A Phase 2A Partially-Blind Placebo Controlled Trial to Evaluate the Safety and Immunogenicity of Live Attenuated, Intranasal B. Pertussis Vaccine (BPZE1) in Healthy Adults
Phase: PHASE2
Conditions: Pertussis, Pertussis Immunisation
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

A Phase 2A Partially-blind Placebo Controlled Trial to Evaluate the 
Safety and Immunogenicity of Live Attenuated, Intranasal  
B. pertussis vaccine (BPZE1) in Healthy Adults  
DMID Protocol Number: 17-0010 
DMID Funding Mechanism: Vaccine and Treatment Evaluation Units 
Pharmaceutical Support Provided by: ILiAD Biotechnologies 
Other Identifying Numbers: Clinical Trials #_____________ 
IND Sponsor: NIAID/DMID 
Principal Investigator: C. Buddy Creech, MD, MPH 
DMID Scientific Lead: Kristina T. Lu, PhD 
DMID Medical Officer: Francisco Leyva, MD, PhD, ScM 
DMID Clinical Project Manager: Tena Knudsen, RN 
DMID Medical Monitor: Mo Elsafy, MD, MSc 
Draft or Version Number: 5.0 
25 June 2019 
 

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STATEMENT OF COMPLIANCE 
 
The study trial will be carried out in accordance with Good Clinical Practice (GCP) and as 
required by the following: 
1. United States Code of Federal Regulations (CFR) 45 CFR Part 46: Protection of Human 
Subjects 
2. Food and Drug Administration (FDA) Regulations, as applicable: 21 CFR Part 50 
(Protection of Human Subjects), 21 CFR Part 54 (Financial Disclosure by Clinical 
Investigators), 21 CFR Part 56 (Institutional Review Boards), 21 CFR Part 11, and 21 
CFR Part 312 (Investigational New Drug Application), 21 CFR 812 (Investigational 
Device Exemptions) 
3. International Council for Harmonisation: Good Clinical Practice (ICH E6); 62 Federal 
Register 25691 (1997); and future revisions 
4. Belmont Report: Ethical Principles and Guidelines for the Protection of Human Subjects 
of Research, Report of the National Commission for the Protection of Human Subjects of 
Biomedical and Behavioral Research 
5. National Institutes of Health (NIH) Office of Extramural Research, Research Involving 
Human Subjects, as applicable 
6. National Institute of Allergy and Infectious Diseases (NIAID) Clinical Terms of Award, 
as applicable 
7. Applicable Federal, State, and Local Regulations and Guidance 
 

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SIGNATURE PAGE 
The signature below provides the necessary assurance that this trial will be conducted according 
to all stipulations of the protocol, including all statements regarding confidentiality, and 
according to local legal and regulatory requirements and applicable US federal regulations and 
ICH E6 Good Clinical Practice (GCP) guidelines. 
 
I agree to conduct the study in compliance with GCP and applicable regulatory requirements. 
 
I agree to conduct the study in accordance with the current protocol and will not make changes to 
the protocol without obtaining the sponsor’s approval and IRB/IEC approval, except when 
necessary to protect the safety, rights, or welfare of subjects. 
 
 
Site Investigator: 
 
Signed: 
 
Date: 
 
 
C. Buddy Creech, MD, MPH 
Associate Professor of Pediatrics 
Division of Pediatric Infectious Diseases 
Director, Vanderbilt Vaccine Research Program 
Vanderbilt University School of Medicine 
 
 
 

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TABLE OF CONTENTS 
Statement of Compliance .................................................................................................................2 
Signature Page .................................................................................................................................3 
Table of Contents .............................................................................................................................4 
List of Tables ...................................................................................................................................8 
List of Figures ..................................................................................................................................9 
List of Abbreviations .....................................................................................................................10 
Protocol Summary .........................................................................................................................12 
1 
Key Roles ..............................................................................................................................15 
2 
Background Information and Scientific Rationale ...............................................................18 
2.1 
Background Information ...........................................................................................18 
2.1.1 Phase 1a Clinical Study Results ....................................................................21 
2.1.2 Phase 1b Clinical Study Results ...................................................................21 
2.2 
Rationale ...................................................................................................................23 
2.3 
Potential Risks and Benefits .....................................................................................23 
2.3.1 Potential Risks ..............................................................................................23 
2.3.1.1 Risks of Study Participation..............................................................23 
2.3.1.2 Risk to Study Subjects ......................................................................23 
2.3.1.3 Risks to the Environment or Potential for Interaction with wild-type 
B. pertussis strains ............................................................................25 
2.3.2 Known Potential Benefits .............................................................................26 
3 
Objectives and Outcome Measures .......................................................................................27 
3.1 
Study Objectives .......................................................................................................27 
3.2 
Study Outcome Measures .........................................................................................27 
3.2.1 Primary Outcome Measures ..........................................................................27 
3.2.2 Secondary Outcome Measures ......................................................................27 
4 
Study Design .........................................................................................................................29 
5 
Study Enrollment and Withdrawal ........................................................................................30 
5.1 
Subject Inclusion Criteria .........................................................................................30 
5.2 
Subject Exclusion Criteria ........................................................................................32 
5.3 
Treatment Assignment Procedures ...........................................................................33 
5.3.1 Randomization Procedures ...........................................................................33 
5.3.2 Masking Procedures ......................................................................................34 
5.3.3 Reasons for Withdrawal ................................................................................34 
5.3.4 Handling of Withdrawals ..............................................................................35 
5.3.5 Subject Replacement .....................................................................................36 
5.3.6 Termination of Study ....................................................................................36 

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Study Intervention/Investigational Product ..........................................................................37 
6.1 
Study Product Description ........................................................................................37 
6.1.1 Acquisition ....................................................................................................38 
6.1.1.1 BPZE1 vaccine and Placebo .............................................................38 
6.1.1.2 Additional Components ....................................................................38 
6.1.2 Formulation, Packaging, and Labeling .........................................................39 
6.1.3 Product Storage and Stability........................................................................40 
6.2 
Dosage, Preparation and Administration of Study Intervention/Investigational 
Product ......................................................................................................................40 
6.3 
Modification of Study Intervention/Investigational Product for a Participant .........41 
6.4 
Accountability Procedures for the Study Intervention/Investigational Product(s) ...41 
6.5 
Assessment of Subject Compliance with Investigational Product ............................42 
6.6 
Concomitant Medications/Treatments ......................................................................42 
7 
Study Schedule......................................................................................................................43 
7.1 
Screening Visit (Visit 00, Screening, Clinic Visit [Window: Day -30 to Day -1]) ..43 
7.2 
Enrollment/Vaccination (Visit 1, Clinic Visit [Day 1]) ............................................44 
7.3 
Follow-up Visits........................................................................................................45 
7.3.1 Visit 2, Day 8, Clinic Visit (Window: Day 8 ± 2 days) ................................45 
7.3.2 Visit 3, Day 15, Clinic Visit (Window: Day 15 ± 3 days) ............................46 
7.3.3 Visit 4, Day 29, Clinic Visit (Window: Day 29 ± 3 days) ............................46 
7.3.4 Visit 4a (If needed, based on B. pertussis positive culture on Day 29), 
Day 46, Clinic Visit (Day 46 ± 3 days) ........................................................47 
7.3.5 Visit 4b (if needed, based on positive B. pertussis culture from Day 46), 
Day 50, Phone Call (Day 50 ± 3 days) .........................................................47 
7.4 
Final Study Visit, Visit 5, Day 181, Clinic Visit (Day 181 ± 21 days) ....................47 
7.5 
Early Termination Visit ............................................................................................48 
7.6 
Unscheduled Visit .....................................................................................................49 
8 
Study Procedures/Evaluations ..............................................................................................50 
8.1 
Clinical Evaluations ..................................................................................................50 
8.2 
Laboratory Evaluations .............................................................................................51 
8.2.1 Clinical Laboratory Evaluations ...................................................................51 
8.2.2 Special Assays or Procedures .......................................................................52 
8.2.3 Specimen Preparation, Handling, and Shipping ...........................................52 
8.2.3.1 Instructions for Specimen Preparation, Handling, and Storage ........52 
8.2.3.2 Specimen Shipment ..........................................................................52 
9 
Assessment of Safety ............................................................................................................53 
9.1 
Specification of Safety Parameters ...........................................................................53 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters .................................................................................................................53 

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9.2.1 Adverse Events .............................................................................................53 
9.2.2 Reactogenicity ...............................................................................................55 
9.2.3 Serious Adverse Events ................................................................................56 
9.2.4 New Onset Chronic Medical Conditions ......................................................58 
9.2.5 Adverse Events of Special Interest ...............................................................58 
9.2.6 Procedures to be Followed in the Event of Abnormal Laboratory Test 
Values or Abnormal Clinical Findings .........................................................58 
9.3 
Reporting Procedures ................................................................................................58 
9.3.1 Serious Adverse Events ................................................................................58 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored 
IND ...............................................................................................................59 
9.3.3 Reporting of Pregnancy ................................................................................60 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events .........................60 
9.5 
Halting Rules ............................................................................................................61 
9.6 
Safety Oversight (ISM plus SMC) ............................................................................62 
9.6.1 Independent Safety Monitor (ISM) ...............................................................62 
9.6.2 Safety Monitoring Committee (SMC) ..........................................................62 
10 
Clinical Monitoring ...............................................................................................................64 
10.1 
Site Monitoring Plan .................................................................................................64 
11 
Statistical Considerations ......................................................................................................65 
11.1 
Study Hypotheses......................................................................................................65 
11.2 
Sample Size Considerations ......................................................................................65 
11.3 
Planned Interim Analyses .........................................................................................67 
11.3.1 Interim Safety Review ..................................................................................67 
11.3.2 Interim Immunogenicity Review ..................................................................67 
11.4 
Final Analysis Plan ...................................................................................................68 
11.4.1 Analysis Populations .....................................................................................69 
11.4.1.1 Safety Population ..............................................................................69 
11.4.1.2 Immunogenicity Population ..............................................................69 
11.4.1.3 Per Protocol Population ....................................................................69 
11.4.2 Safety Data ....................................................................................................70 
11.4.3 Immunogenicity Data ....................................................................................70 
11.4.4 Missing Values and Outliers .........................................................................71 
12 
Source Documents and Access to Source Data/Documents .................................................72 
13 
Quality Control and Quality Assurance ................................................................................73 
14 
Ethics/Protection of Human Subjects ...................................................................................74 
14.1 
Ethical Standard ........................................................................................................74 
14.2 
Institutional Review Board .......................................................................................74 
14.3 
Informed Consent Process ........................................................................................75 

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14.4 
Exclusion of Women, Minorities, and Children (Special Populations) ....................76 
14.5 
Subject Confidentiality .............................................................................................77 
14.6 
Study Discontinuation ...............................................................................................78 
14.7 
Costs, Subject Compensation, and Research Related Injuries ..................................78 
14.8 
Future Use of Stored Specimens ...............................................................................79 
15 
Data Handling and Record Keeping .....................................................................................80 
15.1 
Data Management Responsibilities...........................................................................80 
15.2 
Data Capture Methods ..............................................................................................80 
15.3 
Types of Data ............................................................................................................81 
15.4 
Timing/Reports .........................................................................................................81 
15.5 
Study Records Retention ...........................................................................................82 
15.6 
Protocol Deviations ...................................................................................................82 
16 
Publication Policy .................................................................................................................83 
17 
Literature References ............................................................................................................84 
Appendix A: Schedule of Events ...................................................................................................85 
Appendix B: Clinical Safety Laboratory Adverse Event Grading .................................................88 
 
 
 
 

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LIST OF TABLES 
Table 1: Treatment Groups and Intranasal Vaccine to be Administered ...................................... 13 
Table 2: Treatment Groups and Intranasal Vaccine to be Administered ...................................... 29 
Table 3:  Probability (%) of Observing at Least One Adverse Event ........................................... 66 
Table 4: Minimum Detectable Difference in Proportion of Subjects Attaining Seroconversion 
comparing Placebo versus the BPZE1 (107 or 109 CFU) ............................................................ 66 
 
 

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LIST OF FIGURES 
Figure 1: Schematic of Study Design ........................................................................................... 14 
  
 

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LIST OF ABBREVIATIONS 
AE 
Adverse Event/Adverse Experience 
AEFI 
Adverse Event Following Immunization 
AESI 
Adverse Event of Special Interest 
ALT 
Alanine aminotransferase 
aPV 
Acellular Pertussis Vaccine 
B. pertussis 
Bordetella pertussis 
CFR 
Code of Federal Regulations 
CI 
Confidence Interval 
CRF 
Case Report Form 
DCC 
Data Coordinating Center 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases  
DNT 
Dermonecrotic Toxin 
DSMB 
Data and Safety Monitoring Board 
eCRF 
Electronic Case Report Form 
FDA 
Food and Drug Administration 
FHA 
Filamentous Hemagglutinin 
FIM 
Fimbriae 
FWA 
Federalwide Assurance 
GCP 
Good Clinical Practice 
GMT 
Geometric Mean Titer 
GMFR 
Geometric Mean Fold Rise 
GMTR 
Geometric Mean Titer Ratio 
ICH 
International Council for Harmonization  
IEC 
Independent or Institutional Ethics Committee 
IND 
Investigational New Drug Application 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
MedDRA  
Medical Dictionary for Regulatory Activities 
MOP 
Manual of Procedures 
N 
Number (typically refers to subjects) 
NOCMC 
New Onset Chronic Medical Condition 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
OHRP 
Office for Human Research Protections 
PI 
Principal Investigator 

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PRN 
Pertactin 
PT 
Pertussis Toxin 
QA 
Quality Assurance 
QC 
Quality Control 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SMC 
Safety Monitoring Committee 
SOP 
Standard Operating Procedure 
TCT 
Tracheal Cytotoxin 
US 
United States 
SWFI 
Sterile Water for Injection 
wPV 
Whole cell Pertussis Vaccine 
 

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PROTOCOL SUMMARY 
Title: 
A Phase 2A Partially-blind Placebo Controlled Trial to Evaluate 
the Safety and Immunogenicity of Live Attenuated, Intranasal B. 
pertussis vaccine (BPZE1) in Healthy Adults  
Phase: 
2a 
Population: 
Approximately fifty healthy males and non-pregnant females, 
18-49 years of age, in the US.  
Number of Sites: 
One Vaccine and Treatment Evaluation Unit (VTEU) site, 
Vanderbilt University Medical Center 
Study Duration: 
Approximately 12 months 
Subject Participation 
Duration: 
Approximately 6 months 
Description of Agent or 
Intervention: 
Lyophilized, live-attenuated Bordetella pertussis vaccine 
(BPZE1) reconstituted with water for injection (SWFI) and 
administered as a single intranasal dose of either 107 colony 
forming units (CFU) or 109 CFU 
Placebo: The same constituents, in the same quantities as 
BPZE1, absent attenuated Bordetella pertussis, reconstituted 
with water for injection (SWFI) 
Objectives: 
 
Primary: 
1. To assess the safety and tolerability of a single intranasal 
dose of either 107 or 109 colony forming units (CFU) of 
lyophilized BPZE1 vaccine.  
Secondary: 
1. To assess the humoral immunogenicity of lyophilized 
BPZE1 vaccine at Day 15, Day 29 and Day 181 
following receipt of one intranasal dose of 107 or 109 
CFU of BPZE1. 

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2. To assess mucosal immunogenicity of lyophilized 
BPZE1 vaccine at Day 29 and Day 181 following receipt 
of one intranasal dose of 107 or 109 CFU of BPZE1. 
3. To evaluate nasal clearance of BPZE1 by culture at Day 
29 (and if still positive, at Day 46) following receipt of 
one intranasal dose of lyophilized BPZE1 vaccine of 107 
or 109 CFU of BPZE1. 
Description of Study Design: 
 
This is a phase 2a, single center, randomized, partially blind, 
placebo controlled, clinical trial evaluating a single intranasal 
dose of either 107 CFU or 109 CFU of BPZE1 in healthy adults 
(18-49 years of age, inclusive). The study will evaluate a 
lyophilized formulation of the product, defining the optimal dose 
for subsequent clinical trials. Since it is anticipated that the 
VaxINator device will be the primary mechanism of delivery in 
subsequent clinical trials, the majority of volunteers will receive 
vaccine or placebo via VaxINator device. Five individuals will 
receive 109 CFU of BPZE1 by needleless tuberculin syringe in 
order to compare immunogenicity to previous Phase 1 clinical 
trials of BPZE1.  
Fifty healthy adults, 18-49 years of age will be randomized to 
one of four treatment groups in a 3:3:3:1 ratio, as described in 
the table below. 
 
Table 1: Treatment Groups and Intranasal Vaccine to be 
Administered 
Group 
N 
Intranasal 
Vaccination 
Device Used for 
Administration 
1 
15 
BPZE1 
107 CFU 
VaxINator 
2 
15 
BPZE1 
109 CFU 
VaxINator 
3 
15 
Placebo 
VaxINator 
4 
5 
BPZE1 
109 CFU 
Needleless Tuberculin Syringe 
Estimated Time to Complete 
Enrollment: 
12 months 

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Figure 1: Schematic of Study Design 
 
 
 
 
 
 
 
 

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1 
KEY ROLES 
For questions regarding this protocol, contact Tena Knudsen, BSN (NIAID/DMID) at 
knudsent@niaid.nih.gov. 
Individuals: 
Site Principal 
Investigator: 
 
                    
 
 
DMID Clinical Project 
Manager: 
 
 
C. Buddy Creech, MD, MPH 
Associate Professor of Pediatrics, Division of Pediatric 
Infectious Diseases 
Director, Vanderbilt Vaccine Research Program 
CCC-5319 Medical Center North 
1161 21st Avenue South 
Nashville, Tennessee 37232 
Phone: 615/343-0332 
buddy.creech@vanderbilt.edu 
 
Tena Knudsen, BSN 
Respiratory Diseases Branch 
DMID, NIAID, NIH 
5601 Fishers Lane, Room 8E36, MSC 9825 
Rockville, MD 20852-9825 
Phone: 240/627-3290 
Email: knudsent@niaid.nih.gov  
 
DMID Scientific Lead: 
Kristina T. Lu, PhD 
Respiratory Diseases Branch 
DMID, NIAID, NIH  
5601 Fishers Lane, Room 8E35, MSC 9825 
Rockville, MD 20852-9825 
Phone: 240/627-3307 
Email: lukr@mail.nih.gov  
 
DMID Medical Officer  
 
 
 
Francisco Leyva, MD, PhD, ScM 
Respiratory Diseases Branch 
DMID, NIAID, NIH  

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Clinical Materials 
Services 
 
  
 
 
 
 
 
  
 
 

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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
Bordetella pertussis (B. pertussis) is a gram-negative bacterium and a causative agent of 
pertussis colonizing infection, a precursor to the disease more commonly known as whooping 
cough. It is a microbe that is transmitted from person to person via airborne droplets from the 
cough or sneeze of an infected host. A recent publication modeling pertussis cases and deaths 
estimated there were 24.1 million pertussis cases and 160,700 deaths in children younger than 5 
years in 2014 worldwide.[1]Despite the dramatic decline in Whooping Cough cases and deaths 
in industrialized nations during the 20th Century due to public health vaccine initiatives, recent 
decades have witnessed a sharp increase in cases and it is generally agreed that there is a critical 
need for a new and more effective vaccine targeting B. pertussis.  
 
Two types of pertussis vaccines are available worldwide: whole cell vaccine (wPV) and acellular 
vaccine (aPV). Although aPV overcomes several potential adverse effects of wPV (e.g. fever and 
seizures) and is the only pertussis vaccine marketed in the U.S., the protective efficacy of aPV 
appears to be lower than wPV with induced immunity waning in about 4-12 years. Additionally, 
the high cost of aPV limits its use in developing countries.  
 
Acellular pertussis vaccines (aPV) have several deficiencies that have contributed to pertussis 
outbreaks across the US and around the world. Young infants are especially vulnerable, but are 
not fully protected until they complete a 3 dose aPV vaccination series, at about 7 months of age. 
The clinical and regulatory path to a vaccine to prevent pertussis in the most vulnerable infants, 
however, will be lengthy and a more immediate impact can be made with an adult and adolescent 
vaccine that prevents transmission of B. pertussis to vulnerable infants.  
 
In addition to rapidly waning immunity after aPV immunization, preclinical baboon studies 
suggest that aPV does not prevent nasopharyngeal colonization and, hence, does not prevent 
transmission from adults and adolescents to vulnerable infants.[2] Multiple published studies in 
various countries have demonstrated high rates of subclinical (asymptomatic) B. pertussis 
colonizing infections. In a study that took nasal swabs from 629 asymptomatic school children in 
a Chinese province with 99% aP vaccination coverage, 4.8% were confirmed to be PCR positive 
for B. pertussis.[3] Based on data on controls in the adult and adolescent aP clinical trials by the 
APERT Study Group, the incidents of asymptomatic pertussis in the U.S. was predicted to be 5 - 
10 million cases per year in a year when the total number of diagnosed cases was only 7,000.[4] 
Surveillance studies using seroprevalence of pertussis to identify recently infected asymptomatic 

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individuals demonstrate high rates of circulating B. pertussis, including 9.3% in the 
Netherlands[5] and 14.1% in Italy[6]. Mathematical models predict that asymptomatic B. 
pertussis colonization is the most probable cause of the surge in pertussis rates.[7] Thus, a need 
exists for an adult and adolescent vaccine that induces potent and durable systemic immunity, 
but also mucosal immunity to prevent nasopharyngeal colonization and transmission to infants.  
 
Since the best-known method of inducing both potent systemic and mucosal immunity is by 
natural B. pertussis infection, BPZE1 was constructed 
 
 
 
 
 
Over 20 peer-reviewed publications have demonstrated the 
preclinical safety and efficacy of the live-attenuated vaccine[8-11], and two Phase 1 clinical 
studies [12, 13] have demonstrated safety and immunogenicity at various doses in healthy adults. 
Despite the low dose and low volume used in the original Phase-1a (low dose) study, BPZE1 
demonstrated the ability to transiently colonize healthy adult volunteers and, upon colonization, 
induce B. pertussis targeted immune responses in all colonized individuals[12]. In the Phase 1b 
study, discussed in detail below, 80% of subjects were transiently colonized with BPZE1 and 
78% met criteria for serologic response, including all subjects who received high-dose 
investigational BPZE1 liquid vaccine.  
 
The investigational BPZE1
 
 
 
Preclinical studies, including a 1010 CFU dose (10x higher than the highest dose in this study) in 
baboons, showed no safety concerns.[14] In the non-human primate model, BPZE1 also 
demonstrated the ability to reduce the total burden of B. pertussis colonization by 99.9% 
compared to non-vaccinated controls and 99.8% relative to previous studies with aPV. 
 
This study allows the new lyophilized formulation of the investigational BPZE1 vaccine to be 
evaluated in US adults and provides an opportunity to characterize the immunological response 
of two different dosages delivered by nasal immunization. The main advantage of the lyophilized 
BPZE1 formulation over the liquid formulation is long-term stability and storage at refrigeration 
temperature instead of storage at very low freezing temperature. For the Phase 1 clinical studies, 
the liquid BPZE1 formulation required freezing at -70° C to maintain long-term stability. Since 
the lyophilized product is reconstituted before administration, both formulations are delivered as 

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liquids and the lyophilized product simply provides improved storage temperature and time. 
 
Thus, the primary purpose of this Phase 2a study is to evaluate the safety 
and immunogenicity of the lyophilized formulation. 
 
The working cell bank (WBC) for both the liquid (Phase 1b) and lyophilized formulations were 
grown in THIJS medium.[15] Thus, the BPZE1 B. pertussis strain is the same active ingredient 
for both formulations, and the same WCB is used to inoculate the first series of shake-flask pre-
cultures.  
 
The lyophilized formulation consists of BPZE1 bacteria suspended in a lyophilization buffer 
(Section 6). 
 
 
 
 
 
 
 
 
 
.  
 
 
Two Phase 1 clinical studies have been performed at the Karolinska University Hospital 
(Stockholm, Sweden). Both studies were single center, dose-escalating, placebo-controlled 
studies of liquid BPZE1, given as a single intranasal dose to healthy adult volunteers. A Phase 1a 
clinical study[12], NCT01188512, has been completed using a BPZE1 low dose liquid 
suspension (i.e. 103, 105 and 107 CFU) formulation in Sweden, which had no vaccine related 
serious adverse events but only induced 42% of the healthy adult male subjects in the 107 CFU 
group to transiently colonize with BPZE1. To improve the percentage of subjects transiently 
colonizing with BPZE1 after vaccination, a Phase 1b study [13], NCT02453048, was performed 
in Sweden to evaluate a higher dose liquid suspension (i.e., 107, 108 and 109 CFU) in a greater 
volume (i.e., 400 μL per nostril). The Phase 1b study included both adult males and females and 
excluded subjects with high baseline anti-pertactin antibody titers from randomized study 
groups. The randomized portion of both Phase 1 studies included 48 volunteers, 12 in each dose 
group and in one placebo group. The 12 subjects in the placebo group were evenly distributed 
over the whole study period in both studies to enable evaluation of apparent differences in the 
frequency of adverse events or immunogenicity over time. 

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2.1.1 
Phase 1a Clinical Study Results 
The investigational BPZE1 liquid vaccine was safe in the 103, 105 and 107 CFU doses used for 
nasal administration and BPZE1 colonization was dose-dependent, with 42% of the subjects in 
the 107 CFU group having positive BPZE1 colonization. 
 
There were no vaccine related serious adverse events (SAE). There were no detectable 
differences in adverse events (AE) between placebo and the different dosage groups. The most 
frequent AEs were rhinorrhea (n=43 episodes in 26 subjects), sneezing (n=41 episodes in 19 
subjects), nasal congestion (n=38 episodes in 29 subjects), headache (n=34 episodes in 21 
subjects), and tiredness (n=32 episodes in 23 subjects). Many of the volunteers had 
uncomplicated upper respiratory tract infections (i.e., common cold) during the follow up period, 
which may explain at least a portion of their symptoms. The efficacy results, included:  
• Colonization of mucosal surface: The vaccine strain BPZE1 was isolated in a total of 
seven subjects, one in each of the 103 and 105 dose group and five subjects in the 107 dose 
group. The colonization was first detected on day 4-14 and lasted to day 11-28 after 
vaccination 
• Immunogenicity: Anti-pertussis IgG appeared only in the seven subjects who were 
colonized at the nasopharyngeal mucosa. The antibodies appeared after day 14 and 
remained at high levels after the 5-6 months follow up.  
 
In summary, this Phase 1a study showed that BPZE1 used for nasal administration in humans 
can colonize the nasopharyngeal mucosa was both immunogenic and safe, without eliciting any 
symptoms of either mild or severe whooping cough. The results of this study suggested the 
possibility that a higher degree of colonization and immune response can be achieved by using a 
higher vaccine dose, higher vaccine volume, an improved method for application, or by 
excluding subjects with high baseline anti-pertactin antibody titers.  
2.1.2 
Phase 1b Clinical Study Results 
The BPZE1 investigational liquid vaccine was safe in the higher doses and volumes used for 
nasal administration. Proportion of subjects having transiently BPZE1 colonized nasopharyngeal 
mucosa was not dose-related at 107 CFU or higher, but instead all dosage groups achieving 75% 
colonization rate or greater, and an overall combined colonization rate was observed for subjects 
from all randomized groups of over 80%. A dose response trend was observed, however, in the 
rate of colonization and clearance, with faster colonization rate and faster clearance rate being 
correlated with higher dosage. The serum antibody responses were strongly correlated to positive 
colonization, but there was no dose response. A small non-randomized group of subjects with 

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pre-existing high antibody titers for pertactin had a 33% rate of BPZE1 colonization after 
immunization with the 109 CFU dose. 
 
There were no SAEs and any differences in AEs within randomized study groups, between 
placebo and the different dosages, were not detectable. The most frequent AEs were sneezing 
(n=55 episodes in 24 subjects), rhinorrhea (n=35 episodes in 22 subjects), nasal congestion 
(n=31 episodes in 19 subjects), cough (n=21 episodes in 16 subjects), and fatigue (n=21 episodes 
in 16 subjects). Efficacy results, included: 
• Colonization of mucosal surface: The vaccine strain BPZE1 was isolated in a total of 29 
randomized subjects, 10 in the 107 CFU dose group, 9 in the 108 CFU dose group, and 10 
subjects in the 109 CFU dose group. Colonization was first detected on day 4-14 and 
lasted through day 11-28 after vaccination. 
• Immunogenicity: Serologic responses (IgG and IgA) were measured to PT, PRN, FIM, 
and FHA and reported individually. In addition, responses were grouped by either:  
o ≥2-fold rise in PT-specific IgG or IgA that was at least 4x greater than the 
minimum limit of detection (MLD) of the assay, or 
o ≥2-fold rise in at least 2 antibodies targeting PRN, FIM, or FHA (IgG or IgA) that 
were at least 4x greater than the MLD 
Greater than 92% of subjects in the 107 CFU dose group and 100% of subjects in the 109 
CFU dose group had a two-fold rise in at least one antibody titer (either PT, PRN, FIM, 
or FHA, IgG or IgA) that was at least 4x greater than the MLD (i.e., seropositive). All 
subjects receiving the 109 CFU BPZE1 dose were seropositive for at least 2 or more 
antibodies (IgG or IgA) or PT (IgG or IgA). No placebo subjects had 2 seropositive 
antibodies (IgG or IgA) or seropositive for PT antibodies (IgG or IgA). In the vast 
majority of subjects, an increase in serum antibody titers (IgG/IgA) occurred at day 14 or 
21, peaked at day 28, and remained at high levels at the 6 months follow up visit 
(sometimes rising further at 6-months). 
• Despite 33% BPZE1 colonization rate in the non-randomized pre-existing high pertactin 
group, 66% of the subjects experienced a seropositive response to ≥ 2 antibodies (IgG or 
IgA) or PT (IgG or IgA); in other words, multiple subjects generated an immunological 
response despite unrecognized BPZE1 colonization. 
 
In summary, the investigational BPZE1 liquid vaccine was safe in the doses used for nasal 
administration. Colonization occurred in > 80% of the subjects across the three dosage groups 
and BPZE1 groups, especially the 109 CFU group, had a high proportion of subjects that reached 
the pre-specified threshold of seropositivity. Additionally, subjects with high baseline antibodies 
were capable of further seropositive antibody boosting, but at a lower rate and lower percent 
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2.2 
Rationale 
The availability of a cost-effective pertussis vaccine that provides improved efficacy, durable 
protection, and the potential to stop transmission would represent a breakthrough in the 
prevention of pertussis infections and may lead to a reduction in the population reservoir of B. 
pertussis in undiagnosed individuals. The ability to prevent colonization and transmission could 
even facilitate global eradication of B. pertussis disease. 
 
Intranasal administration provides an opportunity to generate a locally effective mucosal 
antibody response at the site of exposure. Preclinical data, as described above, supports the 
ongoing development of BPZE1 (either 107 CFU or 109 CFU), with the transition to a lyophilized 
formulation being the goal of this study. The hypothesis for this trial is that lyophilized BPZE1, 
administered intranasally, will be sufficiently safe and immunogenic. Given the safety profile of 
liquid BPZE1 in previous clinical trial recipients, we hypothesize that lyophilized BPZE1 will 
share similar safety characteristics. Thus, the goals of this clinical trial are to assess the safety, 
reactogenicity and immunogenicity of investigational, lyophilized BPZE1 vaccine administered 
intranasally by VaxINator device at a dose of either 107 CFU or 109 CFU to healthy adult 
volunteers. To aid in the transition from earlier Phase 1 studies, we will enroll a small number of 
subjects who will receive lyophilized BPZE1 by needleless tuberculin syringe, as this was the 
method of administration used previously.  
2.3 
Potential Risks and Benefits 
2.3.1 
Potential Risks 
Potential risks include risks of study participation (in general), specific risks to study subjects, 
and theoretic risks to the environment through the introduction of an attenuated B. pertussis 
strain into human hosts. 
2.3.1.1 
Risks of Study Participation 
The risks of study participation include exposure to the study product, maintenance of 
confidentiality, and side effects of phlebotomy. All risks will be minimized to every extent 
possible.  
2.3.1.2 
Risk to Study Subjects 
B. pertussis colonization is strictly limited to respiratory epithelium without dissemination of the 
bacteria outside the respiratory tract, which also excludes systemic bacteremia of the BPZE1 
strain.  

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B. pertussis is spread mainly by aerosol formed by coughing of infected persons. The coughing 
is induced by the tracheal cytotoxin (TCT), which is more than 95% reduced in BPZE1. The 
BPZE1 strain is not expected to induce coughing, therefore transmission is highly unlikely. 
Bordetella species have fastidious growth requirements and have limited survival time outside 
the human body.  
 
B. pertussis has not been shown to be allergenic in any preclinical or clinical studies to date, nor 
have any of the excipients in the lyophilized formulation. BPZE1 has been shown to protect 
against airway inflammation induced by allergens or viral infections in a murine model. [7] 
BPZE1 has also been shown to protect against infection with wild type B. pertussis infection 3 
hours after immunization in a murine model.[8] However, there remains a theoretical risk of 
allergic reaction, as is present with any vaccine product. 
 
BPZE1 vaccine or a placebo will be administered nasally via the VaxINator atomization device 
attached to a syringe to healthy adult volunteers under strictly controlled conditions. The 
VaxINator atomizes the liquid vaccine as it exits the syringe. There are no additional risks to 
study subjects based on the use of the VaxINator device, which is described in more detail in 
Section 6; similarly, there are no independent risks associated with the use of the tuberculin 
syringe used to drop BPZE1 vaccine into the nose. 
 
To minimize the risk of transmission, the volunteers will stay at the study center for 2 hours after 
administration of the vaccine. In addition, volunteers with household or professional contact with 
infants below one year of age or household/professional contact with individuals with 
immunodeficiency will be excluded from participation in the study. The attenuated BPZE1 
bacteria colonize the upper respiratory tract similarly to wild-type B. pertussis. Colonization of 
live organism will be assessed from a nasopharyngeal swab performed 28 days after 
administration to ensure all subjects are cleared of colonization. If a subject is not clear at day 
29, a repeat nasal swab and culture will be performed at day 46. Chronic carriage of BPZE1 has 
not been reported (i.e., the vast majority of subjects have been clear at day 29 and no subject has 
had positive cultures at day 46) and is therefore not expected. Any subject who remains positive 
at day 46 will be provided a short course of azithromycin, which is clinically used to eradicate B. 
pertussis from the nasopharynx.  Azithromycin will be prescribed by a study physician and filled 
by the hospital pharmacy or the patient’s desired pharmacy at no cost to the subject. 
 
Furthermore, in animal studies, when female mice were vaccinated with BPZE1 shortly before 
mating, no negative effect on either the pregnancy or their offspring was observed. The offspring 
were protected against B. pertussis challenge, based on evaluation of the lungs at necropsy.[16]  
 

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2.3.1.3 
Risks to the Environment or Potential for Interaction with wild-type B. pertussis 
strains 
Detailed information about the 
 BPZE1 is available in the 
environment risk assessment located in the investigator’s brochure.  
 
To avoid accidental exposure to the environment and to study staff, actions should be taken to 
minimize generation of aerosols, since the bacteria are strictly respiratory tract organisms. Staff 
members should wear eye-protective glasses during the vaccination. Persons handling BPZE1 
should wear gloves and must practice standard hand hygiene before touching their skin or eyes.  
 
 
 
 
 
 
 
There is no known animal vector or reservoir for B. pertussis. BPZE1 bacteria are not invasive and 
have no selective advantage in the environment. The potential for exchange of genetic material is 
virtually inexistent, since B. pertussis does not harbor plasmids or conjugative transposons. In 
addition, B. pertussis Tohama I (background used for the BPZE1 strain) does not harbor intact 
prophage genomes and is therefore incapable of producing functional phage particles.  
 
 
 
 
 

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proliferation power of the BPZE1 cells in the trachea and lungs and no weight 
  
 
  
 
 
 
 
 
 
 
 
 
 
 
 
Based on these data, BPZE1 has been classified as a Biosafety level 1 organism by French 
authorities Republique Francaise Ministere De L'enseignement Superieur Et De La Recherche 
(French Ministry of Higher Education and Research), http://www.enseignementsup-
recherche.gouv.fr/. Germany, Belgium, and Sweden have accepted the French Authority’s BSL1 
rating for the purpose of manufacturing and clinical studies.  
 
No cross-contamination between the volunteers was observed in the previous phase 1 clinical 
trials of BPZE1, nor was any risk to the family members of study participants observed.  In 
summary, the preliminary risk assessment for this study suggests there is an extremely low risk 
for potential environmental impact associated with administering the BPZE1 to study subjects. 
2.3.2 
Known Potential Benefits 
The benefits of the study lie primarily in the opportunities to science and humanity. No direct 
personal benefit from participation in the study can be guaranteed, as the vaccine may or may not 
confer protection in humans.  

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3 
OBJECTIVES AND OUTCOME MEASURES 
3.1 
Study Objectives 
Primary: 
1. To assess the safety and tolerability of a single intranasal dose of either 107 or 109 colony 
forming units (CFU) of lyophilized BPZE1 vaccine.  
Secondary: 
1. To assess the humoral immunogenicity of lyophilized BPZE1 vaccine at Day 15, Day 29 
and Day 181 following receipt of one intranasal dose of 107 or 109 CFU of BPZE1. 
2. To assess mucosal immunogenicity of lyophilized BPZE1 vaccine at Day 29 and Day 
181 following receipt of one intranasal dose of 107 or 109 CFU of BPZE1. 
3. To evaluate nasal clearance of BPZE1 by culture at Day 29 (and if still positive, at Day 
46) following receipt of one intranasal dose of lyophilized BPZE1 vaccine of 107 or 109 
CFU of BPZE1. 
3.2 
Study Outcome Measures 
3.2.1 
Primary Outcome Measures 
1. Occurrence of solicited local and systemic reactogenicity from the time of the study 
vaccination through 14 days post-vaccination 
2. Occurrence of unsolicited non-serious adverse events from the time of the study 
vaccination through 28 days post-vaccination 
3. Occurrence of serious adverse events (SAEs) and new onset chronic medical conditions 
(NOCMCs) from the time of the study vaccination through approximately Day 181 post 
vaccination 
4. Occurrence of adverse events of special interest through approximately Day 29 post 
vaccination 
3.2.2 
Secondary Outcome Measures 
1. Humoral Immune Response 
o Geometric mean titers (GMTs) by serum IgG and IgA ELISA to PT, FHA, PRN, 
FIM 2/3 on Days 1, 15, 29, and 181  

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o Geometric mean fold rise (GMFR) by serum IgG and IgA ELISA to PT, FHA, 
PRN, FIM 2/3 on Days 15, 29 and 181 relative to baseline (Day 1) 
o The proportion of subjects that achieve seroconversion (defined as ≥2-fold rise 
from baseline) to individual pertussis antigens (PT, FHA, PRN, and FIM 2/3) by 
serum IgG or IgA ELISA at Days 15, 29, 181 or any time point relative to 
baseline (Day 1)  
o Proportion of subjects achieving seroconversion (serum IgG or IgA ELISA) to 
one or more pertussis antigens at Days 15, 29, 181 or any time point, relative to 
baseline (Day 1) 
o Proportion of subjects achieving seroconversion (serum IgG/IgA ELISA) to two 
or more pertussis antigens at Days 15, 29, 181 or any time point, relative to 
baseline (Day 1) 
2. Mucosal Immune Response 
o Geometric mean titer ratios (GMTRs) and ratio of antigen-specific IgA ELISA 
(PT-IgA, FHA-IgA, PRN-IgA, FIM-IgA) to total IgA by nasal aspirate at 
screening, and Days 29 and 181. 
o Fold rise of the ratio of antigen-specific IgA (PT-IgA, FHA-IgA, PRN-IgA, FIM-
IgA) to total IgA by nasal aspirate from the screening visit to Days 29 and 181 
o The proportion of subjects that achieve seroconversion (defined as ≥2-fold rise of 
the ratio from baseline) to individual pertussis antigens (PT, FHA, PRN, and FIM 
2/3) by nasal aspirate IgA ELISA at Days 29 and 181 or any time point relative to 
baseline (screening) 
o Proportion of subjects that achieve seroconversion (≥2-fold rise of the ratio from 
baseline) to at least 1 antigen (PT, FHA, PRN, or FIM 2/3) by nasal aspirate IgA 
ELISA from the screening visit to Days 29 and/or 181. 
3. Proportion of subjects with detectable B. pertussis from nasopharyngeal cultures at Day 
29.  
o For those who remain positive at Day 29, the subjects with detectable B. pertussis 
at Day 46 will also be evaluated and reported. Detection of B. pertussis in culture 
will be assumed to be due to study-product administration.  
 
 

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4 
STUDY DESIGN 
This is a phase 2a, single center, randomized, partially blind, placebo controlled, clinical trial 
evaluating a single intranasal dose of either 107 CFU or 109 CFU of BPZE1 in healthy adults 
(18-49 years of age, inclusive). The study will evaluate a lyophilized formulation of the product, 
with the goal of testing for the optimal dose for subsequent clinical trials. Since it is anticipated 
that the VaxINator device will be the primary mechanism of delivery in subsequent clinical 
trials, the majority of volunteers will receive vaccine or placebo via VaxINator device. Five 
individuals will receive 109 CFU of BPZE1 by needleless tuberculin syringe in order to compare 
immunogenicity to previous Phase 1 clinical trials of BPZE1.  
 
We anticipate that enrollment will require approximately 12 months, with each subject 
participating for 6 months. Fifty healthy adults, 18-49 years of age will be randomized to one of 
four treatment groups in a 3:3:3:1 ratio, as described in the table below. Safety will be assessed 
through an Independent Safety Monitor (ISM) and a Safety Monitoring Committee (SMC) 
described in Section 9.6. 
 
Table 2: Treatment Groups and Intranasal Vaccine to be Administered 
Group 
N 
Intranasal Vaccination 
Device Used for Administration 
1 
15 
BPZE1 107 CFU 
VaxINator 
2 
15 
BPZE1 109 CFU 
VaxINator 
3 
15 
Placebo 
VaxINator 
4 
5 
BPZE1 109 CFU 
Needleless Tuberculin Syringe 
 
 

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5 
STUDY ENROLLMENT AND WITHDRAWAL 
Approximately 50 healthy male and non-pregnant female subjects 18-49 years of age, inclusive, 
who meet all eligibility criteria will be enrolled in this study from one VTEU site. Enrollment 
will occur over an approximately 6-month period, and the target population will be the general 
Nashville community. Information regarding this study may be mailed or emailed to subjects 
who have previously participated in vaccine trials conducted at the VTEU site; other forms 
and/or mechanisms of recruitment may also be used. The local IRB will approve all materials 
prior to use.  
 
Subject eligibility criteria must be assessed by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site PI or sub-investigator.  
 
No exemptions are granted on Subject Inclusion or Exclusion Criteria in DMID-sponsored 
studies. Questions about eligibility should be directed toward the DMID Medical Officer. 
Subjects otherwise eligible with grade 1 abnormalities in hematology or chemistry values may be 
considered for enrollment if, in the opinion of the investigator (or clinician on the 1572), the 
abnormalities are not clinically significant and do not pose additional risk to the study or the 
volunteer. 
5.1 
Subject Inclusion Criteria 
Subjects eligible to participate in this study must meet all inclusion criteria: 
1. Provide written informed consent prior to initiation of any study procedures 
2. Able to understand and comply with planned study procedures and be available for all 
study visits 
3. Males or non-pregnant females, 18-49 years of age, inclusive 
4. In good health1 
 
1As determined by medical history and physical examination to evaluate acute or currently ongoing chronic medical 
diagnoses or conditions, defined as those that have been present for at least 90 days that would affect the 
assessment of the safety of subjects or the immunogenicity of study vaccinations. Chronic medical diagnoses or 
conditions should be stable for the last 60 days. This includes no change in chronic prescription medication, dose, 
or frequency because of deterioration of the chronic medical diagnosis or condition in the 60 days prior to 
enrollment. Any prescription change that is due to change of health care provider, insurance company, etc., or that 
is done for financial reasons, if it is in the same class of medication, will not be considered a deviation of this 
inclusion criterion. Any change in prescription medication due to improvement of a disease outcome, as determined 
by the site principal investigator or appropriate sub-investigator, will not be considered a deviation of this inclusion 
criterion. Subjects may be on chronic or as needed (prn) medications if, in the opinion of the site principal 
investigator or appropriate sub-investigator, they pose no additional risk to subject safety or assessment of 
reactogenicity and immunogenicity and do not indicate a worsening of medical diagnosis or condition. Similarly, 

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medication changes after enrollment and study vaccination are acceptable provided there was no deterioration in 
the subject’s chronic medical condition that necessitated a medication change, and there is no additional risk to the 
subject or interference with the evaluation of responses to study vaccination. Note: Topical and inhaled medications 
(with the exception of inhaled or nasal corticosteroids within 30 days prior to enrollment), herbals, vitamins, and 
supplements are permitted. 
5. Oral temperature is less than or equal to 100°F 
6. Pulse is 45 to 100 bpm, inclusive2 
 
2Pulse can be 45 to 50 bpm, inclusive, if no other symptoms are present. Otherwise, pulse should be 50-100 bpm. 
 
7. Systolic blood pressure is 85 to 150 mm Hg, inclusive 
8. Diastolic blood pressure is 55 to 95 mm Hg, inclusive 
9. White blood cell count is 3,900 cells/µL or greater3 
10. Hemoglobin is 13.0 g/dL or greater (men) or 11.8 g/dL or greater (women)3 
11. Platelet count is 135,000 cells/µL or greater3 
12. Alanine aminotransferase is less than 45 U/L (women) or 62 U/L (men)3 
13. Serum creatinine is less than or equal to 1.25 mg/dL (men) or 1.11 mg/dL (women)3 
 
3 Abnormalities in white blood count, hemoglobin, platelet count, alanine aminotransferase, and serum creatinine 
that are suspected to be due to laboratory anomalies may be repeated once to ensure accuracy; additionally, 
otherwise eligible subjects with grade 1 abnormalities in these values may be considered for enrollment if, in the 
opinion of the investigator (or clinician on the 1572), the abnormalities are not clinically significant and do not 
pose additional risk to the study or the volunteer.  
 
14. Negative serum HIV antibody assay 
15. Women of childbearing potential4 must use an acceptable contraception method5 from 
30 days before study vaccination until 60 days after vaccination. 
 
4Not sterilized via tubal ligation, bilateral oophorectomy, salpingectomy, hysterectomy, or successful Essure® 
placement (permanent, non-surgical, non-hormonal sterilization) with documented radiological confirmation test at 
least 90 days after the procedure, and still menstruating or <1 year has passed since the last menses if menopausal. 
 
5Includes, non-male sexual relationships, abstinence from sexual intercourse with a male partner, monogamous 
relationship with vasectomized partner who has been vasectomized for 180 days or more prior to the subject 
receiving study vaccination, barrier methods such as condoms or diaphragms/cervical caps with spermicide, 
effective intrauterine devices, NuvaRing®, and licensed hormonal methods such as implants, injectables or oral 
contraceptives (“the pill”). 
 
16. Women of childbearing potential must have a negative urine or serum pregnancy test 
within 24 hours prior to study vaccination. 

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5.2 
Subject Exclusion Criteria 
1. Have immunosuppression as a result of an underlying illness or treatment, or use of 
anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study 
vaccination 
2. Have known or suspected active chronic autoinflammatory condition 
3. Have known active neoplastic disease (excluding non-melanoma skin cancer) or a history 
of any hematologic malignancy 
4. Have a history of persistent asthma, major anatomic nasopharyngeal abnormality, or 
sinus polyp disease due to chronic sinusitis6 
6 If a patient has a history of nasopharyngeal surgery such as, but not limited to rhinoplasty, tonsillectomy 
or sinus surgery, adequate healing time per the judgment of the investigator must occur prior to 
enrollment. 
 
5. Have known hepatitis B or hepatitis C infection 
6. Have a history of alcohol or drug abuse within 5 years prior to study vaccination 
7. Currently untreated or clinically unstable (in the opinion of the investigator) 
schizophrenia, bipolar disease, or other psychiatric diagnosis that may interfere with 
subject compliance or safety evaluations 
8. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement 
for danger to self or others within 5 years prior to study vaccination 
9. Have received corticosteroids (including oral, parenteral, inhaled, nasal, or intra-articular) 
of any dose within 30 days prior to study vaccination 
10. Individual with PT serum IgG antibodies ≥ 20 IU/mL and/or PRN serum IgG antibodies 
≥ 125 IU/mL 
11. Unwilling to refrain from smoking tobacco for 28 days post vaccination 
12. Receipt of immunoglobulin or blood derived products within 90 days of enrollment 
13. Receipt of a vaccine against pertussis in the past 2 years 
14. Receipt of a live vaccine within 30 days of study vaccination or an inactivated vaccine 
within 14 days of study vaccination 
15. Planned vaccination with a licensed vaccine within 28 days of study vaccination 
16. History of severe allergic reaction (e.g., anaphylaxis) or Bell’s palsy, or Guillain-Barré 
syndrome, after a previous dose of any diphtheria toxoid-tetanus toxoid-, or pertussis-
containing vaccine, or encephalopathy within 7 days of administration of a previous 
pertussis containing vaccine. 
17. History of a progressive neurologic disorder 
18. In close contact7 with children less than 1 year of age or contact with persons with known 
immunocompromising conditions 
 

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7 Close contact includes sharing a household, serving as a healthcare worker, or working professionally in settings 
with repeated exposures. 
 
19. Receipt of B. pertussis-active antibiotics8 within 7 days prior to vaccination 
 
8 B. pertussis active antibiotics include macrolides, fluoroquinolones, trimethoprim-sulfamethoxazole, 
tetracyclines 
 
20. Known hypersensitivity to any component of the study vaccine. 
21. Hypersensitivity to azithromycin, which may be used in the event of ongoing BPZE1 
colonization 
22. Any condition that, in the opinion of the investigator, might interfere with objectives of 
the study or safety to the individual 
23. Acute illness, including temperature >100˚F within one week prior to vaccination9 
9  Enrollment may be postponed if acute illness occurs; subjects must remain within the screening window, 
however, and must be rescreened if >30 days elapses prior to enrollment. 
5.3 
Treatment Assignment Procedures 
5.3.1 
Randomization Procedures 
Once consented and upon entry of demographic data and confirmation of eligibility for this 
study, the subject will be enrolled. Approximately 50 subjects will be assigned randomly to 1 of 
4 treatment arms, in a 3:3:3:1 ratio. The first treatment arm (n=15) will be vaccinated with one 
dose of 107 CFU BPZE1 using the VaxINator device. The second treatment arm (n=15) will be 
vaccinated with one dose of 109 CFU BPZE1 using the VaxINator device. The third treatment 
arm (n=15) will be vaccinated with one dose of placebo via VaxINator device. The fourth 
treatment arm (n=5) will be vaccinated with one dose of 109 CFU BPZE1 using a needleless 
tuberculin syringe.  
 
Enrollment of subjects will be done online using the enrollment module of AdvantageEDCSM. 
The randomization code will be prepared by statisticians at the DCC and included in the 
enrollment module for this study. AdvantageEDCSM will assign each subject to a treatment arm 
after the demographic and eligibility data have been entered into the system. A designated 
individual at the participating VTEU site will be provided with a code list for emergency 
unblinding purposes, which will be kept in a secure location. 
 
Instructions for use of the enrollment module are included in the AdvantageEDCSM User’s 
Guide. Manual back-up procedures and instructions are provided for use in the event that the 

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participating VTEU site temporarily loses access to the Internet or the online enrollment system 
is unavailable. 
5.3.2 
Masking Procedures 
This is a partially-blind clinical study as the method of immunization (i.e., VaxINator vs. 
needleless syringe) will be known. After reconstitution and transfer to a 1-mL syringe, the 107 
CFU dose investigational vaccine and the placebo are transparent and indistinguishable while  
the 109 CFU dose can be discriminated from the placebo and the 107 CFU dose due to its opaque 
appearance. Therefore, syringes will be wrapped with a colored sleeve to avoid potential 
unblinding of study personnel or the subject. Investigators, the subject, and study personnel will 
remain blinded to allocation of BPZE1 or placebo. 
 
The randomization scheme will be generated by the DCC and provided to unblinded study 
personnel (i.e., research pharmacists performing study vaccination preparations and unblinded 
study vaccine administrators) at the participating VTEU site. 
 
The unblinded study vaccine administrator is a study personnel member credentialed to 
administer vaccines and may also participate in dose preparation, but will not be involved in 
study-related assessments or have subject contact for data collection following study vaccine 
administration. 
 
Laboratory personnel performing immunogenicity assays will be blinded to study assignment. 
Upon request by the participating VTEU site, and at the discretion of DMID, unmasking may 
occur after all data have been locked at Day 181, analyzed, interpreted, and the final CSR has 
been completed. 
 
The Safety Monitoring Committee (SMC) may receive data in aggregate and presented by 
treatment arm. The SMC may also be provided with expected and observed rates of the expected 
AEs in an unblinded fashion and may request the treatment arm be unblinded for an individual 
subject if required for safety assessment. The SMC will review grouped and unblinded data in 
the closed session only.  
5.3.3 
Reasons for Withdrawal  
Subjects may voluntarily withdraw their consent for study participation at any time and for any 
reason, without penalty or loss of benefits to which they are otherwise entitled.  
 

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An investigator may also withdraw a subject from receiving the study product for any reason. 
Follow-up safety evaluations will be conducted, if the subject agrees. If a subject withdraws or is 
withdrawn prior to completion of the study, the reason for this decision must be recorded in the 
case report forms (CRFs).  
 
The reasons, might include, but are not limited to the following: 
1. Subject no longer meets eligibility criteria 
2. Subject meets individual halting criteria  
3. Subject becomes noncompliant 
4. Medical disease or condition, or new clinical finding(s) for which continued 
participation, in the opinion of the investigator might compromise the safety of the 
subject, interfere with the subject's successful completion of this study, or interfere with 
the evaluation of responses 
5. Subject lost to follow-up 
6. Subject becomes pregnant, if applicable 
7. Determined by a physician’s discretion to require additional therapy not indicated in the 
protocol to ensure subject’s health and well-being (or treatment failure, if applicable) 
 
The investigator should be explicit regarding study follow-up (e.g. safety follow-up) that might 
be performed. If the subject consents, every attempt will be made to follow all AEs through 
resolution. The procedures that collect safety data for the purposes of research must be inclusive 
in the original informed consent or the investigator may seek subsequent informed consent using 
an IRB/IEC-approved consent form with the revised procedures. 
 
The investigator will inform the subject that already collected data will be retained and analyzed 
even if the subject withdraws from this study.  
5.3.4 
Handling of Withdrawals  
The primary reason for withdrawal from this study will be recorded on the appropriate data 
collection form. Subjects will be encouraged to complete the Early Termination Visit. The Early 
Termination Visit procedures are listed in Section 7.5. 
 
Although subjects are free to withdraw at any time or may be withdrawn by the site principal 
investigator or appropriate sub-investigator at any time, subjects will be encouraged to remain in 
this study for follow-up safety assessments (may be conducted by phone rather than in person) 
continuing through approximately 6 months after study vaccination. These subjects will also be 

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encouraged to provide a venous blood sample for serological assessment at approximately 28 
days after study vaccination. See the protocol-specific Manual of Procedures (MOP) for alternate 
follow-up requirements.  
 
Every attempt will be made to follow all adverse events, including systemic reactions, 
unsolicited non-serious adverse events, and serious adverse events ongoing at the time of early 
withdrawal through resolution. 
 
For subjects who fail to appear for a follow-up safety assessment, extensive effort (e.g., three 
documented contact attempts via phone calls, e-mails, etc., made on separate occasions and 
followed by a certified letter) will be made to locate them. Any questions to determine their 
health status will be made by phone or visit by the subject. These efforts will be documented in 
the subject’s study records. 
5.3.5 
Subject Replacement 
Subjects who withdraw, are withdrawn/terminated from this study, or are lost to follow-up after 
signing the informed consent form and randomization but before receipt of study vaccine may be 
replaced. Subjects who are vaccinated, but do not complete the Day 15 visit, will be replaced. 
Subjects who are vaccinated and subsequently, after the Day 15 visit, withdraw, are withdrawn 
or terminated from this study, or are lost to follow-up will not be replaced. Subjects who were 
previously screened and found to be ineligible due to initial serum concentrations of PRN IgG ≥ 
20 IU/ml in protocol version 3.0 may be rescreened per the revised exclusion criterion of 
protocol version 4.0.   
5.3.6 
Termination of Study  
Although the sponsor has every intention of completing this study, it reserves the right to 
terminate the study at any time for clinical or administrative reasons. Reasons for termination 
include, but are not limited to, study closure due to SMC recommendation and at the discretion 
of DMID. 
If the study is prematurely terminated by the sponsor, any regulatory authority, or the 
investigator for any reason, the investigator will promptly inform the study subjects and assure 
appropriate therapy or follow-up for the subjects, as necessary. The investigator will provide a 
detailed written explanation of the termination to the IRB/IEC. 
 
 

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Placebo 
The placebo consists of the same constituents in the same quantities as the BPZE1 
investigational vaccines, absent the attenuated B. pertussis cells. 
VaxINator atomization device 
BPZE1 investigational vaccine or placebo will be administered nasally via the VaxINator 
atomization device attached to a 1 mL syringe. The VaxINator is manufactured by Teleflex 
Medical. Teleflex has a Master File (MF #16712) for the VaxINator. The VaxINator is a conical 
shape plastic component that Luer-locks onto a standard syringe. The VaxINator atomizes the 
liquid vaccine as it exits the syringe. The conical shape forms a plug in the nostril and high 
applied-pressure atomizes the liquid into a fine mist. 
6.1.1 
Acquisition 
6.1.1.1 
BPZE1 vaccine and Placebo 
Individual boxes of packaged and labeled investigational vaccine/placebo contain investigational 
vaccine/placebo. The boxes shall be shipped from 
 
 for distribution to the clinical site. All boxes of investigational 
vaccine/placebo will be shipped at below -20 C with temperature logging devices and should be 
stored at -10˚C or below after arrival to the site. 
6.1.1.2 
Additional Components 
VaxINator kits are supplied in individual sterile pouches containing a VaxINator atomization 
device, a 1-mL single-use syringe, and other accessories not utilized in the current protocol that 
will be discarded. VaxINator kits will be shipped from 
 
 for distribution to the clinical site. 
 
The following additional components will be sourced by 
 for distribution to 
the clinical site:  
1) 18G x 1 ½”, sterile needles in a box containing 100 units in total 
2) 1-mL, sterile, tuberculin syringes in a box containing 100 units in total  
 
All boxes of components and VaxINators (excluding investigational vaccine or placebo) should 
be stored at ambient temperature.  
Upon request by DMID, BPZE1 vaccine, Placebo and VaxINator atomization device and 
additional components will be transferred to the following address: 

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The placebo consists of the same constituents in the same quantities as the BPZE1 investigational 
vaccines, absent the attenuated B. pertussis cells. After reconstitution and transfer to a 1-mL 
syringe, the 109 CFU dose can be discriminated from the placebo and the 107 CFU dose due to the 
opaque appearance of the 109 CFU dose BPZE1 investigational vaccine, while the placebo and the 
107 CFU dose investigational vaccine are transparent. The 107 CFU dose investigational vaccine 
and the placebo are indistinguishable after reconstitution.  
The study product will be labeled according to manufacturer specifications and include the 
statement “Caution: New Drug – Limited by Federal Law to Investigational Use.”  
Further details are included in the Bordetella pertussis vaccine (BPZE1) IB, as well as in the 
protocol-specific MOP. 
6.1.3 
Product Storage and Stability 
BPZE1 vaccine and Placebo 
Both BPZE1 and placebo product will be stored at -10˚C or less until ready for use in order to 
maintain viability of BPZE1 prior to reconstitution with SWFI. The exposure of reconstituted 
BPZE1 to room temperature prior to vaccination should not exceed 60 minutes. A detailed 
description regarding the storage, reconstitution and handling the investigational vaccines is 
given in the Manual of Procedures (MOP). 
 
VaxINator devices are stored at ambient temperature. 
 
Additional details of product storage and stability can be found in the MOP. 
 
6.2 
Dosage, Preparation and Administration of Study 
Intervention/Investigational Product 
See the protocol-specific MOP for detailed information on the preparation, labeling, storage, and 
administration of study vaccine for each treatment arm. Study vaccine preparation will be 
performed by the participating site research pharmacist. 
For those randomized to the VaxINator groups, a full-dose (i.e., 800 µL containing 107 or 109 
CFU) will be drawn into one 1 mL syringe from the vial, the VaxINator tip connected, and 400 
μL volume will be administered to each nostril. The VaxINator provides a uniform, controlled 
delivery, which allows the clinician to accurately deliver 400 μL of vaccine to the initial nostril 
and then administer the remaining 400 μL to the opposite nostril.  

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For those randomized to administration via needleless tuberculin syringe, the same procedures 
will be followed, deleting the connection of a VaxINator tip.  
The dose of BPZE1 active ingredient or placebo will be administered to the volunteer within 60 
minutes of reconstitution. 
For the 109 CFU vaccine dose, one mL of SWFI is used to reconstitute the 1 mL vial of 
lyophilized BPZE1, but only 800 μL of vaccine are used for administration into both nostrils. 
The SWFI will be drawn into 1 mL syringes, the 1 mL SWFI will be injected into the lyophilized 
BPZE1 vial for reconstitution, 900 μL vaccine dose will be withdrawn into a separate 1 mL 
syringe, the plunger will be expelled to the 0.8-mL mark, the VaxINator shall be attached to the 
syringe, and 400 μL volume will be administered to each nostril. The VaxINator provides a 
uniform, controlled delivery, which allows the vaccinator to accurately deliver 400 μL of vaccine 
to the initial nostril and then administer the remaining 400 μL to the opposite nostril. 
For the 107 CFU vaccine dose, three 1 mL SWFI-filled syringes and two placebo cryovials are 
used to reconstitute and dilute the 1 mL vial of lyophilized 109 BPZE1.  The SWFI will be 
injected into each of the placebo cryovials and the 1 mL vial of lyophilized BPZE1.  Two serial 
dilutions (10-fold and 100-fold) of the reconstituted 109 CFU vaccine will be done to prepare a 
107 CFU vaccine dose. A 900 μL vaccine dose will be withdrawn into a separate 1 mL syringe, 
the plunger will be expelled to the 0.8-mL mark, the VaxINator shall be attached to the syringe, 
and 400 μL volume will be administered to each nostril. The VaxINator provides a uniform, 
controlled delivery, which allows the vaccinator to accurately deliver 400 μL of vaccine to the 
initial nostril and then administer the remaining 400 μL to the opposite nostril.To avoid 
accidental exposure to the environment and to study staff, actions should be taken to minimize 
generation of aerosols, since the bacteria are strictly respiratory tract organisms. Staff members 
should wear eye-protective glasses during the vaccination. Persons handling BPZE1 should wear 
gloves and must practice standard hand hygiene before touching their skin or eyes. 
6.3 
Modification of Study Intervention/Investigational Product for a 
Participant 
No modifications of the investigational product are anticipated as this is a single dose study.  
6.4 
Accountability Procedures for the Study Intervention/Investigational 
Product(s) 
Study products will be stored and shipped from the DMID-CMS, 
, to the 
participating VTEU. Once received, study products will be stored in and dispensed by the 
Investigational Pharmacy. Unused product will be handled in accordance with the MOP.  

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The United States Food and Drug Administration (FDA) requires accounting for the disposition 
of all investigational products. The Investigator is responsible for ensuring that a current record 
of product disposition is maintained and product is dispensed only at an official study site by 
authorized personnel as required by applicable regulations and guidelines. Records of product 
disposition, as required by federal law, consist of the date received, date administered, quantity 
administered, and the subject number to whom the drug was administered.  
The Investigational Pharmacist will be responsible for maintaining accurate records of the 
shipment and dispensing of the investigational product. The pharmacy records must be available 
for inspection by the DMID monitoring contractors, and is subject to inspection by a regulatory 
agency (e.g., FDA) at any time. An assigned Study Monitor will review the pharmacy records. 
Used and unused vials of vaccine and placebo will be retained until clinical trial accountability is 
completed, monitored and released for disposition as applicable. At study termination, all unused 
investigational product and VaxINator devices will be handled in accordance with the MOP 
following complete drug accountability and monitoring. 
6.5 
Assessment of Subject Compliance with Investigational Product 
Subjects will be directly observed at the time of dosing by a member of the clinical research 
team who is licensed to administer the study product. Administration will be documented on the 
appropriate data collection form and entered into the eCRF. 
6.6 
Concomitant Medications/Treatments 
Administration of medications, therapies, or vaccines will be recorded on the appropriate data 
collection form. Concomitant medications will include all current medications and medications 
taken within 30 days prior to signing the informed consent form through approximately 28 days 
after study vaccination (or through the early termination visit if prior to that time). Prescription 
and over-the-counter drugs will be included, as well as herbals, vitamins and supplements. In 
addition, receipt of non-study vaccines will be solicited through approximately 181 days after 
study vaccination and reported in the eCRF. 
Use of new medication should prompt evaluation for the presence of a new diagnosis of chronic 
medical disease or condition. Medications that might interfere with the evaluation of the 
investigational product should not be used during the study-reporting period unless clinically 
indicated as part of the subject’s health care. Use of bronchodilator medication due to wheezing 
(new prescription or daily use) will be used along with signs and symptoms to define a medically 
significant wheezing adverse event. 

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7 
STUDY SCHEDULE 
7.1 
Screening Visit (Visit 00, Screening, Clinic Visit [Window: Day -30 to 
Day -1]) 
Potential subjects will be screened for eligibility up to 30 days prior to the administration of the 
study vaccine. The following activities will be performed: 
• Subjects will be provided with a description of this study (purpose and study procedures) 
and asked to read and sign the informed consent form. The informed consent form will be 
signed prior to performing any study procedures, including any screening procedures.  
• Demographic information will be obtained by interview of subjects. 
• Eligibility criteria will be reviewed with subjects. 
• Complete medical history will be obtained by interview of subjects to ensure eligibility. 
• All concomitant medications taken and vaccines received within 30 days prior to signing 
the informed consent form will be reviewed with subjects and reported in the electronic 
Case Report File (eCRF).  
• Vital signs, including oral temperature, pulse, and blood pressure, will be obtained to 
ensure eligibility. Subjects must not eat or drink anything hot or cold, or smoke within 10 
minutes prior to taking oral temperature. 
• A physical examination will be performed by a study clinician licensed to make medical 
diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
• A urine or serum pregnancy test will be performed on all women of childbearing 
potential and must be negative to ensure eligibility. 
• Approximately 10 mL of venous blood will be collected for complete blood count, serum 
chemistries and HIV antibody testing. 
• Approximately 5 mL of venous blood will be collected for screening antibody assays (PT 
and PRN serum IgG).  
• Nasal samples will be obtained for baseline mucosal antibody assays. 

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7.2 
Enrollment/Vaccination (Visit 1, Clinic Visit [Day 1]) 
• Subject’s willingness to participate will be reconfirmed and documented in the subject’s 
study records prior to performing any further study procedures, including administration 
of the study vaccination. 
• Eligibility criteria will be reviewed with subjects prior to study vaccination to ensure 
continued eligibility. 
• Complete medical history and any updates obtained by interview of subjects since the 
screening visit will be reviewed with subjects prior to study vaccination to ensure 
continued eligibility. 
• All concomitant medications will be reviewed with subjects prior to study vaccination for 
accuracy and completeness. Any new concomitant medications taken since the screening 
visit will be reviewed with subjects and assessed for continued eligibility prior to study 
vaccination. Medications reported in the eCRF are limited to those taken within 30 days 
prior to study vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, will be obtained prior 
to study vaccination. Vital signs assessed on Day 1 will be considered as baseline. 
Subjects must not eat or drink anything hot or cold, or smoke within 10 minutes prior to 
taking oral temperature. 
• A limited physical examination will be performed, focused on the nasopharynx, 
oropharynx, and lungs prior to study vaccination; in addition, a targeted physical 
examination will be performed, if indicated, based on review of interim medical history, 
by a study clinician.  
• A urine or serum pregnancy test will be performed within 24 hours prior to study 
vaccination on all women of childbearing potential. Results must be negative and known 
prior to randomization and first study vaccination. 
• Approximately 10 mL of venous blood will be collected immediately prior to study 
vaccination for baseline antibody assays. 
• For subjects who consent to additional future use samples, approximately 24 mL of 
venous blood will be collected immediately prior to study vaccination for PBMC 
isolation and possible future research. 

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• Subjects will be enrolled in AdvantageEDCSM and assigned randomly to a treatment arm 
prior to study vaccination. 
• Pre-administration reactogenicity assessments will be performed prior to vaccination to 
establish baseline. Subjects will then receive a single dose of study vaccine via intranasal 
application. Subjects will be observed in the clinic for two (2) hours to record post-
administration reactogenicity and any AE/SAEs will be recorded on the appropriate data 
collection form prior to discharge from the clinic. 
• Subjects will be provided with a memory aid and other study-related materials to record 
daily oral temperature, solicited mucosal and systemic reactions, any unsolicited AEs, 
and concomitant medications. Subjects will be encouraged to take their oral temperature 
around the same time each day. Subjects must not eat or drink anything hot or cold, or 
smoke within 10 minutes prior to taking oral temperature. Subjects will be instructed on 
how to use their memory aid and how to measure and record AEs prior to discharge from 
the clinic. Subjects will be instructed to notify the study center if they develop any severe 
reactions after study vaccination. If the site principal investigator or appropriate sub-
investigator deems the reaction severe enough, further instructions will be given to the 
subject on the proper course of action, including a return to the clinic for immediate 
evaluation if appropriate. 
7.3 
Follow-up Visits  
7.3.1 
Visit 2, Day 8, Clinic Visit (Window: Day 8 ± 2 days) 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects. 
• All concomitant medications and vaccines will be recorded on the appropriate data 
collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A limited physical examination will be performed, focused on the nasopharynx, 
oropharynx, and lungs; in addition, a targeted physical examination will be performed, if 
indicated, based on review of interim medical history, by a study clinician. 

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• Approximately 10 mL of venous blood will be collected for safety laboratory studies, to 
include a complete blood count (white blood cell count, platelet count, hemoglobin 
concentration) and serum chemistries (ALT and creatinine) 
• For subjects who consent to additional future use samples, approximately 24 mL of 
venous blood will be collected for PBMC isolation and possible future research. 
7.3.2 
Visit 3, Day 15, Clinic Visit (Window: Day 15 ± 3 days) 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• All concomitant medications and vaccines will be recorded on the appropriate data 
collection form. 
• Memory aid information will be reviewed with subjects. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A limited physical examination will be performed, focused on the nasopharynx, 
oropharynx, and lungs; in addition, a targeted physical examination will be performed, if 
indicated, based on review of interim medical history, by a study clinician. 
• Approximately 10 mL of venous blood will be collected for antibody assays. 
7.3.3 
Visit 4, Day 29, Clinic Visit (Window: Day 29 ± 3 days) 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• All concomitant medications and vaccines will be recorded on the appropriate data 
collection form. 
• All AE/SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination may be performed, if indicated based on review of 
interim medical history, by a study clinician. 
• Approximately 10 mL of venous blood will be collected for antibody assays. 

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• For subjects who consent to additional future use samples, approximately 24 mL of 
venous blood will be collected for PMBC isolation and possible future research. 
• Nasal samples will be obtained for B. pertussis culture and immunological assays.  If 
culture is positive for B. pertussis, the isolate will be cryopreserved as detailed in the 
MOP. 
7.3.4 
Visit 4a (If needed, based on B. pertussis positive culture on Day 29), Day 46, 
Clinic Visit (Day 46 ± 3 days) 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• All concomitant vaccines will be recorded on the appropriate data collection form. 
• All SAEs will be recorded on the appropriate data collection form. 
• A targeted physical examination may be performed, if indicated based on review of 
interim medical history, by a study clinician. 
• Nasal samples will be obtained for B. pertussis culture (if necessary-positive from day 
29) and immunological assays.  If culture is positive for B. pertussis, the isolate will be 
cryopreserved as detailed in the MOP. 
7.3.5 
Visit 4b (if needed, based on positive B. pertussis culture from Day 46), Day 50, 
Phone Call (Day 50 ± 3 days) 
• If a participant’s sample from Visit 4a is positive for B. pertussis, a study investigator 
will prescribe a 5-day course of azithromycin (500 mg orally once per day). 
7.4 
Final Study Visit, Visit 5, Day 181, Clinic Visit (Day 181 ± 21 days) 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• All concomitant vaccines will be recorded on the appropriate data collection form. 
• All SAEs will be recorded on the appropriate data collection form. 

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• A targeted physical examination may be performed, if indicated based on review of 
interim medical history, by a study clinician. 
• Approximately 10 mL of venous blood will be collected for antibody assays. 
• For subjects who consent to additional future use samples, approximately 24 mL of 
venous blood will be collected for PBMC isolation and possible future research. 
• Nasal samples will be obtained for immunological assays 
7.5 
Early Termination Visit 
The following activities will be performed at the early termination visit for subjects who 
withdraw or are withdrawn/terminated from this study: 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects (if within 15 days after study 
vaccination). 
• All concomitant medications will be recorded on the appropriate data collection form (if 
prior to 29 days after study vaccination). 
• All non-study vaccines will be recorded on the appropriate data collection form (if prior 
to 181 days after study vaccination).  
• All AE/SAEs will be recorded on the appropriate data collection form. Only SAEs will 
be recorded after 28 days following study vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated based on clinical history. Subjects must not eat or drink anything hot or cold, or 
smoke within 10 minutes prior to taking oral temperature. 
• A targeted physical examination may be performed, if indicated based on review of 
interim medical history, by a study clinician.  
• Post-administration reactogenicity assessments will be performed (if within 15 days after 
study vaccination). 

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• Venous blood will only be obtained if the early termination visit occurs: 1) prior to Visit 
2, 2) after day 14 post-vaccination but before Visit 3, or 3) at least 28 days following 
Visit 3.  
• Approximately 10 mL of venous blood will be collected for antibody assays.  
• For subjects who consent to additional future use samples, approximately 24 mL 
of venous blood will be collected for PBMC isolation and possible future 
research. 
7.6 
Unscheduled Visit  
Unscheduled visits may occur at any time during this study. Any of the following activities may 
be performed: 
• Interim medical history, including an assessment for new medical conditions, will be 
obtained by interview of subjects and any changes since the previous clinic visit or 
contact will be noted. 
• Memory aid information will be reviewed with subjects (if within 15 days after study 
vaccination). 
• All concomitant medications will be recorded on the appropriate data collection form (if 
prior to 29 days after study vaccination). 
• All non-study vaccines will be recorded on the appropriate data collection form (if prior 
to 181 days after study vaccination).  
• All AE/SAEs will be recorded on the appropriate data collection form. Only SAEs will 
be recorded after 28 days following study vaccination. 
• Vital signs, including oral temperature, pulse, and blood pressure, may be obtained if 
indicated based on clinical history. Subjects must not eat or drink anything hot or cold, or 
smoke within 10 minutes prior to taking oral temperature. 
• A targeted physical examination may be performed, if indicated based on review of 
interim medical history, by a study clinician.  
• Post-administration reactogenicity assessments will be performed (if within 15 days after 
study vaccination). 

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8 
STUDY PROCEDURES/EVALUATIONS 
8.1 
Clinical Evaluations 
Complete medical history will be obtained by interview of subjects at the screening visit and will 
be updated on Day 1 prior to the study vaccination. Subjects will be queried regarding a history 
of significant medical disorders of the head, eyes, ears, nose, throat, mouth, cardiovascular 
system, lungs, gastrointestinal tract, liver, pancreas, kidney, urologic system, nervous system, 
blood, lymph nodes, endocrine system, musculoskeletal system, skin, and genital/reproductive 
tract. A history of any allergies, persistent asthma, nasopharyngeal symptoms (e.g., sinusitis) 
cancer, immunodeficiency, psychiatric illness, substance abuse, and autoimmune disease will be 
solicited. At follow-up visits after study vaccination, an interim medical history will be obtained 
by interview of subjects noting any changes since the previous clinic visit or contact. The interim 
medical history should include an assessment for new medical conditions. 
Medications history (concomitant medications) will include a review of all current medications 
and medications taken within 30 days prior to signing the informed consent form through 
approximately 28 days after study vaccination or through the early termination visit (if prior to 
28 days after the last study vaccination), whichever occurs first. All medications will be recorded 
on the appropriate data collection forms. Prescription and over-the-counter drugs will be 
included as well as herbals, vitamins, and supplements. In addition, receipt of non-study vaccines 
will be solicited from 28 days prior to enrollment through approximately 181 days after study 
vaccination and reported in the eCRF. Use of new medication should prompt evaluation for the 
presence of a new diagnosis of chronic medical disease or condition. Assessment of eligibility 
will include a review of all permitted and prohibited medications per the Subject Inclusion and 
Exclusion Criteria (see Sections 5.1and 5.2). In addition, the site principal investigator or 
appropriate sub-investigator may identify other medications that should not be used due to a risk 
to subject safety or assessment of reactogenicity and immunogenicity. 
At the screening visit, a physical examination will be performed by a study clinician licensed to 
make medical diagnoses and listed on the Form FDA 1572 as the site principal investigator or 
sub-investigator. On Visit 1, prior to study vaccination, and on Visits 2 and 3, a limited physical 
examination will be performed, focused on the nasopharynx, oropharynx, and lungs; in addition, 
a targeted physical examination will be performed, if indicated, based on review of interim 
medical history, by a study clinician. At follow-up visits 4, 4a and 5, a targeted physical 
examination may be performed, if indicated based on subject’s interim medical history, by a 
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Vital signs (oral temperature, pulse, and blood pressure) will be collected at the screening visit 
and prior to study vaccination. Vital signs assessed on Day 1 prior to vaccination will be 
considered as baseline. Subjects must not eat or drink anything hot or cold, or smoke within 10 
minutes prior to taking oral temperature. 
Reactogenicity assessments will include an assessment of solicited adverse events occurring 
from the time of each study vaccination through 14 days after study vaccination, which includes 
an assessment of mucosal reactions that include: runny nose, stuffy nose/congestion, nasal 
pain/irritation, epistaxis, sneezing, sinus pressure/pain, sore throat, cough, and shortness of 
breath, as well as systemic reactions that include fever, chills, fatigue, malaise, myalgia, 
arthralgia, headache, and hypersensitivity. Pre-administration reactogenicity assessments will be 
performed prior to study vaccination to establish baseline, then the study vaccination will be 
given. 
Subjects will be observed in the clinic for approximately 2 hours after study vaccination. Post-
administration reactogenicity assessments will be performed, and any AE/SAEs will be recorded 
on the appropriate data collection form prior to discharge from the clinic.  
All subjects will complete a subject memory aid from the time of study vaccination through 14 
days after study vaccination. Subject memory aids will be reviewed with the subjects for adverse 
events (solicited injection site and systemic reactions and unsolicited AEs) at follow-up visits 
occurring through approximately 14 days after study vaccination. 
8.2 
Laboratory Evaluations 
8.2.1 
Clinical Laboratory Evaluations 
Urine or serum pregnancy tests will be performed locally by the site at the screening visit and 
within 24 hours prior to study vaccination on all women of childbearing potential. Results must 
be negative and known prior to randomization on Day 1 to be eligible for participation in this 
study. 
Clinical screening laboratory parameters to be evaluated to confirm study eligibility and receipt 
of study vaccination will include complete blood counts, serum chemistries and a negative serum 
HIV antibody assay. A venous blood sample will be collected from each subject at the screening 
visit.  

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8.2.2 
Special Assays or Procedures 
The following assays will be performed in the Vanderbilt Vaccine Research Program 
Laboratory. Details regarding each of the following assays are found in the MOP. 
 
Venous blood for baseline pertussis antibodies 
Venous blood will be obtained at screening to confirm PT IgG< 20 IU/mL and PRN IgG< 125 
IU/mL serum. Standard ELISA will be used to measure serum concentrations of antibodies. 
 
Venous blood for immune response to vaccination 
Venous blood will be taken prior to and after vaccination to measure serum IgG and IgA by 
ELISA to four pertussis antigens (PT, FHA, PRN, FIM) 
 
Nasal aspiration samples to evaluate mucosal immune response 
Nasal aspiration samples will be used to measure mucosal IgA levels by ELISA to PT, FHA, 
PRN and FIM following vaccination. Procedures for collection of nasal aspiration samples are 
described in the Manual of Procedures.  
 
B. pertussis Culture 
Standard microbiologic techniques will be used to assess the presence of B. pertussis by culture 
at Day 29 following vaccination. 
8.2.3 
Specimen Preparation, Handling, and Shipping 
8.2.3.1 
Instructions for Specimen Preparation, Handling, and Storage 
Instructions for specimen preparation, handling, and storage are included in the protocol-specific 
MOP. 
8.2.3.2 
Specimen Shipment 
Specimen shipment will occur at intervals during the course of this study following all applicable 
International Air Transport Association (IATA) requirements and according to the specifics for 
storage temperature and documentation as detailed in the protocol-specific MOP. 
Specimens for pertussis-specific immunology studies will remain at the Vanderbilt VTEU site as 
experiments are being conducted. Any specimens remaining once the protocol-defined studies 
are completed will be shipped to the DMID CAR. 
Further instructions for specimen shipment are included in the protocol-specific MOP. 

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9 
ASSESSMENT OF SAFETY 
9.1 
Specification of Safety Parameters 
Safety will be assessed by the frequency and severity of: 
1. Serious adverse events (SAE) occurring from the time of vaccination through 
approximately 6 months after vaccination. 
2. Solicited Adverse Events – reactogenicity events occurring from the time of each study 
vaccination through 14 days after study vaccination: 
a) Mucosal reactions including runny nose, stuffy nose/congestion, nasal pain/irritation, 
epistaxis, sneezing, sinus pressure/pain, sore throat, cough, and shortness of breath, 
cough, rhinorrhea, and sneezing 
b) Systemic reactions including fever, chills, fatigue, malaise, myalgia, arthralgia, 
headache, and hypersensitivity. 
3. Unsolicited Adverse Events – study vaccine-related non-serious adverse events occurring 
from the time of study vaccination through approximately 28 days after study 
vaccination. 
4. New onset chronic medical conditions (NOCMCs) and adverse events of special interest 
(AESI) – NOCMC’s will be assessed throughout the follow-up period, while medically 
attended wheezing events (an AESI) will be collected through Day 29 post-vaccination.  
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
9.2.1 
Adverse Events 
Adverse Event (AE): International Council for Harmonisation (ICH) E6 defines an AE as any 
untoward medical occurrence in a patient or clinical investigation subject administered a 
pharmaceutical product regardless of its causal relationship to the study treatment. An AE can 
therefore be any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medicinal (investigational) product. 
The occurrence of an AE may come to the attention of study personnel during study visits and 
interviews of a study recipient presenting for medical care, or upon review by a study monitor. 

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AEs, including solicited mucosal (vaccination site) and systemic (subjective and quantitative) 
reactions, not meeting the protocol-defined criteria for SAEs will be captured on the appropriate 
data collection form and eCRF. Information to be collected for unsolicited non-serious AEs 
includes event description, date of onset, licensed study physician’s assessment of severity and 
relationship to study product and alternate etiology (if not related to study product), date of 
resolution of the event, seriousness and outcome. AEs occurring during the collection and 
reporting period will be documented appropriately regardless of relationship. AEs will be 
followed through stabilization or resolution. 
Any medical condition that is present at the time that the subject is screened will be considered 
as baseline and not reported as an AE; however, if the severity of any pre-existing medical 
condition increases, it will be recorded as an AE. 
AEs must be graded for severity and assessed for relationship to study product (see definitions 
below). Adverse events characterized as intermittent require documentation of onset and duration 
of each episode. The start and stop date of each reported AE will be recorded on the appropriate 
data collection form and eCRF. 
FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether or not considered drug related. 
Severity of Event: AEs will be assessed by a licensed study physician listed on the Form FDA 
1572 as the site principal investigator or appropriate sub-investigator using a protocol-defined 
grading system (see Sections 9.2.2 and Appendix B). For events not included in the protocol-
defined grading system, the following guidelines will be used to quantify severity: 
• 
Mild (Grade 1): Events require minimal or no treatment and do not interfere with the 
subject’s daily activities. 
• 
Moderate (Grade 2): Events result in a low level of inconvenience or concern with 
therapeutic measures. Moderate events may cause some interference with functioning and 
daily activities. 
• 
Severe (Grade 3): Events interrupt the subject’s daily activities and may require systemic 
drug therapy or other treatment. Severe events are usually incapacitating. 
Relationship to Study Product: The licensed study physician’s assessment of an AE's 
relationship to study product is part of the documentation process, but it is not a factor in 
determining what is or is not reported in this study. If there is any doubt as to whether a clinical 
observation is an AE, the event should be reported. The relationship to study product must be 

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• Death, 
• a life-threatening adverse event*, 
• inpatient hospitalization or prolongation of existing hospitalization, 
• a persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions, or 
• a congenital anomaly/birth defect. 
• Important medical events that may not result in death, be life-threatening, or require 
hospitalizations may be considered serious when, based upon appropriate medical 
judgment they may jeopardize the patient or subject and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition. Examples of such 
medical events include allergic bronchospasm requiring intensive treatment in an 
emergency room or at home, blood dyscrasias or convulsions that do not result in 
inpatient hospitalization, or the development of drug dependency or drug abuse. 
• All events described as Guillain-Barré syndrome will also be considered SAEs. 
* Life-threatening adverse event. An adverse event is considered “life-threatening” if, in the 
view of either the site principal investigator or sponsor, its occurrence places the patient or 
subject at immediate risk of death. It does not include an adverse event that, had it occurred in a 
more severe form, might have caused death. 
SAEs will be: 
• 
Assessed for severity and relationship to study product and alternate etiology (if not 
related to study product) by a licensed study physician listed on the Form FDA 1572 as 
the site principal investigator or sub-investigator. 
• 
Recorded on the appropriate SAE form and eCRF. 
• 
Followed through resolution by a licensed study physician listed on the Form FDA 1572 
as the site principal investigator or sub-investigator. 
• 
Reviewed and evaluated by an Independent Safety Monitor (ISM), the SMC (periodic 
review unless related), DMID, and the IRB. 

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9.2.4 
New Onset Chronic Medical Conditions 
The development of new onset chronic medical conditions will be assessed through Day 181. 
These are defined as new medical conditions, not present at the time of screening or enrollment, 
that require ongoing medical care and intervention.  
9.2.5 
Adverse Events of Special Interest  
Medically attended wheezing events will be considered adverse events of special interest given 
the route of study product administration and the nature of the study product (i.e., a live 
attenuated bacterial vaccine). Though subjects with a history of persistent asthma will be 
excluded from the study, it is possible that new onset wheezing episodes could occur. These will 
be evaluated by a study investigator and relatedness to study product will be assessed. Medically 
attended wheezing events will be evaluated through Day 29.  
9.2.6 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings 
Solicited mucosal site and systemic reactogenicity events will be documented and reported from 
the time of study vaccination through 14 days (Day 15) after study vaccination. 
Unsolicited, non-serious AEs will be documented from the time of study vaccination through 
approximately 28 days (Day 29) after study vaccination. 
SAEs will be documented from the time of study vaccination through approximately 6 months 
post vaccination.  
In addition, if a subject has an abnormal laboratory test value (e.g., complete blood count or 
serum chemistries), it will be recorded as an AE and will be graded per toxicity table and will be 
followed up until resolution.  
9.3 
Reporting Procedures 
9.3.1 
Serious Adverse Events 
SAEs will be followed until resolution even if this extends beyond the study-reporting period. 
Resolution of an AE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic.  

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adverse reaction as soon as possible, but in no case later than 7 calendar days after the sponsor’s 
initial receipt of the information. Relevant follow up information to an IND safety report will be 
submitted as soon as the information is available. Upon request from FDA, DMID will submit to 
FDA any additional data or information that the agency deems necessary, as soon as possible, 
but in no case later than 15 calendar days after receiving the request. 
All serious events designated as “not related” to study product(s), will be reported to the FDA at 
least annually in a summary format. 
9.3.3 
Reporting of Pregnancy 
Pregnancies occurring in study subjects will be reported via AdvantageEDCSM on the Pregnancy 
Report form. With the subject’s permission, all protocol-required venous blood samples will be 
obtained, and the subject will continue to be followed for safety for the duration of this study. 
Efforts will be made to follow all pregnancies reported during the course of this study until 
approximately 1 to 2 months post-pregnancy pending the subject’s permission. 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events 
AEs will be collected, assessed, and followed through resolution from the time of study 
vaccination through approximately 28 days after study vaccination. 
SAEs will be collected, assessed, and followed from the time of study vaccination through 
resolution even if this extends beyond the study-reporting period (approximately Day 181). 
Resolution of an AE/SAE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic. 
If the site principal investigator or appropriate sub-investigator becomes aware of an acute 
febrile illness and the site principal investigator or appropriate sub-investigator decides to bring 
the subject in for an evaluation to determine etiology, then the site principal investigator or 
appropriate sub-investigator, at their own discretion, can determine the specific testing that 
should be performed. 
Follow-up procedures, evaluations, and outcomes will be recorded on the appropriate data 
collection form. 

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9.5 
Halting Rules 
Further enrollment and study vaccinations will be halted for SMC review/recommendation if any 
of the following are reported: 
• Any subject experiences ulceration, abscess, or necrosis in the nose that is considered 
related to study product administration, through the subject’s last study visit. 
• Any subject experiences laryngospasm, bronchospasm, or anaphylaxis within 1 day after 
administration of study product that is considered related to study product. 
• Two or more subjects experience generalized urticaria within 3 days after administration 
of study product that is considered related to study product. 
• Any subject experiences a study product-related SAE from the time of study vaccination 
through the subject’s last study visit. 
• Any subject experiences acute weakness of limbs and/or cranial nerve innervated muscles 
(description of potential signal of GBS) after administration of study product, through the 
subject’s last study visit. 
This study will also be halted for SMC review/recommendation if, within 8 days after 
administration of each study vaccination, any of the following occurs: 
• Two or more subjects experience the same severe (Grade 3) mucosal Reactogenicity 
adverse event (table in section 9.2.2).  
• Two or more subjects experience the same severe (Grade 3) Subjective Systemic 
Reactogenicity adverse event. Grading scales for solicited reactions are included in 
Section 9.2.2. 
• Two or more subjects experience the same severe (Grade 3) Quantitative Systemic 
Reactogenicity adverse event. Grading scales for solicited reactions are included in 
Section 9.2.2. 
• Two or more subjects experience the same severe (Grade 3) study related laboratory 
adverse event (HGLT in MedDRA coding) 
• Two or more subjects experience the same severe (Grade 3) study related unsolicited 
systemic adverse event (HGLT in MedDRA coding) 

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DMID retains the authority to suspend additional enrollment and study interventions/ 
administration of study product during the entire study, as applicable. 
9.6 
Safety Oversight (ISM plus SMC) 
9.6.1 
Independent Safety Monitor (ISM) 
The ISM is a physician with relevant expertise whose primary responsibility is to provide 
independent safety monitoring in a timely fashion. The ISM will review SAEs in real time and 
other AEs as needed and provide an independent medical assessment and recommendation to 
DMID. The participating VTEU site will have an ISM with experience in infectious diseases or 
internal medicine, in close proximity to the participating VTEU site, and have the authority to 
readily access study participant records. 
9.6.2 
Safety Monitoring Committee (SMC) 
Safety oversight will be conducted by a SMC that is an independent group of experts that 
monitors subject safety and advises DMID. The SMC members will be separate and independent 
of study personnel participating in this study and should not have scientific, financial or other 
conflict of interest related to this study. The SMC will consist of members with appropriate 
expertise to contribute to the interpretation of the data from this study. 
The SMC will review study progress and participant, clinical, safety, and reactogenicity data at 
the following time points: 
• Organizational meeting: Prior to start of the study  
• Data Review Meeting (DRM): After 15-day reactogenicity data are available for the first 
25 subjects 
• Ad hoc meetings: When a halting rule is met, for immediate concerns regarding 
observations during this study, or as needed. 
Final review meeting: Approximately 6 months after final clinical database lock to review the 
cumulative unblinded safety data for this trial. The data will be provided in a standard summary 
format. The SMC may be asked to provide recommendations in response to questions posed by 
DMID. The SMC will operate under the rules of a DMID-approved charter that will be approved 
at the organizational meeting of the SMC. At this time, each data element that the SMC needs to 
assess will be clearly defined. Procedures for SMC reviews/meetings will be defined in the 
charter. The SMC will review applicable data to include, but not limited to, study progress and 

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participant, clinical, safety, and reactogenicity data. Reports may include enrollment and 
demographic information, medical history, concomitant medications, physical assessments, and 
solicited and unsolicited AE/SAEs. Additional data may be requested by the SMC, and interim 
statistical reports may be generated as deemed necessary and appropriate by DMID. The SMC 
may receive data in aggregate and presented by treatment arm. The SMC may also be provided 
with expected and observed rates of the expected AEs in an unblinded fashion, and may request 
the treatment assignment be unblinded for an individual subject if required for safety assessment.  
The SMC will review grouped and unblinded data in the closed session only. The SMC will meet 
and review this data at scheduled time points or ad hoc as needed during this study as defined in 
the SMC charter. As an outcome of each review/meeting, the SMC will make a recommendation 
as to the advisability of proceeding with study vaccinations (as applicable), and to continue, 
modify, or terminate this study. 
DMID or the SMC chair may convene the SMC on an ad hoc basis according to protocol criteria 
or if there are immediate concerns regarding observations during the course of the study. The 
DMID Medical Monitor is empowered to stop enrollment and study vaccinations if adverse 
events that meet the halting criteria are reported. The DMID Medical Monitor and the ISM will 
be responsible for reviewing SAEs in real time. The SMC will review SAEs on a regular basis 
and ad hoc during the study. 
 

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10 
CLINICAL MONITORING 
10.1 
Site Monitoring Plan 
 
Site monitoring will be conducted to ensure that human subjects protections, study and 
laboratory procedures, study intervention administration, and data collection processes are of 
high quality and meet sponsor, ICH/GCP guidelines and applicable regulations, and that this 
study is conducted in accordance with the protocol, protocol-specific MOP and applicable 
sponsor standard operating procedures. DMID, the sponsoring agency, or its designee will 
conduct site-monitoring visits as detailed in the clinical monitoring plan. Site visits will be made 
at standard intervals as defined by DMID and may be made more frequently as directed by 
DMID. Monitoring visits will include, but are not limited to, review of regulatory files, 
accountability records, eCRFs, informed consent forms, medical and laboratory reports, and 
protocol compliance. Site monitors will have reasonable access to the participating VTEU site, 
study personnel, and all study documentation according to the DMID-approved site monitoring 
plan. Study monitors will meet with the site principal investigators to discuss any problems and 
actions to be taken and document visit findings and discussions. 
 

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11 
STATISTICAL CONSIDERATIONS 
The goals of this study are to assess the immunologic response, safety, and tolerability of 
lyophilized BPZE1 administered intranasally at 107 CFU compared to 109 CFU, in healthy 
volunteers 18-49 years of age. Subjects in three of four treatment arms (45 of 50 expected 
subjects) will be vaccinated using the VaxINator device. In addition, the study will enroll a small 
number of subjects (5 expected) who will receive lyophilized BPZE1 administered by needleless 
tuberculin syringe to allow for historical, informal comparisons to previous Phase 1 data. 
This study is designed as a randomized, placebo controlled, partially-blind (i.e., the method of 
immunization [VaxINator vs. needleless syringe] will be known) clinical trial evaluating the 
safety and immunogenicity of a single dose of either 107 CFU or 109 CFU of BPZE1 delivered 
by intranasal VaxINator device or needleless tuberculin syringe.  
11.1 
Study Hypotheses 
This Phase 2a study is not designed to test a formal null hypothesis. Rather, it is intended to 
obtain preliminary estimates of the safety and immune response induced by the BPZE1 
lyophilized vaccine in healthy adults. See Section 3 for Study Objectives and Outcome 
Measures. 
11.2 
Sample Size Considerations 
The study population for this clinical trial includes healthy male and non-pregnant female 
subjects 18-49 years of age, inclusive, who meet all eligibility criteria. Study subjects will be 
recruited from the general population at the Vanderbilt University Medical Center VTEU site 
This study is planned to enroll approximately 50 healthy adults randomized into four treatment 
groups in a 3:3:3:1 ratio, as described in Table 2 (Section 4). The tables below describe the 
probability of observing safety events and the minimum detectable difference in seroconversion 
rates for the planned sample size.  
 
Table 3 indicates the probability of observing one or more safety events (such as solicited local 
or systemic reactogenicity events, or an unsolicited non-serious AE of a particular type) for a 
treatment arm with the assumed event frequency and for the given sample size. For example, for 
any of the treatment arms with a sample size of 15 subjects, there is a 79.4% probability of 
detecting an AE that occurs at a frequency of 10%, and a 14% probability if the AE occurs at a 
rate of 1%. 
 

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Table 3: Probability (%) of Observing at Least One Adverse Event 
Event 
Frequency 
N = 5 
N=15 
≥0.01% 
Rare 
<0.1 
<0.1 
≥0.1% 
Uncommon 
0.5 
1.5 
 ≥1% 
Common 
4.9 
14.0 
≥10% 
Very Common 
41.0 
79.4 
 
One of the secondary outcome measures is the proportion of subjects that achieve seroconversion 
to individual pertussis antigens (PT, FHA, PRN, and FIM 2/3) by serum IgG or IgA ELISA at 
Days 15, 29 and 181 for humoral immune response. Table 4 shows the minimum detectable 
differences with 80% power between the proportion of subjects attaining seroconversion in the 
BPZE1 by VaxINator group versus Placebo using a two-sided Fisher’s Exact test and alpha = 
0.05.  
Table 4: Minimum Detectable Difference in Proportion of Subjects Attaining 
Seroconversion comparing Placebo versus the BPZE1 (107 or 109 CFU) 
Placebo 
by 
VaxINator 
BPZE1 (107 
or 109 CFU) 
by 
VaxINator 
 
Assumed  
Proportion 
of Subjects 
Attaining 
Seroconversion 
(Placebo) 
 
Proportion 
of Subjects 
Attaining 
Seroconversion 
(BPZE1) 
Minimum Detectable 
Difference 
(Treatment – Placebo) 
N=15 
N=15 
0.00 
0.41 
0.41 
0.01 
0.43 
0.42 
0.05 
0.54 
0.49 
0.10 
0.63 
0.53 
0.20 
0.76 
0.56 
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11.3 
Planned Interim Analyses 
Interim analyses would only be used to terminate this trial in the event that unanticipated safety 
events deemed to be of sufficient concern require such action by the sponsor. These interim 
assessments will not be made based on testing a formal statistical hypothesis; therefore, p-value 
adjustment will not be made to any analyses. 
11.3.1 
Interim Safety Review 
An ISM and SMC will be convened by DMID to review study progress and participant, clinical, 
safety, and reactogenicity data, as described in Section 9.6. An interim safety review may include 
enrollment and demographic information, medical history, concomitant medications, physical 
assessments, clinical laboratory values, dosing compliance, solicited and unsolicited AE/SAEs, 
and B. pertussis colonization data. Additional data may be requested by the SMC, and interim 
statistical reports may be generated as deemed necessary and appropriate by DMID. The SMC 
may receive data in aggregate and presented by treatment arm, including expected and observed 
rates of the expected AEs. The SMC will review grouped data in the closed session only. The 
SMC will meet and review this data at scheduled time points or ad hoc as needed during this trial 
as defined in the SMC charter. As an outcome of each review/meeting, the SMC will make a 
recommendation as to the advisability of proceeding with study vaccinations (as applicable), and 
to continue, modify, or terminate this trial. 
Additionally, this trial will be monitored to determine if any of the halting rules described in 
Section 9.5 are met. 
A preliminary report prepared with data through Day 46 (or Day 29 if no subjects remain 
colonized with B. pertussis at the Day 29 visit) will be prepared as described in Section 11.4. 
This report will include analyses of safety data. 
11.3.2 
Interim Immunogenicity Review 
A preliminary report of safety and immunogenicity and B. pertussis colonization data through 
Day 46 (or Day 29 if no subjects remain colonized with B. pertussis at the Day 29 visit) will be 
prepared. The analyses presented in this report will be released while subjects remain in the trial 
for long-term safety and immunogenicity follow-up, and will be included in the final analysis of 
these data. 

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11.4 
Final Analysis Plan 
The results obtained from this study will be used to inform the dose recommended for a 
subsequent Phase 2b study. Clinical, safety, reactogenicity, immunogenicity and B. pertussis 
colonization data through approximately 45 days after study vaccination, or 28 days after study 
vaccination if no subjects remain colonized with B. pertussis at Day 29, will represent the 
primary clinical database for this trial. Once the last subject completes the visit that occurs 
approximately 45 days after study vaccination (or approximately 28 days after study vaccination, 
if no subjects remain colonized with B. pertussis at the Day 29 visit), the primary clinical 
database will be cleaned, monitored, and frozen. Analyses of safety, reactogenicity, 
immunogenicity, and B. pertussis colonization data by treatment arm are planned, including 
descriptive summaries of 15-day reactogenicity data, 28-day adverse event data (including SAE, 
NOCMC and AESI), 28 and 45-day nasal clearance data, and humoral and mucosal 
immunogenicity data through Day 46 (or Day 29 if no subjects remain colonized with B. 
pertussis at the Day 29 visit). As it is anticipated that subjects will remain in long term 
immunogenicity and safety follow-up at the time of these analyses, the presentation of these data 
will not include any unblinded listings of individual subject data and aggregate summaries will 
be in a format that prevents inadvertent unblinding of the sponsor or clinical investigators to any 
individual subject’s treatment assignment or immune response data. 
 
A preliminary report will be prepared by the DCC after the primary clinical database is cleaned, 
monitored, and frozen and all immunogenicity and colonization data through Day 46 (or Day 29 
if no subjects remain colonized with B. pertussis at the Day 29 visit) are received. These analyses 
may be made available to DMID and the vaccine company partner and to the lead principal 
investigator for planning subsequent trials. These analyses will not be used to make any 
decisions concerning the conduct of this trial. A formal statistical analysis plan that defines the 
analyses of data to be included in the preliminary report for early release and the CSR will be 
developed and finalized prior to the to the Day 46 data freeze (or Day 29 if no subjects remain 
colonized with B. pertussis at the Day 29 visit). 
 
Analysis of Day 181 humoral and mucosal immunogenicity data may be performed and released 
as the data are available from the research laboratories. Any such analyses would be considered 
the final analysis for the endpoint, and included in the CSR.  
 
Once the last subject completes the last visit, the final clinical database will be cleaned, 
monitored and locked. Unblinded analyses of safety, reactogenicity, colonization, and available 
immunogenicity data will then be performed by the DCC and will be included in the CSR. All 
safety analyses will be performed using the safety population, all immunogenicity analyses will 
be performed using the immunogenicity and the per-protocol (PP) populations, whereas all 

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colonization data will be summarized using per-protocol population. The analysis populations 
are defined below (Section 11.4.1.)  
 
All analyses will be presented by treatment group (i.e. group by BPZE1 109 CFU, BPZE1 107 
CFU, or Placebo). In addition, results for BPZE1 at dose of 109 CFU will be further grouped by 
vaccination device (group by VaxINator or needleless tuberculin syringe) to allow for 
comparison of immunogenicity results with previous results from Phase 1 clinical trials of 
BPZE1 (these previous trials used needleless tuberculin syringe as device for administration).  
11.4.1 
Analysis Populations 
11.4.1.1 
Safety Population  
The Safety Population will consist of all subjects who have received the study vaccine and for 
whom any data on safety are available. Subjects will be classified according to the treatment 
received. The primary safety analysis will be done on this population.  
11.4.1.2 
Immunogenicity Population 
The Immunogenicity Population will include all subjects who have received the study 
vaccination and contributed both pre-vaccination samples and either at least one post vaccination 
sample for humoral immunogenicity testing for which valid results were reported, or at least one 
post vaccination nasal sample for which valid results were reported. Subjects will be classified 
according to the treatment received.  
11.4.1.3 
Per Protocol Population 
The Per Protocol (PP) Population will include all subjects in the immunogenicity population, 
with the following exclusions: 
• Data from all available visits for subjects found to be ineligible at baseline. 
• Data from all visits subsequent to major protocol deviations, such as: 
• Receipt of non-study licensed live vaccine within 30 days before or after the study 
vaccination, 
• Receipt of non-study licensed inactivated vaccine within 14 days before or after the 
study vaccination, 
• Receipt of immunosuppressive therapy (e.g., corticosteroids) within 30 days before 
or after the study vaccination. 

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• Data from any visit that occurs substantially out of window. Whether a visit that occurs 
substantially out of window will be determined jointly by the principal investigator, the 
Sponsor, and the DCC.  
 
For analyses using the PP population, subjects will be grouped based on study vaccinations 
received. 
11.4.2 
Safety Data 
Summaries and analysis of safety data will be presented for the Safety Analysis Population. All 
summaries and analyses will be presented by treatment group. 
Solicited AEs will be summarized by severity for each day after the study vaccination (Days 1-
14 post study vaccination) and as the maximum severity over all 15 days. Additionally, solicited 
AEs will be analyzed by taking the most severe response over the follow-up period, 
dichotomizing into a binary variable (none versus mild, moderate, or severe) and using standard 
techniques, such as exact confidence intervals, to summarize the proportion of subjects reporting 
each symptom, any local symptom, and any systemic symptom. Summaries of solicited AEs will 
be presented separately for each study vaccination as well as overall study vaccinations by 
treatment arm. The proportion of subjects reporting symptoms may be compared between 
treatment arms using Chi-square or Fisher’s exact test. 
Unsolicited AEs will be coded by Medical Dictionary for Regulatory Activities (MedDRA) for 
preferred term and system organ class (SOC). SAEs will be reported by detailed listings showing 
the event description, MedDRA preferred term and SOC, relevant dates (study vaccinations and 
AEs), severity, relatedness, and outcome for each event. Non-serious unsolicited AEs will be 
summarized as number and percentage of subjects reporting at least one event in each 
MedDRA preferred term and SOC, cross tabulated by severity and relationship to study 
product. Additionally, the proportion of subjects and exact 95% confidence intervals of AEs in 
aggregate and by MedDRA categories will be computed. 
Subjects will be tested for B. pertussis colonization by culture at Day 29 and, if positive, again at 
Day 46. The proportion of subjects with positive B. pertussis culture will be reported for the PP 
population at each time point with corresponding 95% confidence intervals. 
11.4.3 
Immunogenicity Data 
Summaries and analysis of immunogenicity data will be presented for the immunogenicity and 
PP populations by treatment group. 

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Immune responses in terms of toxin-specific serum IgG and IgA antibody titers (PT, FHA, PRN, 
FIM 2/3) will be summarized at each time point. Analyses will include the Geometric Mean 
Titers (GMTs) along with corresponding 95% confidence intervals and Geometric Mean Fold 
Rise (GMFR), as well as the proportion of subjects achieving seroconversion (defined as 2-fold 
increase over baseline of GMT) for each antigen (PT, FHA, PRN, and FIM 2/3) and the 
proportion of subjects that achieved seroconversion for ≥ 1 and ≥ 2 antigens (PT, FHA, PRN, 
and FIM 2/3).  
Immune responses in terms of the ratio of toxin-specific (PT, FHA, PRN, FIM 2/3) nasal 
mucosal IgA antibody to total nasal mucosal IgA antibody will be summarized at each time 
point. Analyses will include the geometric mean titer ratios (GMTR) of antigen-specific IgA to 
total IgA and corresponding 95% confidence intervals, along with fold change of the GMTR 
from baseline. The proportion of subjects achieving seroconversion, defined as ≥2-fold rise of 
the GMTR from baseline, to each antigen (PT, FHA, PRN, and FIM 2/3) will be reported as well 
as the proportion of subjects that achieve seroconversion by nasal aspirate ELISA to one or more 
antigens (PT, FHA, PRN, and FIM 2/3) at either or both of Day 29 and Day 181. 
11.4.4 
Missing Values and Outliers 
All attempts will be made to collect all data per protocol. As missing data are expected to be 
minimal, no imputation will be performed for missing values. Any data point that appears to be 
erroneous or inexplicable based on clinical judgment will be investigated as a possible outlier. If 
data points are identified as outliers, sensitivity analyses will be performed to examine the 
impact of including or excluding the outliers. Any substantive differences in these analyses will 
be reported. 
 

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12 
SOURCE DOCUMENTS AND ACCESS TO SOURCE 
DATA/DOCUMENTS 
The participating VTEU site will maintain appropriate medical and research records for this 
study, in compliance with ICH E6, Section 4.9, and regulatory and institutional requirements for 
the protection of confidentiality of subjects. The participating VTEU site will permit authorized 
representatives of the DMID, its designees, and appropriate regulatory agencies to examine (and 
when required by applicable law, to copy) clinical study records for the purposes of quality 
assurance reviews, audits, monitoring and evaluation of the study safety and progress. These 
representatives will be permitted access to all source data, which include, but are not limited to, 
hospital records, clinical and office charts, laboratory notes, memoranda, evaluation checklists, 
pharmacy dispensing records, recorded data from automated instruments, copies or transcriptions 
certified after verification as being accurate and complete, microfiches, photographic negatives, 
microfilm or magnetic media, x-rays, and subject files and records kept at the pharmacy, at the 
laboratories, and medico-technical departments involved in the study. Source data are all 
information, original records of clinical findings, observations, or other activities in a clinical 
study necessary for the reconstruction and evaluation of the study. Interview of subjects is 
sufficient for obtaining medical history. Solicitation of medical records from the subject’s 
primary care provider is not required unless needed. 
 
 

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13 
QUALITY CONTROL AND QUALITY ASSURANCE 
Following a written DMID-accepted site quality management plan, the participating VTEU site 
is responsible for conducting routine quality assurance (QA) and quality control (QC) activities 
to internally monitor study progress and protocol compliance. The site principal investigators 
will provide direct access to all study-related sites, source data/data collection forms, and reports 
for monitoring and auditing by the sponsor, and inspection by local and regulatory authorities. 
The site principal investigators will ensure all study personnel are appropriately trained and 
applicable documentations are maintained on site. The DCC will implement quality control 
procedures beginning with the data entry system and generate data quality control checks that 
will be run on the database. Any missing data or data anomalies will be communicated to the 
participating VTEU site for clarification and resolution. 
DMID-designated clinical monitors will verify that the clinical trial is conducted and data are 
generated, documented (recorded), and reported in compliance with the protocol, ICH/GCP 
guidelines, and the applicable regulatory requirements. Clinical monitoring reports will be 
submitted to DMID. 
 
 
 
 

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14 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 
Ethical Standard 
The site’s principal investigator will obtain IRB approval for this protocol to be conducted at the 
research site and send supporting documentation to the DMID before initiating recruitment of 
subjects. The investigator will submit applicable information to the IRB on which it relies for the 
review, to conduct the review in accordance with 45 CFR 46, ICH E6 GCP, and as applicable, 21 
CFR 56 (Institutional Review Boards) and 21 CFR 50 (Protection of Human Subjects), other 
federal, state, and local regulations. The IRB must be registered with OHRP as applicable to the 
research. DMID must receive the documentation that verifies IRB-approval for this protocol, 
associated informed consent documents, and upon request any recruitment material and handouts 
or surveys intended for the subjects, prior to the recruitment and enrollment of subjects.   
Any amendments to the protocol or consent materials will be approved by the IRB before they 
are implemented. IRB review and approval will occur at least annually throughout the enrollment 
and follow-up of subjects, and may cease if annual review is no longer required by applicable 
regulations. The investigator will notify the IRB of deviations from the protocol and reportable 
SAEs, as applicable to the IRB policy. 
Each institution engaged in this research will hold a current Federalwide Assurance (FWA) 
issued by the Office of Human Research Protection (OHRP) for federally funded research. 
14.2 
Institutional Review Board 
Informed consent, or IRB approved waiver of consent or altered consent process, is required for 
all subjects participating in a DMID-sponsored clinical trial, in compliance with applicable 
regulations and guidelines, including but not limited to 45 CFR 46, 21 CFR 50, 56, 312/812 for 
FDA-regulated studies, and ICH E6 Guidelines. Prior to the beginning of the trial, the 
investigator should have the IRB’s written approval, favorable opinion of the written informed 
consent form(s), and any other written information to be provided to the subjects. The informed 
consent documents should meet the requirements of 21 CFR 50.20 and contain the information 
required of 21 CFR 50.25(a) and 21 CFR 50.25 (b). The relevant IRB will have the final 
authority for ensuring the adequacy of the information in the informed consent document.  
 
Prior to enrollment of subjects into this study, the approved protocol and informed consent form 
will be reviewed and approved by the appropriate IRB listed on its FWA. The responsible 
official for the IRB will sign the IRB letter of approval of the protocol prior to the start of this 
study and a copy will be provided to DMID. The IRB FWA number will be provided to DMID. 

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Should amendments to the protocol be required, the amendments will be written by the sponsor 
and provided to the site principal investigator for submission to the IRB. A single IRB will be 
used for this study. 
14.3 
Informed Consent Process 
The site principal investigators will choose subjects in accordance with the eligibility criteria 
detailed in Section 5.1. Before any study procedures are performed, subjects must sign an 
informed consent form that complies with the requirements of 21 CFR Part 50 and 45 CFR 46 
and the local IRB. Study personnel may employ recruitment efforts prior to obtaining study 
consent if a patient-specific screening consent is on record or if the IRB has agreed that chart 
review is allowed without a fully executed screening consent. In cases where there is not a 
patient-specific screening consent on record, site clinical staff may pre-screen via chart review 
and refer potential subjects to the research staff. Research staff would obtain written consent per 
the standard informed consent process before conducting protocol-specific screening activities.  
Informed consent is a process that is initiated prior to an individual agreeing to participate in a 
study and continuing throughout the individual’s study participation. Before any study 
procedures are performed, including pre-screening of subjects for eligibility, subjects will 
receive a comprehensive explanation of the proposed study procedures and study 
interventions/products. This will include the nature, risks and possible benefits of this study, 
alternate therapies, any known AEs, the investigational status of the study interventions/products, 
and the other elements that are part of obtaining proper informed consent. Subjects will also 
receive a detailed explanation of the proposed use and disclosure of their protected health 
information, including specifically their serum and urine samples. Subjects will be allowed 
sufficient time to consider participation in this research study, after having the nature, risks and 
possible benefits of this study explained to them, and can discuss this study with their family, 
friends or legally authorized representative or think about it prior to agreeing to participate.  
 
Informed consent forms describing in detail the study interventions/products, study procedures, 
risks and possible benefits will be given to subjects. The informed consent form must not include 
any exculpatory statements. Informed consent forms will be IRB-approved and subjects will be 
asked to read and review the appropriate document. Upon reviewing the appropriate document, 
the site principal investigator (or designee) will explain this research study to subjects and 
answer any questions that may arise. Subjects must sign the informed consent form, and written 
documentation of the informed consent process is required prior to starting any study procedures 
specifically for this study, including determining eligibility and administering study product. 
 

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By signing the informed consent form, subjects agree to complete all study procedures required 
by this study, unless the subject withdraws voluntarily, or is withdrawn or terminated from this 
study for any reason. The rights and welfare of subjects will be protected by emphasizing to 
subjects that the quality of their medical care will not be adversely affected if they decline to 
participate in or withdraw from this study.  
 
DMID will provide the site principal investigators, in writing, any new information that 
significantly impacts the subjects' risk of receiving the investigational products. This new 
information will be communicated by the site principal investigators to subjects who consent to 
participate in this study in accordance with IRB requirements. The informed consent document 
will be updated and subjects will be re-consented per IRB requirements, if necessary. Subjects 
will be given a copy of all informed consent forms that they sign. 
14.4 
Exclusion of Women, Minorities, and Children (Special Populations) 
This study will be inclusive of all adults who meet the Subject Inclusion Criteria (see Section 
5.1) and do not meet the Subject Exclusion Criteria (see Section 5.2), regardless of religion, 
sex, or ethnic background.  
 
Women of childbearing potential who are not sterilized via tubal ligation, bilateral 
oophorectomy, salpingectomy, hysterectomy, or successful Essure® placement (permanent, non-
surgical, non-hormonal sterilization) with documented radiological confirmation test at least 90 
days after the procedure, and still menstruating or <1 year has passed since the last menses if 
menopausal must agree to practice highly effective contraception that may include, non-male 
sexual relationships, abstinence from sexual intercourse with a male partner, monogamous 
relationship with a vasectomized partner who has been vasectomized for 180 days or more prior 
to the subject receiving the study vaccination, barrier methods such as condoms or 
diaphragms/cervical cap with spermicide, effective intrauterine devices, NuvaRing®, and 
licensed hormonal methods such as implants, injectables or oral contraceptives (the “pill”), with 
use of a highly effective method of contraception for a minimum of 30 days prior to study 
product exposure and agree to practice highly effective contraception for the duration of study 
product exposure, including 60 days after vaccination. A highly effective method of 
contraception is defined as one that results in a low failure rate (i.e., less than 1% per year) when 
used consistently and correctly. In addition to contraceptive use, all women of childbearing 
potential will be required to have a negative urine or serum pregnancy test within 24 hours prior 
to study vaccination. If a female subject becomes pregnant while participating in this study, we 
will ask her permission to follow-up with her about her health and the health of her baby through 
pregnancy outcome.  
 

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Children will not be included in this study. 
14.5 
Subject Confidentiality 
Subject confidentiality is strictly held in trust by the site principal investigators, other study 
personnel, the sponsor, and their agents. This confidentiality is extended to cover testing of 
biological samples, in addition to the clinical information relating to participating subjects. 
Subjects will have code numbers and will not be identified by name.  
 
The study protocol, documentation, data, and all other information generated will be held in 
strict confidence. No information concerning this study or the data will be released to any 
unauthorized third party without prior written approval of the sponsor.  
 
All information provided by the sponsor and all data and information generated by the 
participating VTEU site as part of this study (other than a subject’s medical records) will be kept 
confidential by the site principal investigators and other study personnel to the extent permitted 
by law. This information and data will not be used by the site principal investigator or other 
study personnel for any purpose other than conducting this study. These restrictions do not apply 
to: (1) information which becomes publicly available through no fault of the site principal 
investigator or other study personnel; (2) information which is necessary to disclose in 
confidence to an IRB solely for the evaluation of this study; (3) information which is necessary 
to disclose to provide appropriate medical care to a study subject; or (4) study results which may 
be published as described in Section 16. The study monitor, applicable regulatory authorities, 
such as the FDA, or other authorized representatives of the sponsor may inspect all documents 
and records required to be maintained by the site principal investigators. This includes, but is not 
limited to, medical records (office, clinic, or hospital) and pharmacy records for the subjects in 
this study. The participating VTEU site will permit access to such records.  
 
To protect privacy, we have received a Certificate of Confidentiality.  With this Certificate, the 
researchers cannot be forced to release information that may identify the research subject, even 
by a court subpoena, in any federal, state, or local civil, criminal, administrative, legislative, or 
other proceedings.  The researchers will use the Certificate to resist any demands for information 
that would identify the subject, except as explained below. 
 
The Certificate cannot be used to resist a demand for information from personnel of the United 
States Government that is used for auditing or evaluation of federally funded projects, like this 
study, or for information that must be released in order to meet the requirements of the Federal 
Food and Drug Administration (FDA). 
 

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A Certificate of Confidentiality does not prevent the subject from voluntarily releasing 
information about themselves or their involvement in this research.  If any person or agency 
obtains a written consent to receive research information, then the researchers may not use the 
Certificate to withhold that information.   
 
The Certificate of Confidentiality does not prevent the researchers from reporting without the 
subject’s consent, information that would identify the subject as a participant in the research 
project regarding matters that must be legally reported, including child and elder abuse, sexual 
abuse, or wanting to harm themselves or others.   
14.6 
Study Discontinuation 
If the study is prematurely terminated by the sponsor, any regulatory authority, or the 
investigator for any reason, the investigator will promptly inform the study subjects and assure 
appropriate therapy or follow-up for the subjects, as necessary. The investigator will provide a 
detailed written explanation of the termination to the IRB. If any subject’s private information 
will continue to be collected for this study, the IRB must approve a consent form with the study 
procedures, any risks and discomforts, and applicable elements, and the investigator or designee 
will re-consent the subjects as approved by the IRB. 
 
14.7 
Costs, Subject Compensation, and Research Related Injuries 
There is no cost to subjects for the research tests, procedures, and study product while taking part 
in this trial. Procedures and treatment for clinical care may be billed to the subject, subject’s 
insurance or third party. Subjects may be compensated for their participation in this trial. 
Compensation will be in accordance with the local IRB’s policies and procedures, and subject to 
IRB approval. 
If it is determined by the site principal investigator that an injury occurred to a subject as a direct 
result of the tests or treatments that are done for this trial, then referrals to appropriate health care 
facilities will be provided to the subject. Study personnel will try to reduce, control, and treat any 
complications from this trial. Immediate medical treatment may be provided by the participating 
site. No financial compensation will be provided to the subject by the NIAID, NIH to the subject, 
or by the participating site, for any injury suffered due to participation in this trial. 

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14.8 
Future Use of Stored Specimens 
Subjects will be asked for permission to keep any remaining serum from venous blood samples 
for possible use in future research studies, such as examining additional immunological 
assessments or testing for antibodies against other viruses or bacteria.  
 
Additionally, subjects will be asked for permission to collect approximately 24 mL of additional 
venous blood for PBMC isolation and possible future research during Visits 1, 2, 4, 5 or, if 
applicable, during an early termination visit. 
 
Samples will be stored indefinitely at a central clinical storage facility and may be shared with 
investigators at the participating VTEU site and with other investigators at other institutions. The 
samples will not be sold or used directly for production of any commercial product. No human 
genetic tests will be performed on the samples. Each sample will be encoded (labeled) only with 
a barcode and a unique tracking number to protect subject confidentiality.  
 
There are no benefits to subjects in the collection, storage and subsequent use of their specimens 
for future research. Reports about future research done with subjects’ samples will not be kept in 
their health records.  
 
Subjects will be given the option to decide if they want their samples to be used for future 
research or have their samples destroyed at the end of this study. The subject’s decision can be 
changed at any time by notifying the study doctors or nurses in writing; however, if a subject 
originally consents to future use and subsequently changes his/her decision, any data from a 
previously collected sample may still be used for this research. 
 

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15 
DATA HANDLING AND RECORD KEEPING 
The site principal investigator is responsible to ensure the accuracy, completeness, legibility, and 
timeliness of the data reported.  
Data collection forms will be derived from the eCRF and provided by the DCC to record and 
maintain data for each subject enrolled in this study. All data collection forms should be 
completed in a neat, legible manner to ensure accurate interpretation of data. Black or blue ink is 
required to ensure clarity of reproduced copies. When making a change or correction, cross out 
the original entry with a single line and initial and date the change. Do not erase, overwrite, or 
use correction fluid or tape on the original.  
Data reported in the eCRF derived from the data collection forms should be consistent with the 
data collection forms or the discrepancies should be explained.  
The sponsor and/or its designee will provide guidance to the site principal investigators and other 
study personnel on making corrections to the data collection forms and eCRF. 
15.1 
Data Management Responsibilities 
All data collection forms and laboratory reports must be reviewed by the clinical team and data 
entry personnel, who will ensure that they are accurate and complete. Adverse events must be 
recorded on the appropriate data collection form, assessed for severity and relationship, and 
reviewed by the site principal investigator or appropriate sub-investigator.  
Data collection is the responsibility of the study personnel at the participating VTEU site under 
the supervision of the respective site principal investigators. During this study, the site principal 
investigators must maintain complete and accurate documentation for this study.  
The DCC for this study will be responsible for data management, quality review, analysis, and 
reporting of the study data. 
15.2 
Data Capture Methods 
Clinical (including, but not limited to, AE/SAEs, concomitant medications, medical history, and 
physical assessments), reactogenicity, serologic immunogenicity, and mucosal immunogenicity 
data will be entered into a 21 CFR 11-compliant Internet Data Entry System provided by the 
DCC. The data system includes password protection and internal quality checks, such as 
automatic range checks, to identify data that appear inconsistent, incomplete, or inaccurate. 

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Clinical and reactogenicity data will be entered directly from the data collection forms completed 
by the study personnel. 
15.3 
Types of Data 
Data for this study will include clinical, safety, and outcome measures (e.g., reactogenicity, 
serologic immunogenicity/mucosal immunogenicity [antibody assays], and shedding of BPZE1 
from nasal samples). 
15.4 
Timing/Reports 
A preliminary analysis will be performed when all safety, reactogenicity, immunogenicity and B. 
pertussis colonization data through Day 46 (or Day 29 if no subjects remain colonized with B. 
pertussis at Day 29) are available. Analyses will be made available to DMID, the lead principal 
investigator, and vaccine company partner for planning subsequent clinical trials. These analyses 
will not be used to make any decisions concerning the conduct of this trial. As it is anticipated 
that subjects will remain in long term follow-up at the time of these analyses, blinded 
investigators who will not be involved in the analysis will be responsible for assessing SAEs and 
AESIs until all subjects have completed the final follow-up visit.  
 
Other interim statistical reports may be generated as deemed necessary and appropriate by 
DMID. Safety and immunogenicity summary reports may be generated for the SMC, as well.  
See Section 9.3 for further reporting details. 
 
The final clinical study report (CSR) will be completed when all clinical, safety, and 
reactogenicity data through approximately 180 days after the study vaccination are available. 
Once the last subject completes the last visit, the final clinical database will be cleaned, 
monitored and locked. Unblinded analyses of safety, reactogenicity, colonization, and available 
immunogenicity data will then be performed by the DCC and will be included in the CSR.  
 
After full analysis and final reporting is complete, and upon request and DMID approval, the 
DCC will provide the participating VTEU site with a summary of results by treatment arm 
and/or subject treatment assignments. In this regard, the participating VTEU site requesting such 
information to share with study subjects must do so in compliance with their respective IRB 
guidelines. 

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15.5 
Study Records Retention 
Study records and reports, including, but not limited to, eCRFs, source documents, informed 
consent forms, and study drug disposition records, shall be maintained for 2 years after a 
marketing application is approved for the drug for the indication for which it is being 
investigated; or, if no application is to be filed or if the application is not approved for the drug, 
until 2 years after the investigation is discontinued and FDA has been so notified. The 
participating VTEU site must contact DMID for authorization prior to the destruction of any 
study records. Informed consent forms for future use will be maintained as long as the sample 
exists. 
15.6 
Protocol Deviations 
A protocol deviation is any noncompliance with the study protocol, GCP or protocol-specific 
MOP requirements. The noncompliance may be either on the part of the subject, the site 
principal investigators, or other study personnel. As a result of deviations, corrective actions are 
to be developed by the site and implemented promptly.  
 
These practices are consistent with ICH E6:  
4.5  
Compliance with Protocol, Sections 4.5.1, 4.5.2, and 4.5.3  
5.1  
Quality Assurance and Quality Control, Section 5.1.1  
5.20  Noncompliance, Sections 5.20.1, and 5.20.2.  
 
It is the responsibility of the site principal investigators and other study personnel to use 
continuous vigilance to identify and report deviations within five working days of identification 
of the protocol deviation, or within five working days of the scheduled protocol-required 
activity. All deviations must be promptly reported to DMID, via the DCC’s AdvantageEDCSM. 
 
All protocol deviations, as defined above, must be addressed in study subject data collection 
forms. A completed copy of the DMID Protocol Deviation Form must be maintained in the 
Regulatory File, as well as in the subject's chart. Protocol deviations must be sent to the local 
IRB/IEC per its guidelines. The site principal investigator and other study personnel are 
responsible for knowing and adhering to their IRB requirements. 
 

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CONFIDENTIAL 
16 
PUBLICATION POLICY 
All investigators funded by the NIH must submit or have submitted for them to the National 
Library of Medicine’s PubMed Central (http://www.ncbi.nlm.nih.gov/pmc/) an electronic 
version of their final, peer-reviewed manuscripts upon acceptance for publication, to be made 
publicly available no later than 12 months after the official date of publication. The NIH Public 
Access Policy ensures the public has access to the published results of NIH funded research. It 
requires investigators to submit final peer-reviewed journal manuscripts that arise from NIH 
funds to the digital archive PubMed Central upon acceptance for publication. Further, the policy 
stipulates that these papers must be accessible to the public on PubMed Central no later than 12 
months after publication. 
Refer to: 
• 
NIH Public Access Policy, http://publicaccess.nih.gov/ 
• 
NIH Office of Extramural Research (OER) Grants and Funding, 
http://grants.nih.gov/grants/oer.htm 
As of January 2018, all clinical trials supported by the NIH must be registered on 
ClinicalTrials.gov, no later than 21 days after the enrollment of the first subject. Results of all 
clinical trials supported by the NIH, generally, need to be submitted no later than 12 months 
following the primary completion date. A delay of up to 2 years is available for trials that meet 
certain criteria and have applied for certification of delayed posting. 
As part of the result posting a copy of this protocol (and its amendments) and a copy of the 
Statistical Analysis Plan will be posted on ClincialTrials.gov. 
For this trial, the responsible party is DMID which will register the trial and post results.  
The responsible party does not plan to request certification of delayed posting. 
Refer to: 
• 
Public Law 110-85, Section 801, Clinical Trial Databases 
• 
42CFR11 
• 
NIH NOT-OD-16-149 
 

DMID Protocol 17-0010  
Version 5.0 
Intranasal BPZE1 Pertussis Vaccine in Healthy Adults 
25 June 2019 
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DMID/NIAID/NIH 
 
 
 
 
 
 
 
 
 
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17 
LITERATURE REFERENCES 
1. 
Yeung, K.H.T., et al., An update of the global burden of pertussis in children younger 
than 5 years: a modelling study. Lancet Infect Dis, 2017. 17(9): p. 974-980. 
2. 
Warfel, J.M., L.I. Zimmerman, and T.J. Merkel, Acellular pertussis vaccines protect 
against disease but fail to prevent infection and transmission in a nonhuman primate 
model. Proc Natl Acad Sci U S A, 2014. 111(2): p. 787-92. 
3. 
Zhang, Q., et al., Prevalence of asymptomatic Bordetella pertussis and Bordetella 
parapertussis infections among school children in China as determined by pooled real-
time PCR: a cross-sectional study. Scand J Infect Dis, 2014. 46(4): p. 280-7. 
4. 
Ward, J.I., et al., Efficacy of an acellular pertussis vaccine among adolescents and adults. 
N Engl J Med, 2005. 353(15): p. 1555-63. 
5. 
de Greeff, S.C., et al., Seroprevalence of pertussis in The Netherlands: evidence for 
increased circulation of Bordetella pertussis. PLoS One, 2010. 5(12): p. e14183. 
6. 
Palazzo, R., et al., Evidence of increased circulation of Bordetella pertussis in the Italian 
adult population from seroprevalence data (2012-2013). J Med Microbiol, 2016. 
7. 
Althouse, B.M. and S.V. Scarpino, Asymptomatic transmission and the resurgence of 
Bordetella pertussis. BMC Med, 2015. 13: p. 146. 
8. 
Mielcarek, N., et al., Live attenuated B. pertussis as a single-dose nasal vaccine against 
whooping cough. PLoS Pathog, 2006. 2(7): p. e65. 
9. 
Mielcarek, N., et al., Dose response of attenuated Bordetella pertussis BPZE1-induced 
protection in mice. Clin Vaccine Immunol, 2010. 17(3): p. 317-24. 
10. 
Skerry, C.M., et al., A live attenuated Bordetella pertussis candidate vaccine does not 
cause disseminating infection in gamma interferon receptor knockout mice. Clin Vaccine 
Immunol, 2009. 16(9): p. 1344-51. 
11. 
Feunou, P.F., et al., Genetic stability of the live attenuated Bordetella pertussis vaccine 
candidate BPZE1. Vaccine, 2008. 26(45): p. 5722-7. 
12. 
Thorstensson, R., et al., A phase I clinical study of a live attenuated Bordetella pertussis 
vaccine--BPZE1; a single centre, double-blind, placebo-controlled, dose-escalating study 
of BPZE1 given intranasally to healthy adult male volunteers. PLoS One, 2014. 9(1): p. 
e83449. 
13. 
Thorstensson, R., et al., A phase Ib (high dose), double-blind, single center, dose-
escalating, placebocontrolled, randomized study of a live attenuated B. pertussis strain 
given as a single intranasal dose to healthy adult volunteers. 2017: on file at ILiAD 
Biotechnologies. 
14. 
Locht, C., et al., Live attenuated pertussis vaccine BPZE1 protects baboons against B. 
pertussis disease and infection. J Infect Dis, 2017. 
15. 
Thalen, M., et al., Effect of relevant culture parameters on Pertussis Toxin expression by 
Bordetella pertussis. Biologicals, 2006. 34(3): p. 213-20. 
16. 
Feunou, P.F., N. Mielcarek, and C. Locht, Reciprocal interference of maternal and infant 
immunization in protection against pertussis. Vaccine, 2016. 34(8): p. 1062-9. 
 

DMID Protocol 17-0010  
Version 5.0 
Intranasal BPZE1 Pertussis Vaccine in Healthy Adults 
25 June 2019 
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DMID/NIAID/NIH 
 
 
 
 
 
 
 
 
 
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CONFIDENTIAL 
APPENDIX B: CLINICAL SAFETY LABORATORY ADVERSE 
EVENT GRADING  
 
Hematology 
Protocol 
Range 
Reference 
Range 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
WBC 103/uL (Decrease) 
≥3.9 
3.9 – 10.7 
2.5 – 3.8 
1.5 – 2.4 
<1.5 
WBC 103/uL (Increase) 
≥3.9 
3.9 – 10.7 
10.8 – 15.0 
15.1 – 20.0 
>20.0 
Hemoglobin g/dL 
(Decrease, Male) 
≥13.0 
13.0 – 18.1 
11.0 – 12.9 
9.5 – 10.9 
<9.5 
Hemoglobin g/dL 
(Decrease, Female) 
≥11.8 
11.8 – 16.0 
10.1 – 11.7 
8.5 – 10.0 
<8.5 
Platelet count 103/uL 
(Decrease) 
≥135 
135 – 371 
125 – 134 
100 – 124 
<100 
Platelet count 103/uL 
(Increase) 
≥135 
135 – 371 
372 – 550 
551 – 750 
>750 
 
Chemistry 
Protocol 
Range 
Reference 
Range 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
ALT U/L (Increase, 
Female) 
<45 
0-44 
45 – 100 
101 – 200 
>200 
ALT U/L (Increase, Male) 
<62 
0-61 
62 – 138 
139 – 275 
>275 
Creatinine mg/dL 
(Increase, Female) 
≤1.11 
0.57-1.11 
1.12 – 1.60 
1.61 – 2.00 
>2.00 or requires 
dialysis 
Creatinine mg/dL 
(Increase, Male) 
≤1.25 
0.72-1.25 
1.26 – 1.70 
1.71 – 2.00 
>2.00 or requires 
dialysis