silent change stabilizes RSV cDNA n E. coli 
SH 
4495 
A 
 
T 
T 
T64 
Silent change stabilizes RSV cDNA n E. coli 
SH 
4497-8 
AG 
 
GA 
GA 
stop codon 
Silent change stabilizes RSV cDNA n E. coli 
SH 
4499-4610 
 
 
112 nt deletion 
112 nt deletion 
ncr 2 
deletion of SH 3' ncr to  
stabilize RSV cDNA in E. coli 
F 
6314 
A 
 
C 
C 
E218A 
"cp" mutation 
F 
7229 
C 
 
T 
T 
T523I 
"cp" mutation 
M2-2 
8161 
T 
 
C 
C 
silence ATG1 
removal of potential translation initiation 
sites of M2-2 ORF 
M2-2 
8167 
T 
 
C 
C 
silence ATG3 
M2-2 
8179 
T 
 
C 
C 
silence ATG7 
M2-2 
8189-429 
 
 
241 nt deletion 
241 nt deletion 
 
Deletion of M2-2 ORF 
L 
9454 
G 
 
A 
A 
C319Y 
"cp" mutation 
L 
9455 
T 
 
C 
C 
L 
10325 
C 
 
C 
T 
N609 
silent mutation in Seed and Lot RSV#009B 
L 
13566 
C 
 
T 
T 
H1690Y 
"cp" mutation 
L 
13568 
T 
 
C 
C 
1 Genomic position in reference to wt recombinant RSV strain A2 D46 (19); Genbank accession number KT992094). The term “D46” refers to a specific 
sequence and cDNA of strain A2. The LID cp ΔM2-2 cDNA contains a 112-nt deletion of the 3' ncr of the SH gene together with several nt changes 
immediately upstream, which stabilize the cDNA in E. coli, and contains a 241-nt deletion of the M2-2 ORF that is a major attenuating mutation. The 
numbering of all sequence positions is based on the complete sequence of wt recombinant RSV strain A2 D46 that was described by Collins et al (19), which is 
the parent of the present vaccine strains. All sequences are positive-sense. 
2 ncr, non-coding region  
The attenuating elements of LID cp ΔM2-2 (deletion of M2-2 ORF and the "cp" mutations) are indicated with light shading.

 
  
 
IMPAACT 2012, Version 1.0 
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Table 10: Viral titers of nasopharyngeal swab samples from AGMs inoculated with LID ΔM2-2, LID M2-2 1030s, 
or LID cp M2-2a 
RSV Vaccine 
candidate 
AGM ID 
NP virus titer (log10 PFU/mL) on indicated daysb   
Duration of 
sheddingc 
Peak 
virus  
titer 
Sum of 
daily titers d 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
12 
LID M2-2 
7806 
- 
1.4 
1.7 
2.7 
2.6 
4.0 
3.9 
1.4 
- 
2.7 
- 
9 
4.0 
21.4 
7705 
- 
- 
- 
2.7 
2.3 
3.6 
2.4 
1.2 
- 
- 
- 
5 
3.6 
14.3 
7747 
- 
- 
1.3 
0.7 
- 
1.5 
1.3 
- 
- 
- 
- 
5 
1.5 
7.2 
7674 
- 
0.7 
- 
- 
- 
2.3 
1.8 
1.5 
- 
- 
- 
7 
2.3 
8.8 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
6.5 
2.9 
12.9 
LID M2-2 1030s 
8033 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
7720 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
7844 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
7847 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
0 
0.35 
3.9 
LID cp M2-2 
8120 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
8528 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0 
0.35 
3.9 
8336 
- 
- 
- 
0.7 
- 
- 
- 
- 
- 
- 
- 
4 
0.7 
4.2 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
1.3 
0.5 
4.0 
 
a AGMs were inoculated by the combined intranasal and intratracheal routes with 106 PFU of the indicated virus in a 1 mL inoculum per site (total dose: 2x106 
PFU per animal). The AGM study was approved by the Animal Care and Use Committee of NIAID, NIH. 
b Combined nasopharyngeal (NP) swabs were placed in 2 mL of L-15 medium with sucrose phosphate buffer as stabilizer. Virus titrations were performed on 
Vero cells at 37°C (LID M2-2, LID M2-2 1030s) or 32°C (LID cp M2-2).  The lower limit of detection was 0.7 log10 PFU/mL. Samples with no detectable 
virus are represented as “-“.  Peak titers for each animal are underlined.  
c The period of days from the first to the last day on which virus was detected, including negative days (if any) in between.  
d A value of 0.35 was used for samples with no detectable virus. 
e The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve).   
 
 
 
 
 
 

 
  
 
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Table 11: Viral titers of tracheal lavage samples from AGMs inoculated with LID ΔM2-2, LID M2-2 1030s, or LID 
cp M2-2 a 
 
RSV vaccine candidate 
AGM ID 
Tracheal lavage virus titer (log10 PFU/mL) on 
indicated daysb 
Duration of 
shedding c 
Peak virus  
titer 
Sum of 
daily 
titers d 
 
2 
4 
6 
8 
10 
12 
 
 
 
LID M2-2 
7806 
2.5 
3.4 
4.6 
- 
- 
- 
7 
4.6 
12.6 
7705 
1.6 
- 
3.3 
1.5 
- 
- 
9 
3.3 
8.5 
7747 
1.8 
1.0 
6.0 
2.3 
- 
- 
9 
6.0 
12.5 
7674 
- 
1.3 
2.7 
2.3 
1.0 
- 
9 
2.7 
8.7 
 
Mean: 
 
 
 
 
 
 
9.0 
4.2 
10.6 
LID M2-2 1030s 
8033 
- 
- 
- 
- 
- 
- 
0 
0.7 
4.2 
7720 
- 
- 
- 
- 
- 
- 
0 
0.7 
4.2 
7844 
- 
- 
- 
- 
- 
- 
0 
0.7 
4.2 
7847 
- 
- 
- 
- 
- 
- 
0 
0.7 
4.2 
 
Mean: 
 
 
 
 
 
 
0 
0.7 
4.2 
LID cp M2-2 
8120 
2.1 
- 
- 
- 
- 
- 
3 
2.1 
5.6 
8528 
1.0 
- 
- 
- 
- 
- 
3 
1.0 
4.5 
8336 
 
1.0 
- 
- 
- 
- 
5 
1.0 
4.5 
 
Mean: 
 
 
 
 
 
 
3.7 
1.4 
4.9 
a 
AGMs were inoculated by the combined intranasal and intratracheal routes with 106 PFU of the indicated virus in a 1 mL inoculum per site (total dose: 2 x 
106 PFU per animal).  
b 
On days 2, 4, 6, 8, 10, and 12, tracheal lavage was performed with 3 mL of PBS. Virus titrations were performed on Vero cells at 37°C (LID M2-2, LID 
M2-2 1030s) or 32°C (LID cp M2-2).  The lower limit of detection was 1.0 log10 PFU/mL of lavage solution.  Samples with no detectable virus are 
represented as “-“.  Peak titers for each animal are underlined.  
c 
The period of days from the first to the last day on which virus was detected, including negative days (if any) in between. 
d 
A value of 0.7 was used for samples with no detectable virus. 
e   The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve).   
 

 
  
 
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Table 12:  Neutralizing antibody titers of AGMs inoculated with LID ΔM2-2, LID M2-2 1030s, or LID cp M2-2a 
RSV Vaccine candidate 
AGM ID 
Neutralizing antibody titers  
(PRNT60, reciprocal log2) on indicated daysb 
0 
21 
28 
LID M2-2 
7806 
<3.3 
7.2 
7.2 
7705 
<3.3 
8.8 
8.2 
7747 
<3.3 
8.3 
8.4 
7674 
<3.3 
6.7 
6.2 
 
Mean: 
<3.3 
7.8 
7.5 
LID M2-2 1030s 
8033 
<3.3 
5.4 
6.6 
7720 
<3.3 
<3.3 
<3.3 
7844 
<3.3 
<3.3 
4.3 
7847 
<3.3 
6.8 
6.8 
 
Mean: 
<3.3 
4.7 
5.2 
LID cp M2-2 
8120 
<3.3 
10.0 
10.5 
8528 
<3.3 
10.1 
9.8 
8336 
<3.3 
8.1 
7.7 
 
Mean: 
<3.3 
9.4 
9.3 
a  AGMs were inoculated i.n. and i.t. with 106 PFU of the indicated virus in a 1 mL inoculum per site (total dose = 106.3 PFU per animal). 
b On days 0, 21, and 28 p.i., serum was obtained. Neutralizing antibody titers were determined in a 60% plaque reduction neutralization assay. The lower limit of detection 
was 3.3 (1:10). 
 
 

 
  
 
IMPAACT 2012, Version 1.0 
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Table 13:  Viral Titers of Nasopharyngeal Swab Samples from AGMs Inoculated with CTM LID cp M2-2, Lot 
RSV#009B 
Group 
Virus Test Articlea
ID 
NP virus titer (log10 PFU/mL) on indicated daysb 
Peak virus 
titer 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
12 
TM1 
16RSV06-tox 
LID cp M2-2  
Lot RSV#009Bc  
8565 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0.35 
8564 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0.35 
8431 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0.35 
8797 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
- 
0.35 
  
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
0.35 
a Monkeys were inoculated i.n. and i.t. with 105.7 PFU of  LID cp ∆M2-2 in a 1.25 mL inoculum per site (total dose =1x106 PFU/AGM) 
b Virus titrations were performed on Vero cells at 32°C. The lower limit of detection was 0.7 log10 PFU/mL. Samples with no detectable virus are represented as “-“. The results 
show that LID cp ∆M2-2 is strongly restricted in the URT of AGMs. 
c CTM vial numbers 0019 to 0026, 1267 to 1274, and 2483 to 2490. 
 
 
 
Table 14:  Viral Titers of Tracheal Lavage samples from AGMs Inoculated with CTM LID cp M2-2, Lot RSV#009B 
Group 
Virus Test Articlea 
ID 
TL virus titer (log10 PFU/mL) on indicated dayb 
Peak virus 
titer 
2 
4 
6 
8 
10 
12 
 
TM1 
16RSV06-tox 
LID cp M2-2  
Lot RSV#009Bc  
8565 
- 
- 
- 
1.0 
- 
- 
1.0 
8564 
- 
- 
- 
- 
- 
- 
0.7 
8431 
- 
- 
- 
- 
- 
- 
0.7 
8797 
- 
- 
- 
- 
- 
- 
0.7 
Mean: 
 
 
 
 
 
 
0.8 
a Monkeys were inoculated i.n. and i.t. with 105.7 PFU of  LID cp ∆M2-2 in a 1.25 mL inoculum per site (total dose =1x106 PFU/AGM) 
b Virus titrations were performed on Vero cells at 32°C. The lower limit of detection was 1.0 log10 PFU/mL. Samples with no detectable virus are represented as “-“. LID cp 
∆M2-2 is strongly restricted in the LRT of AGMs. 
c CTM vial numbers 0019 to 0026, 1267 to 1274, and 2483 to 2490. 
 
 

 
  
 
IMPAACT 2012, Version 1.0 
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Table 15:  Viral Titers of Tracheal Lavage samples from AGMs Inoculated with CTM LID cp M2-2, Lot RSV#009B 
Group 
Virus Test Article 
ID 
 
RSV Neutralization Titer 
 (Log2 of reciprocal) on days 
0 
21 
28 
TM1 
16RSV06-tox 
LID cp M2-2  
Lot RSV#009Bc  
8565 
< 3.3 
10.0 
9.9 
8564 
< 3.3 
7.0 
7.6 
8431 
< 3.3 
7.2 
7.6 
8797 
< 3.3 
9.2 
9.8 
 
 
Mean: 
- 
8.4 
8.7 
a CTM vial numbers 0019 to 0026, 1267 to 1274, and 2483 to 2490. On days 0, 21, and 28, serum was obtained. Neutralizing antibody titers were determined in a 60% plaque 
reduction neutralization assay. The lower limit of detection was 3.3 (1:10). Samples below the lower limit of detection are recorded as “-“.  
 
 
 

 
  
 
IMPAACT 2012, Version 1.0 
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Appendix II: Schedule of Events: Screening, Acute Phase, and Post-Acute Phase  
 
 
 
 
ACUTE PHASE 
 
 
 
POST-ACUTE 
PHASE 
 
 
Screening 
Day 0 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 6 
Day 7 
Day 8 
Day 9 
Day 10 
Day 11 
Day 12 
Day 13 
Day 14 
Day 15 
Day 16 
Day 17 
Day 18-27 
(contact 
each day) 
Day 28 
Day 29 
Day 30-55 
Day 56  
Illness 
Visit  
Early DC 
In person visit 
X 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
 
X 
X 
X 
Non-visit contact 
 
 
X 
X 
 
X 
 
X 
 
X 
X 
 
X 
 
X 
 
X 
X 
 
X 
 
X 
Per 6.5 
 
 
 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
History 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Interim History 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Per 6.5 
X 
X 
X 
Physical exam (full) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical assessment 
(focused PE) 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
 
 
X 
 
Administer study 
product 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood for: 
immunologic assays 
5mL 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5mL 
 
5mL 
Blood for: cellular 
immune assay 
(viable PBMCs) 
3mL 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3mL 
 
3mL 
Nasal wash for: 
 RSV antibody 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
Nasal wash for: 
viral detection & 
quantification 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
 
 
X 
X 
Request 
adventitious agent 
assay 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Total blood volume 
8mL 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
8mL 
-- 
8mL 

IMPAACT 2012, Version 1.0 
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08 August 2016 
Appendix III: Schedule of Events: RSV Pre-season Sampling, seasonal 
surveillance, and Post-season Sampling 
 
 
 
Pre-RSV season 
Weekly contact 
Post-RSV season 
Illness 
Visit 
Early 
DC 
Visit Period 
Oct 1st to Oct 31st  Nov 1st to Mar 31st 
Apr 1st to Apr 30th 
 
 
Clinical assessment 
(focused PE) 
 
 
 
X 
 
Interim history 
 
X 
 
X 
X 
LABORATORY EVALUATIONS 
Blood for: 
immunologic assays 
5 mL 
 
5 mL 
 
5 mL 
Blood for: cellular 
immune assay (viable 
PBMCs) 
3 mL 
 
3 mL 
 
3 mL 
Nasal wash for: 
viral detection & 
quantification 
 
 
 
X 
 
Request adventitious 
agent assay  
 
 
 
X 
 
TOTAL BLOOD 
VOLUME 
8 mL 
-- 
8 mL 
-- 
8 mL 
 

IMPAACT 2012, Version 1.0 
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08 August 2016 
Appendix IV: Definitions of Solicited Adverse Events 
 
Event 
Defined 
Fever 
Temporal temperatures ≥100.0oF unconfirmed by rectal temp -or-  
Rectal temperature of ≥100.4oF. 
Acute Otitis Media1 
Loss of tympanic membrane landmarks, accompanied by erythema and loss 
of mobility. May or may not be associated with fever or other respiratory 
symptoms. Confirmed with tympanometry if possible. 
Upper Respiratory Tract Illness (URI) 
Rhinorrhea 
Two or more consecutive days of clear or purulent discharge from the 
nares.  
Note: Not associated with crying, change of room temperature, or eating 
and drinking. 
Pharyngitis 
Two or more consecutive days of pharyngeal erythema accompanied by 
exudate, and/or enlarged tender lymph nodes.  
Note: May be associated with sore throat, or painful or difficult swallowing. 
Cough without LRI  
Two or more consecutive days of 3 or more episodes of cough during a 15-
minute timed observation period, or cough awakens child from sleep.  
Note: Not associated with eating, drinking or choking. 
Hoarseness 
An unnaturally deep or rough quality of voice. 
Lower Respiratory Tract Illness (LRI) 
Wheezing2,3 
Sustained, high pitched, musical breath sounds, especially during the 
expiratory phase, which do not clear with cough. 
Pneumonia1,2,3 
Rales and crackles, originating in the lower respiratory tract, usually 
accompanied by tachypnea, which do not clear with cough. May be 
confirmed by x-ray showing areas of consolidation. 
Laryngotracheobronchitis 
(croup) 1,2,3 
Barking cough, hoarseness, and inspiratory stridor 
Rhonchi2,3 
Coarse breath sounds which are not transmitted noises from the upper 
airway and do not clear with cough. 
Rales2,3 
Abnormal lung sound heard through a stethoscope. Rales may be sibilant 
(whistling), dry (crackling) or wet (more sloshy) depending on the amount 
and density of fluid refluxing back and forth in the air passages. 
1 Diagnosis must be made by a medical professional 
2 Must be sustained over 20 minutes. 
3 Clinical assessment must be made by a medical professional and confirmed by a second medical professional, if possible. 
 
NOTE: Solicited AEs will only be recorded on CRFs according to criteria defined in Section 7.2

IMPAACT 2012, Version 1.0 
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08 August 2016 
Appendix V: RSV Seasonality in Baltimore 
 
 
 
 
 
 
 
 
All specimens collected and tested at Johns Hopkins Hospital through 10 March 2016 
 
 

IMPAACT 2012, Version 1.0 
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Appendix VI: Sample Informed Consent Form 
 
DIVISION OF AIDS  
INTERNATIONAL MATERNAL PEDIATRIC ADOLESCENT AIDS CLINICAL TRIALS 
(IMPAACT) NETWORK  
 
For protocol: 2012 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of a Single Dose of a 
Recombinant Live-Attenuated Respiratory Syncytial Virus Vaccine, LID cp ΔM2-2, Lot RSV#009B, 
Delivered as Nose Drops to RSV-Seronegative Infants 6 to 24 Months of Age 
 Version 1.0, dated 08 August 2016 
 
SHORT TITLE FOR IMPAACT 2012: Safety and Immunogenicity of a Single Dose of the LID cp ΔM2-
2 Vaccine  
 
INTRODUCTION 
 
You are being asked to allow your baby to take part in this research study to test a vaccine to prevent 
Respiratory Syncytial Virus (RSV) illness in infants. This study is sponsored by the National Institutes of 
Health (NIH).  The doctor in charge of this study at this site is: [Site: insert name of site investigator].  
Before you decide if you want your baby to be a part of this study, we want you to know about the study.  
 
This is a consent form. It gives you information about this study.  The clinical research staff will talk with 
you about this information.  You are free to ask questions about this study at any time.  If you agree to 
allow your baby to take part in this study, you will be asked to sign this consent form. You will get a copy 
to keep. 
 
WHY IS THIS STUDY BEING DONE? 
 
The study is being done to look at the safety (side effects) and antibody (germ fighters) response of 
infants to a single dose of a vaccine against a virus called respiratory syncytial virus, or RSV. The study 
will measure the amount of vaccine virus that sheds (comes) from each infant receiving the vaccine. It 
will tell us how stable and strong the vaccine was during the study.  This research study is testing an 
experimental vaccine. This vaccine has not been licensed by the U.S. Food and Drug Administration 
(FDA). 
 
 
RSV is a virus (germ) that can cause breathing problems in infants and children. Symptoms of 
infection with RSV may include: 
 
Fever 
 
Runny nose 
 
Sore throat 
 
Ear infection 
 
Cough  
 
Croup (barky cough with hoarseness) 
 
RSV can cause serious lung infections such as pneumonia and wheezing. At this time, there is no 
approved vaccine to prevent RSV illness. 
 
 
 

 
  
 
IMPAACT 2012, Version 1.0 
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Doctors who develop vaccines at the NIH have made a live virus vaccine that may help prevent RSV 
illness in babies and children. A live virus vaccine contains a weakened, live virus that is designed to help 
your body respond in a way that will protect you from getting sick from the virus.  This is called an 
“immune response.” The investigational RSV vaccine to be used in this study contains a live, weakened 
form of RSV and is given as nose drops. This vaccine has not been evaluated in humans. However, other 
RSV vaccines very similar to this one have been tested in both adults and children. There were not many 
side effects, and there was an immune response. 
  
We are asking you to allow your baby to participate in this study. If you agree, we will give your baby 
either 1 dose of vaccine or 1 dose of placebo. The placebo is a mixture of water, salt, vitamins, and sugar 
that is gentle on human tissue, sterile, and approved for use on people. The placebo has no vaccine in it. 
This is the first time this vaccine will be evaluated in people. Up to 33 babies who have not already had 
an illness caused by RSV virus will take part in the study. Your baby was chosen to be in this study 
because your baby is at least 6 months (180 days) old and less than 25 months (750 days) old, has not had 
RSV, and is otherwise healthy.  
 
WHAT DOES MY BABY HAVE TO DO IF HE/SHE IS IN THIS STUDY? 
 
The vaccine/placebo will be given to your baby by squirting it inside his/her nose like nose drops. The 
amount is very small (a few drops). Your baby will need to lie down on his/her back for one minute after 
receiving the vaccine/placebo. Approximately 2 of each 3 enrolled babies will receive the RSV vaccine 
and approximately 1 of each 3 enrolled babies will receive nose drops without vaccine (placebo).  
Whether your baby receives the vaccine or nose drops without the vaccine (placebo) will be decided 
randomly by computer, like flipping a coin. Neither you nor the study doctors or study nurses will know 
whether your baby received the vaccine or placebo until the study ends. However, this information can be 
made available to the study doctor if needed in an emergency. 
 
At the first visit, your baby’s blood will be tested to see if your baby has had RSV in the past.  You will 
be told the result of the test. If your baby goes on the study, you will be told whether he/she got vaccine or 
placebo after the end of the study in the spring. You will not receive information about your baby’s 
response to the vaccine, but you will receive a summary of the overall response to the vaccine for 
everyone in the study. 
 
If you agree to allow your baby to take part in this study, you will be asked some questions to be sure 
he/she can participate in this study.   
 
Your baby cannot take part in this study if he/she already has antibodies against RSV, lives with people 
who have weak immune systems, or is not well.  Your baby cannot take part in this study if he/she lives 
with or is in a daycare room with babies younger than 6 months of age, unless you are willing to keep 
your baby out of daycare for 28 days after he/she receives vaccine or placebo. Your baby should not 
receive any vaccines, including rotavirus vaccine for at least 14 days and other live vaccines for at least 
28 days after receiving the study vaccine or placebo. We ask that you talk with the study staff before your 
baby receives a routine vaccine for the 28 days after the study vaccine or placebo.  We ask that your baby 
does not take part in any other experimental vaccine or drug studies for 8 weeks after they receive vaccine 
or placebo. We will ask you to review and sign this study consent prior to administering the 
vaccine/placebo to your baby. At that time, we will ask you to answer questions to see how well you 
understand the study. 
 
Your baby will participate in this study until April of the year after he/she started the study. For the first 8 
weeks after receiving vaccine or placebo, your baby will be followed closely. During this part of the 
study, there will be about 9 days when your baby is seen by the clinical research staff and 21 days when 

 
  
 
IMPAACT 2012, Version 1.0 
Page 87 of 94 
08 August 2016 
your baby will not be seen but you will be contacted by telephone or email by the clinical research staff. 
Your baby will also be followed from November 1st until March 31st (the winter season after your baby 
receives vaccine or placebo).  During this winter time, we will contact you [site: insert the methods of 
contact] each week to ask about your baby's health and arrange for follow-up visits if needed.  
 
Your baby will participate in this study until April of the year after he/she started the study. Study visits 
will last about 30 minutes, except on the day when your baby is screened and on the day he/she is given 
the vaccine or placebo; those 2 visits may take about 1 to 2 hours each. 
 
 
If your baby has RSV symptoms, such as runny nose, sore throat, cough, fever or difficulty 
breathing, he/she might need to be seen for an evaluation, sometimes as quickly as within 24 
hours. 
 
Study visits, except the visit where your baby receives vaccine or placebo, may take place at your 
home, at your baby's pediatric practice, or at one of the research sites. The visit where your baby 
receives vaccine or placebo must take place at your baby's pediatric practice or at one of the 
research sites where emergency equipment is available. 
 
For temperature measurements, you will be asked to use a temporal thermometer, which is used 
on your baby’s forehead. You will measure forehead temperatures 3 times in a row, following the 
directions. The highest of the 3 readings will be recorded on a chart we will give to you. If your 
baby has a forehead temperature ≥100.0oF, you will be asked to check your baby’s rectal 
temperature within 20 minutes. Forehead and rectal thermometers will be provided to you for use 
during the study. 
 
Screening Visit 
 
The purpose of the screening visit is to find out if your baby may enter the study. It will take about 1 to 2 
hours and will include: 
 
the study staff telling you about the study and asking you questions to be sure you understand the 
study. 
 
reviewing and signing the study consent form. 
 
reviewing your baby's medical history and completing a physical examination. If the physical 
examination results are not normal, the clinical research staff will tell you and refer your baby for 
follow-up care with your baby's primary medical provider. 
 
answering questions about the health of your baby and people living in your house. 
 
collecting a small amount of blood (about 2 teaspoons) to test for antibodies (germ fighters) 
against RSV and how your baby’s cells are reacting to RSV. If your baby had been screened for 
IMPAACT 2011, we may not need to collect this sample. 
 
if requested, giving written permission to review your and your baby’s medical records. 
 
if your baby is found to be eligible for the study, your baby will be asked to return for a series of 
study visits, beginning with the visit when we will give your baby the vaccine or placebo. 
 
Day Vaccine/Placebo is Given 
 
We will confirm that your baby has not been ill recently and will check your baby’s temperature, 
pulse, and how fast your baby is breathing. 
 
Your baby will have a nasal wash. To do this, we will gently squirt less than 2 tablespoons of salt 
water inside your baby’s nose and then collect it when it comes back out of the nose. This is done 
before the vaccine or placebo is given to check for other viruses and to check for antibodies in the 
nose. 
 
Your baby will receive 1 dose of vaccine or placebo given as nose drops using a small medicine 
dropper. Your baby will be lying on his/her back while we give the nose drops and will remain 
lying down for about 1 minute afterwards. Your baby can be in your lap during this time. 

 
  
 
IMPAACT 2012, Version 1.0 
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08 August 2016 
 
After the nose drops are given, we will watch your baby in the clinic for 30 minutes.  
 
We will provide you with the dates of the remaining visits and telephone/email contact days.  
 
You will be given a forehead thermometer, a rectal thermometer, and a temperature card to record 
your baby's temperature daily for 29 days (including the day the vaccine/placebo is given to your 
baby), and at any other time you are concerned about fever. 
 
Monitoring for 56 Days after Vaccine/Placebo is Given 
 
Your baby will have study visits on Days 3, 5, 7, 10, 12, 14, 17, and 28  
(each ± 1 day), after the vaccine or placebo is given. Each visit will take about 30 minutes, and 
we will: 
o Check your baby's temperature, pulse, and breathing rate. 
o Do a brief clinical assessment. 
o Ask about your baby's health since the last visit. 
o Give your baby a nasal wash using less than 2 tablespoons of salt water, as described 
above, to check for the virus that’s in the study vaccine and other viruses. 
o Because study visits will be less frequent after the first month, on Day 28, we will review 
when you should contact the study staff in the event your baby becomes ill during the 
following month. 
 
The study nurse will contact you on Days 1, 2, 4, 6, 8, 9, 11, 13, 15, 16, and daily from Days 18 
to 27, and on Day 29.  The study staff will ask you to report your baby’s temperatures and any 
illness your baby has had since the last visit or contact.  The contact may be by telephone, text, or 
email, whichever you prefer.   
 
Your baby will have a follow-up visit about 56 days after the study nose drops were given. At this 
visit, we will ask about your baby’s health since the last visit and take a small amount of blood 
(about 2 teaspoons) from your baby's vein to measure antibodies (germ fighters) against RSV and 
how your baby’s blood cells are making antibodies. We will also give your baby a nasal wash 
using less than 2 tablespoons salt water to check for antibodies in the nose. 
 
We also ask you to call us right away to tell us about any illness that your baby has from the day 
he/she receives the nose drops up to the follow-up visit (8 weeks). 
 
A study nurse or study doctor will be available by telephone to answer your questions 24 hours a 
day during the 28 days after your baby receives the vaccine or placebo. 
 
If your baby becomes ill, you may be asked to bring him/her to the clinic for an examination, 
sometimes as quickly as within 24 hours. We may do a nasal wash at that time to look for the 
RSV vaccine virus or any other virus that may be in your baby's nose. 
 
Monitoring Before, During, and After RSV Season 
 
Your baby will also be followed during the winter RSV season (November 1st - March 31st) after 
receiving the study nose drops. We will be in contact with you each week to inquire about your 
baby's health. If your baby has a fever, a respiratory illness, or an ear infection that requires 
medical care, we will work with you to schedule a visit so that we can perform a nasal wash and 
clinical assessment. 
 
We will collect a small amount of blood (about 2 teaspoons) once in October before the winter 
RSV season and once in April after the winter RSV season to look at the antibodies (germ 
fighters) against natural RSV infection and how your baby’s blood cells are making antibodies. 
HOW MANY PEOPLE WILL TAKE PART IN THIS STUDY? 
There will be up to 33 babies taking part in this study. 

 
  
 
IMPAACT 2012, Version 1.0 
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08 August 2016 
HOW LONG WILL MY BABY BE IN THIS STUDY? 
Your baby will be in this study through next April, which is between 7 and 13 months from now, 
depending on which month of the year he/she started the study.    
WHAT ARE THE RISKS OF THE STUDY? 
Risks of the Vaccine 
 
If the vaccine is not weakened enough, it may cause a runny nose, sore throat, cough, or other 
signs of a cold. It is also possible that it may cause a sinus infection, croup, ear infection, fever, 
wheezing, or pneumonia (infection of the lungs). In another study with a similar vaccine, mild 
respiratory illnesses were observed frequently in babies who received either vaccine or placebo.  
Runny nose occurred more often in babies who got the vaccine than those who got the placebo. 
 
Study investigators have used the same placebo for studies of RSV, parainfluenza, and influenza 
vaccines in several hundred babies and children over the past 20 years.  They have not noticed 
side effects with this placebo.   
 
There is no specific medicine to treat RSV illness. If any symptoms of RSV occur, such as runny 
nose, sore throat, cough, or difficulty breathing, your baby will receive prompt medical care. 
 
The way the vaccine was made is very safe and was designed to minimize the possibility of other 
ingredients. However, as with all biological products, there is a small chance that it contains 
unidentified material. There is a very small chance that such material may cause illness, including 
possibly serious illness. 
 
There may be other side effects of the vaccine that are not yet known. If new information about 
possible side effects of this vaccine becomes available, we will let you know. 
 
It is possible that the vaccine virus could be spread from your baby to other people in the home or 
daycare and may make them sick. It could be spread to young babies and people with weakened 
immune systems. We have not seen this type of spread when other vaccines like this one have 
been studied.  
 
The vaccine could cause a severe allergic reaction. A severe reaction can cause hives, throat 
swelling, rapid heart rate, weakness, difficulty breathing, or death. Such reactions are rare. 
 
Risks of Nasal Washes 
Nasal washes may cause brief discomfort or pain that is like the feeling of getting salt water in the nose 
and may rarely cause a nosebleed. 
 
Risks of Having Blood Drawn 
Blood drawing can cause bleeding, pain, bruising, or infection at the place where the blood is taken. 
Sometimes, blood drawing can cause your baby to feel lightheaded or to faint. It sometimes takes more 
than 1 try to get blood from a small baby. 
WHY WOULD THE DOCTOR TAKE MY BABY OFF THIS STUDY EARLY? 
The study doctors or the sponsor have the right to end your baby's participation in the study at any time 
without your consent for any of the following reasons: 
 
It would be dangerous for your baby to continue; 
 
You do not follow study procedures as directed by the study doctors;  
 
New information becomes available regarding the safety of the vaccine; 
 
If it is in your baby's best interest; 

 
  
 
IMPAACT 2012, Version 1.0 
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08 August 2016 
 
You do not consent to continue in the study after being told of changes in the research that may 
affect your baby; 
 
The study sponsor, the International Maternal Pediatric Adolescent AIDS Clinical Trials Network 
(IMPAACT), the Institutional Review Board (IRB), the Office for Human Research Protections 
(OHRP), the National Institute of Allergy and Infectious Diseases (NIAID), or the United States 
Food and Drug Administration (FDA) decide to end the study. (An IRB is a committee that 
watches over the safety and rights of research participants.) 
 
WHAT HAPPENS IF MY BABY IS INJURED? 
 
If your baby suffers physical injury from this study, the study doctor will provide or will refer your baby 
for immediate medical treatment.  The study doctor will also provide referrals to appropriate health care 
facilities. The cost for this treatment will be charged to you or your insurance company.  There is no 
program for compensation either through this institution or the National Institutes of Health (NIH). No 
financial compensation by the doctors that gave your baby the vaccine or placebo will be made for any 
discomfort suffered because of participation in this study. You will not be giving up any of your legal 
rights by signing this consent form. 
ARE THERE BENEFITS TO TAKING PART IN THIS STUDY? 
 
Your baby is not expected to receive any direct benefit from enrollment in the study. 
 
Your baby's involvement in the study may help find a vaccine that works well to prevent serious 
RSV illness. Such a vaccine may be of future benefit to babies and children in this country and in 
the rest of the world. 
WHAT OTHER CHOICES DO I/DOES MY BABY HAVE BESIDES THIS STUDY? 
There are no licensed vaccines to protect against RSV illness at this time. There is no other similar study 
or licensed vaccine that we can offer your baby. You may choose to not allow your baby to take part in 
this study. 
WHAT ABOUT CONFIDENTIALITY? 
To help us protect your privacy, we have obtained a Certificate of Confidentiality from the National 
Institutes of Health.  With this Certificate, the researchers cannot be forced to disclose information that 
may identify you, even by a court subpoena, in any federal, state, or local civil, criminal, administrative, 
legislative, or other proceedings.  The researchers will use the Certificate to resist any demands for 
information that would identify you, except as explained below.  The Certificate cannot be used to resist a 
demand for information from personnel of the United States Government that is used for auditing or 
evaluation of federally funded projects or for information that must be disclosed in order to meet the 
requirements of the federal Food and Drug Administration (FDA).   
 
You should understand that a Certificate of Confidentiality does not prevent you or a member of your 
family from voluntarily releasing information about you or your baby’s participation in this research.  If 
an insurer, employer, or other person obtains your written consent to receive research information, then 
the researchers may not use the Certificate of Confidentiality to withhold that information. 
 
The Certificate of Confidentiality does not prevent the researchers from disclosing voluntarily, without 
your consent, information that would identify you as a participant in the research project under certain 
circumstances such as child abuse.  

 
  
 
IMPAACT 2012, Version 1.0 
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08 August 2016 
 
Your baby’s name, birth date, and Social Security number are not routinely given to anyone unless required by 
law. All of the information you give us during this study will be put in locked file cabinets and/or in 
password-protected computer files. The only people who will have access to this information will be those 
who are involved in the study. 
 
There will be people involved in the study who need to see your baby’s health information. These people 
may include the researchers, study and laboratory personnel, and other clinical research staff. Others who may 
see your baby’s information are the groups of people who make sure that the study is being done as it 
should be: Hospital Institutional Review Boards (IRBs), the Center for Immunization Research (CIR), 
the National Institute of Allergy and Infectious Diseases (NIAID; NIH) Intramural Data and Safety 
Monitoring Board and others who need to see your baby’s information to make sure that the study is going 
as planned.  
 
Other groups of people who may be involved in the study and may need to see your baby’s 
information are: 
 
The government agency “Office for Human Research Protections,” that makes sure that we are 
conducting the research as planned, and the U.S. FDA 
 
The sponsor of the study and people with whom the sponsor may contract for the study, such as 
study monitors. 
 
At the end of the study, whatever we learn from the research may be used in a medical journal or used for 
teaching. Your baby’s name or other details about his/her health will not be used in a manner such that 
anyone can personally identify your baby. 
WHAT ARE THE COSTS TO ME? 
There are no costs to you or your baby for him/her being in the study. The costs for vaccine/placebo, 
study visits, or study procedures are covered by the sponsor (NIH/NIAID).  However, taking part in this 
study may lead to added costs to you or your baby and your/your baby’s insurance company if medical 
complications arise or if your baby’s doctor decides extra tests are needed.  In some cases, it is possible 
that your/your baby’s insurance company will not pay for these costs, because your baby is taking part in 
a research study.   
WILL MY BABY RECEIVE ANY COMPENSATION? 
You will be paid for your baby's participation in this study at the following rate [Site: insert payment 
schedule and amount.] 
 
You will also be paid during the winter RSV surveillance period as follows [Site: insert payment schedule 
and amount.] 
 
[Optional, depending on site: If you stop your baby from taking part in the study early, you will only be 
paid for the days of the study that your baby completed. Your baby may also receive age-appropriate 
books or small toys. If needed, bus tokens or parking passes will be given to you.] 
 
You may be required to provide your Social Security number to be paid. If your payment for study 
participation exceeds $600 per year, this information must be reported to the Internal Revenue Service. 

 
  
 
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08 August 2016 
WHAT ARE MY BABY’S RIGHTS AS A RESEARCH PARTICIPANT? 
Taking part in this study is completely voluntary.  You may choose not to have your baby take part in this 
study or leave this study at any time.  Your decision will not have any impact on your baby’s participation 
in other studies and will not result in any penalty or loss of benefits to which you or your baby are 
otherwise entitled. 
 
A study physician, physician assistant, nurse practitioner, or study nurse will inform you of any 
significant abnormal physical findings and will make appropriate referrals back to your baby’s primary 
care giver, if necessary. 
 
We will tell you about new information from this or other studies that may affect your baby’s health, 
welfare, or willingness to stay in this study. You may be asked to sign a revised consent form if this 
occurs. If you want the results of the study, let the clinical research staff know. 
 
At the end of the study, you will be told in writing whether your baby was given the vaccine or the 
placebo. 
WHAT ARE MY RESPONSIBILITIES? 
 
If you decide to withdraw your baby from the study early, we ask that you notify the study nurse 
or study doctor. 
 
If your baby comes off the study early, we will ask you to bring him/her into the clinic for an 
early discontinuation visit so that we can do a final blood draw (about 1 teaspoon) and nasal 
wash. 
 
Any baby who has received the study product will be encouraged to remain in the study so that 
safety information can be collected. 
 
It is important that you do not enroll your baby in other studies where your baby receives 
vaccines or medications for 8 weeks after he/she receives vaccine/placebo. 
 
WHAT DO I DO IF I HAVE QUESTIONS OR PROBLEMS? 
For questions about this study or a research-related injury, contact: 
 
[Site: Insert name of the investigator or other study staff] 
 
[Site: insert telephone number of above] 
For questions about your baby’s rights as a research participant, contact: 
 
 
[Site: insert name or title of person on the Institutional Review Board (IRB) or other 
organization] 
 
[Site: insert telephone number of above]   
 
 
 

 
  
 
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GENETIC TESTING 
 
Some of the blood tests done for this study will look at how your baby’s genes (DNA) affect his or her 
response to RSV.  Researchers may use your baby’s samples for limited genetic testing.  For example, 
researchers may do “genetic variations” research. They may look at genes that affect how your baby 
fights infections.  Our genes are passed to us from our birth parents. Genes are the basic “instruction 
book” for the cells that make up our bodies. The differences in people’s genes can help explain why some 
people get a disease while others do not. The researchers will not contact you or your baby’s regular 
health care provider with the results of these tests.  This is because these tests are often done with 
experimental procedures and the results should not be used to make decisions about your baby’s health 
care.  However, if the researchers decide that a result is important information for your baby’s health care, 
the study doctor will be notified.  If you would like to be contacted with this sort of information, you must 
notify the study staff of any changes of your address and phone number. Your baby’s name will not be 
available to the laboratory or to the scientists who may be doing limited genetic testing. 
 
You may decide that you do not want your baby’s blood used for limited genetic testing.  Your baby can 
still be in this study even if you make this decision.  Please read the following statement carefully and 
then mark your initials in the appropriate space provided. 
 
I allow my baby’s blood to be used for limited genetic testing, including future limited genetic testing, as 
part of this study. 
 
Yes:  
Initials _________ 
Date ____________ 
 
No:  
Initials _________ 
Date ____________ 
 
 
STORAGE AND FUTURE USE OF UNUSED SPECIMENS  
 
If you agree, any unused blood or nasal wash samples taken from your baby will be stored indefinitely 
(with protectors of identity) once this study is complete. These unused blood and nasal wash samples may 
be used for future laboratory studies to learn more about RSV and other viruses. This information may 
lead to other new virus vaccines in the future. 
 
 
Your baby’s unused blood or nasal wash samples, if any, will be used only for laboratory studies 
and will not be sold or used directly to make products that will be for sale. 
 
The samples will be coded so that your baby’s name cannot be easily identified. 
 
Reports about studies done with your baby’s unused samples will not be put in your baby’s health 
or study records.  
 
There will be no direct benefit to your baby in using the samples as noted above, but from 
studying the unused samples of babies taking part in the studies, we may learn more about the 
RSV germ or other viruses that cause illness in babies and children. 
 
Results from future studies using your baby’s unused samples may be included in medical papers 
and meeting reports, but your baby’s name will not be used. 
 
You can change your mind at any time about allowing your baby’s unused samples to be used for future 
laboratory studies. If you do change your mind, contact the study doctor or study nurse and let him/her 
know. Then the samples will no longer be used for laboratory studies and will be destroyed. 
 
 

 
  
 
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PERMISSION FOR STORAGE AND FUTURE USE OF UNUSED SPECIMENS 
Your choice will not have any effect on your baby’s taking part in this study. 
 
I will allow the use of my baby’s unused blood or nasal wash samples to be stored indefinitely and to be 
used in future laboratory studies for the purposes described above. Your baby’s name will not be 
available to the laboratory or to the scientists who may be doing any future tests. (Please check one and 
initial below) 
 
Yes:  
Initials _________ 
Date ____________ 
 
No:  
Initials _________ 
Date ____________ 
 
 
If NO, your baby’s study samples will be destroyed after the study is completed.  
 
 
 

 
  
 
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08 August 2016 
SIGNATURE  
 
If you have read this consent form (or had it explained to you), all your questions have been answered, 
and you agree to take part in this study, please sign your name below. 
 
 
____________________________                 
Study Participant’s Name (print) 
 
 
 
 
 
_____________________________              ____________________________________ 
Participant’s Legal Guardian (print) 
 
Legal Guardian’s Signature and Date 
 
 
 
 
____________________________                ____________________________________ 
Clinical Research Staff Conducting 
 
Clinical Research Staff Signature and Date 
 
 
Consent Discussion (print)  
 
 
____________________________                ____________________________________ 
Witness’ Name (print)  
 
            Witness’ Signature and Date 
 
 
 
(As appropriate) 
 
 
____________________________ 
 
__________________________________ 
Second Parent/Guardian’s Name 
 
Signature and Date 
(As appropriate)