Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of a Single Dose of a Recombinant Live-attenuated Respiratory Syncytial Virus Vaccine, RSV 6120/∆NS1, Lot RSV#018A, or RSV 6120/F1/G2/∆NS1, Lot RSV#016A, Delivered as Nose Drops to RSV-seropositive Children 12 to 59 Months of Age and RSV-seronegative Infants and Children 6 to 24 Months of Age
Phase: PHASE1
Conditions: RSV Infection
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

1 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of a Single 
Dose of a Recombinant Live-attenuated Respiratory Syncytial Virus Vaccine, RSV 
6120/∆NS1, Lot RSV#018A, or RSV 6120/F1/G2/∆NS1, Lot RSV#016A, Delivered as Nose 
Drops to RSV-seropositive Children 12 to 59 Months of Age and RSV-seronegative Infants 
and Children 6 to 24 Months of Age 
Sponsored by: 
National Institute of Allergy and Infectious Diseases (NIAID) 
 
Center for Immunization Research (CIR) Protocol Number: CIR 330 
WCG IRB Protocol Number: WCG IRB # 20181405 
FDA Investigational New Drug (IND) Number: 018219 
Project Assurance: FWA #00000287 
 
IND Sponsor: 
Office of Clinical Research Policy and Regulatory Operations (OCRPRO) 
Division of Clinical Research (DCR) 
National Institute of Allergy and Infectious Diseases (NIAID) 
National Institutes of Health (NIH) 
5601 Fishers Lane 
Room 4B11, MSC 9820 
Bethesda, MD 20892 
 
 
Respiratory Syncytial Virus (RSV) 6120/∆NS1 
RSV 6120/F1/G2/∆NS1 
Provided by: 
Laboratory of Infectious Diseases (LID), NIAID, NIH 
 
Medical Monitor: 
Kaeryn Lewis, MD 
 
Principal Investigator: 
Ruth Karron, MD 
 
Version 5.0 
June 5, 2023 
 
 
 
 
The information contained herein is the property of the NIAID and may not be reproduced, published 
or disclosed to others without the written authorization of the NIAID. 

2 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
TABLE OF CONTENTS 
LIST OF TABLES ......................................................................................................................................................... 5 
LIST OF FIGURES ....................................................................................................................................................... 6 
LIST OF APPENDICES ............................................................................................................................................... 6 
STUDY STAFF ROSTER ............................................................................................................................................. 7 
LIST OF ABBREVIATIONS ........................................................................................................................................ 9 
PROTOCOL SUMMARY .......................................................................................................................................... 12 
1. 
BACKGROUND AND SCIENTIFIC RATIONALE ....................................................................................... 14 
1.1. 
BACKGROUND ............................................................................................................................................. 14 
1.1.1. Epidemiology, Disease Burden, and the Need for a Vaccine ............................................................... 14 
1.2. 
PRIOR RESEARCH ........................................................................................................................................ 16 
1.2.1. Experimental Vaccines against RSV .................................................................................................... 16 
1.2.2. Preclinical Studies ................................................................................................................................ 17 
1.2.2.1. 
Replication of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 (Experimental Lots) in vitro ........................ 17 
1.2.2.2. 
Replication of experimental lot and CTM of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ ΔNS1 in African green 
monkeys (AGMs) ............................................................................................................................................................. 19 
1.2.3. Previous Clinical Experience ............................................................................................................... 20 
1.3. 
RATIONALE ................................................................................................................................................. 20 
1.4. 
CLINICAL DEVELOPMENT PLAN .................................................................................................................. 21 
1.5. 
HYPOTHESES ............................................................................................................................................... 21 
2. 
OBJECTIVES ................................................................................................................................................... 21 
2.1. 
PRIMARY OBJECTIVES................................................................................................................................. 21 
2.2. 
SECONDARY OBJECTIVES ............................................................................................................................ 22 
2.3. 
EXPLORATORY OBJECTIVE ......................................................................................................................... 22 
3. 
STUDY DESIGN ............................................................................................................................................. 22 
4. 
STUDY POPULATION ................................................................................................................................... 24 
4.1             INCLUSION CRITERIA FOR RSV-SEROPOSITIVE CHILDREN ....................................................................... 25 
4.2. 
EXCLUSION CRITERIA FOR RSV-SEROPOSITIVE CHILDREN ........................................................................ 25 
4.3. 
INCLUSION CRITERIA FOR RSV-SERONEGATIVE INFANTS & CHILDREN ..................................................... 27 
4.4. 
EXCLUSION CRITERIA FOR RSV-SERONEGATIVE INFANTS & CHILDREN .................................................... 27 
4.5. 
CO-ENROLLMENT CONSIDERATIONS .......................................................................................................... 29 
4.6. 
RE-ENROLLMENT CONSIDERATIONS ........................................................................................................... 29 
4.7. 
RECRUITMENT PROCESS ............................................................................................................................. 29 
4.8. 
PARTICIPANT RETENTION ........................................................................................................................... 30 
4.9. 
PARTICIPANT WITHDRAWAL OR TERMINATION FROM THE STUDY .............................................................. 30 
5. 
STUDY PRODUCT ......................................................................................................................................... 30 
5.1. 
STUDY PRODUCT REGIMENS ....................................................................................................................... 31 
5.2. 
STUDY PRODUCT FORMULATION ................................................................................................................ 31 
5.2.1. Vaccines ............................................................................................................................................... 31 
5.2.2. Diluent for RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 .................................................................. 32 
5.2.3        Placebo for RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1.............................................................. 32 
5.3. 
STUDY PRODUCT STORAGE ......................................................................................................................... 32 
5.4. 
STUDY PRODUCT PREPARATION ................................................................................................................. 33 
5.4.1. Diluent .................................................................................................................................................. 33 
5.4.2. Placebo ................................................................................................................................................. 33 
5.4.3. Live RSV 6120/∆NS1 or RSV 6120/F1/G2/ΔNS1 .............................................................................. 33 

3 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
5.5. 
STUDY PRODUCT INOCULATION PROCEDURE ............................................................................................. 34 
5.6. 
STUDY PRODUCT ACQUISITION ................................................................................................................... 34 
5.7. 
STUDY PRODUCT ACCOUNTABILITY ........................................................................................................... 35 
5.8. 
DISPOSITION OF USED/UNUSED STUDY PRODUCT ...................................................................................... 35 
5.9. 
FINAL DISPOSITION OF STUDY PRODUCTS .................................................................................................. 35 
5.10. 
CONCOMITANT MEDICATIONS .................................................................................................................... 35 
6. 
STUDY VISITS AND PROCEDURES ........................................................................................................... 36 
6.1. 
CONSENTING PROCESS ................................................................................................................................ 36 
6.2. 
SCREENING VISIT ........................................................................................................................................ 37 
6.3. 
RANDOMIZATION ........................................................................................................................................ 38 
6.4. 
ENROLLMENT ............................................................................................................................................. 39 
6.4.1. Enrollment - Inoculation (Day 0) ......................................................................................................... 39 
6.5. 
STUDY PHASES ........................................................................................................................................... 41 
6.5.1. RSV-Seropositive Children (Group 1) ................................................................................................. 42 
6.5.1.1. 
Acute Phase: In-person Study Visit Days ..................................................................................................... 42 
6.5.1.2. 
Acute Phase: Non-Visit Study Day Contacts ................................................................................................ 42 
6.5.1.3. 
Study Day 11 ................................................................................................................................................ 43 
6.5.1.4. 
Post-Acute Phase .......................................................................................................................................... 44 
6.5.1.5. 
Study Day 28 Visit ........................................................................................................................................ 44 
6.5.2. RSV-Seronegative Infants and Children .............................................................................................. 44 
6.5.2.1. 
Acute Phase: In-person Study Visit Days ..................................................................................................... 44 
6.5.2.2. 
Acute Phase: Non-Visit Study Day Contacts ................................................................................................ 46 
6.5.2.3. 
Study Day 29 ................................................................................................................................................ 46 
6.5.2.4. 
Post-Acute Phase .......................................................................................................................................... 46 
6.5.2.5. 
Study Day 56 Visit ........................................................................................................................................ 47 
6.5.2.6. 
Period after Day 56 through October 31st ..................................................................................................... 48 
6.6. 
RSV SURVEILLANCE SEASON (RSV-SERONEGATIVE SUBJECTS) ................................................................ 48 
6.6.1. Pre-RSV Season Surveillance Study Visit ........................................................................................... 48 
6.6.2. Weekly Contact for Surveillance during the RSV Season ...................................................................... 48 
6.6.3. Post-RSV Season Surveillance Study Visit .......................................................................................... 50 
6.7. 
ILLNESS VISIT ............................................................................................................................................. 50 
6.8. 
EARLY DISCONTINUATION STUDY VISIT .................................................................................................... 51 
6.9. 
LABORATORY PROCEDURES ....................................................................................................................... 53 
6.9.1. Specimen Collection ............................................................................................................................. 53 
6.9.2. Virus Detection .................................................................................................................................... 53 
6.9.3. Specimen Collection ............................................................................................................................. 53 
6.9.4. Specimen Preparation, Testing, Storage, and Shipping ........................................................................ 54 
6.9.5. Research Laboratories .......................................................................................................................... 54 
6.9.6. Plan for Use and Storage of Biological Samples .................................................................................. 54 
6.9.7. Biohazard Containment ........................................................................................................................ 54 
7. 
SAFETY ASSESSMENT, MONITORING, AND REPORTING ................................................................... 55 
7.1. 
SAFETY-RELATED ROLES AND RESPONSIBILITIES....................................................................................... 55 
7.1.1. Principal Investigator ........................................................................................................................... 55 
7.1.2. Safety Review and Communications Plan (SRCP) .............................................................................. 55 
7.1.3. Sponsor Medical Monitor ..................................................................................................................... 55 
7.1.4. Data Safety Monitoring Board ............................................................................................................. 55 
7.1.5. Sponsor Reporting ................................................................................................................................ 55 
7.2. 
SAFETY-RELATED RECORDING ON CRFS ................................................................................................... 56 
7.3. 
SERIOUS ADVERSE EVENT REPORTING ....................................................................................................... 60 
7.4. 
REPORTING OF UNANTICIPATED PROBLEMS TO OFFICE OF CLINICAL RESEARCH   POLICY AND REGULATORY 
OPERATIONS (OCRPRO) ...................................................................................................................... 60 
8. 
PARTICIPANT MANAGEMENT ................................................................................................................... 61 
8.1. 
MANAGEMENT OF ADVERSE EVENTS .......................................................................................................... 61 

4 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
8.1.1. Solicited Adverse Events ...................................................................................................................... 62 
8.1.2. Serious Adverse Events ........................................................................................................................ 63 
8.1.3. Unexpected Adverse Event .................................................................................................................. 63 
8.2. 
GRADING THE SEVERITY OF ADVERSE EVENTS .......................................................................................... 63 
8.2.1. AE Grading .......................................................................................................................................... 64 
8.2.2. Fever Grading ....................................................................................................................................... 64 
8.3. 
PAUSING AND STOPPING RULES .................................................................................................................. 64 
9. 
STATISTICAL CONSIDERATIONS .............................................................................................................. 66 
9.1. 
GENERAL DESIGN ISSUES ........................................................................................................................... 66 
9.1.1. General Design ..................................................................................................................................... 66 
9.1.2. Description of the Statistical Methods to be Employed ....................................................................... 66 
9.2. 
OUTCOME MEASURES ................................................................................................................................. 66 
9.2.1. Primary Outcome Measures ................................................................................................................. 66 
9.2.2. Secondary Outcome Measures ............................................................................................................. 67 
9.3. 
SAMPLE SIZE AND ACCRUAL ...................................................................................................................... 67 
9.3.1. Sample Size and Randomization .......................................................................................................... 67 
9.4. 
MONITORING .............................................................................................................................................. 71 
9.4.1. Site Monitoring Plan ............................................................................................................................ 71 
9.4.2. Monitoring by the NIAID Intramural Data and Safety Monitoring Board ........................................... 71 
9.5. 
ANALYSES .................................................................................................................................................. 72 
9.5.1. Assessment of Primary Objectives ....................................................................................................... 72 
9.5.2. Assessment of Secondary Objectives ................................................................................................... 73 
10 
DATA HANDLING AND RECORD KEEPING ............................................................................................. 73 
10.1. 
DATA MANAGEMENT RESPONSIBILITIES .................................................................................................... 73 
10.2. 
ESSENTIAL AND SOURCE DOCUMENTS AND ACCESS TO SOURCE DATA ...................................................... 73 
10.3. 
CLINICAL INVESTIGATOR’S BROCHURES .................................................................................................... 74 
10.4. 
QUALITY CONTROL AND QUALITY ASSURANCE ......................................................................................... 74 
11 
CLINICAL SITE MONITORING .................................................................................................................... 74 
12 
HUMAN SUBJECTS PROTECTIONS ........................................................................................................... 75 
12.1. 
INSTITUTIONAL REVIEW BOARD/ETHICS COMMITTEE REVIEW AND APPROVAL ......................................... 75 
12.2. 
VULNERABLE PARTICIPANTS ...................................................................................................................... 76 
12.3. 
INFORMED CONSENT ................................................................................................................................... 76 
12.4. 
POTENTIAL BENEFITS ................................................................................................................................. 76 
12.5. 
POTENTIAL RISKS ....................................................................................................................................... 76 
12.5.1. Venipuncture ........................................................................................................................................ 76 
12.5.2. Nasal Wash or Swab ............................................................................................................................. 77 
12.5.3. Nasosorption SAM Strips ..................................................................................................................... 77 
12.5.4. Topical Anesthetic Cream .................................................................................................................... 77 
12.5.5. Receipt of Study Product ...................................................................................................................... 77 
12.6. 
REIMBURSEMENT/COMPENSATION ............................................................................................................. 77 
12.7. 
PRIVACY AND CONFIDENTIALITY ................................................................................................................ 78 
12.8. 
MANAGEMENT OF INCIDENTAL FINDINGS ................................................................................................... 78 
13 
ADMINISTRATIVE PROCEDURES ............................................................................................................. 78 
13.1. 
REGULATORY OVERSIGHT .......................................................................................................................... 78 
13.2. 
PROTOCOL REGISTRATION .......................................................................................................................... 78 
13.3. 
STUDY IMPLEMENTATION ........................................................................................................................... 79 
13.4. 
PROTOCOL DEVIATION REPORTING ............................................................................................................ 79 
13.5. 
CLINICALTRIALS.GOV ................................................................................................................................. 79 
14 
PUBLICATIONS .............................................................................................................................................. 79 

5 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
15 
REFERENCES ................................................................................................................................................. 80 
 
LIST OF TABLES 
 
Table 1: Inoculation schedule ......................................................................................................... 12 
Table 2: Screening Visit Procedures ............................................................................................... 38 
Table 3: Enrollment Visit Procedures ............................................................................................. 40 
Table 4: Acute Phase In-Person Visit Procedures – RSV-Seropositive Children .......................... 42 
Table 5: Acute Phase Non-Visit Contact Procedures – RSV-Seropositive Children ..................... 43 
Table 6: Day 11 Non-Visit Procedures – RSV-Seropositive Children ........................................... 43 
Table 7: Day 28 Visit Procedures – RSV-Seropositive Children ................................................... 44 
Table 8: Acute Phase In-Person Visit Procedures – RSV-Seronegative Children ......................... 45 
Table 9: Acute Phase Non-Visit Contact Procedures – RSV-Seronegative Children .................... 46 
Table 10: Day 29 Non-Visit Procedures – RSV-Seronegative Children ........................................ 46 
Table 11: Day 56 Visit Procedures – RSV-Seronegative Children ................................................ 47 
Table 12: Pre-RSV Season Surveillance Study Visit Procedures ................................................... 48 
Table 13: RSV Season Surveillance Procedures ............................................................................ 49 
Table 14: Post-RSV Seasonal Surveillance Study Visit Procedures .............................................. 50 
Table 15: Illness Visit Timeframe .................................................................................................. 51 
Table 16: Illness Visit Procedures .................................................................................................. 51 
Table 17: Early Discontinuation Procedures .................................................................................. 52 
Table 18: AE CRF Recording Requirements for RSV-Seropositive Participants .......................... 57 
Table 19: AE CRF Recording Requirements for RSV-Seronegative Participants ......................... 58 
Table 20: Grading for Adverse Events ........................................................................................... 64 
Table 21: Grading for Fever ........................................................................................................... 64 
Table 22: Pausing and Stopping Rules ........................................................................................... 65 
Table 23: The Probability of Observing LRI Events in Seronegative Vaccine Recipients ............ 68 
Table 24: Percent of Participants Experiencing LRI or AEs with Exact 90% CI ........................... 69 
Table 25: Magnitude of Difference in Responses Detectable with 80% Power ............................. 69 
Table 26: Viral titers of nasopharyngeal swab samples from AGMs inoculated with RSV ΔNS1 
mutants, or with recombinant wt RSV rA2 .................................................................. 84 
Table 27: Viral titers of tracheal lavage samples from AGMs inoculated with RSV ΔNS1 mutants, 
or with Recombinant wt RSV rA2 ................................................................................ 85 
Table 28: Serum PRNT60 Titers in AGMs Inoculated with RSV ΔNS1 mutants, or with 
Recombinant wt RSV rA2 ............................................................................................ 86 
Table 29: Schedule of Events: Group 1: RSV-Seropositive Participants ....................................... 87 
Table 30: Schedule of Events: Group 2: RSV-Seronegative Participants ...................................... 88 
Table 31: Schedule of Events: RSV Seasonal Surveillance ........................................................... 89 
Table 32: Definitions of Solicited Adverse Events ......................................................................... 90 
 

6 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
LIST OF FIGURES 
 
Figure 1.1-1: Group 1 – Study Overview ....................................................................................... 13 
Figure 1.1-2: Group 2 – Study Overview ....................................................................................... 13 
Figure 1.2-1. Multicycle replication of experimental lots in Vero cells or in an in-vitro model of 
normal human bronchial/tracheal epithelial cells ......................................................... 18 
Figure 5.2-1: Investigational Product Label Sample ...................................................................... 32 
Figure 5.2-2: Investigational Product Label Sample ...................................................................... 32 
Figure 9.3-1: Power Curves for Assessment of Vaccine Virus Shedding in RSV seronegative 
Vaccinees ...................................................................................................................... 70 
Figure 6.6-1: RSV Seasonality in Baltimore .................................................................................. 91 
 
 
 
 
 
 
LIST OF APPENDICES 
 
Appendix I: Tables Referenced in the Background and PRECLINICAL Sections ........................ 84 
Appendix II: Schedule of Events: Screening, Acute Phase, and Post-Acute Phase ....................... 87 
Appendix III: Schedule of Events: RSV Seasonal Surveillance ..................................................... 89 
Appendix IV: Definitions of Solicited Adverse Events .................................................................. 90 
 

7 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
STUDY STAFF ROSTER 
PRINCIPAL INVESTIGATOR 
Ruth A. Karron, MD 
Center for Immunization Research (CIR) 
Johns Hopkins Bloomberg School of Public Health (BSPH) 
624 N. Broadway, Room 217 
Baltimore, MD 21205 
Phone: 410-614-0319 
Administrative Coordinator: 410-955-1624  
Email: rkarron@jhu.edu 
 
CLINICAL INVESTIGATORS  
Kawsar Talaat, MD 
CIR, JHSPH 
624 N. Broadway, Room 249 
Baltimore, MD 21205 
Phone: 410-502-9627 
Email: ktalaat1@jhu.edu 
 
 
Kristi Herbert, CRNP-P 
 
CIR, JHSPH 
624 N. Broadway, Room 207 
Baltimore, MD 21205 
Phone: 410-955-4362  
Email: kherber1@jhu.edu 
 
Elizabeth Schappell, RN, MSN, CCRP 
CIR, JHSPH 
624 N. Broadway, Room 205 
Baltimore, MD 21205 
Phone: 410-614-9114  
Email: eschappe@jhu.edu 
 
Jocelyn San Mateo, CRNP-F, CCRP 
CIR, JHSPH 
624 N. Broadway, Room 201 
Baltimore, MD 21205 
Phone: 410-614-4306 
Email: jsanmat1@jhu.edu  
Beulah Sabundayo, PharmD, MPH 
CIR, JHSPH 
624 N. Broadway, Room 200 
Baltimore, MD 21205 
Phone: 410-502-7451 
Email: bsabund1@jhu.edu 
 
Suzanne Woods, CRNP-P, CCRP 
CIR, JHSPH 
624 N. Broadway, Room 212 
Baltimore, MD 21205 
Phone: 410-614-1880 
Cell: 443-813-0697 
Email: swoods12@jhu.edu 
 
SCIENTIFIC INVESTIGATORS  
 
Ursula Buchholz, PhD 
LID, NIAID, NIH 
Building 50, Room 6503 
50 South Drive, MSC 8007 
Bethesda, MD 20892 
Phone: 301-594-1533 
Email: ubuchholz@niaid.nih.gov 
 
 
 
 
 
 
 
 

8 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
LINICAL TRIAL SITES 
Center for Immunization Research 
Johns Hopkins Bloomberg School of 
Public Health  
624 N. Broadway, Room 117 
Baltimore, MD 21205 
Phone: 410-502-3333 
 
CLINICAL LABORATORIES 
 
Johns Hopkins Hospital 
Laboratories of Pathology 
600 N. Wolfe Street 
Baltimore, MD 21287 
Phone: 410-955-3142 
 
Center for Immunization Research East  
Johns Hopkins Bayview Medical Center  
Campus 
301 Mason Lord Drive 
Baltimore, MD 21224 
Phone: 410-550-2725 
 
Center for Immunization Research South 
 
9811 Mallard Drive Suite 210 
Laurel MD 20708 
 
Quest Diagnostics  
1901 Sulphur Spring Road 
Baltimore, MD 21227 
Phone: 410-247-0001 
 
Investigational Drug Service 
Sasha Beselman, Pharm.D. 
Rachel Sands, Pharm D. 
Clinical Pharmacist 
Johns Hopkins Hospital 
 
Phone: 410-955-6337 
VIROLOGY/IMMUNOLOGY 
LABORATORIES 
CIR Laboratories 
615 N. Wolfe Street, Room E5402 
Baltimore, MD 21205 
Phone: 410-955-7230 
 
INSTITUTIONAL REVIEW BOARD 
 
Western Copernicus GroupInstitutional Review Board® 
(WCG IRB) 
1019 39th Avenue SE Suite 120 
Puyallup, WA 98374-2115 
Phone: 1-855-818-2289  
Email:researchquestions@wcgirb.com 
 
INSTITUTIONAL BIOSAFETY OFFICE (IBC) 
2024 E. Monument Street, Room B-200 
Baltimore, MD 21287 
 
 
SPONSOR REPRESENTATIVE 
John Tierney, RN, MPM 
OCRPRO, DCR, NIAID, NIH 
5601 Fishers Lane 
Room 4B11, MSC 9820 
Bethesda, MD 20892 
Phone: 301-451-5136  
Email: jtierney@niaid.nih.gov 
 
SPONSOR MEDICAL MONITOR 
Kaeryn Lewis, MD 
Medical Monitor 
Clinical Monitoring Research Program 
Leidos Biomedical Research, Inc. 
5705 Industry Lane, Room   
Frederick, Maryland 21704 
Phone 301-473-2314  
Email: kaeryn.lewis@nih.gov 
 
 
 

9 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
LIST OF ABBREVIATIONS 
 
ACIP 
AE 
Advisory Committee on Immunization Practices (CDC) 
adverse event 
AGM 
African green monkey 
cDNA 
complementary deoxyribonucleic acid 
CFR 
Code of Federal Regulations 
CI 
confidence interval 
CIR 
Center for Immunization Research 
CRF 
case report form 
CRL 
Charles River Laboratories 
CSO 
Clinical Safety Office 
CTM 
clinical trial material 
DCR 
Division of Clinical Research 
DNA 
deoxyribonucleic acid 
DSMB 
Data and Safety Monitoring Board 
ELISA 
enzyme-linked immunosorbent assay 
F protein 
fusion protein  
FDA 
GCP 
Food and Drug Administration 
Good Clinical Practice 
GMT 
geometric mean titer 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
human immunodeficiency virus 
IB 
investigator’s brochure 
ICD 
informed consent document 
ICH 
International Council for Harmonisation of Technical Requirements for Pharmaceuticals 
for Human Use 
IgA, IgG, IgE 
immunoglobulin A, G, E 
i.n. 
intranasally 
IND 
investigational new drug 
IRB 
institutional review board 

10 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
i.t. 
intratracheally 
JHSPH 
Johns Hopkins Bloomberg School of Public Health 
JHU 
Johns Hopkins University 
LID 
Laboratory of Infectious Diseases 
LRI 
lower respiratory tract illness 
LRT 
lower respiratory tract 
MOP 
manual of procedures 
mRNA 
messenger ribonucleic acid 
NHP 
nonhuman primate 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
OCRPRO 
Office of Clinical Research Policy and Regulatory Operations 
OHRP 
Office for Human Research Protections 
ORF 
open reading frame 
PE 
physical examination 
PFU 
plaque-forming units 
PI 
principal investigator 
PRNT 
plaque reduction neutralization 
r 
recombinant 
RE 
regulatory entity 
RNA 
ribonucleic acid 
rRT-PCR 
reverse transcription polymerase chain reaction 
RSV 
respiratory syncytial virus 
RT-qPCR 
quantitative reverse transcription polymerase chain reaction 
SAE 
serious adverse event 
SAM 
synthetic absorptive matrix 
SERF 
Safety Expedited Report Form 
SMM 
sponsor medical monitor 
SOP 
standard operating procedure 
SUSAR 
serious, unexpected, suspected adverse reaction 
TL 
tracheal lavage 

11 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
ts 
temperature sensitivity 
UP 
unanticipated problem  
URI 
upper respiratory tract illness 
URT 
upper respiratory tract 
US 
United States 
VAR 
vaccine administration record 
WIRB 
Western Institutional Review Board 
wt 
wild-type 
 
 

12 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
PROTOCOL SUMMARY 
 
Phase I Placebo-Controlled Study of the Infectivity, Safety and Immunogenicity of a Single 
Dose of a Recombinant Live-attenuated Respiratory Syncytial Virus Vaccine, RSV 
6120/∆NS1, Lot RSV#018A, or RSV 6120/F1/G2/∆NS1, Lot RSV#016A, Delivered as Nose 
Drops to RSV-seropositive Children 12 to 59 Months of Age and RSV-seronegative Infants 
and Children 6 to 24 Months of Age 
SHORT TITLE  
RSV 6120/∆NS1 or RSV 6120/F1/G2/∆NS1 
PURPOSE           
To assess whether the RSV 6120/∆NS1 or RSV 6120/F1/G2/∆NS1 vaccines will 
each be safe, infectious 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 study product in RSV-seropositive 
and RSV-seronegative participants. Participants will be randomized to receive RSV 
6120/∆NS1 vaccine, RSV 6120/F1/G2/∆NS1 vaccine, or placebo in a 2:2:1 ratio 
STUDY POPULATION Group 1: Healthy RSV-seropositive children ≥ 12 months to < 60 months  
Group 2: Healthy RSV-seronegative infants and children ≥ 6 months to <25 months 
SAMPLE SIZE 
Group 1: Approximately 25 RSV-seropositive children^ 
Group 2: Approximately 35-50 RSV-seronegative infants and children  
STUDY PRODUCT 
Single intranasal dose of RSV 6120/∆NS1, RSV 6120/F1/G2/∆NS1, or placebo 
Table 1: Inoculation schedule 
  
 
 
 
 
 
 
 
 
Group 1: RSV-seropositive: RSV serum neutralizing antibody titer ≥ 1:40; determined prior to 
inoculation and within the calendar year of inoculation. 
^If 3 or more RSV-seropositive children (group 1) with the same treatment assignment have respiratory 
or febrile illness that coincides with shedding of > 102.5 PFU of vaccine virus/mL of nasal wash as 
detected by culture, then we will evaluate the safety, infectivity, and immunogenicity of that vaccine 
virus in an additional 15 RSV-seropositive children before determining whether to continue its 
evaluation in group 2 
Group 2: RSV-seronegative: RSV serum neutralizing antibody titer < 1:40; determined ≤ 42 days prior 
to inoculation  
*plaque-forming units (PFU) 
Group # 
N 
Study Product 
Dose 
1 
10 
RSV 6120/∆NS1 
106.0 PFU* 
10 
RSV 6120/F1/G2/∆NS1 
105.8 PFU* 
5 
Placebo 
0 
2 
14-20 
RSV 6120/∆NS1 
105.0 PFU* 
14-20 
RSV 6120/F1/G2/∆NS1 
105.0 PFU* 
7-10 
Placebo 
0 

13 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Our intent is to randomize (2:2:1; RSV 6120/∆NS1: RSV 6120/F1/G2/∆NS1: placebo) for evaluation of each 
vaccine and placebo. Evaluation of a vaccine will proceed to group 2 if there are not more than 2 solicited 
adverse events (AEs) and no lower respiratory tract illnesses (LRIs) associated with that vaccine in 
RSV-seropositive children. If 3 or more RSV-seropositive children (group 1) with the same treatment 
assignment have respiratory or febrile illness that coincides with shedding of > 102.5 PFU of vaccine 
virus/mL of nasal wash as detected by culture, then we will evaluate the safety, infectivity, and 
immunogenicity of that vaccine virus in an additional 15 RSV-seropositive children before determining 
whether to continue its evaluation in group 2, or whether to discontinue the evaluation of this candidate.  
STUDY DURATION 
Participants will be enrolled in the study , outside of the RSV season. RSV-seropositive participants will be 
followed for 28 days after inoculation, and RSVseronegative participants will remain on the study until they 
complete the post–RSV season visit.  
Figure 1.1-1: Group 1 – Study Overview 
 
Figure 1.1-2: Group 2 – Study Overview 
 
(nasal rinse or nasal swab may be obtained at all timepoints; post-surveillance specimens to be 
collected beginning April 1st) 
 

14 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
1. BACKGROUND AND SCIENTIFIC RATIONALE  
1.1. 
BACKGROUND 
1.1.1. 
Epidemiology, Disease Burden, and the Need for a Vaccine 
In the United States (US) alone, respiratory syncytial virus (RSV) is responsible for 75,000 to 
125,000 hospitalizations of infants yearly (1), and in 2015, RSV was estimated to have caused at 
least 33 million cases of acute lower respiratory infection (ALRI) in children under 5 years old, 
resulting in an estimated 3.2 million RSV LRI hospitalizations and approximately 59,600 RSV-
attributable deaths (2, 3). 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 airway disease during childhood (4, 5). 
RSV is an enveloped ribonucleic acid (RNA) virus that is a member of the newly organized 
Pneumoviridae family, genus Orthopneumovirus (6). RSV has a single negative-sense strand RNA 
genome of 15.2 kilobases encoding 10 messenger ribonucleic acids (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 nucleoprotein 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.  
A formalin-inactivated vaccine against RSV was evaluated clinically in the 1960s and did not confer 
protection; instead, disease enhancement occurred at a high rate following natural infection of 
vaccinees with wild-type (wt) RSV (7). 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 (8, 9).  
Currently, no licensed immunization against RSV is available to protect healthy older infants and 
toddlers. 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. Recently, the extended half-life RSV mAb nirsevimab was licensed by the FDA to provide 
passive immunity to healthy infants through their first RSV season and to high-risk infants through 
their second RSV season (https://www.fda.gov/news-events/press-announcements/fda-approves-
new-drug-prevent-rsv-babies-and-toddlers).  Additionally, licensure of the RSVpreF maternal 
vaccine is expected in Augut 2023. Both of these products are expected to provide passive immunity 
to younger infants but do not provide active, durable immunity and are not expected to protect older 
infants and toddlers against RSV disease.     
Attempts at developing RSV vaccines at the National Institute of Allergy and Infectious Diseases 
(NIAID) have focused largely on live-attenuated approaches to provide active immunity to older 
inants and young children (10). Importantly, throughout a period of over 20 years, a number of live-

15 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
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 (11). 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 (i.n.), induce protective 
mucosal immunity in the respiratory tract (as well as systemic immunity), infect in the presence of 
maternally-derived RSV serum antibody, and have been well tolerated and immunogenic when 
administered to infants as young as four weeks of age (12).  
Human RSV has a single serotype with two antigenic subgroups, A and B. The two 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 (13). Consistent with this level of antigenic 
relatedness, F protein expressed by a recombinant (r) 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 (14). 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 vaccines to be evaluated in this study were derived using a recombinant deoxyribonucleic 
acid (rDNA)–based technique called reverse genetics (15). The technique of reverse genetics has 
been used to produce a number of licensed vaccines; among them is FluMist® (MedImmune). 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. 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 trial material [CTM]) does not contain any rDNA. The RSV vaccine 
candidates to be tested under this protocol are derivatives of strain A2, subgroup A. 
The vaccines to be evaluated in this study, RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1, are cDNA 
derived live-attenuated RSV vaccine candidates that contain a deletion of the NS1 gene (ΔNS1) (16). 
The RSV NS1 protein functions as viral interferon antagonist and as an inhibitor of RSV induced 
apoptosis, and deletion of NS1 is attenuating (16-24). NS1 also has a suppressive effect on the 
maturation of dendritic cells. RSV with the NS1 deletion is more potent in activating CD8+ T cells 
and Th17 cells, and its activating effect on IL4-producing CD4+ T cells is decreased (25, 26). 
Genetic stability of live-attenuated RNA viruses is a concern in clinical vaccine development. The 
deletion of an entire gene, the NS1 gene, should be refractory to genetic or phenotypic reversion. The 
envelope glycoproteins F and G represent the major RSV neutralization and protective antigens. 
RSV 6120/F1/G2/∆NS1 was designed for increased expression of the F and G proteins (27); to 
achieve this, both glycoprotein genes were moved in the RSV genome from their natural 7th (G) and 
8th (F) position to the first (F1) and second (G2) genome promoter proximal position in RSV 
6120/F1/G2/∆NS1. 

16 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
In historic studies in chimpanzees, the only experimental animal model in which replication and 
virulence of RSV approaches that in humans, replication of recombinant RSV lacking the NS1 gene 
was reduced compared to recombinant wt RSV by about 2,200 fold and 17,000 fold in the upper 
respiratory tract (URT) and lower respiratory tract (LRT), respectively, with low mean peak titers of 
1.6 and 1.2 log10 PFU per mL in nasopharyngeal swabs and tracheal lavage (TL) fluid (16). Despite 
the low level of replication, the virus was immunogenic and, importantly, protective against wt RSV 
challenge infection in chimpanzees. In preclinical studies, both vaccine candidates were significantly 
restricted in replication compared to recombinant wt RSV in a primary human airway epithelial cell 
model. In preclinical studies in African green monkeys (AGM), replication in the upper and lower 
respiratory tract of both vaccine candidates was reduced compared to recombinant wt RSV. RSV 
6120/F1/G2/∆NS1, expressing the glycoprotein genes from promoter proximal positions, was further 
restricted in replication than RSV 6120/∆NS1, but comparable to RSV 6120/∆NS1 in its 
immunogenicity in the AGM model. 
1.2. 
PRIOR RESEARCH 
1.2.1. 
Experimental Vaccines against RSV 
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 (11). 
The vaccines to be evaluated in this protocol, RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1, have 
not been previously evaluated in humans. However, over recent years, NIAID's ongoing RSV 
vaccine development program has evaluated several live-attenuated pediatric RSV vaccine 
candidates sequentially in adults, RSV seropositive children, and RSV seronegative infants and 
children. In one approach, RSV was attenuated by temperature sensitivity (ts) mutations, in 
combination with deletion of the SH gene (12, 28-30). In another approach, RSV was attenuated by 
deletion of the transcription processivity factor M2-2 (NCT02601612, NCT02952339, 
NCT03102034, NCT03099291, NCT02890381, NCT03227029). In addition, several live-attenuated 
RSV vaccine candidates with a deletion of another RSV interferon antagonist, NS2, were evaluated 
sequentially in adults, children, and infants. Results from the most recent study of the lead candidate 
with NS2 deletion (ref Luongo), RSV∆NS2/∆1313/I1314L (NCT01893554), are summarized below.  
In a recent Phase I trial (NCT01893554), the RSV∆NS2/∆1313/I1314L candidate vaccine was 
evaluated in RSV-seropositive children 12-59 months of age at a dose of 106 PFU (10V, 5P), and no 
shedding or immune response was detected, indicative of attenuation. Next, it was evaluated in 
seronegative children 6-24 months of age at two sequential dose levels.  
At the lower dose of 105 PFU (15V, 7P), the vaccine was poorly infectious (7% and 80% of 
recipients shed virus detected by culture and quantitative reverse transcription polymerase chain 
reaction [RT-qPCR], respectively) and immunogenicity was low. 73% of the vaccinees and 57% of 
placebo recipients experienced a respiratory or febrile illness during the first 28 days following 

17 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
receipt of vaccine. Specifically, mild rhinorrhea was reported in 11 of 15 (73%) vaccinees and 4 of 7 
(57%) of placebo recipients, frequently associated with shedding of rhinovirus. Mild cough was 
reported in two participants of each cohort, associated with shedding of adventitious viruses in all 
cases.  
The vaccine was subsequently evaluated at the higher dose of 106 PFU in RSV seronegative infants 
and children 6-24 months of age (20V/10P). In this cohort, 80% and 90% of vaccine recipients shed 
vaccine detected by culture and RT-qPCR, respectively [median peak titers (MPT) 1.7 log10 
PFU/mL; 3.6 log10 copies/mL]. During the first 28 days following receipt of vaccine, 55% of 
vaccinees and 70% of placebo recipients experienced a respiratory or febrile illness, mostly mild 
rhinorrhea [10 of 20 (50%) of vaccinees; 4 of 10 (40%) of placebo recipients]. Cough was reported 
in one vaccinee (5%) and 3 placebo recipients (30%), and a single case of otitis media was present in 
a placebo recipient. Fever was also less frequent in vaccinees [2 of 20 (10%), highest grade: Grade 2] 
than in placebo recipients [4 of 10 (40%), highest grade: Grade 3]. Other respiratory viruses 
(rhinovirus, parainfluenza virus type 3, adenovirus, enterovirus, bocavirus) were detected 
concurrently with respiratory symptoms in most cases. Among vaccine recipients, mild rhinorrhea 
without a detectable adventitious agent was observed in five participants (25%) during the time 
period of vaccine virus shedding. In placebo recipients, mild rhinorrhea without a detectable 
adventitious agent was observed in 2 of 10 participants (20%) during the same period. Overall, this 
shows that the vaccine was well tolerated. Ninety percent of the vaccine recipients developed an 
RSV F immunoglobulin G (IgG) antibody response, and 80% developed an RSV neutralizing 
antibody response, detected on Day 56 after receipt of vaccine. The serum antibody responses to 
RSV ∆NS2/∆1313/I1314L were durable over the RSV surveillance season, and the vaccine 
candidate primed for strong anamnestic responses to wt RSV. Based on these results, RSV 
∆NS2/∆1313/I1314L was selected as an appropriate candidate for further clinical evaluation. 
1.2.2. 
Preclinical Studies 
1.2.2.1. 
Replication of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 
(Experimental Lots) in vitro 
Multicycle replication of experimental lots of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 was 
compared to that of recombinant RSV 6120, a control virus with a phenotype approaching that of wt 
RSV. Two experimental systems were used, namely Vero cells which do not express type I 
interferon, and a 3D mucociliary human tissue model consisting of normal differentiated primary 
human tracheal/bronchial airway epithelial cells (EpiAirway, Mattek, Inc.). To evaluate replication 
in Vero cells, triplicate 25 cm flasks of subconfluent Vero cells were infected at an MOI of 0.01 with 
RSV 6120/ΔNS1, RSV 6120/F1/G2/ΔNS1, or RSV 6120. After 2 hours of adsorption, the inoculum 
was gently removed, and the cultures were washed and incubated with fresh medium at 37°C. To 
evaluate replication in primary human airway cells, cultures of pseudostratified differentiated airway 
cells were incubated in an air-liquid interface at 37°C in 12-mm transwell inserts with 0.4 µM pore 
sizes in Dulbecco’s Modified Eagle’s Medium (DMEM), supplemented with a proprietary mix of 
epidermal growth factors and other proprietary factors. Prior to infection, the apical surfaces of the 
cultures were washed with 500 µL PBS to remove any accumulated mucus. Triplicate wells were 
infected at an MOI of 0.1 with 100 µl per well of RSV 6120/ΔNS1, RSV 6120/F1/G2/ΔNS1, or RSV 
6120. After 2 hours of adsorption, the inoculum was gently removed, and the apical surfaces of the 
cultures were washed 3 times with 300 µL PBS. Apical washes were collected daily on days 1-7 
post-infection to assay viral replication. For these washes, 350 µL serum-free DMEM was applied to 

18 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
the apical surface of the cultures. Plates were returned to the incubator for 30 minutes of incubation. 
Apical medium was then collected, snap frozen, and stored at -80°C. Apical medium supernatants, as 
well as the supernatants from the Vero replication study, were titered by plaque assay on Vero cells, 
incubated at 37°C, 5% CO2, as described above. The lower limit of detection of the samples was 0.7 
log10 PFU per mL.  
Recombinant wt RSV 6120 replicated efficiently in Vero cells and in differentiated primary human 
tracheal/bronchial epithelial cells (Figure 1.2-1). In Vero cells, which do not produce type I 
interferon, peak titers of RSV 6120/ΔNS1 and the control virus RSV 6120/F1/G2/ΔNS1 were less 
than 10-fold lower than those of wt RSV. However, in interferon competent mucociliary normal 
human tracheal/bronchial epithelial cells, RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 were highly 
restricted in replication compared to the parental wt like virus, and titers were about 1000-fold (RSV 
6120/ΔNS1) and 10,000-fold (RSV 6120/F1/G2/ΔNS1) lower than those of the control virus. Thus, 
in interferon competent cells, the deletion of the NS1 gene is much more restrictive than in Vero 
cells. RSV 6120/F1/G2/ΔNS1 with glycoprotein genes shifted to promoter proximal positions was 
further restricted than RSV 6120/ΔNS1 with NS1 deletion alone. 
 
Figure 1.2-1. Multicycle replication of experimental lots in Vero cells (left) or in an in-vitro model of 
normal human bronchial/tracheal epithelial cells (right). Cultures were infected at a multiplicity of infection of 
0.01 (Vero cells) or 0.1 (epithelial cells), and supernatants were harvested on indicated days, snap frozen, and 
titered later on Vero cells.  
 

19 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
1.2.2.2. 
Replication of experimental lot and CTM of RSV 6120/ΔNS1 and RSV 
6120/F1/G2/ ΔNS1 in African green monkeys (AGMs) 
Replication and immunogenicity of RSV 6120/ΔNS1 was evaluated in nonhuman primates (NHP), 
in African green monkeys (AGMs). AGMs are semi-permissive for RSV infection. The first NHP 
study was done to evaluate an Experimental Lot of RSV 6120/ΔNS1, and a second study was done to 
evaluate the CTM of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 (Lots RSV#016A and 
RSV#018A). Male and female AGMs which were seronegative for RSV were inoculated (i.n.) and 
intratracheally (i.t.) with RSV 6120/ΔNS1 or RSV 6120/F1/G2/ΔNS1. A total dose of 2 x 106 PFU 
(corresponding to 106.3 PFU) per animal was used for all inoculations, except for CTM RSV 
6120/F1/G2/ΔNS1, Lot RSV#018A, which was used at a dose of 1 x 106 PFU (corresponding to 106.0 
PFU per animal). Nasopharyngeal (NP) swabs were collected daily on Days 0 through 10 and on 
Days 12 and 14 (Table 26), TL samples were collected every other day from Day 2 through Day 14 
(Table 27), and virus shedding was analyzed by immunoplaque assay (Table 28). Serum RSV 
neutralizing antibody titers were determined by a complement-enhanced 60% plaque reduction 
neutralization assay (Table 28). Results from studies following the same protocol, performed in 
animals from the same group and origin, inoculated with recombinant wt RSV A2 at the same dose, 
were included for comparison. Studies were approved by the Animal Care and Use Committee of 
NIAID, National Institutes of Health (NIH).  
In both studies, shedding of RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 from the URT and LRT 
was detectable over several days (Table 26, Table 27). In both studies, the mean peak titers of RSV 
6120/ΔNS1 in the NP samples were significantly reduced compared to that of wt RSV A2 (3.0 and 
2.2 log10 PFU per mL for the Experimental lot and the CTM of RSV 6120/ΔNS1, compared to 4.2 
log10 PFU per mL for RSV A2; significantly lower than RSV A2; P<0.001 and P<0.01, ANOVA, 
Tukey post-hoc analysis). The mean peak titer of RSV 6120/F1/G2/ΔNS1 was even further reduced 
to 1.5 log10 PFU per mL (P<0.05 compared to the mean peak titer of RSV in the CTM of RSV 
6120/ΔNS1, P<0.0001 compared to RSV A2). The mean peak titers in the LRT, measured by 
titration of TL samples, were 3.3 and 3.5 log10 PFU per mL for the Experimental lot and the CTM of 
RSV 6120/ΔNS1, compared to 4.1 log10 PFU per mL for RSV A2. Replication of the CTM of RSV 
6120/F1/G2/ΔNS1 was lower than that of RSV 6120/ΔNS1, at mean peak titers of 2.2 log10 PFU per 
mL (P<0.05; ANOVA, Tukey post-hoc analysis).  
In both studies, the RSV 6120/ΔNS1 virus induced serum neutralizing antibodies that were 
comparable to or slightly higher than wt RSV A2 (Table 28). The titers induced by RSV 
6120/F1/G2/ΔNS1 were slighly but not significantly lower than those of wt RSV. These results show 
that RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1, administered i.n. and i.t., are immunogenic in 
AGMs. 
These results indicate that RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 are restricted for replication 
in the upper and lower respiratory tract of AGMs. At the dose of 1 x 106 PFU per site, replication of 
RSV 6120/ΔNS1 in the upper and lower respiratory tract was detected in all AGMs, and RSV 
6120/ΔNS1 was immunogenic. At a dose of 5 x 105 PFU per site, replication of RSV 
6120/F1/G2/ΔNS1 was significantly reduced compared to wt RSV in the upper and lower respiratory 
tract, but the difference in immunogenicity to wt RSV was not significant, despite the low level of 
replication. In summary, it is anticipated that the investigational live vaccine candidates RSV 
6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 will be well-tolerated in RSV seropositive participants upon 
intranasal application, and well-tolerated, infectious, and immunogenic in RSV seronegative 
participants. 

20 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
1.2.3. 
Previous Clinical Experience 
To our knowledge, RSV vaccine viruses containing NS1 deletions have not been previously tested in 
humans. Data from recent clinical evaluations of other live-attenuated RSV vaccine candidates 
containing deletion mutations are described in Section 1.2.1.  
1.3. 
RATIONALE 
In previous Phase I studies of live-attenuated RSV vaccines in RSV-seronegative infants and 
children 6 to 24 months of age, RSV strains based on two different attenuation strategies have 
emerged as the most promising candidates: (1) candidates that are attenuated by deletion of the RSV 
interferon antagonist NS2, and (2) candidates attenuated by deletion of the RNA regulatory protein 
M2-2. Results from chimpanzee studies suggest that the deletion of the second RSV interferon 
antagonist, NS1, may represent a third powerful attenuation strategy for RSV vaccine candidates, 
yielding a level of attenuation similar to that of the deletion of M2-2. RSV 6120/ΔNS1 was designed 
to be attenuated by a 529 nucleotide deletion of the NS1 gene. In addition to its attenuating effect, 
the NS1 deletion also is expected to increase immunogenicity due to the immune-stimulatory effects 
of interferon. To further increase immunogenicity, RSV 6120/F1/G2/ΔNS1 was generated. This 
candidate has the RSV glycoprotein genes, encoding the major neutralizing antigens F and G, moved 
to the first and second genome promoter proximal position, increasing their level of expression. 
Based on preclinical results in primary human airway epithelial cells and African green monkeys, 
this genome rearrangement is expected to be further attenuating in the natural human host. This 
Phase 1 study will evaluate the suitability of the deletion of NS1 and of a genome rearrangement to 
generate safe and immunogenic live-attenuated RSV vaccines.  
The rationale for sequential evaluation in RSV seropositive children, followed by evaluation in RSV 
seronegative children and infants is as follows. This will be the first-in-human testing of RSV 
vaccine candidates containing a deletion of the NS1 gene. Based on chimpanzee studies and pre-
clinical testing results, the NS1 deletion is expected to be highly attenuating, similar to deletion of 
the M2-2 open reading frame (ORF), and it is expected that replication of RSV 6120/ΔNS1 and RSV 
6120/F1/G2/ΔNS1 will be highly restricted or absent in RSV-seropositive children. If these 
candidates are found to be safe and restricted in RSV-seropositive children, testing will proceed to 
RSV-seronegative children.  
This protocol will evaluate safety, infectivity, replication, and immunogenicity of both candidates, 
with particular attention to vaccine virus infectivity and replication (i.e., percentage of participants 
shedding virus, peak vaccine virus titer in nasal washes or nasal swabs, as well as duration of 
shedding), which are the most quantifiable metrics for the level of attenuation. Based on results from 
this study, a comparison of infectivity and immunogenicity of these candidates will be possible. It is 
anticipated that the lead candidate emerging from this study will move forward to expanded studies 
to further evaluate safety and immunogenicity.  

21 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
The primary immunogenicity endpoints to be evaluated in this study are serum RSV neutralizing 
antibody titers and serum antibody titers to the RSV F protein (measured by ELISA). Neutralizing 
antibody is a well-established and important surrogate marker of protection from RSV disease. 
Antibodies to the F protein are also associated with cross-subgroup neutralization and protection. 
These assays will be performed at the Johns Hopkins University (JHU) Center for Immunization 
Research (CIR) laboratory. 
1.4. 
CLINICAL DEVELOPMENT PLAN 
The investigational RSV vaccines RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1 will be evaluated 
sequentially in RSV-seropositive children followed by evaluation in RSV-seronegative infants and 
children. The main purpose of this study is to determine whether these vaccines are well tolerated 
and restricted in replication in RSV-seropositive children, and well tolerated, infectious, 
immunogenic, and genetically stable in RSV-seronegative infants and children.  
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 (14). These assays will be performed at 
the JHU CIR laboratory. 
1.5. 
HYPOTHESES 
RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1 will be well-tolerated in RSV-seropositive and RSV-
naïve participants. In RSV-seronegative participants, these vaccines are expected to be safe and 
result in infection, limited vaccine replication, and the induction of neutralizing antibody responses 
to RSV.  
2. OBJECTIVES 
2.1. 
PRIMARY OBJECTIVES 
Safety  
To estimate and compare (each vaccine group to placebo):  
1. the frequency and severity of study product–related solicited and unsolicited AEs in RSV-
seropositive participants (group 1) from Day 0 through the 10th day following inoculation  
2. the frequency and severity of study product–related solicited and unsolicited AEs in RSV-
seronegative participants (group 2) from Day 0 through the 28th day following inoculation 
3. the frequency of study product–related serious adverse events (SAEs) and lower respiratory tract 
illness (LRIs) in RSV-seropositive participants (group 1) from Day 0 through the 28th day following 
inoculation  
4. the frequency of study product–related SAEs in RSV-seronegative participants (group 2) from Day 0 
through the 56th day following inoculation  
  

22 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Infectivity  
1. To determine the peak titer of vaccine virus shed and duration of virus shedding by each participant  
2. To assess the proportion of vaccinated participants infected* with study vaccine. For the 
seronegative cohort, the primary aim is to determine whether >90% of vaccinees are infected with 
vaccine virus 
 
* Infected with vaccine virus is defined as shedding vaccine virus, detected by RT-qPCR, and/or ≥4-
fold rise in RSV-specific serum antibodies, detected by ELISA against the RSV F protein and/or 
RSV plaque reduction neutralization (RSV-PRNT) assay. 
 
Immunogenicity 
1. To characterize serum antibody responses to the study product in each study arm 28 days after 
inoculation of RSV-seropositive vaccine recipients (group 1)  
2. To characterize serum antibody responses to the study product in each study arm 56 days after 
inoculation of RSV-seronegative vaccine recipients (group 2)  
2.2. 
SECONDARY OBJECTIVES 
1. To characterize the frequency and severity of symptomatic medically attended respiratory and febrile 
illness in RSV-seronegative vaccine and placebo recipients (group 2) who experience natural 
infection with wt RSV during the subsequent RSV season  
2. To characterize serum antibody responses in RSV-seronegative vaccine and placebo recipients 
(group 2) who experience natural infection with wt RSV during the subsequent RSV season 
3. To characterize mucosal antibody responses to each vaccine in RSV-seropositive (group 1) and a 
subset of RSV-seronegative (the first 13 group 2 enrolled prior to March 2020) participants  
4. To identify differences in infectivity and immunogenicity between the vaccines. 
 
2.3. 
EXPLORATORY OBJECTIVE 
Study samples may be used in comparative assays with samples from other RSV vaccine studies 
initiated by the Laboratory of Infectious Diseases (LID), NIAID, NIH. 
3. STUDY DESIGN 
The study will be double-blind, randomized, and placebo-controlled. Participants in groups 1 and 2 
will be randomized at a ratio of 2:2:1 to the three arms of the study; two active vaccine arms and one 
placebo arm. 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 (28-30) and statistical considerations (see Section 9).  

23 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
The vaccine will be evaluated in a stepwise fashion beginning with RSV-seropositive children 
(group 1) and proceeding sequentially in RSV-seronegative children (group 2). For the purpose of 
this study, RSV-seropositive is defined as having a serum neutralizing antibody titer of ≥ 1:40, and 
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 (12, 29, 31). 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. 
Our intent is to randomize 2:2:1; RSV 6120/∆NS1: RSV 6120/F1/G2/∆NS1: placebo. Evaluation of 
each vaccine will proceed to group 2 if there are not more than 2 solicited adverse events (AEs) and 
no LRIs associated with that vaccine in RSV-seropositive children. If 3 or more RSV-seropositive 
children (group 1) with the same treatment assignment have respiratory or febrile illness that 
coincides with shedding of >102.5 PFU of vaccine virus/mL of nasal wash as detected by culture, 
then we will evaluate the safety, infectivity, and immunogenicity of the vaccine virus in an additional 
15 RSV-seropositive children before determining whether to continue with group 2, or a decision 
will be made to discontinue the evaluation of this candidate.  
Enrollment occurs at the time of inoculation with study product, Day 0, and will  avoid the time 
during which wt RSV typically circulates in the community, through ongoing local and national 
(CDC NREVSS) surveillance(Appendix IV).  
For group 1, RSV-seropositive children, the total duration of participation in the study is 28 days, 
consisting of an Acute Phase, from the day of inoculation (Study Day 0) through Study Day 10, and 
a Post-Acute Phase, from Study Day 11 through Study Day 28 (Table 29).  
For group 2, RSV-seronegative children, the duration of participation in the initial phase of the study 
is 56 days. The initial phase consists of two components: an Acute Phase, from the day of 
inoculation (Study Day 0) through Study Day 28, and a Post-Acute Phase, from Study Day 29 
through Study Day 56 (Table 30).  

24 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
During the Acute Phase of the study, the participants’ parents/guardians will be contacted daily. 
These contacts will consist either of an in-person evaluation of interim medical history, clinical 
assessment, and nasal rinse or swab, or an interim medical history conducted by a mutually agreed 
upon communication method. During the Acute Phase, the participants will be evaluated for AEs and 
SAEs. Participants who have a febrile or respiratory illness or otitis media will have a study visit to 
perform a clinical assessment and a nasal rinse or swab. Children will not be enrolled in this study 
while a Stay at Home Order is in place. However, in the event that a Stay at Home Order is initiated 
within the first 28 days following inoculation, if a child develops a respiratory illness, parents may 
be asked to obtain the nasal swabs, and clinical assessments may be completed via remote or in-
person research visits. During remote research visits parents will also be asked to measure the child’s 
temperature, heart rate and respiratory rate. Parents will be informed of these possibilities prior to 
enrollment, and will be trained in the methods used to collect an anterior nasal swab specimen. The 
nasal rinse or swab will be tested for RSV and for other respiratory pathogens that are considered 
adventitious agents. During the Post-Acute Phase, the RSV-seropositive participants' 
parents/guardians will be instructed to contact the study staff if any SAEs or LRIs occur and will 
have a scheduled follow-up visit on the 28th day after inoculation, and the RSV-seronegative 
participants’ parents/guardians will be instructed to contact the study staff if any SAEs occur and 
will have a scheduled follow-up visit on the 56th day after inoculation. The schedules of evaluations 
during the Acute Phase and Post-Acute Phase are shown in Appendix II, Table 29, and Table 30. 
For the RSV-seronegative participants (group 2), the study has a third phase that assesses the 
incidence and severity of RSV-associated illness occurring during the RSV season following 
inoculation. During the RSV Season Surveillance Period, beginning on the date of seasonal 
enrollment pause and ending on March 31, study staff will make weekly contact with the 
participants’ parents/guardians to identify medically attended episodes of fever, URI, LRI, or otitis 
media. Participants who have such an episode will have an in-person evaluation of interim medical 
history, clinical assessment, and a nasal rinse or swab to evaluate for RSV and other respiratory 
pathogens that are considered adventitious agents (Appendix III, Table 31). As noted above, if a Stay 
at Home Order is initiated during the RSV Surveillance Period, parents may be asked to obtain nasal 
swabs and clinical assessments may be completed via remote or in-person research visits. 
The RSV-seronegative participants will also have a study visit during the post-RSV season  to collect 
a blood sample to assess the durability of the vaccine response and to assess the immune response to 
naturally occurring wt RSV infection. For 13 participants enrolled prior to March 2020,  a pre-RSV 
season blood draw and nasal rinse and/or nasosorption sample (using the synthetic absorptive matrix 
(SAM) strip) for immunological assays were collected. The samples will be used to assess the 
durability of the vaccine response.  The RSV-seronegative participants may have an overlap between 
the initial phases and the RSV surveillance phase of the study, depending upon the date of 
inoculation.  
4. STUDY POPULATION 
In group 1, approximately 25 RSV-seropositive children ≥ 12 months to < 60 months of age will be 
enrolled. In group 2, approximately 35 to 50 RSV-seronegative infants and children ≥ 6 months to 
<25 months of age will be enrolled. These numbers were chosen based upon experience with phase I 
evaluation of other live attenuated respiratory virus candidate vaccines (11,23,25) and statistical 
considerations (Section 9). Placebo recipients are needed in pediatric studies to distinguish the 

25 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
background respiratory illnesses that occur in infants and children from those attributable to study 
vaccine. Infants and children will be selected for participation according to the described study-
specific inclusion and exclusion criteria. 
4.1. 
INCLUSION CRITERIA FOR RSV-SEROPOSITIVE CHILDREN  
Potential RSV-seropositive participants must meet all of the following inclusion criteria to be 
enrolled in this study: 
4.1.1. ≥ 12 months of age and < 60 months of age at the time of inoculation 
4.1.2. Screening serum specimen for RSV-neutralizing antibody is obtained within the calendar year 
of inoculation  
4.1.3. Seropositive for RSV antibody, defined as serum RSV-neutralizing antibody titer ≥ 1:40 
4.1.4. Pre-inoculation serum sample for RSV-neutralizing antibody specimen is obtained no more 
than 56 days prior to inoculation 
4.1.5. In good health based on review of the medical record, history, and physical examination (PE) 
at the time of inoculation 
4.1.6. Received routine immunizations appropriate for age based on the Advisory Committee on 
Immunization Practices (ACIP) Recommended Immunization Schedule for Children and 
Adolescents Aged 18 Years or Younger 
4.1.7. Growing normally for age as demonstrated on a standard growth chart and has a current 
height and weight above the 3rd percentile for age  
4.1.8. Expected to be available for the duration of the study 
4.1.9. Parent/guardian is willing and able to provide written informed consent  
4.2. 
EXCLUSION CRITERIA FOR RSV-SEROPOSITIVE CHILDREN 
Potential RSV-seropositive participants who meet any of the following exclusion criteria will be 
excluded from participation in this study: 
4.2.1. 
Born at less than 34 weeks gestation 
4.2.2. Maternal history of positive human immunodeficiency virus (HIV) test  
4.2.3. Evidence of chronic disease 
4.2.4. Known or suspected impairment of immune function 
4.2.5. Bone marrow/solid organ transplant recipient 
4.2.6. Major congenital malformations, including congenital cleft palate or cytogenetic 
abnormalities 
4.2.7. Suspected or documented developmental disorder, delay, or other developmental problem  
4.2.8. Cardiac abnormality requiring treatment 
4.2.9. Lung disease or reactive airway disease 
4.2.10. More than one episode of wheezing in the first year of life 

26 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
4.2.11. Wheezing episode or received bronchodilator therapy within the past 12 months 
4.2.12. Wheezing episode or received bronchodilator therapy after the age of 12 months 
4.2.13. Previous receipt of supplemental oxygen therapy in a home setting 
4.2.14. Previous receipt of an investigational RSV vaccine  
4.2.15. Previous receipt or planned administration of anti-RSV antibody product including ribavirin, 
RSV Ig or RSV mAb 
4.2.16. Previous receipt of immunoglobulin or any antibody products within the past 6 months 
4.2.17. Previous receipt of any other blood products within the past 6 months 
4.2.18. Previous anaphylactic reaction 
4.2.19. Previous vaccine-associated adverse reaction that was Grade 3 or above 
4.2.20. Known hypersensitivity to any vaccine component 
4.2.21. Member of a household that contains an infant who is less than 12 months of age at the date of 
inoculation through the 10th day after inoculation  
4.2.22. Member of a household that, at the date of inoculation through the 10th day after inoculation, 
contains an immunocompromised individual including but not limited to: 
• a person who is HIV-infected 
• a person who has cancer and has received chemotherapy within the 12 months prior to 
enrollment 
• a person living with a solid organ or bone marrow transplant 
4.2.23. Will attend a daycare facility that does not separate children by age and contains an infant 
who is < 12 months of age at the date of inoculation through the 10th day after inoculation 
4.2.24. Receipt of any of the following prior to enrollment: 
• any inactivated 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, or 
• salicylate (aspirin) or salicylate-containing products within the past 28 days 
4.2.25. Scheduled administration of any of the following after planned inoculation: 
• inactivated vaccine or live-attenuated rotavirus vaccine within the 14 days after, or 
• any live vaccine other than rotavirus within the 28 days after, or 
• another investigational vaccine or investigational drug within the 28 days after 
4.2.26. 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 medications except the permitted concomitant medications listed in 
Section 5.0  
4.2.27. Any of the following events at the time of enrollment: 
• fever (temporal or rectal temperature of ≥100.4°F), or  
• upper respiratory signs or symptoms (rhinorrhea, cough, or pharyngitis) or 
• nasal congestion significant enough to interfere with successful inoculation, or  
• otitis media 

27 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
4.3. 
INCLUSION CRITERIA FOR RSV-SERONEGATIVE INFANTS & CHILDREN 
Potential RSV-seronegative participants must meet all of the following inclusion criteria to be 
enrolled in this study: 
4.3.1. ≥ 6 months of age and < 25 months of age at the time of inoculation 
4.3.2. Screening and pre-inoculation serum specimens for RSV-neutralizing antibody are obtained 
no more than 42 days prior to inoculation  
4.3.3. Seronegative for RSV antibody, defined as serum RSV-neutralizing antibody titer < 1:40 
4.3.4. In good health based on review of the medical record, history, and PE at the time of 
inoculation 
4.3.5. Received routine immunizations appropriate for age based on the ACIP Recommended 
Immunization Schedule for Children and Adolescents Aged 18 Years or Younger 
4.3.6. Growing normally for age as demonstrated on a standard growth chart, AND 
• If < 1 year of age: has a current height and weight above the 5th percentile for age  
• If ≥ 1 year of age: has a current height and weight above the 3rd percentile for age  
4.3.7. Expected to be available for the duration of the study 
4.3.8. Parent/guardian is willing and able to provide written informed consent  
4.4. 
EXCLUSION CRITERIA FOR RSV-SERONEGATIVE INFANTS & CHILDREN 
Potential RSV-seronegative participants who meet any of the following criteria will be excluded from 
this study: 
4.4.1. Born at less than 34 weeks gestation 
4.4.2. Born at less than 37 weeks gestation, and at the date of inoculation less than 1 year of age 
4.4.3. Maternal history of a positive HIV test 
4.4.4. Evidence of chronic disease 
4.4.5. Known or suspected infection or impairment of immunological functions 
4.4.6. Bone marrow/solid organ transplant recipient 
4.4.7. Major congenital malformations, including congenital cleft palate or cytogenetic 
abnormalities 
4.4.8. Suspected or documented developmental disorder, delay, or other developmental problem 
4.4.9. Cardiac abnormality requiring treatment 
4.4.10. Lung disease or reactive airway disease 
4.4.11. More than one episode of wheezing in the first year of life 
4.4.12. Wheezing episode or received bronchodilator therapy within the past 12 months 
4.4.13. Wheezing episode or received bronchodilator therapy after the age of 12 months 
4.4.14. Previous receipt of supplemental oxygen therapy in a home setting 
4.4.15. Previous receipt of an investigational RSV vaccine  

28 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
4.4.16. Previous receipt or planned administration of anti-RSV product including ribavirin, RSV Ig, 
or RSV mAb within the past 6 months  
4.4.17. Previous receipt of immunoglobulin or any antibody products within the past 6 months 
4.4.18. Previous receipt of any blood products within the past 6 months  
4.4.19. Previous anaphylactic reaction 
4.4.20. Previous vaccine-associated adverse reaction that was Grade 3 or above 
4.4.21. Known hypersensitivity to any study product component 
4.4.22. Member of a household that contains an infant who is less than 6 months of age at the date of 
inoculation through the 28th day after inoculation 
4.4.23. Member of a household that, at the date of inoculation through the 28th day after inoculation, 
contains an immunocompromised individual including but not limited to: 
• 
a person who is HIV-infected 
• 
a person who has cancer and has received chemotherapy within the 12 months prior to 
enrollment 
• 
a person living with a solid organ or bone marrow transplant 
4.4.24. Attends a daycare facility that does not separate children by age and contains an infant < 6 
months of age at the date of inoculation through the 28th day after inoculation 
4.4.25. Receipt of any of the following prior to enrollment: 
• 
any inactivated influenza vaccine within 3 days prior, or 
• 
any inactivated 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, or 
• 
salicylate (aspirin) or salicylate-containing products within the past 28 days 
4.4.26. Scheduled administration of any of the following after planned inoculation 
• 
inactivated vaccine or live-attenuated rotavirus vaccine within the 14 days after, or 
• 
any live vaccine other than rotavirus within the 28 days after, or 
• 
another investigational vaccine or investigational drug within the 56 days after 
4.4.27. 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 medications except the permitted concomitant medication 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. 
4.4.28. Any of the following events at the time of enrollment: 
• 
fever (temporal or rectal temperature of ≥100.4°F), or  
• 
upper respiratory signs or symptoms (rhinorrhea, cough, or pharyngitis) or 
• 
nasal congestion significant enough to interfere with successful inoculation, or  
• 
otitis media 

29 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
• 
contact with a person diagnosed with COVID-19 disease or active SARS-CoV-2 infection 
within the past 10 days 
4.5. 
CO-ENROLLMENT CONSIDERATIONS 
Co-enrollment in an investigational vaccine or investigational drug study is not allowed during this 
study.  
4.6. 
RE-ENROLLMENT CONSIDERATIONS 
RSV-seropositive participants who receive placebo may re-enroll in the study following completion 
of the Study Day 28 visit if they continue to meet all eligibility requirements. RSV-seronegative 
participants who receive placebo may re-enroll in the study following completion of the initial RSV 
surveillance period if they continue to meet all eligibility requirements.  Subjects will not be allowed 
to enroll in the study more than twice.   
4.7. 
RECRUITMENT PROCESS 
CIR research staff will recruit participants from pediatric practices and clinics in the greater 
Baltimore/Washington area. Upon referral by the participants’ primary care provider or the 
provider’s staff, CIR staff will contact parents/guardians in person or remotely as appropriate,via 
telephone, email, or text message, send them  IRB-approved informational brochures, and obtain 
contact information. Study staff will follow up with interested parents/guardians to elaborate upon 
the details and requirements of the screening and study process.  
CIR research staff may also recruit potential participants by sending IRB-approved documents 
through mail, email, or electronic medical record messaging (e.g. MyChart) to children of local 
pediatric practices and clinics, and to households in local zip codes containing age-appropriate 
children. In addition, CIR staff may recruit participants through group gatherings such as health fairs 
and by social media posting IRB-approved recruitment materials.  
If parents/guardians are interested in having their child participate and the child meets the minimum 
inclusion and exclusion criteria, then the study staff will schedule a screening visit to determine the 
child’s eligibility.  

30 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
4.8. 
PARTICIPANT RETENTION 
Study staff will make every effort to retain participants in the study, thereby minimizing potential 
biases associated with loss to follow-up.  
4.9. 
PARTICIPANT WITHDRAWAL OR TERMINATION FROM THE STUDY 
Participants 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 terminate participation in the study early for any of the following 
reasons: 
• 
Withdrawal of consent – applies to a parent/guardian who verbally or in writing withdraws 
consent for the participant to continue 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. 
• 
Investigator discretion – 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 ended for the following reasons: 
• 
Research is 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, CIR, the institutional review board (IRB), the Office for Human Research 
Protections (OHRP), NIAID, or the US Food and Drug Administration (FDA) may decide to end 
the study. 
 
For any participant who withdraws or who is terminated from the study prior to completion of 
follow-up, study staff will document the reason for the withdrawal or termination. In the event that 
the circumstances that led to a participant’s withdrawal or termination change, the study staff will 
contact the principal investigator (PI) to discuss options for resumption of follow-up. Withdrawn 
participants will not be replaced. 
5. STUDY PRODUCT 
The unblinded dispenser should consult standard operating procedures (SOPs) and the study manual 
of procedures (MOP). Refer to Figure 1.1-1 and Figure 1.1-2 for an overview of the study design and 
to the investigator’s brochure (IB) for further information about the study product. 
 
The products that will be administered in this study are: 
 
• 
Group 1: Live Recombinant Respiratory Syncytial Virus RSV 6120/∆NS1, approximately 
106.0 PFU or RSV 6120/F1/G2/∆NS1 approximately 105.8 per 1.0 mL vaccine 
• 
Group 2: Live Recombinant Respiratory Syncytial Virus RSV 6120/∆NS1 or RSV 

31 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6120/F1/G2/∆NS1, approximately 105.0 PFU per 0.5 mL vaccine 
• 
Group 1: Placebo for the RSV vaccine will be Lactated Ringer’s Solution for Injection, 
USP 1.0 mL 
• 
Group 2: Placebo for the RSV vaccine will be Lactated Ringer’s Solution for Injection, 
USP 0.5 mL 
5.1. 
STUDY PRODUCT REGIMENS 
Enrolled study participants will receive a single dose of the indicated vaccine or placebo, 
administered as nose drops.  
5.2. 
STUDY PRODUCT FORMULATION  
5.2.1. 
Vaccines  
The RSV 6120/∆NS1 vaccine is provided in a sterile 2.0-mL cryovial, each containing 0.6 mL of 
vaccine (Lot RSV#018A). The vaccine virus concentrate is diluted by trained research personnel to a 
dose of approximately 106.0 PFU (group 1) in 1.0 mL or 105.0 PFU (group 2) in a 0.5-mL volume. 
The vaccine vial is labeled as shown in Figure 5.2-1.  
The RSV 6120/F1/G2/∆NS1 vaccine is provided in a sterile 2.0-mL cryovial, each containing 0.6 
mL of vaccine (Lot RSV#016A). The vaccine virus concentrate is used undiluted at a dose of 
approximately 10 5.8PFU (group 1) in 1.0 mL or is diluted by trained research personnel to a dose of 
105.0 PFU (group 2) in a 0.5-mL volume. The vaccine vial is labeled as shown in Figure 5.2-2. 
 
 
 
 

32 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Figure 5.2-1: Investigational Product Label Sample 
 
RSV 6120/ΔNS1: titer 6.2 log10 PFU per mL 
  
Live recombinant Respiratory Syncytial Virus  
RSV 6120/ΔNS1 
VERO GROWN VIRUS VACCINE 
Date: 
20NOV2017 
Vial#:XXXX 
Lot: 
RSV#018A 
CAUTION:NEW DRUG LIMITED 
BY FEDERAL (USA) LAW 
TO INVESTIGATIONAL USE 
Store at –80°C ± 15°C 
Charles River Laboratories, Malvern, PA 
(enlarged sample) 
Figure 5.2-2: Investigational Product Label Sample 
 
RSV 6120/F1/G2/ΔNS1: titer 5.8 log10 PFU per mL 
  
Live recombinant Respiratory Syncytial Virus  
RSV 6120/F1/G2/ΔNS1 
VERO GROWN VIRUS VACCINE 
Date: 
26OCT2017 
Vial#:XXXX 
Lot: 
RSV#016A 
CAUTION:NEW DRUG LIMITED 
BY FEDERAL (USA) LAW 
TO INVESTIGATIONAL USE 
Store at –80°C ± 15°C 
Charles River Laboratories, Malvern, PA 
(enlarged sample) 
 
5.2.2. 
 Diluent for RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 
The diluent for RSV 6120/∆NS1 and RSV 6120/F1/G2/ΔNS1is Lactated Ringer’s Solution for Injection, 
USP.  
5.2.3        Placebo for RSV 6120/ΔNS1 and RSV 6120/F1/G2/ΔNS1 
The placebo for RSV 6120/∆NS1 and RSV 6120/F1/G2/ΔNS1is is Lactated Ringer’s Solution for Injection, 
USP. 
5.3. 
STUDY PRODUCT STORAGE 
Vaccine will be stored in a secure freezer at -80°C ± 15°C. It must remain frozen until the 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.  

33 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Lactated Ringer’s Solution for Injection, USP should be stored at room temperature as recommended 
by the supplier until the day before study product preparation. Vaccine diluent/placebo must be 
transferred to a secure 2°C to 8°C refrigerator at least 24 hours before use.  
Procedures for managing the vaccine and diluent/placebo shipment are in the CIR JHU Pharmacy 
SOP. 
5.4. 
 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 Pharmacy SOP. 
The unblinded dispenser will prepare the correct dose of study product for each participant in a 
biological safety cabinet (BSC) or compounding aseptic containment isolator (CACI) using aseptic 
technique. If necessary to preserve blinding, all prepared syringes will be as described in the MOP. 
5.4.1. 
Diluent  
The diluent is Lactated Ringer’s Solution for Injection, USP. 
5.4.2. 
Placebo 
Placebo is Lactated Ringer’s Solution for Injection, USP.  
Placebo will be drawn up in a sterile syringe to a volume of 1.0 mL for group 1 and to a volume of 
0.5 mL for group 2 and labeled per instructions in the MOP. If necessary to preserve blinding, all 
prepared syringes will be as described in the MOP. 
5.4.3. 
Live RSV 6120/∆NS1 or RSV 6120/F1/G2/ΔNS1 
Diluent will be prepared prior to removal of vaccine from the freezer. The MOP will be followed for 
proper handling of the study product.  
When manipulating the undiluted study product, the smallest gauge needle possible will be used to 
avoid loss of study product in the needle and syringe hub. For participants in group 1, the frozen 
study product will be thawed and diluted prior to administration with Lactated Ringer’s Solution for 
Injection, USP to a dose of approximately 106.0 PFU (RSV 6120/∆NS1) or administered undiluted at 
the indicated dose of 105.8 PFU (RSV 6120/ F1/G2/ΔNS1) in 1.0 mL. For participants in group 2, the 
frozen study product will be thawed and diluted prior to administration with Lactated Ringer’s 
Solution for Injection, USP to a dose of approximately 105.0 PFU in 0.5mL.  
The diluted study product will be drawn up to a volume of 1.0 mL (group 1) or 0.5 mL (group 2) in a 
sterile syringe and labeled per instructions in the MOP. The labeled filled syringes will be 
transported to the clinical site for administration in a Credo medical transport cooler or cooler with 
ice or cold packs with the lid closed, to maintain temperature at 2ºC to 8ºC. Vaccine must be 
administered within 4 hours of being removed from the freezer. Placebo must be administered within 
4 hours of being removed from the refrigerator. 

34 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Samples of undiluted (if available) and diluted study product will be aliquoted from the remaining 
vaccine that has been prepared. The samples will be snap-frozen as per the MOP and stored at -80°C 
± 15°C separate from the concentrated study product. Titration of vaccine will be completed to 
confirm the titer of the vaccine administered to the participants.  
The CIR JHU Pharmacy SOP provides detailed instructions on study product storage, handling, and 
preparation. Details on labeling study product and transporting it to the clinic can be found in the 
MOP. 
5.5. 
STUDY PRODUCT INOCULATION PROCEDURE 
All study participants will receive a single dose of study product, administered as nose drops. There 
is no nasal preparation prior to administration. While the participant is supine, a volume of 1.0 mL 
(group 1) or 0.5 mL (group 2) of study product will be delivered as nose drops (approximately 0.5 
mL [group 1] or 0.25 mL [group 2] per nostril) using a sterile, needle-less, masked syringe. 
Participant will remain supine for approximately 60 seconds following inoculation. 
5.6. 
STUDY PRODUCT ACQUISITION 
The clinical lots of RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1 were generated by Charles River 
Laboratories (CRL) using the seed virus provided by the NIH.  
 
Lactated Ringer’s Solution for Injection, USP will be used as diluent and placebo. 
Vaccine virus will be stored at a NIAID designated commercial repository until formally requested 
by the PI/designee. Prior to IRB approval and FDA safe to proceed determination, the PI/designee at 
JHU CIR may request that vials of vaccine be transferred from the sponsor to the CIR laboratories or 
Investigational Drug Service pharmacy. However, only after receipt of IRB approval and FDA safe 
to proceed determination will vials of vaccine be used for administration to study participants. The 
initial shipment may contain the full number of vials needed for implementation of the protocol. 
Procedures for ordering and shipping of the study product are in the MOP.  

35 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
5.7. 
STUDY PRODUCT ACCOUNTABILITY 
The unblinded dispenser is responsible for maintaining an accurate inventory and accountability 
record of study-product and Lactated Ringer’s Solution for Injection, USP for this study. A copy of 
the randomization code will be retained by the unblinded dispenser as outlined in the MOP. Without 
written request to unblind, the randomization code may not be released. The unblinded dispenser will 
be responsible for maintaining the blind.   
5.8. 
DISPOSITION OF USED/UNUSED STUDY PRODUCT 
After the unblinded dispenser dilutes the vaccine and draws up the vaccine into a syringe for 
administration, the label will be removed from the vaccine vials used for preparation and placed in 
the accountability log. 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 research personnel as per the 
MOP. 
5.9. 
FINAL DISPOSITION OF STUDY PRODUCTS 
After study completion or termination, all unused study product will be disposed of per the sponsor’s 
instructions.  
5.10. 
CONCOMITANT MEDICATIONS  
Permitted concomitant medications at enrollment (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. 
 
The use of prophylactic antipyretics, decongestants, or antihistamines is discouraged during the 
Acute Phase: day of inoculation day through the 10th day after inoculation for group 1; and day of 
inoculation through the 28th day following inoculation for group 2. However, use of these 
medications for treatment of symptoms is allowed.  
Due to the potentially confounding effect on study immunogenicity results, the following 
concomitant medications should be avoided after inoculation unless deemed clinically necessary.  
• 
Licensed inactivated vaccine or live-attenuated rotavirus vaccine within 14 days of inoculation, 
with the exception of inactivated influenza vaccine 
• 
Licensed live virus vaccine, other than rotavirus vaccine, within 28 days of inoculation 
• 
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 within 28 days of inoculation 
• 
Immunoglobulins and/or any blood products within 28 days of inoculation 
• 
Investigational drug or investigational vaccine within 28 days of inoculation for group 1 and 
within 56 days of inoculation for group 2 

36 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6. STUDY VISITS AND PROCEDURES 
An overview of the study visits, evaluation schedule, and specimen collection is provided in 
Appendix II and Appendix III. This section contains additional information on visit-specific study 
procedures.  
Study visits, except inoculation, may be performed at one of the clinical sites or at a mutually 
agreed-upon location. Inoculation must be performed at one of the CIR clinical sites or pediatric 
offices where emergency supplies are available. All clinical tasks will be performed by a medical 
professional. The PE will include temperature, heart rate, respiratory rate, assessment of ears, eyes, 
nose, and throat (EENT), lungs, heart, and skin, as well as abdominal, musculoskeletal, and, as 
appropriate, developmental exams. A focused clinical assessment will include temperature, heart 
rate, respiratory rate, EENT, lung, heart and lymph nodes assessment. 
All specimens will be obtained, processed, stored and transferred per the MOP.  
In addition to the protocol-specified procedures listed in this section, study staff may complete other 
tasks consistent with 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 visit procedures will be documented in accordance 
with the MOP. Refer to Table 18 and Table 19 and Section 10 for more information on 
documentation requirements and completion of case report forms (CRFs).  
The study’s medical professional will inform parent/guardian of any significant abnormal physical 
findings and, after obtaining parental release of information, will make appropriate referrals to the 
child’s primary caregiver, if necessary. 
6.1. 
CONSENTING PROCESS      
The screening process may be completed under a separate screening protocol and consent or under 
the study protocol and consent. The parent/guardian must complete the informed consent process and 
sign the screening or study informed consent document (ICD) before procedures are performed. The 
consenting process for the study may take place during the screening visit or may be conducted at a 
separate visit prior to inoculation. During the consenting process, the child’s parent will review the 
ICD, be encouraged to ask questions, and complete a comprehension assessment to evaluate consent 
understanding. Study staff will use incorrect answers from the comprehension assessment to identify 
areas of the ICD that need further review with the parent/guardian. This will help ensure that the 
parent/guardian has sufficient understanding of the process before the ICD is signed. Parent/guardian 
will be offered a copy of the ICD. 
During the consenting process, the parent will be given the opportunity to provide permission for use 
of their child’s picture in recruitment flyers, articles, or presentations. In addition, the parent will be 
given the opportunity to provide permission for storing their child’s specimens for future respiratory 
virus vaccine studies and for research purposes. 
As needed, a parent/guardian will complete a Health Insurance Portability and Accountability Act 
(HIPAA) medical record release to allow study staff to review medical history and immunization 
records, and to allow review of AEs during the study. Only those portions of the medical record that 

37 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
are pertinent to the study will be maintained in the study chart. 
6.2. 
SCREENING VISIT 
The screening process will include reviewing the medical record, obtaining a medical history from 
parent/guardian, and conducting a PE. The PE will occur on the day of screening or the day of 
inoculation. The medical record review should include review of history related to the study 
eligibility criteria, the immunization record, and growth chart data. The medical history from the 
parent should include demographics, prior diagnoses, current medications, signs and symptoms, 
developmental status, age of household members, day care attendance, and use of medications prior 
to inoculation. 
 
The screening visit will also include obtaining a serum sample to test for the presence of RSV 
antibodies. Approximately 5 mL of blood is sufficient quantity for the screening and pre-inoculation 
assays. The procedure will be performed and documented per the MOP. As in previous phase I trials 
of other live-attenuated RSV vaccines (12, 28-30), other screening laboratory tests will not be 
performed on the participant. 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 (32).  
For the RSV-seropositive participants, the screening serum sample must be obtained in the same 
calendar year as study inoculation. The pre-inoculation serum antibody sample must be obtained no 
more than 56 days prior to inoculation.  
For the RSV-seronegative participants, the screening serum sample must be obtained no more than 
42 days prior to inoculation. Study staff should consider potential randomization and inoculation 
dates when scheduling the participant screening visits. 

38 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 2: Screening Visit Procedures 
Screening Visit Procedures  
Administrative Tasks 
 
• Conduct and document consenting process 
• Confirm parent/guardian’s informed consent comprehension 
• Obtain release of medical records as required per HIPAA 
• Review and document medical history 
• Assess eligibility 
Clinical Tasks 
• Obtain, review, and document medical history 
• Perform complete PE, or PE may be deferred until study Day 0 
• Address any concerns 
• Document findings 
• Review participant’s immunization record  
• Review participant’s medical records to determine age-
appropriate developmental assessment, and participant’s weight 
and length 
Laboratory 
Tasks 
Blood 
Collect blood for: 
screening1 and pre-inoculation2 sample for RSV serum antibody 
testing  
Follow-up Preparation 
• Review enrollment visit  
1. Obtained within the same calendar year as inoculation for RSV-seropositive participants and no 
more than 42 days prior to inoculation for RSV-seronegative participants 
2. Obtained no more than 56 days prior to inoculation for RSV-seropositive participants and no 
more than 42 days prior to inoculation for RSV-seronegative participants 
 
6.3. 
RANDOMIZATION 
Randomization numbers will be assigned by the unblinded study personnel and will be forwarded to 
the Data Safety Monitoring Board (DSMB) Executive Secretary. Unblinded study personnel will label 
the syringes per the MOP. The vaccine administrator and verifier will initial the label and place the 
label in the participant’s study chart. The vaccination time will be documented on the vaccine 
administration record (VAR).  All syringes will be disposed of by the study staff following the 
enrollment visit.  

39 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
A copy of the randomization code will be retained by the unblinded dispenser. Without the PI's 
written request to unblind, the randomization code will not be released to the clinical staff until the 
acute monitoring phase is complete for each participant within the randomization group. Pediatric 
participants will be permuted block-randomized, in groups of five, to ensure that the 2:2:1 ratio of 
treatment arm 1, to treatment arm 2, to control participants will be maintained across time. This 
allows for maintenance of the blind during each participant’s acute monitoring phase but also 
provides early information regarding safety, which is appropriate and common practice for phase 1 
respiratory virus vaccine studies (11, 23, 25). Detailed procedures for randomization and unblinding 
will be placed in the pharmacy study files and are included in the MOP. 
If the need arises to unblind a specific participant’s assignment in the event of a serious illness prior to 
completion of the acute monitoring phase, procedures for early unblinding will be followed. In the 
event that unblinding is required, only that specific participant’s assignment will be unblinded. 
Whenever possible, the PI and sponsor 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 be notified of the event in real time. 
In group 1, approximately 25 RSV-seropositive infants and children will be enrolled in the study and 
will receive RSV 6120/ΔNS1 (106.0 PFU), RSV 6120/F1/G2/ΔNS1 (105.8 PFU), or placebo in a 2:2:1 
ratio (10:10:5). As noted in Section 3.0, each vaccine may be evaluated in up to 15 additional 
seropositive children if 3 or more RSV-seropositive children (group 1) with the same treatment 
assignment have respiratory or febrile illness that coincides with shedding of > 102.5 PFU of vaccine 
virus/mL of nasal wash as detected by culture. In group 2, approximately 35 to 50 RSV-seronegative 
infants and children will be enrolled in the study and will receive RSV 6120/ΔNS1 (105.0 PFU), RSV 
6120/F1/G2/ΔNS1 (105.0 PFU), or placebo in a 2:2:1 ratio (20:20:10).  
In the case of children from the same household, all eligible children will be assigned to the same 
study product regimen. This will be done to reduce the potential cross-contamination that could result 
if children living together were to receive different study products. All enrolled children will be 
included in safety analyses. 
The PI, and the Scientific Investigator(s) of the Laboratory of Infectious Diseases will be unblinded to 
all data at the completion of the Post-Acute Phase of follow-up (Day 28 for RSV-seropositive and 
Day 56 for RSV-seronegative) 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.  
6.4. 
ENROLLMENT  
6.4.1. 
Enrollment - Inoculation (Day 0) 
Inoculation with study product will avoid the time during which wt RSV circulates in the 
community, through ongoing local and national (CDC NREVSS) surveillance.. For the purpose of 
this protocol, enrollment will correspond to inoculation with investigational product, which occurs 
on study Day 0, and must be completed at a CIR office site or pediatric practice where emergency 
supplies are available. Prior to inoculation, the clinical staff will provide an authorized request for 
study product to the unblinded dispenser. The request must include the information outlined in the 
MOP.  

40 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
The ICD will be obtained from the parent/guardian of each child who participates in this study prior 
to the performance of any study procedures or inoculation. If not previously obtained, then the ICD 
will be completed on the day of enrollment. The child’s parent/guardian will be encouraged to ask 
questions and complete a comprehension assessment to evaluate understanding of the study. This 
will help ensure that the parent/guardian has sufficient understanding of the study process before the 
ICD is signed. 
If the participant is noted to have any of the following on enrollment day, inoculation must be 
deferred: 
• 
fever (temporal or rectal temperature of ≥ 100.4°F), or  
• 
URI or LRI symptoms or signs (including but not limited to rhinorrhea, cough, pharyngitis), or 
• 
nasal congestion significant enough to interfere with successful inoculation, or  
• 
otitis media 
• 
contact with a person diagnosed with COVID-19 disease or SARS-CoV-2 infection within the 
preceding 10 days  
 
If the inoculation for an RSV-seropositive participant has to be deferred to the following calendar 
year, or if the 42-day window from screening to inoculation is exceeded for an RSV-seronegative 
participant, then the infant or child must be rescreened. 
 
Table 3: Enrollment Visit Procedures 
Enrollment Visit Procedures (Day 0)   
Administrative Tasks 
 
• Confirm study ICD is completed and signed and dated by both 
parent/guardian and study staff who completed the consenting process 
• Complete eligibility determination and confirmation 
• Complete paper-based eligibility checklist 
• Obtain release of medical records as required per HIPAA 
Clinical Tasks 
• Obtain interim history from parent/guardian 
• Perform PE: 
o 
Complete PE if deferred at screening, OR 
o 
Focused PE if physical completed at screening 
• Address any concerns  
• Document findings  
• Confirm eligibility 
Laboratory 
Tasks 
Blood  
If insufficient volume obtained at screening, collect blood for: 
• Pre-inoculation RSV antibody titer*  
Nasal Wash 
or Swab 
and/or 
Nasosorption 
SAM Strip 
Collect nasosorption strip and/or nasal wash for Group 1 and the first 
13 participants enrolled in Group 2: 
• RSV antibody assays 
Note: The nasosorption must be performed prior to the nasal wash or 
swab. 
The nasal wash or swab must be obtained prior to administering the 
study product. 
Collect nasal wash or swab for: 
• RSV viral detection and quantification 

41 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Study Product 
Administration 
• Administer study product and maintain participant in a supine position 
for a minimum of 1 minute 
• Observe for a minimum of 30 minutes after inoculation to evaluate for 
immediate hypersensitivity reactions 
Follow-up Preparation 
• 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 and schedule next in-person visit 
• Offer parent/guardian safety seat education materials, and a safety seat 
educator appointment 
• Offer parent lactation services by a certified lactation counselor, if 
appropriate 
*Within same calendar year as screening for RSV-seropositive participants, and no more than 42 days from 
screening for RSV-seronegative participants 
The parent/guardian will record the infant/child’s temperatures and signs of illness on the 
temperature card and will report these to study personnel during an in-person or remote research visit 
or non-visit day contact. New rectal thermometers will be given and temporal artery thermometers 
will be provided to parent/guardian for use during the study. For temperature measurements, 
parent/guardian 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 (33). 
The parent/guardian will measure temporal artery temperatures following the manufacturer’s 
directions. If any temporal artery temperature is ≥ 100.0°F, parent/guardian will be asked to measure 
a rectal temperature within 20 minutes (33). For study-specific management and grading of 
temperatures, see Section 8, Table 21. 
6.5. 
STUDY PHASES  
Refer to Figure 1.1-1 and Figure 1.1-2 for timelines of study visits. The Acute Phase begins with 
inoculation and ends at midnight on the 10th day after inoculation for RSV-seropositive participants, 
and at midnight on the 28th day after inoculation for RSV-seronegative participants. During the Acute 
Phase of the study, a study healthcare professional will be available by telephone 24 hours a day for 
consultation with parent/guardian regarding any illnesses that may occur. 
Study personnel will have daily contact with RSV-seropositive participants’ parents/guardians for the 
first 10 days after inoculation, and for the RSV-seronegative participants during 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 participants (33-36).  
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 illness (Section 6.7, Illness Visit). 
 

42 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.5.1. 
RSV-Seropositive Children (Group 1) 
6.5.1.1. 
Acute Phase: In-person Study Visit Days  
An in-person study visit and clinical assessment performed by a healthcare professional will be 
completed during visits on Days 3, 4, 5, 6, 7, and 10 after inoculation, with a visit window of ± 1 
day. If an in-person visit is moved by ± 1 day, then the non-visit day contact will be completed in 
place of the original interim visit date. 
 
Table 4: Acute Phase In-Person Visit Procedures – RSV-Seropositive Children 
Days 3, 4, 5, 6, 7 and 10 In-person Visit Procedures (each visit ± 1 days) 
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
 
medications and/or immunizations 
 
signs and symptoms of illness 
 
highest temperature reading, indicate temperature method  
• Perform focused clinical examination 
• Address any concerns 
• Review safety data 
• Document findings  
Laboratory 
Tasks 
Nasal wash   
Collect nasal wash  for: 
• RSV viral detection and quantification 
Follow-up Preparation 
• Schedule non-visit day contact and follow-up, in-person visits 
Day 10 only:  
• Review SAE and LRI criteria with participants and how to contact 
study personnel during Post-Acute Phase  
During an Acute Phase study visit, if the participant is diagnosed with or suspected of having URI, 
LRI, otitis media or fever (as defined in Appendix IV), testing of the nasal wash specimen for 
adventitious agents will be performed as described in the MOP.  
6.5.1.2. 
Acute Phase: Non-Visit Study Day Contacts 
In the 10 days following inoculation, parental contact will be made on days that an in-person visit is 
not completed: Days 1, 2, 8, and 9 (each visit ± 1 day). 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 (Section 6.8). 

43 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 5: Acute Phase Non-Visit Contact Procedures – RSV-Seropositive Children 
Days 1, 2, 8, and 9 Contact Procedures (each visit ± 1 day) 
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
• medications and/or immunizations 
• signs and symptoms of illness 
• highest temperature reading, indicate temperature method 
• Address any concerns 
• Review safety data 
• Document findings 
Follow-up Preparation 
• Schedule an illness appointment if necessary 
 
6.5.1.3. 
Study Day 11  
There will be a non-visit contact on Day 11 to obtain interim history through midnight on the 10th 
day following inoculation. If the Day 10 Visit takes place on Day 11, it is not necessary to have an 
additional contact with the family on Day 11. 
 
Table 6: Day 11 Non-Visit Procedures – RSV-Seropositive Children 
Day 11 Non-Visit Procedures  
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
 
medications and/or immunizations 
 
signs and symptoms of illness 
 
highest temperature reading, indicate temperature method 
• Address any concerns 
• Review safety data 
• Document findings 
Follow-up Preparation 
• Review SAE and LRI criteria with participants and how to contact 
study personnel during Post-Acute Phase  
 
 
 

44 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.5.1.4. 
Post-Acute Phase  
The Post-Acute Phase begins on the 11th day after inoculation and ends on the 28th day after 
inoculation. During the Post-Acute Phase, parent/guardian will be instructed to monitor for and 
contact the study staff if their child has symptoms that are suggestive of a SAE or LRI. If the parent 
reports an SAE or LRI that may meet the study pause or stop criteria (Section 8.3), then an Illness 
Visit will be scheduled (Section 6.7). 
6.5.1.5. 
Study Day 28 Visit 
The Day 28 Visit should be conducted between 28 and 35 days following inoculation. Because the 
Post-Acute Phase ends as the 28th day following inoculation, only events through that time should be 
evaluated as having occurred during the Post-Acute Phase.  
Table 7: Day 28 Visit Procedures – RSV-Seropositive Children 
Day 28 Visit (+7 Days) 
Administrative Tasks 
• Provide study compensation 
Clinical Tasks 
• Document the participant’s interim history as reported from the 
parent/guardian from midnight of the 10th day through the 28th day 
following inoculation, including:  
 
visits to medical provider/hospitalizations 
 
medications and/or immunizations 
 
signs and symptoms of illness meeting SAE or LRI criteria 
• Address any concerns 
• Review safety data 
• Document findings 
Laboratory 
Tasks 
Blood 
Collect blood for: 
• Serum antibodies to RSV 
Nasal Wash 
and/or 
Nasosorption 
SAM Strip  
Collect nasosorption strip for: 
• RSV antibody assays 
Note: The nasosorption must be performed prior to the nasal wash. 
Collect nasal wash   for: 
• RSV antibody assays 
6.5.2. 
RSV-Seronegative Infants and Children 
6.5.2.1. 
Acute Phase: In-person Study Visit Days  
An in-person study visit and clinical assessment performed by a healthcare professional will be 
completed during visits on Study Days 3, 5, 7, 10, 12, 14, 17 and 28 after inoculation with a visit 
window of ± 1 day. If an in-person visit is moved by ± 1 day, then the non-visit day contact is 
completed in place of the original interim visit date. The relevant CDC and JHU institutional 
guidance regarding in-person visits and use of personal proctective equipment will be followed. If a 
Stay at Home Order is initiated following inoculation, or if the child has a respiratory or febrile 

45 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
illness, the parent may be asked to obtain the nasal swab and the clinical assessment may be done via 
a remote or in-person research visit. Transport of specimens from the home to the research 
laboratory will be arranged as needed. 
Table 8: Acute Phase In-Person Visit Procedures – RSV-Seronegative Children 
Days 3, 5, 7, 10, 12, 14, 17, and 28 In-person Visit Procedures (each visit ± 1 days) 
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
 
medications and/or immunizations 
 
signs and symptoms of illness 
 
highest temperature reading, indicate temperature method  
• Perform focused clinical examination 
• Address any concerns 
• Review safety data 
• Document findings  
Laboratory 
Tasks 
Nasal Wash 
or Swab  
Nasosorption 
SAM Strip 
 
 
 
Collect nasal wash or swab for: 
• RSV viral detection and quantification 
In addition to above, Day 28 only for the first 13 participants 
enrolled prior to March 2020: Collect nasosoption strip and/or 
nasal wash for: 
• RSV antibody assays 
 
Follow-up Preparation 
• 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  
• If Day 28 visit is conducted on or after seasonal enrollment pause:  
• 
Review plans for weekly contact during the RSV-season 
surveillance period (Section 6.6.2) 
 
 

46 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.5.2.2. 
Acute Phase: Non-Visit Study Day Contacts 
In the 28 days following inoculation, parental contact will be made on days that an in-person visit is 
not completed: Days 1, 2, 4, 6, 8, 9, 11, 13, 15, 16, and 18–27 (each visit ± 1 day). On non-visit days, 
study staff will contact the parent/guardian and will document the parent/guardian-provided 
temperatures and signs of illness. Participants with illness may have additional visits to assess the 
severity of the illness (Section 6.7). 
 
Table 9: Acute Phase Non-Visit Contact Procedures – RSV-Seronegative Children 
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 ± 1 day) 
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
 
medications and/or immunizations 
 
signs and symptoms of illness 
 
highest temperature reading, indicate temperature method 
• Address any concerns 
• Review safety data 
• Document findings 
Follow-up Preparation 
• Schedule an illness appointment if necessary 
 
6.5.2.3. 
Study Day 29  
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 takes place on Day 29, it is not necessary to have an 
additional contact with the family on Day 29. 
 
Table 10: Day 29 Non-Visit Procedures – RSV-Seronegative Children 
Day 29 Non-Visit Procedures  
Clinical Tasks 
• Document the participant’s previous days’ interim history as 
reported from the parent/guardian, include:  
 
medications and/or immunizations 
 
signs and symptoms of illness 
 
highest temperature reading, indicate temperature method 
• Address any concerns 
• Review safety data 
• Document findings 
Follow-up Preparation 
• Review SAE criteria with participants and how to contact study 
personnel during Post-Acute Phase  
 
6.5.2.4. 
Post-Acute Phase  
The Post-Acute Phase begins on the 29th day after inoculation and ends on the 56th day after 
inoculation. During the Post-Acute Phase, parent/guardian will be instructed to monitor for and 
contact the study staff if their infant or child has symptoms that are suggestive of a SAE. If the 

47 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
parent reports an SAE that may meet the study pause or stop criteria (Section 8.3) then schedule an 
Illness Visit (Section 6.7). 
6.5.2.5. 
Study Day 56 Visit 
The Day 56 Visit should be conducted between 56 and 63 days following inoculation. Because the 
Post-Acute Phase ends on the 56th day following inoculation, only events through that time should be 
evaluated as having occurred during the Post-Acute Phase. If a Stay at Home Order is put in place, 
any required blood draws will be collected in-person as soon as possible after the Stay at Home 
Order is lifted. 
Table 11: Day 56 Visit Procedures – RSV-Seronegative Children 
Day 56 Visit (+7 Days) 
Administrative Tasks 
• Provide first study compensation payment 
Clinical Tasks 
• Document the participant’s interim history as reported from the 
parent/guardian from midnight of the 28th day through the 56th day 
following inoculation, including:  
 
visits to medical provider/hospitalizations 
 
medications and/or immunizations 
 
signs and symptoms of illness meeting SAE criteria 
• Address any concerns 
• Review safety data 
• Document findings 
Laboratory 
Tasks 
 
Blood 
Collect blood for: 
• Serum antibodies to RSV 
Nasal Wash 
and/or 
Nasosorption 
SAM Strip 
Collect nasosorption strip and/or nasal wash for13 participants 
enrolled prior to March 2020: 
• RSV antibody assays 
• Note: The nasosorption must be performed prior to the nasal wash  
 
Follow-up Preparation 
Review plans for weekly contact during the RSV Season 
Surveillance Period (Section 6.6.2). 
For participants enrolled prior to March 2020, if Day 56 Visit is 
conducted: 
• Prior to October 1 
 
schedule Pre-RSV Season Visit (Section 6.6.1) 
• On or after October 1 
 
Day 56 Visit will also be the Pre-RSV Season Visit 
 
Review plans for weekly contact during the RSV Season 
Surveillance Period (Section 6.6.2) 
 

48 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.5.2.6. 
Period after Day 56 through Start of RSV Surveillance Phase 
During this period, contact with the participant is not required. No clinical data will be recorded on 
CRFs or reported under this protocol except for data as outlined in Table 19 and in Section 7.2. For 
participants enrolled prior to March 2020, contact with the participant may have occurred for the 
Pre-RSV Season Study Visit described in Section 6.6.1. 
6.6. 
RSV SURVEILLANCE SEASON (RSV-SERONEGATIVE SUBJECTS)  
6.6.1. 
Pre-RSV Season Surveillance Study Visit Only for Participants Enrolled Prior 
to March 2020 
For the 13 participants enrolled prior to March 2020, an in-person Pre-RSV Season Study Visit was 
completed between October 1- 31 for collection of a blood sample, nasal wash samples, and/or 
nasosorption strips for RSV antibody assays.The Pre-RSV Season Study Visit is not required if the 
Day 56 Visit is conducted on or after October 1; the samples collected at the Day 56 Visit are 
sufficient for the Pre-RSV Season Study Visit.  If a Stay at Home Order is put in place, any required 
blood draws will be collected in-person as soon as possible after the Stay at Home Order is lifted. 
Table 12: Pre-RSV Season Surveillance Study Visit Procedures: for 1st 13 
participants 
Pre-RSV Season Study Visit (October 1 through October 31 of enrollment year) 
Clinical Tasks 
• Document findings related to study procedure 
Laboratory 
Tasks 
 
Blood 
 
Collect blood for 13 participants enrolled prior to March 2020: 
• Serum antibodies to RSV  
Nasal Wash  
and/or 
Nasosorption 
SAM Strip 
Collect nasosorption strip and/or nasal wash for 13 participants 
enrolled prior to March 2020: 
• RSV antibody assays 
• Note: The nasosorption must be performed prior to the nasal wash  
 
Follow-up Preparation 
• Review plans for weekly contact during the RSV Season 
Surveillance Period (see Section 6.6.2) 
 
6.6.2. Weekly Contact for Surveillance during the RSV Season  
Based on previous data regarding the seasonality of RSV in the Baltimore, MD, area (Appendix IV, 
Figure 6.6-1), JHU will seasonally pause enrollment and start the RSV Surveillance Phase of the 
study based on local and/or national (CDC NREVSS) surveillance for RSV positivity locally in the 
Baltimore, MD area, as detailed in the MOP. The surveillance for RSV-associated disease will be 
conducted between pause of enrollment and March 31 during the first RSV season following receipt 
of study product. For some RSV-seronegative participants, surveillance during the RSV season may 
overlap with the Acute and/or Post-Acute study phases. In this case, all evaluations required for the 
RSV season surveillance and the relevant phases of the study will be conducted.  

49 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
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 via weekly 
telephone or email communication or an in-person or remote visit. (Rhinorrhea and cough need not 
meet the Appendix IV criteria if they are reported as the primary reason for seeking medical 
attention): 
• 
Medically attended fever 
• 
Medically attended URI  
• 
Medically attended otitis media 
• 
Medically attended LRI  
An Illness Visit will be scheduled within 3 days of study staff notification of any of these events 
(Section 6.7). Relevant CDC and JHU institutional guidelines regarding in-person visits and use of 
personal protective equipment to ensure protection of subjects, families, and staff will be followed in 
the context of the SARS-CoV-2 pandemic. If a Stay at Home Order is initiated, the parent may be 
asked to obtain the nasal swab for the RSV Surveillance illness, and the clinical assessment may be 
completed via a remote or in-person research visit. 
 
Table 13: RSV Season Surveillance Procedures 
RSV Season Surveillance  following inoculation 
Clinical Tasks 
• Obtain interim history 
• Review safety data 
• Document findings 
Follow-up Preparation 
• Continue with weekly contacts through March 31 
• Schedule an Illness Visit if warranted  
• Schedule the Post-RSV Season Study Visit (targeted April 1-April 
30th, with allowable window through September 30th if Stay at 
Home Orders require that this study visit be delayed) 
 

50 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.6.3. 
Post-RSV Season Surveillance Study Visit 
The Post-RSV Season Surveillance Visit will occur in the calendar year following receipt of study 
product, ideally  between April 1 and April 30, with an allowable window through September 30th if 
Stay at Home Orders require that this study visit be delayed. 
Table 14: Post-RSV Seasonal Surveillance Study Visit Procedures 
Post-RSV Season Study Visit  
Administrative Tasks 
• Provide study surveillance compensation 
Clinical Tasks 
• Document findings related to study procedures 
Laboratory 
Tasks 
 
Blood 
Collect blood for: 
• Serum antibodies to RSV 
Nasal Wash 
and/or 
Nasosorption 
SAM Strip  
Collect nasosorption strip and/or nasal wash  for 13 participants 
enrolled prior to March 2020: 
• RSV antibody assays 
• Note: The nasosorption must be performed prior to the nasal wash  
 
6.7. 
ILLNESS VISIT 
The timeframe after staff notification in which the Illness Visit must occur depends on the study phase 
and the grading of the fever and respiratory symptoms per Section 8.2 and Table 20. If the Illness 
Visit occurs on a day concurrent with an in-person or remote study visit, a single nasal wash or swab 
collection is required and adventitious agent testing will be requested. All symptoms will be followed 
until resolution or deemed stable or chronic by appropriate medical personnel (e.g., medical doctor or 
nurse practitioner). Illness Visits may occur during any of the study phases. The relevant CDC and 
JHU institutional guidelines regarding in-person visits and use of personal protective equipment to 
ensure protection of subjects, families, and staff will be followed in the context of the SARS-CoV-2 
pandemic. For illness visits, the parent may be asked to obtain the nasal swab. During remote research 
visits parents  be asked to measure the child’s temperature, heart rate, and respiratory rate. As otitis 
media and LRI cannot be detected by remote study visit, medical records from provider visits will be 
requested to document these events if a remote visit is completed or an in-person visit cannot be 
completed in a timely fashion. 
 
If the acute or post-acute phases overlap the surveillance period (between the date of seasonal pause 
of enrollment and March 31), then the timelines for the acute and post-acute phases will be used. 
  
 
 

51 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 15: Illness Visit Timeframe 
Illness Visit Timeframe 
Phase 
Symptoms 
Grade 
Visit Timeframe 
Acute 
Fever, otitis media or 
URI 
1 
Within 3 days  
Acute 
Fever, otitis media or 
URI 
≥ 2 
Within 2 days  
Acute; Post-acute seropositive* 
LRI  
Any 
Within 1 day 
Post-Acute 
SAE that meets study 
pause or stop criteria 
(Section 8.3) 
≥ 2 
Within 3 days 
RSV Season Surveillance 
Medically attended 
fever, otitis media, 
URI or LRI 
≥ 2 
Within 3 days 
*Days 0-28 for RSV-seropositive and RSV-seronegative children  
 
Table 16: Illness Visit Procedures 
Illness Visit Procedures 
Administrative Tasks 
• Complete Adventitious Agent Assay Request for reverse 
transcription polymerase chain reaction (rRT-PCR) on nasal 
swab for adventitious agents 
• Complete medical record release if needed 
Clinical Tasks 
• Document the participant’s interim history as reported from the 
parent/guardian, include:  
 medications and/or immunizations 
 signs and symptoms of illness 
 highest temperature reading, indicate temperature method  
• Perform focused clinical examination 
• Address any concerns 
• Review safety data 
• Document findings  
Laboratory 
Tasks 
Nasal Wash 
or Swab 
Collect nasal wash or swab for: 
• Viral detection and quantification 
Follow-up Preparation 
• Schedule follow-up as appropriate 
6.8. 
EARLY DISCONTINUATION STUDY VISIT 
In the event that a child is unable to continue participation in the study, parent/guardian will be 
encouraged to allow the participant to complete safety monitoring through Day 28 for RSV-
seropositive participants and through Day 56 for RSV-seronegative participants. Every effort should 
be made to schedule a final Early Discontinuation Visit. 

52 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 17: Early Discontinuation Procedures 
Early Discontinuation 
Administrative Tasks 
• Record data on CRF  
Clinical Tasks 
• Document interim history 
• Address any concerns 
• Review safety data 
• Encourage parent/guardian to allow the participant to complete 
safety monitoring through Day 28 for RSV-seropositive 
participants and through Day 56 for RSV-seronegative participants 
• Document reason for early discontinuation 
Laboratory 
Tasks 
 
Blood 
Collect blood for: 
• Serum antibodies to RSV 
Nasal Wash 
or Swab 
and/or 
Nasosorption 
SAM Strip 
If Early Discontinuation Visit is within 28 days of inoculation for 
RSV-seropositive participants or 56 days for 13 RSV-seronegative 
participants enrolled prior to March 2020 (Appendix II), collect 
nasosorption strip and/or nasal wash for: 
• RSV antibody assays 
Note: The nasosorption must be performed prior to the nasal wash or 
swab. 
Collect nasal wash or swab for: 
• Viral detection and quantification 

53 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.9. 
LABORATORY PROCEDURES 
6.9.1. 
Specimen Collection 
Specimens will be collected for this study as indicated in the Schedule of Events (Appendix II and 
Appendix III). Further information on collection of blood, nasosorption and nasal wash or swab 
specimens will also be provided in the MOP.  
6.9.2. 
Virus Detection 
Specimens for quantification of vaccine virus shedding by viral culture and/or rRT-PCR will be 
obtained by nasal wash or swab once before and approximately 6 times after inoculation for RSV-
seropositive participants and approximately 8 times after inoculation RSV-seronegative participants 
as shown in Appendix II & III. These specimens may also be tested for adventitious respiratory 
viruses if needed. Additional nasal wash or swab specimens for detection of RSV and adventitious 
respiratory viruses by culture and/or rRT-PCR will also be obtained from participants who meet 
illness criteria during the initial phase (Day 0 through Day 28 for RSV-seropositive participants; Day 
0 through Day 56 for RSV-seronegative participants) as well as during the RSV Season Surveillance 
Period (from seasonal pause in enrollment – March 31) for all RSV-seronegative participants. If a 
participant becomes ill, then up to 10 additional nasal washes or swabs may be obtained to exclude 
infection with an adventitious virus. Laboratory testing will be performed by personnel who are not 
involved with clinical assessment to maintain the blinding status. 
6.9.3. 
Specimen Collection 
For measurement of RSV serum antibodies, serum specimens will be obtained in RSV-seropositive 
children not more than 56 days prior to inoculation and on Day 28 (+7) post-inoculation; and in 
RSV-seronegative infants and children not more than 42 days prior to inoculation and on Day 56 
(+7) post inoculation. In addition, pre-RSV season serum specimens will be collected for the the first 
13 RSV-seronegative participants enrolled prior to March 2020,  and post-RSV season serum 
specimens will be collected in all RSV-seronegative infants and children. These samples will be used 
to determine whether a four-fold 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. 
Specimens will be obtained by venipuncture, finger-stick, or heel-stick. No more than 5 mL of blood 
will be drawn at each blood draw visit. A maximum of 10 mL of blood will be taken from RSV-
seropositive participants for study purposes. A maximum of 15 mL of blood will be taken from 
RSV-seronegative participants for study purposes from screening through surveillance.  
For RSV-seropositive participants, nasal wash specimens and/or nasosorption strips for measurement 
of secretory immunity will be obtained before inoculation and 28 days after inoculation. For 13 
RSV-seronegative participants enrolled prior to March 2020, nasal wash specimens and/or 
nasosorption strips for measurement of secretory immunity will be obtained before inoculation, 28 
and 56 days after inoculation, and at the pre- and post-RSV season surveillance visits. These 
specimens may be generated from the same nasal wash  obtained for viral culture, but specimen 
processing is different as described in the MOP.  

54 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
6.9.4. 
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 MOP and the JHU CIR SOPs. The frequency of specimen collection 
and testing will be directed by the Schedule of Events (Appendix II and Appendix III).  
6.9.5. 
Research Laboratories 
Quantitation of the amount of vaccine virus shed, assays to measure immune responses before and 
after inoculation, and assessment of nasal washes or swabs for adventitious viral agents will be 
performed at the CIR. Nasosorption assays will be performed at the LID, NIAID. 
Cytokine/chemokine assays may also be performed on nasal washes or swabs from participants 
infected with vaccine virus if sufficient material is available. Selected specimens may be sent to LID, 
NIAID for confirmatory testing.  
6.9.6. 
Plan for Use and Storage of Biological Samples 
All specimens collected as part of this study may, with the parent/guardian’s permission, be stored 
for future research as part of CIR’s approved biospecimen repository for vaccine research. These 
samples and data may be used for future screening for respiratory virus vaccine studies and to learn 
more about RSV infection and other diseases. The parent/guardian or child will not own the blood 
specimens, nasal specimens, or data after it is given to the study. No financial benefit will be 
provided to the parent/guardian or child from any product or idea created by the investigators using 
the data or materials. These samples will not be sold or used to make commercial products. Genetic 
tests will not be performed on these samples unless a separate informed consent is obtained.  
Samples stored in the repository will be labeled with the study identification number 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’ numbers. A master log linking the study participants’ identification numbers to their 
names is maintained by the study’s clinical staff in a password-protected computer system with 
limited access to authorized research team members and will not be shared with the laboratory. 
Study participants, or their parents/guardians, may withdraw consent for future testing of their 
specimens at any time. 
6.9.7. 
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. The procedures for obtaining, shipping, and handling of all specimens for this study will 
follow the current recommendation of the Centers for Disease Control and Prevention (CDC) and the 
NIH. All infectious specimens will be transported using packaging mandated in Title 42 of the Code 
of Federal Regulations (CFR), Part 72 (42 CFR 72) and in accordance with individual carrier 
guidelines (e.g., Federal Express). 

55 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
7. SAFETY ASSESSMENT, MONITORING, AND REPORTING 
Participant safety will be carefully assessed, monitored, and reported at multiple levels throughout 
this study. Sections 7.1-7.4 describe safety-related roles, responsibilities, and procedures. The safety 
monitoring roles of the NIAID Intramural Data Safety and Monitoring Board (DSMB) are briefly 
referenced in Section 7.1.4 and described in detail in Section 9.4.2. 
7.1. 
SAFETY-RELATED ROLES AND RESPONSIBILITIES 
7.1.1. 
Principal Investigator 
The PI is responsible for continuous monitoring of the CIR’s study participants and for alerting the 
Scientific Investigators if unexpected concerns arise. Trained study staff will record safety-related 
data on CRFs as indicated in Section 7.2. The PI is also responsible for prompt reporting to the IRBs 
and other applicable review bodies of any unanticipated problems (UPs) involving risks to 
participants or others.  
7.1.2. 
Safety Review and Communications Plan (SRCP)  
A Safety Review and Communication Plan (SRCP) will be developed for the protocol. The SRCP is 
an internal communications document between the PI and the IND sponsor (OCRPRO) Clinical 
Safety Office, which delineates the safety oversight responsibilities of the PI, the CSO, and other 
stakeholders. The SRCP also includes the overall plan for conducting periodic safety surveillance 
assessments.  
7.1.3. 
Sponsor Medical Monitor 
A medical monitor representing the IND sponsor (OCRPRO) has been appointed for oversight of 
safety in this clinical study. The sponsor medical monitor will be responsible for performing safety 
assessments as outlined in a Safety Review and Communications Plan (SRCP). 
7.1.4. 
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 
the monitoring of this study.  
7.1.5. 
Sponsor Reporting 
A SUSAR is a suspected adverse reaction that is both serious and unexpected, as defined in 21 CFR 
312.32.  Events that meet SUSAR criteria will be reported promptly by the PI to the CSO and will be 
evaluated by the PI and CSO as a possible protocol Stop trigger. SUSARs will be reported to the 
FDA and all participating investigators as IND Safety Reports. The IND Sponsor will also submit a 

56 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
brief report of the progress of the investigation to the FDA on an annual basis as defined in 21 CFR 
312.33. 
Following notification from the PI, OCRPRO, as the IND Sponsor, will report all SAEs to the FDA 
within the required timelines. Fatal and life-threatening events will be reported within 7 calendar 
days of the sponsor’s awareness and all other SAEs will be reported within 15 calendar days of the 
sponsor’s awareness.  
7.2. 
 SAFETY-RELATED RECORDING ON CRFS 
AEs that occur during protocol-specified AE reporting periods (see Table 18 and Table 19) 
following inoculation of study product should be considered AEs. 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 study investigator or designee, elicited or volunteered from the 
parent/guardian or participant, or captured on participant’s temperature cards. Assessment of safety 
will include clinical observation and monitoring of laboratory parameters as necessary. Follow-up 
measures such as history, clinical assessment, and laboratory testing and/or treatment may be 
necessary if a participant experiences an AE. Details of AEs will be properly documented on the 
source documents, recorded on CRFs, and reported to LID investigators and the DSMB in separate 
semi-annual and annual reports. AEs will be provided to the IRB as defined by the IRB policy. 
This study has several periods of AE observation that have different AE CRF recording 
requirements. In addition, for RSV-seronegative participants, there may be a period when no AEs are 
recorded on CRFs if the Day 56 Visit occurs in advance of the start of the RSV Season Surveillance 
Period (date of seasonal enrollment pause). The AEs (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 18 and Table 19.  
Concomitant medications and AEs identified during the acute and post-acute phase will be recorded 
on CRFs. Medically-attended febrile, respiratory or OM during RSV season surveillance and 
concomitant medications related to SAEs identified during the RSV season surveillance period will 
be recorded on CRFs. AEs will be recorded as signs, symptoms, laboratory test results, and 
diagnoses, as shown in Table 18 and Table 19. In addition, the following concomitant medications 
will be recorded on CRFs regardless of whether the identified AE is recorded on the CRF: 
 
• All cough and cold remedies including decongestants, cough suppressants, expectorants 
• All nasal sprays (except saline spray) 
• All antihistamines 
• All antipyretics 
• All prescription medications 
 

57 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 18: AE CRF Recording Requirements for RSV-Seropositive Participants 
Study Phase at the Time of Event 
Onset 
Calendar 
Date 
AEs to Record on CRFs 
Concomitant Medications to 
Record on CRFs 
Day 0 through midnight of 10th day 
following inoculation 
(Acute Phase) 
ANY 
• All SAEs 
• All solicited AEs that meet 
Appendix IV, Table 32 criteria  
• All unsolicited AEs (Grades 1 to 
4) 
•  Exception of diaper rash, 
teething pain, and spitting up 
unless treated with prescription 
medication or non-prescription 
medications with antipyretic 
properties 
Record these medications on the 
CRFs regardless of whether the 
related event is recorded on the 
CRFs: 
 
• All cough and cold remedies 
including decongestants, cough 
suppressants, expectorants 
• All nasal sprays (except saline 
spray) 
• All antihistamines 
• All antipyretics 
• All prescription medications 
 
For SAEs and LRIs: 
• All medications related to the 
recorded event 
From the 11th day after inoculation 
to the 28th day after inoculation 
(Post-Acute Phase) 
ANY 
• All SAEs and LRIs 
 
 
• All medications related to the 
recorded event 
Throughout study 
ANY 
• Unresolved AEs or SAEs with 
onset date from Day 0 to 28th day 
after inoculation 
• All medications related to the 
recorded event 
 
 
 

58 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 19: AE CRF Recording Requirements for RSV-Seronegative Participants 
Study Phase at the Time of Event 
Onset 
Calendar 
Date 
AEs to Record on CRFs 
Concomitant Medications to 
Record on CRFs 
Day 0 through midnight of 28th day 
following inoculation 
(Acute Phase) 
ANY 
• All SAEs 
• All solicited AEs that meet 
Appendix IV, Table 32 criteria  
• All unsolicited AEs (Grades 1 to 
4)  
• Exception of diaper rash, teething 
pain, and spitting up, unless 
treated with prescription 
medication or non-prescription 
medications with antipyretic 
properties 
 
Record these medications on the 
CRFs regardless of whether the 
related event is recorded on the 
CRFs: 
 
• All cough and cold remedies 
including decongestants, cough 
suppressants, expectorants 
• All nasal sprays (except saline 
spray) 
• All antihistamines 
• All antipyretics 
• All prescription medications 
 
For SAEs and LRIs: 
• All medications related to the 
recorded event 
From the 29th day after inoculation 
to the 56th day after inoculation 
(Post-Acute Phase) 
ANY 
• All SAEs  
 
• All medications related to the 
recorded event 
From Day 56 Visit through start of 
RSV Season Surveillance Period 
 
Up to date 
of seasonal 
pause in 
enrollment  
in year of 
inoculation 
• Grade ≥ 3 AEs or SAEs that is 
deemed related to Pre-RSV 
Season Study Visit procedures 
• All medications related to the 
recorded event 
RSV Season Surveillance Period 
Date of 
seasonal 
pause in 
enrollment 
to March 
31  
• Fevers, LRIs, URIs, and/or otitis 
media that are medically 
attended  
• All SAEs  
 
Note: These events do not need to 
meet the Appendix IV, Table 32 
criteria 
For SAEs and LRIs (all grades): 
• All medications related to the 
recorded event 
 
Medications related to recorded 
medically attended illness should 
be documented in source notes 
Post-RSV Season 
Ideally 
April 1 to 
April 30; 
allowable 
through 
Sept 30th 
in the year 
after the 
inoculation 
• Grade ≥ 3 AEs or SAEs that are 
deemed related to Post-RSV 
Season Study Visit procedures 
• All medications related to the 
recorded event 

59 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 19: AE CRF Recording Requirements for RSV-Seronegative Participants 
Study Phase at the Time of Event 
Onset 
Calendar 
Date 
AEs to Record on CRFs 
Concomitant Medications to 
Record on CRFs 
Throughout study 
ANY 
• Unresolved AEs or SAEs with 
onset from Day 0 to midnight on 
the 28th day after inoculation 
• Unresolved SAEs with onset prior 
to the 56th day following 
inoculation 
• Unresolved SAEs with onset 
during RSV Surveillance Period 
or related to the Pre- or Post-RSV 
Season Study Visit 
• All medications related to the 
recorded event 
 
 

60 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
7.3. 
 SERIOUS ADVERSE EVENT REPORTING 
Serious Adverse Events (SAEs) are described in Section 8.1.2.  All SAEs will be reviewed by a study 
physician, recorded on the Safety Expedited Report Form (SERF) and followed through to resolution. 
SAEs that have not resolved by the end of the follow-up period are followed until final outcome is 
known. If it is not possible to obtain a final outcome for an SAE (e.g., the participant is lost to follow-
up), the reason a final outcome could not be obtained will be recorded by the investigator on the AE 
CRF and the SERF. In addition to the SAE Reporting Category identified below, other AEs that must 
be reported in an expedited manner are LRIs as defined in Appendix IV if they occur during study 
Days 0 to 28 in group 1 and group 2. 
 
Deaths and immediately life-threatening SAEs will be reported within 1 business day after the site 
becomes aware of the event. All other SAEs will be reported within 3 business days of site awareness. 
SAEs will be reported either by fax or e-mail to all of the following: 
• 
Sponsor: Office of Clinical Research Policy and Regulatory Operations, NIAID, NIH: Clinical 
Safety Office (CSO): Phone: 301-846-5301, Fax: 301-846-6224, rchspsafety@mail.nih.gov 
• 
DSMB Executive Secretary: Phone 301-846-5301, Fax: 301-846-6224, 
niaiddsmbia@niaid.nih.gov 
• 
LID, NIAID: Ursula Buchholz, PhD, NIAID/NIH, 301-594-1533, Fax: 301-480-1268, 
Email: ubuchholz@niaid.nih.gov 
  
SAEs will also be reported to Western Copernicus Group Institutional Review Board (WCGIRB 
contact information below) based on its reporting requirements: 
• 
Western Copernicus Group Institutional Review Board (WCG IRB), 1019 39th Ave SE, 
Puyallup, WA 98374, Phone: 1-855-818-2289 
 
7.4. 
REPORTING OF UNANTICIPATED PROBLEMS TO OFFICE OF CLINICAL RESEARCH   
POLICY AND REGULATORY OPERATIONS (OCRPRO) 
An unanticipated problem (UP) is defined as any event, incident, experience, or outcome that is: 
1. Unexpected in terms of nature, severity, or frequency in relation to 
a. The research procedures that are described in the IRB-approved research protocol and 
informed consent or other study documents; and 
b. The characteristics of the participant population being studied; and  
2. Related to participation in the research; and 
3. Places participants or others at a greater risk of harm (including physical, psychological, 
economic, or social harm) than was previously known or recognized. Per the investigational new 
drug (IND) sponsor, an AE with a serious outcome will be considered increased risk. 
 
UPs must be reported to the local IRB per their requirements. Non-Serious AEs that are UPs must 
also be reported to the sponsor CSO. Submit the local IRB UP report form to the CSO at the 
following address no later than 7 calendar days of the PI awareness of the event.  
 

61 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
 
SPONSOR CLINICAL SAFETY OFFICE CONTACT INFORMATION:  
Clinical Safety Office 
5705 Industry Lane 
Frederick, MD 21704 
Phone 301-846-5301 
Fax 301-846-6224 
E-mail: rchspsafety@mail.nih.gov 
UPs that are not AEs (UPnonAE) are not routinely reported to the CSO. However, an UPnonAE that 
may, in the opinion of the investigator, involve risk to the participant, affect others in the research 
study, or significantly impact the integrity of research data would be considered a non-serious UP 
and would be reported to the CSO. For example, we will report occurrences of breaches of 
confidentiality, accidental destruction of study records, or unaccounted-for study drug. 
8. PARTICIPANT MANAGEMENT 
8.1. 
MANAGEMENT OF ADVERSE EVENTS 
An AE is any untoward medical occurrence in a participant administered a pharmaceutical product 
that 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 subject’s participation in the research, whether or not related to the 
subject’s participation in the research. This includes exacerbation of pre-existing conditions and 
intercurrent illnesses.  
Causality assessment is based on available information at the time of the assessment of the AE. The 
investigator may revise the causality assessment as additional information becomes available. 
All AEs identified in this study will be source documented, 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 Sections 8.2.1 and 8.2.2 and its relationship to study 
product, assessed by the MD or NP according to the following categories and definitions: 
Related 
There is a reasonable possibility that the adverse event may be related to the 
study drug. 
Not related 
There is not a reasonable possibility that the adverse event may be related to 
the study drug.  
There are two categories of AEs specific to CIR 330: solicited and unsolicited. Solicited AEs are 
described in Section 8.1.1. Unsolicited AEs are all other AEs. However, for infants and children, the 
following common events will not be recorded as AEs unless a prescribed concomitant medication is 
used to treat them: non-infectious rashes, teething pain, and spitting up. The use of medications will 
only be captured in the event of an AE or pre-existing condition. 

62 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
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 study product administration, 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 meet the definitions per Appendix IV, Table 32 Individual symptoms listed in the “events” 
column that fail to meet the criteria in the “definition” column in Appendix IV, Table 32 are 
recorded in source documents but are not recorded on the CRFs. During the Acute Phase of this 
study, Days 0 through 10 for RSV-seropositive participants and Days 0 through 28 for RSV-
seronegative participants, solicited AEs meeting the criteria for reporting will be recorded on CRFs, 
assigned a severity grade (Section 8.2), and assessed for relationship to study product (see Section 
8.1). Solicited AEs are defined in Appendix IV and include the following:  
1. Fever 
2. URI 
a. Rhinorrhea, 
b. Pharyngitis,  
c. Cough without LRI, or 
d. Hoarseness 
3. Otitis Media 
4. LRI 
a. Wheezing, 
b. Pneumonia, 
c. Laryngotracheobronchitis (croup), 
d. Rhonchi, or 
e. Rales 
 
Solicited AEs Elicited by History Unconfirmed by Clinical Assessment 
With the exception of fever, solicited AEs reported by parents/guardians are NOT recorded on CRFs 
if a clinical assessment done on the day of the event(s) does not/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 
If the parent/guardian report of a fever meets the “definition” column criteria in Appendix IV on a 
day on which there was a clinical assessment, the fever will be recorded on CRFs regardless of 
whether the clinical assessment confirmed its presence. 
 
Events elicited by parent/guardian history for days on which there was no clinical exam will be: 
1) Recorded on the CRFs as AEs if the parent/guardian description meets the “definition” column 
criteria in Appendix IV 
2) 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 

63 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
8.1.2. 
Serious Adverse Events 
A SAE is an AE, whether considered related to the study product or not, that meets one or more of 
the following criteria: 
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 participant or may require intervention to prevent one of 
the outcomes listed above. 
 
8.1.3. 
Unexpected Adverse Event 
An AE is unexpected if it is not listed in the IB or Package Insert (for marketed products) or is not 
listed at the specificity or severity that has been observed. 
It is the responsibility of the IND Sponsor to make this determination. 
 
8.2. 
GRADING THE SEVERITY OF ADVERSE EVENTS 
The Investigator will assess all AEs with respect to Seriousness (criteria listed above), Severity 
(intensity or grade), and Causality (relationship to study agent and relationship to research). All AEs 
and fever will be graded using the protocol-defined grading system outlined in Table 20 and Table 
21.  
 
 

64 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
8.2.1. 
AE Grading  
 
Table 20: Grading for Adverse Events 
Severity 
Defined 
Grade 1 Mild 
No medical intervention required; may include over-the-counter medications 
managed by the participant or caregiver for treatment of symptoms 
Grade 2 Moderate 
Moderate symptoms, i.e., symptoms interfering to some degree with usual 
activities. In most cases, symptoms severe enough to necessitate a medical care 
visit would likely meet this criterion; however, if medical care is sought and 
symptoms are assessed as only mild, the event may remain Grade 1. If prescription 
medication is used or recommended for symptoms, the event automatically moves 
to at least Grade 2. 
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 
 
8.2.2. 
Fever Grading 
 
 
 
 
Table 21: Grading for Fever 
Severity 
Defined 
Grade 1 
≥100.4ºF but ≤101.4ºF 
Grade 2 
≥101.5ºF but ≤102.4ºF 
Grade 3 
≥102.5ºF but ≤104.8ºF 
Grade 4 
≥104.9ºF 
Applies to any modality of temperature measurement  
8.3. 
PAUSING AND STOPPING RULES 
If any of the following occur in a participant during the specified period after receiving the study 
product, additional enrollment/inoculations will be temporarily suspended (paused) (Table 22). 
 
 
 

65 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 22: Pausing  Rules 
 
Specified Phase 
Event 
Reporting 
Acute and Post-Acute 
Days 0 through 28 
following inoculation 
for RSV-seropositive 
participants and Days 0 
through 56 following 
inoculation for RSV-
seronegative 
participants 
An SAE that cannot be attributed to 
an etiology or cannot be attributed 
to a cause unrelated to the study 
product. 
A description of the vaccine-associated 
AE(s) or safety issue must be reported by 
the PI or study staff, within one business 
day of the PI's awareness, to the CSO and 
the DSMB by fax or email and the IRB, 
as applicable 
 
Acute Only 
Days 0 through 10 
following inoculation 
for RSV-seropositive 
participants and Days 0 
through 28 following 
inoculation for RSV-
seronegative 
participants 
An LRI per Appendix IV, OR 
A fever of Grade 4 OR 
Any Grade 3 or above solicited AE 
(other than fever) 
Post-Acute  
Days 11 through 28 
following inoculation 
for RSV-seropositive 
participants 
An LRI per Appendix IV 
A description of the vaccine-associated 
AE(s) or safety issue must be reported by 
the PI or study staff, within three business 
days of the PI's awareness, to the CSO and 
the DSMB by fax or email 
 
 The PI and/or CSO will determine if the study should be paused or stopped.  Pausing the study 
requires suspension of enrollment until a decision is made whether or not to continue enrollment and 
study vaccine administration or permanently stop the study.    
The IND sponsor (OCRPRO) will determine if the FDA needs to be notified.  The FDA may stop the 
study at any time following review of any safety concerns. The DSMB and IND sponsor will be 
informed and receive pertinent data of the event by the PI. Follow-up visits for participants already 
inoculated will continue as outlined. 
If the event is determined to have occurred in a participant who received active agent (vaccine), and 
the event meets one of the following pausing rule criteria, then the event will be reviewed by the 
DSMB prior to resuming enrollment or permanently stopping the study.  
1. One or more participants experience 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 develop 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 develop 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 study team 
considers a significant safety issue for participants. 
 
The IND Sponsor, in collaboration with the PI, the DSMB and the Safety Review Team will 
determine if it is safe to resume the study or if the study needs to be stopped. The PI will notify the 

66 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
IRB of the decision on whether the study will be resumed or 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.  
9. STATISTICAL CONSIDERATIONS 
9.1. 
GENERAL DESIGN ISSUES 
9.1.1. 
General Design 
The goal of this Phase I, double-blinded, randomized, placebo-controlled vaccine trial is to assess the 
safety, infectivity, and immunogenicity of the RSV 1620/ΔNS1 and RSV 1620/F1/G2/ΔNS1 vaccine 
candidates in RSV-seropositive and RSV-seronegative pediatric participants and to assess whether 
these two vaccines are good candidates to move forward into a Phase IB study. In Group 1, 
approximately 25 RSV-seropositive infants and children will be enrolled in the study and will receive 
RSV 6120/ΔNS1 (106.0 PFU), RSV 6120/F1/G2/ΔNS1 (105.8 PFU), or placebo in a 2:2:1 ratio 
(10:10:5). In Group 2, approximately 35-50 RSV-seronegative infants and children will be enrolled in 
the study and will receive RSV 6120/ΔNS1 (105 PFU), RSV 6120/F1/G2/ΔNS1 (105 PFU), or placebo 
in a 2:2:1 ratio (20:20:10).  
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 AEs 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 
in Section 9.5.  
The Study Team is interested in whether the vaccines are better than placebo with respect to infectivity 
and immunogenicity; thus, the statistical comparisons between vaccine and placebo groups will consist 
of one-tailed tests. The study will not be powered for the modest difference expected between the two 
vaccines; thus, the groups receiving vaccine will be compared by means of descriptive analyses. Both 
vaccines may be used in a subsequent trial aimed at assessing efficacy, provided that this is not counter 
indicated by safety results.  
9.2. 
 OUTCOME MEASURES 
9.2.1. 
Primary Outcome Measures 
Safety: Types and grades of study product-related: 
• 
Solicited AEs as defined in Appendix IV from Study Days 0-10 for RSV-seropositive and Study 
Days 0-28 for RSV-seronegative participants 
• 
Unsolicited AEs from Study Days 0-10 for RSV-seropositive and Study Days 0-28 for RSV-
seronegative participants 

67 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
• 
SAE (Section 8.1.2) from Study Days 0-28 for RSV-seropositive and Study Days 0-56 for RSV-
seronegative participants 
 
Infectivity: 
• 
Infection with RSV as defined as:  
1) vaccine virus identified in a nasal wash or swab from Study Days 0-10 for RSV-
seropositive and Study Days 0-28 for RSV-seronegative (a binary outcome based on 
nasal washes or swabs done throughout the study period; Day 0 nasal wash or swab 
will be counted as baseline); or  
2) ≥4-fold rise in serum RSV neutralizing antibody titer or serum RSV F (IgG) titer 
from Study Days 0-28 for RSV-seropositive and Study Days 0-56 for RSV-
seronegative participants 
• 
Peak titer of vaccine virus shed from Study Days 0-10 for RSV-seropositive and Study Days 0-
28 for RSV-seronegative participants 
• 
Duration of virus shedding in nasal washes or swabs as determined by a) culture and/or b) rRT-
PCR from Study Days 0-10 for RSV-seropositive and Study Days 0-28 for RSV-seronegative 
participants 
 
Immunogenicity: 
• 
≥4-fold rise in RSV-neutralizing antibody titer from study entry to Study Day 28 for 
RSV-seropositive and from study entry to Study Day 56 for RSV-seronegative participants 
• 
≥4-fold rise in IgG antibody responses to RSV F glycoprotein (by ELISA) from study entry to 
Study Day 28 for RSV-seropositive and from study entry and Study Day 56 for RSV-
seronegative participants 
9.2.2. 
 Secondary Outcome Measures 
 
• 
The frequency and severity of symptomatic, medically attended respiratory and febrile illness in 
the RSV-seronegative (group 2) vaccine and placebo recipients who experience natural infection 
with wt RSV during the RSV season. 
• 
The antibody responses in the RSV-seronegative vaccine and placebo recipients who experience 
natural infection with wt RSV during the RSV season. 
• 
Mucosal antibody titers to vaccine, in nasal wash  or nasosorption samples in Group 1 and the 
first 13 Group 2 participants enrolled prior to March 2020. 
9.3. 
SAMPLE SIZE AND ACCRUAL 
9.3.1. 
Sample Size and Randomization 
In group 1, approximately 25 RSV-seropositive infants and children will be enrolled in the study and 
will receive RSV 6120/ΔNS1 (106.0 PFU), RSV 6120/F1/G2/ΔNS1 (105.8 PFU), or placebo in a 2:2:1 
ratio (10:10:5), and in Group 2, approximately 35-50 RSV-seronegative infants and children will be 
enrolled in the study and will receive RSV 6120/ΔNS1 (105 PFU), RSV 6120/F1/G2/ΔNS1 (105 
PFU), or placebo in a 2:2:1 ratio (20:20:10). In total, approximately 30 vaccine recipients per 
vaccine arm and 15 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 (29, 30, 37). The 2:2:1 randomization ratio will be used to maximize the 

68 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
information obtained regarding the response of children to the RSV 6120/∆NS1 and RSV 
6120/F1/G2/∆NS1 vaccines. In the event that a participant is discontinued early from the study the 
participant will not be replaced. 
 Block randomization, in groups of five, will be used to ensure that the 2:2:1 ratio of treated to 
control participants will be maintained across time. All enrolled children will be included in safety 
analyses. 
Given the small sample size, the study will have limitations with respect to detecting AEs and 
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 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 children infected with 
vaccine virus.  
Table 23 shows the probability of observing 0 events of LRI within the sample of 20 seronegative 
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 23: The Probability of Observing LRI Events in Seronegative Vaccine 
Recipients 
 
N = 20 
True underlying probability of LRI or 
AEs 
Pr (0 events) 
Pr (1+ events) 
Pr (2+ events) 
.01 
0.82 
0.18 
0.02 
.03 
0.54 
0.46 
0.12 
.05 
0.36 
0.64 
0.26 
.1 
0.12 
0.88 
0.61 
.15 
0.04 
0.96 
0.82 
 
 
 
 
 
 

69 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Table 24 presents 90% confidence intervals (CIs) around potential rates of LRI or AEs that might be 
observed in the sample of 20 vaccine recipients. 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 vaccine 
recipients, 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 24: Percent of Participants Experiencing LRI or AEs with Exact 90% CI 
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% 
 
Group sample sizes of 20 in each vaccinated group and 10 in the placebo group would achieve 80% 
power to detect a difference between each vaccine group and the placebo 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 25 presents examples of true group differences which can be detected with 
80% power, and Figure 9.3-1 graphically shows power curves for 90% power, 80% power, and 70% 
power given the sample sizes.  
 
Table 25: 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 
 
 
 

70
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1
          
Version 5.0
Principal Investigator: Ruth A. Karron, MD
June 5, 2023
Figure 9.3-1: Power Curves for Assessment of Vaccine Virus Shedding in RSV seronegative 
Vaccinees
(Alpha=.05; N Vaccine=20; N Placebo=10; 1-Sided Exact Test)
With a sample size of 20 seronegative vaccine recipients (for each vaccine), the 90% CI around a 
sample mean peak titer of 2.5 log10, with a SD of 1.5 (from CIR 291/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%) vaccine recipients 
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 vaccine recipients who shed vaccine virus is not lower 
than 78%.
Response proportion in the Vaccine Group

71 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
9.4. 
MONITORING 
9.4.1. 
Site Monitoring Plan 
As per International Conference on Harmonisation of Technical Requirements for Pharmaceuticals 
for Human Use - Good Clinical Practice (ICH-GCP) 5.18 and FDA 21 CFR 312.50, clinical 
protocols are required to be adequately monitored by the study sponsor. This study monitoring will 
be conducted according to the “NIAID Intramural Clinical Monitoring Guidelines.” Monitors under 
contract to the NIAID/OCRPRO will visit the clinical research site to monitor aspects of the study in 
accordance with the appropriate regulations and the approved protocol. The objectives of a 
monitoring visit will be: 1) to verify the existence of signed informed consent documents and 
documentation of the ICF process for each monitored participant; 2) to verify the prompt and 
accurate recording of all monitored data points, and prompt reporting of all SAEs; 3) to compare 
CRIMSON data abstracts with individual participants’ records and source documents (participants’ 
charts, laboratory analyses and test results, physicians’ progress notes, nurses’ notes, and any other 
relevant original participant information); and 4) to help ensure investigators are in compliance with 
the protocol. The monitors also will inspect the clinical site regulatory files to ensure that regulatory 
requirements (OHRP), FDA, and applicable guidelines (ICH-GCP) are being followed. During the 
monitoring visits, the investigator (and/or designee) and other study personnel will be available to 
discuss the study progress and monitoring visit. 
 
The investigator (and/or designee) will make study documents (e.g., consent forms, CRIMSON data 
abstracts) and pertinent hospital or clinical records readily available for inspection by the local IRB, 
the FDA, the site monitors, and the NIAID staff for confirmation of the study data.  
 
A specific protocol monitoring plan will be discussed with the PI and study staff prior to enrollment. 
The plan will outline the frequency of monitoring visits based on such factors as study enrollment, 
data collection status and regulatory obligations. 
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 Executive Secretary will provide the PI 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 PI will submit the written DSMB 
recommendations to the IRB upon receipt. All SAEs, LRIs, UPs and IND safety reports will be 
reported by the PI to the DSMB at the same time that they are submitted to the IRB and/or IND 
sponsor (OCRPRO). 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 a pausing or halting criteria 
are met and obtain a recommendation concerning continuation, modification, or termination of the 
study.  

72 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
9.5. 
ANALYSES 
9.5.1. 
Assessment of Primary Objectives  
Safety data from all participants who have been inoculated will be summarized together, 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 28 Visit follow-up for RSV-
seropositive participants and for the Day 56 Visit follow-up for RSV-seronegative participants (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 5.10 during the first 28 
days after inoculation for RSV-seropositive participants and during the first 56 days after inoculation 
for RSV-seronegative participants may be excluded from the immunogenicity evaluations after the 
time of the treatment. These participants, however, will 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.  
 
The frequency of solicited AEs and unsolicited AEs, along with 90% CIs, during Study Days 0 to 10 
in RSV-seropositive participants and during Study Days 0 to 28 in RSV-seronegative participants 
and of vaccine-related SAE during Study Days 0 to 28 in RSV-seropositive participants and during 
Study Days 0 to 56 in RSV-seronegative participants will be summarized. In addition, line listing of 
individual clinical solicited AEs and unsolicited AEs, graded by severity, will be prepared for events 
occurring during Study Days 0 to 10 in RSV-seropositive participants and during Study Days 0 to 28 
in RSV-seronegative participants and vaccine-related SAE during Study Days 0 to 28 in RSV-
seropositive participants and during Study Days 0 to 56 in RSV-seronegative participants. 
The proportion of participants with infection defined as recovery of vaccine virus from a nasal wash 
or swab as determined by rRT-PCR, and/or a >4-fold rise in neutralizing antibody titer to RSV, will 
be summarized. A line listing of the individual peak titer of vaccine virus shed and duration of virus 
shedding in nasal washes or swabs by each individual will be prepared. In addition, the geometric 
mean peak titer and mean duration of virus shed will be provided for each study group.  
The proportion of participants that develop 4-fold 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 titer (GMT) 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.  
The study results will be compared with the criteria listed in Section 2.1 to determine if these 
vaccines are promising candidates for further evaluation in expanded phase I studies or phase II 
studies. 
 

73 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
9.5.2. 
Assessment of Secondary Objectives  
The summary of the frequency and severity of symptomatic, medically attended respiratory and 
febrile illness in the RSV-seronegative vaccine and placebo recipients who experience natural 
infection with wt RSV during the RSV season will be presented. In addition, a line listing of the 
individual RSV antibody titers before and after the surveillance period will be prepared, and the 
GMT and median antibody titers will be provided for each study group. The adaptive immune 
responses to vaccine will be summarized for each study group.  
The two vaccine groups will be compared descriptively with respect to peak viral titers and antibody 
titers following vaccination. 
10 DATA HANDLING AND RECORD KEEPING 
10.1. 
DATA MANAGEMENT RESPONSIBILITIES 
The CIR will 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 per 
the MOP.  
Study data will be maintained on source documents with key data points abstracted and entered into 
CRIMSON. Source documents include all recordings of observations or notations of clinical 
activities and all reports and records necessary to confirm the data abstracted for this study. Data 
entry into CRIMSON will be performed by authorized individuals. The Investigator is responsible 
for assuring that the data collected are complete, accurate, and recorded in a timely manner.  
Source documents include all recordings of observations or notations of clinical activities, and all 
reports and records necessary for the evaluation and reconstruction of the clinical trial. Source 
documents may include the participant’s medical records and laboratory reports per the MOP. Data 
will be collected directly from participants during study visits and contacts.  
10.2. 
ESSENTIAL AND SOURCE DOCUMENTS AND ACCESS TO SOURCE DATA 
Study-related documentation will be completed as required by the IRB, the sponsor, and regulatory 
authorities. Continuing review documentation will be submitted 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 RSV 6120/∆NS1 and RSV 6120/F1/G2/∆NS1 vaccines 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. 
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, the FDA, regulatory authorities, IRB, 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.  

74 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Study-related documents will be maintained 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 PI 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 PI. 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 PI wish to assign the study records to another party and/or move them to another 
location, the PI 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 from the sponsor prior 
to destruction or relocation of research records. 
 
10.3. 
 CLINICAL INVESTIGATOR’S BROCHURES 
The IBs comprehensively describe all the available preclinical experience with the experimental 
vaccines. If relevant new information becomes available during the course of the trial, the PI will 
receive revised IBs or amendments to the current versions. 
10.4. 
QUALITY CONTROL AND QUALITY ASSURANCE 
Essential documents and participant research records are participant to continuous quality control 
and quality assurance procedures consistent with the MOP. 
11 CLINICAL SITE MONITORING 
As per ICH-GCP 5.18 and FDA 21 CFR 312.50 clinical protocols are required to be adequately 
monitored by the study sponsor. This study monitoring will be conducted according to the “NIAID 
Intramural Clinical Monitoring Guidelines.” Monitors under contract to the NIAID/OCRPRO will 
visit the clinical research site to monitor aspects of the study in accordance with the appropriate 
regulations and the approved protocol. The objectives of a monitoring visit will be: 1) to verify the 
existence of signed informed consent documents and documentation of the ICF process for each 
monitored subject; 2) to verify the prompt and accurate recording of all monitored data points in 
CRIMSON, and prompt reporting of all SAEs; 3) to compare abstracted information entered into 
CRIMSON with individual subjects’ records and source documents (subjects’ charts, laboratory 
analyses and test results, physicians’ progress notes, nurses’ notes, and any other relevant original 
subject information); and 4) to help ensure investigators are in compliance with the protocol. The 
monitors also will inspect the clinical site regulatory files to ensure that regulatory requirements 
(Office for Human Research Protections-OHRP), FDA, and applicable guidelines (ICH-GCP) are 
being followed. During the monitoring visits, the investigator (and/or designee) and other study 
personnel will be available to discuss the study progress and monitoring visit. 
 
The investigator (and/or designee) will make study documents (e.g., consent forms, CRIMSON data 
abstracts) and pertinent hospital, including CRIMSON, or clinical records readily available for 

75 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
inspection by the local IRB, FDA, the site monitors, and the NIAID staff for confirmation of the 
study data.  
 
A specific protocol monitoring plan will be discussed with the Principal Investigator and study staff 
prior to enrollment. The plan will outline the frequency of monitoring visits based on such factors as 
study enrollment, data collection status and regulatory obligations. 
The sponsor will retain originals of the Form FDA 1572 and copies of other study documents as 
deemed necessary. 
 
Statement of Compliance  
The trial will be conducted in compliance with this protocol, ICH-GCP guidelines, FDA guidelines 
and applicable regulatory requirements. The CIR monitoring will be conducted according to the 
OCRPRO Clinical Trial Management Group’s Monitoring Plan. 
12 HUMAN SUBJECTS PROTECTIONS 
12.1. 
INSTITUTIONAL REVIEW BOARD/ETHICS COMMITTEE REVIEW AND APPROVAL  
Prior to study initiation, the PI will obtain IRB review and approval of this protocol and ICDs in 
accordance with 45 CRF 46. In addition to the initial review and approval, the IRB must review the 
study at least annually. The PI must also promptly report to the IRB any changes in the study and 
any UPs involving risks to participants or others. 
All IRB policies and procedures must be followed, and complete documentation of all 
correspondence to and from the IRBs must be maintained in the essential document files. 
A copy of the study approval (including approval of the ICD) is to be maintained in the 
investigator’s study document binder, and a copy will be supplied to the sponsor.  
During the study, the PI is responsible for providing the IRB with all documents subject to review 
(i.e., protocol amendments, ICD 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. 

76 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
12.2. 
VULNERABLE PARTICIPANTS 
The NIH is mandated by law to ensure that children be included in clinical research when 
appropriate (38, 39). 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 and 
children who take part in this study are considered vulnerable participants per the US CFR, and site 
IRBs/IBCs must consider the potential risks and benefits to child participants as described in 45 CFR 
46 Subpart D (for children). 
12.3. 
 INFORMED CONSENT 
In obtaining and documenting informed consent, the PI must comply with the applicable regulatory 
requirements, ICH-GCP guidelines, and ethical principles. The written ICD must be approved by the 
IRB prior to its use. 
Written informed consent for 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. 
As part of the informed consent process, parents/guardians will also be asked whether they agree to 
storage and future research testing of the biological specimens that 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 study participation.  
12.4. 
 POTENTIAL BENEFITS 
Participants may not receive any direct vaccine-related benefit from enrollment in this study. Some 
children who receive vaccine may be protected against infections with wt RSV that circulates in the 
community. 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.  
Parents/guardian may be offered child safety seat educational material and referral to community 
inspection stations by study staff, and may be offered certified lactation counseling services, if 
appropriate. 
12.5. 
POTENTIAL RISKS  
12.5.1. Venipuncture 
Risks occasionally associated with venipuncture include pain, bleeding, and bruising at the site of 
venipuncture, lightheadedness, infection, and rarely syncope. Before each blood draw, we will offer 
to use anesthetic skin cream to decrease the pain. 

77 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
12.5.2. Nasal Wash or Nasal Swab 
Risks occasionally associated with nasal wash or swab  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 are advised to use over-the-counter saline solutions and/or nasal bulb 
suction to clear a young child’s congested nostrils during an URI. The nasal bulb suction and saline-
like solutions are 2 components of our nasal wash procedure. 
12.5.3. Nasosorption SAM Strips 
The risk associated with the Nasosorption SAM Strips is a nasolacrimal reflex which involves slight 
watering of the eyes. 
12.5.4. Topical Anesthetic Cream 
Risks occasionally associated with the use of topical anesthetic cream include temporary skin 
discoloration, skin irritation, rash, hives, and rarely, dizziness or drowsiness. 
12.5.5. Receipt of Study Product 
If a study vaccine (RSV 6120/∆NS1 or RSV 6120/F1/G2/∆NS1) is insufficiently attenuated, 
participants could experience rhinorrhea, cough, fever, otitis media, or LRI. Immediate 
hypersensitivity reactions including urticaria, anaphylaxis, or other immunoglobulin E (IgE)-
mediated responses are possible, as with any vaccine. With any investigational vaccine, there is a 
theoretical possibility of risks about which we have no present knowledge. Parents/guardians will be 
informed of any such risks should further data become available. 
The study participant’s family does not pay for the study product or research visits including 
examinations and laboratory tests that are part of this study, including evaluation of illness, if any. 
NIAID agrees that the funding for appropriate acute care will be provided through the NIAID 
contract (HHS272200900010C) for any side effects that are determined to be related to the 
administration of vaccine.  
12.6. 
  REIMBURSEMENT/COMPENSATION 
Compensation will be in the form of either check or pre-paid card. Participants’ parent/guardian will 
receive compensation at the rate of $50 for the inoculation visit, $30 per each scheduled or 
unscheduled study visit, $5 per each scheduled non-visit day report and weekly surveillance report, 
and a $50 bonus for study completion. In addition, the parent/guardian will receive bus tokens, taxi 
fare, or parking passes as needed for study visits. Parent/guardian will be compensated only for those 
portions of the study that are completed. Compensation will be in accordance with IRB policies and 
procedures and will be subject to IRB approval. 
Participants may receive age-appropriate books or small toys. The total value of the books or toys will 
not exceed $10 per participant.  

78 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
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 9.4 and 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 will be identified by coded number only.  
All local databases must be secured with password protected access systems. Lists, logbooks, 
appointment books, and any other documents that link participant identification numbers to personal 
identifying information should be stored in a separate, locked location in an area with limited access.  
12.8. 
 MANAGEMENT OF INCIDENTAL FINDINGS 
Study clinicians will inform parents/guardians of all clinically meaningful PE findings and 
laboratory tests. When applicable, study 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 
CIR 330 is sponsored by the OCRPRO, Division of Clinical Research (DCR), NIAID, NIH. 
OCRPRO is responsible for regulatory oversight of this study. Safety-related information pertaining 
to the study product will be distributed prior to and during the conduct of the study, in accordance 
with its sponsor obligations.  
NIAID provides 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 9.4. As part of these visits, monitors will inspect study-related documentation to 
ensure compliance with all applicable local and US regulatory requirements. 
13.2. 
PROTOCOL REGISTRATION 
Prior to implementation of this protocol, and any subsequent full version amendments, the CIR must 
have the protocol and the protocol ICDs approved, as appropriate, by their local IRB/IBC, local IBC, 
and any other applicable regulatory entity (RE).  
For any future protocol amendments, upon receiving final IRB/IBC and any other applicable RE 
approvals, CIR should implement the amendment immediately.  

79 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
13.3. 
STUDY IMPLEMENTATION 
This study will be conducted in accordance with the protocol, ICH guidelines, and all applicable 
local and US regulations. Study implementation will also be guided by the study-specific MOP and 
other study implementation materials. 
13.4. 
PROTOCOL DEVIATION REPORTING 
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. 
See MOP for further instructions. 
Deviations will be reported to the IRB 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 one or more study participants and/or significant impacts on the integrity of study data must 
also be reported following procedures specified in the MOP.  
13.5. 
CLINICALTRIALS.GOV 
This protocol is subject to the US Food and Drug Administration Amendments Act of 2007 
(FDAAA), including registration in ClinicalTrials.gov. 
14 PUBLICATIONS 
Publication of the results of this trial will be governed by 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 Cooperative Research and 
Development Agreement (CRADA) or other collaborative agreement in place. 
 
 
 

80 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
15  REFERENCES 
 
1. 
Hall CB, Weinberg GA, Iwane MK, Blumkin AK, Edwards KM, Staat MA, Auinger P, 
Griffin MR, Poehling KA, Erdman D, Grijalva CG, Zhu Y, Szilagyi P. 2009. The burden of 
respiratory syncytial virus infection in young children. N Engl J Med 360:588-98. 
 
2. 
Nair H, Nokes DJ, Gessner BD, Dherani M, Madhi SA, Singleton RJ, O'Brien KL, Roca A, 
Wright PF, Bruce N, Chandran A, Theodoratou E, Sutanto A, Sedyaningsih ER, Ngama M, 
Munywoki PK, Kartasasmita C, Simoes EA, Rudan I, Weber MW, Campbell H. 2010. 
Global burden of acute lower respiratory infections due to respiratory syncytial virus in 
young children: a systematic review and meta-analysis. Lancet 375:1545-55. 
 
3. 
Shi T, McAllister DA, O'Brien KL, Simoes EAF, Madhi SA, Gessner BD, Polack FP, 
Balsells E, Acacio S, Aguayo C, Alassani I, Ali A, Antonio M, Awasthi S, Awori JO, Azziz-
Baumgartner E, Baggett HC, Baillie VL, Balmaseda A, Barahona A, Basnet S, Bassat Q, 
Basualdo W, Bigogo G, Bont L, Breiman RF, Brooks WA, Broor S, Bruce N, Bruden D, 
Buchy P, Campbell S, Carosone-Link P, Chadha M, Chipeta J, Chou M, Clara W, Cohen C, 
de Cuellar E, Dang DA, Dash-Yandag B, Deloria-Knoll M, Dherani M, Eap T, Ebruke BE, 
Echavarria M, de Freitas Lazaro Emediato CC, Fasce RA, Feikin DR, Feng L, et al. 2017. 
Global, regional, and national disease burden estimates of acute lower respiratory infections 
due to respiratory syncytial virus in young children in 2015: a systematic review and 
modelling study. Lancet 390:946-958. 
 
4. 
Hall CB. 2001. Respiratory syncytial virus and parainfluenza virus. N Engl J Med 344:1917-
28. 
 
5. 
Martinez FD. 2003. Respiratory syncytial virus bronchiolitis and the pathogenesis of 
childhood asthma. PediatrInfect DisJ 22:S76-S82. 
 
6. 
Collins PL, Karron RA. 2013. Respiratory syncytial virus and metapneumovirus, p 1086-
1123. In Knipe DM, Howley PM, Cohen JI, Griffin DE, Lamb RA, Martin MA, Racaniello 
VR, Roizman B (ed), Fields Virology, Sixth Edition, Sixth ed, vol 1. Lippincott Williams & 
Wilkins, Philadelphia. 
 
7. 
Kim HW, Canchola JG, Brandt CD, Pyles G, Chanock RM, Jensen K, Parrott RH. 1969. 
Respiratory syncytial virus disease in infants despite prior administration of antigenic 
inactivated vaccine. Am J Epidemiol 89:422-434. 
 
8. 
Collins PL, Murphy BR. 2007. Vaccines against Human Respiratory Syncytial Virus. 
Respiratory Syncytial Virus 14:233-278. 
 
9. 
Acosta PL, Caballero MT, Polack FP. 2015. Brief history and characterization of enhanced 
respiratory syncytial virus disease. Clinical and Vaccine Immunology 23:189-95. 
 

81 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
10. 
Collins PL, Murphy BR. 2005. New generation live vaccines against human respiratory 
syncytial virus designed by reverse genetics. ProcAm ThoracSoc 2:166-173. 
 
11. 
Wright PF, Karron RA, Belshe RB, Shi JR, Randolph VB, Collins PL, O'Shea AF, Gruber 
WC, Murphy BR. 2007. The absence of enhanced disease with wild type respiratory 
syncytial virus infection occurring after receipt of live, attenuated, respiratory syncytial virus 
vaccines. Vaccine 25:7372-8. 
 
12. 
Karron RA, Wright PF, Belshe RB, Thumar B, Casey R, Newman F, Polack FP, Randolph 
VB, Deatly A, Hackell J, Gruber W, Murphy BR, Collins PL. 2005. Identification of a 
recombinant live attenuated respiratory syncytial virus vaccine candidate that is highly 
attenuated in infants. J Infect Dis 191:1093-1104. 
 
13. 
Hendry RM, Burns JC, Walsh EE, Graham BS, Wright PF, Hemming VG, Rodriguez WJ, 
Kim HW, Prince GA, McIntosh K. 1988. Strain-specific serum antibody responses in infants 
undergoing primary infection with respiratory syncytial virus. J Infect Dis 157:640-647. 
 
14. 
Johnson PR, Jr., Olmsted RA, Prince GA, Murphy BR, Alling DW, Walsh EE, Collins PL. 
1987. 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 61:3163-3166. 
 
15. 
Collins PL, Hill MG, Camargo E, Grosfeld H, Chanock RM, Murphy BR. 1995. 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 92:11563-11567. 
 
16. 
Teng MN, Whitehead SS, Bermingham A, St Claire M, Elkins WR, Murphy BR, Collins 
PL. 2000. Recombinant respiratory syncytial virus that does not express the NS1 or M2-2 
protein is highly attenuated and immunogenic in chimpanzees. J Virol 74:9317-21. 
 
17. 
Chatterjee S, Luthra P, Esaulova E, Agapov E, Yen BC, Borek DM, Edwards MR, Mittal A, 
Jordan DS, Ramanan P, Moore ML, Pappu RV, Holtzman MJ, Artyomov MN, Basler CF, 
Amarasinghe GK, Leung DW. 2017. Structural basis for human respiratory syncytial virus 
NS1-mediated modulation of host responses. Nat Microbiol 2:17101. 
 
18. 
Dhar J, Cuevas RA, Goswami R, Zhu J, Sarkar SN, Barik S. 2015. 2'-5'-Oligoadenylate 
Synthetase-Like Protein Inhibits Respiratory Syncytial Virus Replication and Is Targeted by 
the Viral Nonstructural Protein 1. J Virol 89:10115-9. 
 
19. 
Hastie ML, Headlam MJ, Patel NB, Bukreyev AA, Buchholz UJ, Dave KA, Norris EL, 
Wright CL, Spann KM, Collins PL, Gorman JJ. 2012. The human respiratory syncytial virus 
nonstructural protein 1 regulates type I and type II interferon pathways. Mol Cell Proteomics 
11:108-27. 

82 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
20. 
Lo MS, Brazas RM, Holtzman MJ. 2005. Respiratory syncytial virus nonstructural proteins 
NS1 and NS2 mediate inhibition of Stat2 expression and alpha/beta interferon 
responsiveness. J Virol 79:9315-9. 
 
21. 
Spann KM, Tran KC, Chi B, Rabin RL, Collins PL. 2004. Suppression of the induction of 
alpha, beta, and lambda interferons by the NS1 and NS2 proteins of human respiratory 
syncytial virus in human epithelial cells and macrophages [corrected]. J Virol 78:4363-9. 
 
22. 
Spann KM, Tran KC, Collins PL. 2005. Effects of nonstructural proteins NS1 and NS2 of 
human respiratory syncytial virus on interferon regulatory factor 3, NF-kappaB, and 
proinflammatory cytokines. J Virol 79:5353-62. 
 
23. 
Swedan S, Andrews J, Majumdar T, Musiyenko A, Barik S. 2011. Multiple functional 
domains and complexes of the two nonstructural proteins of human respiratory syncytial 
virus contribute to interferon suppression and cellular location. J Virol 85:10090-100. 
 
24. 
Swedan S, Musiyenko A, Barik S. 2009. Respiratory syncytial virus nonstructural proteins 
decrease levels of multiple members of the cellular interferon pathways. J Virol 83:9682-93. 
25. 
Munir S, Hillyer P, Le Nouen C, Buchholz UJ, Rabin RL, Collins PL, Bukreyev A. 2011. 
Respiratory syncytial virus interferon antagonist NS1 protein suppresses and skews the 
human T lymphocyte response. PLoS Pathog 7:e1001336. 
 
26. 
Munir S, Le Nouen C, Luongo C, Buchholz UJ, Collins PL, Bukreyev A. 2008. 
Nonstructural proteins 1 and 2 of respiratory syncytial virus suppress maturation of human 
dendritic cells. J Virol 82:8780-96. 
 
27. 
Krempl C, Murphy BR, Collins PL. 2002. Recombinant respiratory syncytial virus with the 
G and F genes shifted to the promoter-proximal positions. J Virol 76:11931-42. 
 
28. 
Karron RA, Wright PF, Crowe JE, Jr., Clements-Mann ML, Thompson J, Makhene M, 
Casey R, Murphy BR. 1997. Evaluation of two live, cold-passaged, temperature-sensitive 
respiratory syncytial virus vaccines in chimpanzees and in human adults, infants, and 
children. J Infect Dis 176:1428-1436. 
 
29. 
Wright PF, Karron RA, Belshe RB, Thompson J, Crowe JE, Jr., Boyce TG, Halburnt LL, 
Reed GW, Whitehead SS, Anderson EL, Wittek AE, Casey R, Eichelberger M, Thumar B, 
Randolph VB, Udem SA, Chanock RM, Murphy BR. 2000. Evaluation of a live, cold-
passaged, temperature-sensitive, respiratory syncytial virus vaccine candidate in infancy. J 
Infect Dis 182:1331-1342. 
 
30. 
Wright PF, Karron RA, Madhi SA, Treanor JJ, King JC, O'Shea A, Ikizler MR, Zhu Y, 
Collins PL, Cutland C, Randolph VB, Deatly AM, Hackell JG, Gruber WC, Murphy BR. 
2006. The interferon antagonist NS2 protein of respiratory syncytial virus is an important 
virulence determinant for humans. J Infect Dis 193:573-581. 
 
31. 
McFarland EJ, Buchholz UJ, Muresan P, Cunningham CK, Luongo C, Thumar B, Collins P, 
Perlowski C, Valentine M, Schappell E, Gnanashanmugam D, Siberry G, Rexroad V, Barr 

83 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
E, Harding P, DiPerna A, Karron R. High Infectivity of recombinant live-attenuated 
respiratory syncytial virus vaccine RSV LID ΔM2-2 in RSV-seronegativeinfants and 
children, p. In (ed),  
  
32. 
Anonymous. 2007. American Academy of Pediatrics, Committee on Practice and 
Ambulatory Medicine. Recommendations for Preventive Pediatric Health Care. Pediatrics 
120:1376. 
 
33. 
Siberry GK, ener-West M, Schappell E, Karron RA. 2002. Comparison of temple 
temperatures with rectal temperatures in children under two years of age. ClinPediatr(Phila) 
41:405-414. 
 
34. 
Crowe JE, Jr., Bui PT, London WT, Davis AR, Hung PP, Chanock RM, Murphy BR. 1994. 
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 12:691-9. 
 
35. 
Whitehead SS, Juhasz K, Firestone CY, Collins PL, Murphy BR. 1998. Recombinant 
respiratory syncytial virus (RSV) bearing a set of mutations from cold-passaged RSV is 
attenuated in chimpanzees. J Virol 72:4467-4471. 
 
36. 
Gharpure MA, Wright PF, Chanock RM. 1969. Temperature-sensitive mutants of 
respiratory syncytial virus. J Virol 3:414-421. 
 
37. 
Karron RA, Luongo C, Thumar B, Loehr KM, Englund JA, Collins PL, Buchholz UJ. 2015. 
A gene deletion that up-regulates viral gene expression yields an attenuated RSV vaccine 
with improved antibody responses in children. Sci Transl Med 7:312ra175. 
 
38. 
National Institutes of Health. 1994. Guidelines for the Inclusion of Women and Minorities 
as Subjects in Clinical Research. Federal Register., 
  
39. 
National Institutes of Health. 1998. NIH Policy and Guidelines on the Inclusion of Children 
as Participants in Research Involving Human Subjects.  
 
 

 
84 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
APPENDICES 
 
Appendix I: Tables Referenced in the Background and PRECLINICAL Sections 
 
Table 26: Viral titers of nasopharyngeal swab samples from AGMs inoculated with RSV ΔNS1 
mutants, or with recombinant wt RSV rA2 
 
Virus Test 
Articlea 
AGM  
ID 
NP virus titer (log10 PFU/mL) on indicated daysb 
Peak 
virus  
titer 
Sum of 
daily 
titers c 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
12/ 
14 
RSV A2 
CTM d 
RSV#004A 
12RSV01 
7209 
- 
- 
3.1 
4.4 
4.4 
5.2 
3.6 
4.1 
3.4 
2.0 
- 
5.2 
31.3 
7467 
- 
0.7 
2.8 
3.8 
2.1 
3.9 
3.4 
3.2 
1.9 
2.0 
- 
3.9 
24.5 
7468 
- 
1.7 
2.6 
4.0 
3.9 
3.7 
3.6 
2.7 
3.6 
2.6 
- 
4.0 
29.1 
7492 
0.7 
1.2 
2.3 
3.9 
2.9 
3.9 
3.9 
2.4 
1.7 
1.0 
- 
3.9 
24.3 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
4.2 
27.2 
 
6120/ΔNS1 
Exp. lot  
16RSV08-
nontox 
8960 
- 
0.7 
- 
1.7 
2.2 
2.7 
2.8 
1.8 
- 
- 
- 
2.8 
13.7 
8992 
- 
0.7 
- 
0.7 
2.6 
2.7 
2.9 
2.6 
1.9 
0.7 
- 
2.9 
15.8 
8904 
- 
- 
- 
1.7 
1.2 
2.4 
2.8 
1.3 
- 
1.2 
- 
2.8 
12.3 
8951 
- 
- 
- 
2.3 
2.3 
3.1 
3.5 
2.6 
1.5 
- 
- 
3.5 
17.0 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
3.0 
14.7 
 
6120/ΔNS1 
CTM e 
RSV#018A  
18RSV01-tox  
9140 
- 
- 
1.3 
- 
1.0 
1.4 
1.7 
- 
- 
- 
- 
1.7 
7.9 
9147 
- 
0.7 
1.3 
1.9 
2.5 
2.5 
2.8 
2.7 
1.8 
1.7 
- 
2.8 
17.1 
9116 
- 
- 
- 
- 
- 
0.7 
- 
1.6 
- 
- 
- 
1.6 
5.8 
9136 
- 
- 
0.7 
1.4 
1.9 
2.6 
2.0 
2.8 
1.3 
- 
- 
2.8 
14.1 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
2.2 
12.3 
 
6120/F1/G2/ 
∆NS1 CTM f 
RSV#016A  
18RSV01-tox  
9131 
- 
- 
- 
- 
- 
- 
- 
0.7 
0.7 
- 
- 
0.7 
4.6 
9084 
- 
- 
- 
- 
- 
1.0 
- 
1.4 
- 
1.7 
- 
1.7 
6.9 
9135 
- 
- 
- 
1.0 
1.0 
1.3 
1.6 
- 
- 
- 
- 
1.6 
6.8 
9128 
- 
- 
1.0 
- 
- 
2.1 
- 
- 
- 
- 
- 
2.1 
6.3 
 
Mean: 
 
 
 
 
 
 
 
 
 
 
 
1.5 
6.1 
a Monkeys were inoculated i.n. and i.t. with 106 PFU of the indicated virus in a 1 mL inoculum per site (total dose =2x106 
PFU/AGM), except for RSV#016A, which was used at a dose of 5 x 105 PFU in a 1 mL inoculum per site (total dose = 1 x 106 
PFU). 
b Virus titrations were performed on Vero cells. The lower limit of detection was 1.0 log10 PFU/mL. Samples with no detectable 
virus are represented as “-“. Peak titers for each animal are underlined.  
c The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve). A value of 0.35 was use 
for samples with no detectable virus.  
d Lot RSV#004A; vial numbers 263, 886, 1108, 1567. 
e Lot RSV#018A, vial numbers 0021-0026, 1267-1272, 2482-2485 
f Lot RSV#016A, vial numbers .0020-0026, 1267-1274, 2482-2487. 
 

 
85 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
Table 27: Viral titers of tracheal lavage samples from AGMs inoculated with RSV ΔNS1 
mutants, or with Recombinant wt RSV rA2 
Virus Test Articlea 
AGM  ID 
TL virus titer (log10 PFU/mL) on indicated 
dayb 
Peak virus 
titer 
Sum of 
daily  
titers c 
2 
4 
6 
8 
10 
12/14 
 
 
RSV A2 
CTM 
RSV#004A 
12RSV01 d 
7209 
3.0 
3.8 
4.3 
4.5 
2.6 
- 
4.5 
18.9 
7467 
3.0 
3.0 
4.3 
3.4 
2.8 
- 
4.3 
17.2 
7468 
2.5 
1.7 
2.9 
3.0 
2.9 
- 
3.0 
13.7 
7492 
1.9 
3.5 
4.7 
3.3 
- 
- 
4.7 
14.8 
Mean: 
 
 
 
 
 
 
4.1 
16.2 
 
6120/ΔNS1 
Exp. lot  
16RSV08-nontox 
 
8960 
- 
- 
1.5 
2.7 
- 
- 
2.7 
7.0 
8992 
- 
2.9 
3.6 
2.7 
1.0 
- 
3.6 
11.6 
8904 
2.3 
2.7 
3.8 
3.1 
- 
- 
3.8 
13.3 
8951 
- 
3.0 
1.5 
3.3 
- 
- 
3.3 
9.9 
Mean: 
 
 
 
 
 
 
3.3 
10.5 
 
6120/ΔNS1 CTM e 
RSV#018A  
18RSV01-tox  
 
9140 
1.0 
2.9 
4.1 
4.0 
1.3 
- 
4.1 
14.0 
9147 
2.6 
1.6 
2.7 
3.0 
1.5 
- 
3.0 
12.1 
9116 
2.0 
1.0 
2.8 
2.1 
- 
- 
2.8 
9.2 
9136 
2.7 
2.1 
3.9 
2.7 
1.5 
- 
3.9 
13.7 
Mean: 
 
 
 
 
 
 
3.5 
12.2 
 
6120/F1/G2/ 
∆NS1 CTM f 
RSV#016A  
18RSV01-tox  
 
9131 
- 
- 
1.0 
- 
- 
- 
1.0 
4.5 
9084 
- 
1.8 
2.3 
2.3 
1.8 
1.0 
2.3 
9.8 
9135 
1.3 
1.5 
2.4 
2.1 
- 
- 
2.4 
8.7 
9128 
2.4 
1.5 
3.0 
2.4 
1.3 
- 
3.0 
11.3 
Mean: 
 
 
 
 
 
 
2.2 
8.6 
a Monkeys were inoculated i.n. and i.t. with 106 PFU of the indicated virus in a 1 mL inoculum per site (total dose 
=2x106 PFU/AGM), except for RSV#016A, which was used at a dose of 5 x 105 PFU in a 1 mL inoculum per site 
(total dose = 1 x 106 PFU). 
b Virus titrations were performed on Vero cells. The lower limit of detection was 1.0 log10 PFU/mL. Samples with no 
detectable virus are represented as “-“. Underlined value indicates maximum titer for each animal. 
c The sum of daily titers is used as an estimate for the magnitude of shedding (area under the curve). Values of 0.7 
are used for samples with no detectable virus.  
d Lot RSV#004A; vial numbers 263, 886, 1108, 1567. 
e Lot RSV#018A, vial numbers 0021-0026, 1267-1272, 2482-2485 
f Lot RSV#016A, vial numbers .0020-0026, 1267-1274, 2482-2487. 
 
 
 

 
86 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
 
Table 28: Serum PRNT60 Titers in AGMs Inoculated with RSV ΔNS1 mutants, or with 
Recombinant wt RSV rA2 
Virus Test Article 
AGM  ID 
RSV Neutralization Titer      
 (Log2 of reciprocal) on days 
0 
21 
28 
RSV rA2 
wt RSV 
CTM RSV#004A a 
12RSV01 
7209 
<3.3 
7.1 
6.7 
7467 
<3.3 
8.1 
8.7 
7468 
<3.3 
5.9 
5.9 
7492 
<3.3 
7.7 
8.8 
 
Mean: 
 
7.2 
7.5 
6120/ΔNS1 
Experimental lot  
16RSV08-nontox 
8960 
<3.3 
7.9 
8.3 
8992 
<3.3 
8.5 
8.8 
8904 
<3.3 
8.6 
7.3 
8951 
<3.3 
8.4 
8.4 
 
Mean: 
 
8.4 
8.2 
6120/ΔNS1 CTM e 
RSV#018A  
18RSV01-tox  
 
9140 
<3.3 
8.6 
8.1 
9147 
<3.3 
7.1 
7.4 
9116 
<3.3 
6.6 
7.0 
9136 
<3.3 
9.3 
9.2 
 
Mean: 
 
7.9 
7.9 
6120/F1/G2/ 
∆NS1 CTM f 
RSV#016A  
18RSV01-tox  
9131 
<3.3 
4.3 
5.4 
9084 
<3.3 
7.6 
7.6 
9135 
<3.3 
6.4 
7.1 
9128 
<3.3 
8.5 
8.0 
 
Mean: 
 
6.7 
7.0 
a Lot RSV#004A; vial numbers 263, 886, 1108, 1567. 
b Lot RSV#018A, vial numbers 0021-0026, 1267-1272, 2482-2485 
c Lot RSV#016A, vial numbers .0020-0026, 1267-1274, 2482-2487.The lower limit of detection of the 60% Plaque Reduction 
assay is 3.3 (Log2 of the dilution reciprocal). Samples below the lower limit of detection are recorded  
as “-“.  
 
 
 

 
87 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Appendix II: Schedule of Events: Screening, Acute Phase, and Post-Acute Phase  
 
Table 29: Schedule of Events: Group 1: RSV-Seropositive Participants 
 
 
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 12-27 
Day 28  
Illness 
Visit  
Early DC 
Study window  
 
± 1 day 
 
+7 days  
In person visit 
X 
X 
 
 
X 
X 
X 
X 
X 
 
 
X 
 
* 
X 
X 
X 
Non-visit contact 
 
 
X 
X 
 
 
 
 
 
X 
X 
 
X 
* 
 
 
 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
History 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Interim history 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
* 
X 
X 
X 
Physical exam (full) 
(X) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical assessment 
(focused PE) 
 
(X)  
 
X 
X 
X 
X 
X 
 
 
X 
 
* 
 
X 
 
Administer study 
product 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood for: 
immunologic assays 
5mLa 
 
 
 
 
 
 
 
 
 
 
 
 
 
5mL 
 
5mL 
Nasal wash and/or 
nasosorption SAM 
strip for: RSV 
antibody 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
Nasal wash for: 
viral detection & 
quantification 
 
X 
 
 
X 
X 
X 
X 
X 
 
 
X 
 
* 
 
X 
X 
Request adventitious 
agent assay 
 
 
 
 
 
 
 
 
 
 
 
 
 
* 
 
X 
 
Total blood volume 
5mL 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
5mL 
-- 
5mL 
*If a family reports an SAE that may meet the study pause or stop criteria, complete the indicated tasks 
a Blood for RSV immunological assay is obtained not more than 56 days prior to inoculation. Screening RSV antibody 
can be obtained greater than 56 days prior to inoculation so long as it is obtained within the same calendar year. 
 A physical exam can be completed at screening or enrollment.

 
88 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
aBlood for screening is obtained not more than 42 days prior to inoculation.  A physical exam can be completed at screening or enrollment.  
bFor the 13 subjects enrolled prior to March 2020 
c If a Stay at Home Order is initiated or in the case of respiratory illness, the parent may be asked to obtain the nasal swab, and the clinical assessment may be completed via a remote or in-person 
research visit. During remote research visits parents will also be asked to measure the child’s temperature, heart rate and respiratory rate. 
 
Table 30: Schedule of Events: Group 2: RSV-Seronegative Participants 
 
 
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 
Study window  
 
± 1 day 
 
 
+7 days  
In person visitc 
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 
* 
 
 
 
Informed consent 
X 
(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 
* 
X 
X 
X 
Physical exam (full) 
(X) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical assessment 
(focused PE)c 
 
(X)  
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
* 
 
X 
 
Administer study 
product 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood for: 
immunologic assays 
5mLa 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5mL 
 
5mL 
Nasal wash and/or 
nasosorption SAM 
strip for: RSV 
antibodyb 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
X 
 
X 
Nasal wash or swab 
for: 
viral detection & 
quantificationc 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
X 
 
 
X 
 
X 
 
* 
 
X 
X 
Request adventitious 
agent assay 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* 
 
X 
 
Total blood volume 
5mL 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
-- 
5mL 
-- 
5mL 
*If a family reports an SAE that may meet the study pause or stop criteria, complete the indicated tasks  

 
89 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
 
Version 4.0 
Principal Investigator: Ruth A. Karron, MD 
 
August 12, 2022 
Appendix III: Schedule of Events: RSV Seasonal Surveillance 
 
Table 31: Schedule of Events: RSV Seasonal Surveillance 
 
Pre-RSV 
seasona 
Weekly contact 
Post-RSV season 
Illness 
Visit 
Early 
DC 
Visit period 
Oct 1st to Oct 31st  
 
Date of seasonal 
enrollment pause 
to Mar 31st 
Apr 1st to Apr 30th 
Allowable through 
September 30th in the 
event of delays due 
to a Stay at Home 
Order 
 
 
Clinical assessment 
(focused PE)b 
 
 
 
X 
 
Interim history 
 
X 
 
X 
X 
LABORATORY EVALUATIONS 
Blood for: 
immunologic assays 
5 mL 
 
5 mL 
 
5 mL 
Nasal wash  and/or 
nasosorption SAM 
strip for 
immunologic assaya 
X 
 
X 
 
 
Nasal wash or swab 
for 
viral detection & 
quantificationb 
 
 
 
X 
 
Request adventitious 
agent assay  
 
 
 
X 
 
TOTAL BLOOD 
VOLUME 
5 mL 
-- 
5 mL 
-- 
5 mL 
 
a For the 13 subjects enrolled prior to March 2020  
bIf a Stay at Home Order is initiated, the parent may be asked to obtain the nasal swab, and the clinical assessment 
may be completed via a remote or in-person research visit. 
 
 

 
90 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
Appendix IV: Definitions of Solicited Adverse Events 
 
Table 32: Definitions of Solicited Adverse Events 
Event 
Defined 
Fever 
Temporal or rectal temperatures ≥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. 
Pharyngitis1 
Pharyngeal erythema accompanied by exudate or pharyngeal erythema with 
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 (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 assessments 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 8.1.1. 

 
91 
RSV 6120/∆NS1 & RSV 6120/F1/G2/∆NS1 
           
Version 5.0 
Principal Investigator: Ruth A. Karron, MD 
        
June 5, 2023 
 
 
Figure 6.6-1: RSV Seasonality in Baltimore