Institute for Global Health and Infectious Diseases (IGHID) 11911 - Cross-reactive N. Gonorrhoeae Immune Responses Induced by a N. Meningitidis Vaccine
Phase: PHASE4
Conditions: Neisseria Gonorrhoeae Infection
Sponsor: University of North Carolina, Chapel Hill

 
IGHID 11911 - Cross-reactive N. gonorrhoeae Immune Responses 
Induced by a N. meningitidis Vaccine 
 
 
 
NCT number NCT04094883 
Document Date 06/24/2020 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
Page 1 
 
 
 
 
IGHID 11911 - Cross-reactive N. gonorrhoeae Immune Responses Induced by a N. 
meningitidis Vaccine 
 
Funding Sponsor:  North Carolina Translational & Clinical Sciences (NC TraCS) Institute    
 Funding Mechanism: Duke/UNC CTSA Consortium Collaborative Translational Research Grant 
 
 
Protocol Date: 06/24/2020 
 
Protocol Version: 1.2 
 
 
Principal Investigator:  
Joseph Alex Duncan, MD, PhD  
Associate Professor of Medicine 
Division of Infectious Diseases  
University of North Carolina 
Chapel Hill, NC 27599-7030  
Phone: 919-966-6547 
Fax: 919-843-1015  
E-mail: jaduncan@med.unc.edu 
 
Co-Investigator:  
 
Marcia Hobbs, PhD 
Professor of Medicine and 
Microbiology Immunology 
Division of Infectious Diseases 
University of North Carolina 
Chapel Hill, NC 27599-7030  
Phone: 919-966-5050   
Fax: 919-843-1015  
E-mail: 
marcia_hobbs@med.unc.edu 
 
 
 
 
 
 
 
 
 
 
 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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IGHID 11911 - Cross-reactive N. gonorrhoeae Immune Responses Induced by a N. 
meningitidis Vaccine 
 
Protocol Date: 06/24/2020 
Protocol Version: 1.2 
 
 
 
SIGNATURE PAGE  
The signature below documents the review and approval of this protocol and provides the necessary 
assurances that this study will be conducted according to the protocol, including all statements regarding 
confidentiality, and according to national, regional, and local legal and regulatory requirements. 
 
 
 
____________________________________________ 
Site Principal Investigator Name (Print)  
 
 
 
____________________________________________ 
Site Principal Investigator Signature  
 
 
 
 
____________________________________________ 
Date  

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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TABLE OF CONTENTS 
Page 
TABLE OF CONTENTS……………………………………………………………………………….3 
LIST OF ABBREVIATIONS…………………………………………………………………………..5 
PROTOCOL SUMMARY……………………………………………………………………………...6 
1.0 
HYPOTHESIS AND STUDY OBJECTIVES…………………………………………………7 
1.1 
Hypothesis………………………………………………………………………..…………….7 
1.2 
Primary Objectives……………………………………………………………………………..7 
1.3 
Secondary Objective………………………………………………..…………………………..7 
2.0 
INTRODUCTION……………………………………………………………………..……….7 
2.1 
Background……………………………………………………………………………….…….7 
2.2 
Rationale……………………………………………………………………………….……….8 
3.0 
STUDY DESIGN……………………………………………………………………………….9 
4.0 
SELECTION AND ENROLLMENT OF PARTICIPANTS…………………………………..9 
4.1 
Inclusion Criteria……………………………………………………………………………….9 
4.2 
Exclusion Criteria……………………………………………………………………………....9 
4.3 
Study Recruitment and Enrollment Procedures……………………………………………….10 
5.0 
STUDY TREATMENT ……………………………………………………………………….11 
5.1 
Regimens, Administration, and Duration……………………………………………………..11 
5.2 
Study Product Formulation and Preparation…………….……………………………………11 
5.3 
Pharmacy: Study Product Acquisition/Distribution and Accountability……………………..11 
5.4 
Concomitant Medications…………………………………………………………………….11 
6.0 
CLINICAL AND LABORATORY EVALUATIONS……………………………………….12 
6.1 
Schedule of Events……………………………………………………………………………12 
6.2 
Timing of Evaluations………………………………………………………………………..12 
6.3 
Instructions for Evaluations…………………………………………………………………..14 
7.0 
CLINICAL MANAGEMENT ISSUES………………………………………………………17 
7.1 
Dose Modification…………………………………………………………………………….17 
7.2 
Toxicity Management…………………………………………………………………………17 
7.3 
Pregnancy……………………………………………………………………………………..17 
8.0 
ADVERSE EVENT REPORTING……………………………………………………………18 
8.1 
Definitions……………………………………………………………………………………..18 
8.2 
Reporting………………………………………………………………………..……………..19 
9.0 
STATISTICAL CONSIDERATIONS…………………………………………………………19 
9.1 
General Design Issues………………………………………………………….……………….19 
9.2 
Endpoints……………………………………………………………………………………….20 
9.3 
Sample Size and Accrual……………………………………………………………………….21 
9.4 
Monitoring…………………………………………………………………..………………….21 
9.5 
Analyses………………………………………………………………………………..……….22 
10.0 
STUDY MANAGEMENT……………………………………….…………………………….22 
10.1 
Institutional Review Board (IRB) Review and Informed Consent…………………………….22 
10.2 
Registration Procedures………………………………………………………………..……….23 
10.3 
Criteria for Discontinuation…………………………………………………………………….23 
10.4 
Deviations from the Protocol……………………………………………………………..…….24 
10.5 
Clinical Site Monitoring and Record Availability……………………………………….……..24 
10.6 
Required Documentation…………………………………………………………………….….24 
10.7  
Adherence to the Protocol……………………………………………………………………….24 
 
 
 
 
 
 
 
 
 
 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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10.8 
Amendments to the Protocol……………………………………………………………….24 
10.9 
Data Management………………………………………………………………………….24 
10.10  Record Retention………………………………………………….………………………..25 
10.11 Obligations of Investigators………………………………………………..………………25 
10.12 Participant Confidentiality………………………………………………………………....25 
10.13 Study Discontinuation……………………………………………………………………..26 
11.0 
BIOHAZARD CONTAINMENT……………………………………………………...….26 
12.0 
REFERENCES…………………………………………………………………………….26 
  
 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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LIST OF ABBREVIATIONS 
Abbreviation 
Definition 
4CMenB 
4 component Neisseria meningitidis serogroup B vaccine (trade name Bexsero™) 
AE 
Adverse Event 
CI 
Confidence Interval 
CTRC 
Clinical and Translational Research Center 
ID 
Identification 
Ig 
Immunoglobulin 
IL 
Interleukin 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
LOS 
Lipo-oligosaccharide 
MAb 
Monoclonal Antibody 
MBRB 
Medical Biomolecular Research Building 
mL 
Milliliter 
Ng 
Neisseria gonorrhoeae 
Nm 
Neisseria meningitidis 
OMV 
Outer Membrane Vesicles 
PBMC 
Peripheral Blood Mononuclear Cells 
TGF 
Transforming growth factor 
Th 
T helper cell 
Treg 
Regulatory T cell 
UNC 
University of North Carolina at Chapel Hill 
UNCH 
UNC Hospitals 
 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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PROTOCOL SUMMARY 
Protocol Title  
 
Cross-reactive N. gonorrhoeae Immune Responses Induced by a N. 
meningitidis Vaccine 
Study Design 
 
Single arm, prospective clinical trial to assess immunologic responses induced 
by immunization with 4CMenB vaccine that cross react with N. gonorrhoeae. 
Participants will receive two-doses of an FDA-approved vaccine that provides 
protection from N. meningitidis infection according to the recommended dosing 
schedule. The participants will provide samples of blood as well as mucosal 
surface derived samples (urine and/or swabs) at four separate visits. 
Study Population  
 
Individuals 18-25 years of age that are not pregnant, HIV negative, have no 
history of congenital immunologic disorder, and are not taking immune 
suppressive medications 
Number of subjects  
15 
Number of sites  
1 
Clinical Samples  
 
Blood, pharyngeal swab, urine (male subjects), vaginal swab (female subjects) 
Estimated Start of 
Enrollment 
07/31/2019 
 
Estimated Time to 
Completion 
12/31/2020 
 
Protocol Duration 
Each individual will participate in the study for between 6 and 8 weeks. 
Regimen or 
Intervention 
Each participant will receive two doses of 4CMenB vaccine 5 weeks between 
doses. 
 
 
 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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1.0 
HYPOTHESIS AND STUDY OBJECTIVES 
1.1 
Hypothesis 
Immunization with 4CMenB induces cross reactive immune responses against N. gonorrhoeae 
 
1.2 
Primary Objectives 
1. Determine whether immunization with 4CMenB induces serum antibodies against N. 
gonorrhoeae surface antigens 
2. Determine whether polyfunctional CD4+ lymphocytes directed against N. gonorrhoeae 
surface antigens are induced by immunization with 4CMenB  
 
1.3 
Exploratory Objectives 
1. Determine whether immunization with 4CMenB induces mucosal surface antibodies against 
N. gonorrhoeae surface antigens 
2. Determine whether immunization with 4CMenB induces serum and/or mucosal surface 
antibodies against N. gonorrhoeae porB or mtrE 
3. Determine whether  immunization with 4CMenB induces serum and/or mucosal surface 
antibodies against N. gonorrhoeae antignens other than porB and mtrE 
4. Determine whether polyfunctional CD8+ lymphocytes directed against N. gonorrhoeae 
surface antigens are induced by immunization with 4CMenB 
5. Determine quantity and phenotype of CD4+ Tfh cells  lymphocytes induced by immunization 
with 4CMenB 
6. Determine inflammatory cytokine production by N. gonorrhoeae surface antigen-stimulated  
PBMC from 4CMenB-immunized humans.   
7. Identify and clone B-cell receptors that recognize N. gonorrhoeae antigens after 
immunization with 4CMenB 
 
2.0 
INTRODUCTION 
2.1 
Background 
N. gonorrhoeae poses global public health threats because of emerging antibiotic resistance.  
The WHO estimates over 60 million new cases of N. gonorrhoeae (Ng) infection occur each year 
worldwide, making it one of the most common bacterial sexually transmitted infections1. Most 
cases of Ng infection are limited to the lower genital tract, causing urethritis in men and cervicitis 
in women. Antibiotic resistance in Ng presents a major problem in the control of this pathogen2. In 
2018, three cases of extensively drug resistant Ng were identified in Australia and the United 
Kingdom after treatment failure with currently recommended dual antibiotic therapy3. Effective 
preventative strategies, particularly vaccines, are urgently needed to reduce the public health 
burden of Ng infections throughout the world.  
 
Ng infection in humans does not lead to protective adaptive immune responses. Uncomplicated 
Ng infections are associated with increased risk of recurrent infections. Re-infections with the same 
Ng strain occur at a higher frequency than with other circulating strains in the community, 
indicating there is little strain-specific immunity to Ng4. Moreover, male subjects in experimental 
human Ng infection studies show no evidence of immunity upon rechallenge5.    
 
OMV-based meningococcal vaccines appear to provide cross-species protection against Ng 
infection. Effective vaccines for the most prevalent serogroups of N. meningitidis (Nm) have been 
in use in humans for decades. Meningococcal vaccines were first generated against polysaccharide 
capsular antigens of Nm serogroups A,C,Y, and W. Nm serogroup B (NmB) capsule is a poorly 
immunogenic disaccharide in humans which has made the development of vaccines targeting NmB 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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challenging and prevented the use of capsular polysaccharicde antigen for this purpose6. Recently, 
vaccines containing several proteins and lipoproteins have been generated from NmB OMV7,8. 
These vaccines are safe and have been used in mass vaccination campaigns in the setting of NmB 
epidemics in several countries9-11. STI surveillance in the setting of vaccine campaigns in Cuba and 
Norway showed reduced rates of Ng infections, which suggested Nm-OMV-based vaccines may 
offer cross-species protection against Ng12,13. STI education campaigns could have contributed to 
decreased Ng rates in these settings. However, a recent OMV-based vaccine campaign in New 
Zealand demonstrated decreased gonococcal infection rates, and surveillance data showed that 
infection rates for Chlamydia trachomatis (another sexually transmitted bacterial pathogen that 
causes urethritis and cervicitis) did not mirror the decrease in gonorrhea rates. This, and the fact 
that the effect was greater in vaccinated than unvaccinated persons, strongly supports the 
hypothesis that the Nm vaccine provided specific protection from Ng infection14. Data from New 
Zealand suggest that the Nm-OMV-based vaccine offered ~30% protection against Ng infection. 
Researchers at the Uniformed Services University of Health Sciences have demonstrated that mice 
vaccinated with the FDA-approved four component NmB OMV vaccine (4CMenB), delivered 
subcutaneously (SC) or intraperitoneally (IP), substantially accelerated Ng clearance and reduced 
vaginal Ng burdens vs. unvaccinated or adjuvant only-treated animals, supporting the ecologic data 
in humans that suggests Nm-OMV can induce immune responses that protect from Ng. Serum 
pooled from 10 individual humans that were immunized with 3 doses of 4CMenB and one 
individual who received the currently US FDA-approved 2 dose schedule of 4CMenB was recently 
shown to have increased antibody titers against whole Ng bacteria, demonstrating 4CMenB does 
indeed induce immunologic responses that cross react with Ng15. 
 
2.2 
Rationale 
Immunologic responses associated with protection from Nm disease are well-characterized. During 
a Nm outbreak at a US military base, a lack of detectable bactericidal antibodies against the 
outbreak strain of Nm was associated with high-rates of invasive infection in exposed individuals16. 
Bactericidal antibodies to Nm develop naturally during invasive infection and during 
nasopharyngeal carriage. Vaccination with capsular polysaccharide, which induced bactericidal 
antibodies, was shown to protect against invasive Nm disease16-18. In contrast to Nm, infection with 
non-encapsulated Ng weakly induces antibodies; anti-Ng antibody concentrations in vaginal 
secretions and sera are not different in individuals with current or past Ng infection vs. those with 
no history of infection19. There are few published studies of Ng-directed T cell responses in patients 
with Ng, but one small study showed that T cells from 30 Ng-infected patients exhibited weak 
proliferative responses to gonococcal PorB not significantly greater than proliferation without 
antigen20. Interleukin 17 (IL-17), the main cytokine from Th17 lymphocytes that stimulates 
neutrophil (PMN) recruitment and causes local inflammation, increases during Ng infections in 
humans and mice21-23. However, Ng is resistant to PMN killing, leading Russell et al. to suggest 
that stimulation of adaptive Th17 responses to Ng may partially explain the lack of protective 
immunity to Ng in humans22. 
 
Identification of the immunologic correlates of protection against N. gonorrhoeae would provide 
critical information that would allow for development and testing of vaccines for the prevention 
of N. gonorrhoeae infection. Characterization of the anti-gonococcal immune responses that 
develop in response to 4CMenB immunization will provide preliminary data that defines the 
immunologic parameters that can be assessed in the setting of a clinical trial designed to test the 
efficacy of 4CMenB or other vaccines for protection against N. gonorrhoeae infection.  There is 
currently no data regarding the identity of gonococcal outermembrane antigens recognized by 
human antibodies after 4CMenB immunization as well as no documented reports of the cellular 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
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immune response evoked by this vaccine. Consequently, this proposed pilot study will provide a 
crucial first step to defining the immunologic correlates of immunity to N. gonorrhoeae infection. 
Although the 4CMenB vaccine has been FDA approved, it is not currently required for 
enrollment as a student at the University of North Carolina at Chapel Hill. 4CMenB vaccine 
uptake has been estimated at 14%  based on teen National Immunization Survey data; however, 
uptake rates between 14% and 98% have been found at Universities that have implemented 
vaccination campaigns in the setting of NmB disease outbreaks24,25. Thus, it is timely to conduct 
this study while prior MenB immunization rates in the target population are relatively low.   
 
3.0 
STUDY DESIGN  
This study is a single center, single arm, interventional pilot study  in which participants will 
receive two-doses of the 4CMenB vaccine according to the recommended administration 
schedule and will provide blood, pharyngeal swabs, and urine or self-collected vaginal swabs at 
each of four study visits.  
 
The study population will include 15 individuals aged 18-25 years with no contraindication to 
vaccination and no known immune compromising medical condition or medication.  
 
Participants will be seen for informed consent and eligibility screening. Enrolled participants will 
be given 1 dose of 4CMenB at enrollment and a second dose at week 5. Participants will be seen 
at entry, weeks 5, 6, and 7 for blood collection, pharyngeal swabs, and provide urine (male 
participants) or self-collected vaginal swabs collected for secondary screening and baseline 
immunologic testing. 
 
4.0 
SELECTION AND ENROLLMENT OF PARTICIPANTS 
4.1 
Inclusion Criteria 
1. Able to understand and give informed consent 
2. Willingness to undergo all study procedures. 
3. Males or females between the ages of 18 to 25 at screening (inclusive). 
4. Good health, as determined by medical history and targeted physical exam. 
5. Participants agree to abstain from vaccines from study entry to 30 days after receipt of the 
second 4CMenB vaccine. 
6. Female participants of child bearing potential must have a negative urine pregnancy test at 
the screening visit, and prior to receipt of vaccines at study entry and week 5 visits.  Self-
reported history is acceptable documentation of hysterectomy, bilateral oophorectomy, tubal 
ligation, or tubal micro-inserts which eliminate child bearing potential of participant. If 
participating in sexual activity that could lead to pregnancy, women must agree to use a form 
of contraceptive until 28 days after completion of the vaccine series. At least one of the 
following methods must be used appropriately: Condoms (male) with or without spermicidal 
agent, Diaphragm or cervical cap with spermicide, Intrauterine device (IUD), or Hormone-
based contraceptive. Self-report of a monogamous male partner who has a vasectomy is also 
acceptable. 
 
4.2 
Exclusion Criteria 
1. Known allergy/sensitivity or any hypersensitivity to latex or any of the components of the 
study product or its formulation (see section 5.2 for a list of components). 
2. Participants who have received any vaccine directed against N. meningitidis serogroup B 
3. Serious illness or injury requiring hospitalization within 21 days prior to study entry. 

IGHID # 11911 
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4. Current or prior history of a medical condition resulting in impaired immunity (such as HIV 
infection, inborn or acquired immunodeficiency syndromes, all cancers including leukemia or 
lymphoma, or the of use antineoplastic drugs or radiation treatment).  
5. Known active infection with HIV, HCV, or HBV. This information will be obtained verbally 
from the participant. 
6. History of excessive alcohol consumption, drug abuse, psychiatric conditions, social 
conditions or occupational conditions that in the opinion of the investigator would preclude 
compliance with the study. 
7. Hemophilia or other bleeding diatheses. 
8. Receipt of anticoagulants (aspirin or nonsteroidal anti-inflammatory drugs (NSAIDS) are 
acceptable) within 14 days prior to study entry. 
9. Autoimmune disorders; mild autoimmune disorders, such as eczema, are not exclusionary 
and will be determined by the investigator. 
10. Use of any systemic immunomodulatory treatment, systemic corticosteroids, (inhaled and 
topical corticosteroids acceptable), investigational products, interleukins, interferons, growth 
factors, or intravenous immunoglobulin (IVIG) within 45 days prior to study entry. 
11. Pregnant women and nursing mothers or women who are planning to become pregnant or 
breastfeed within 28 days after receipt of their second 4CMenB vaccine.  
12. Have received any licensed vaccine within 30 days prior to study vaccination. 
13. Have donated blood or blood products within 30 days before study vaccination, plan to 
donate blood at any time during the study and up to 30 days after the last blood draw. 
14. Any condition in the opinion of the investigator that would interfere with the proper conduct 
of the trial. 
 
4.3 
Study Recruitment and Enrollment Procedures 
Potential participants will be recruited from the UNC-Chapel Hill University and surrounding 
area population. Potential participants will be recruited through flyers posted on the campus and 
through the UNC email listserv and through Join the Conquest at UNC. Potential participants will 
contact the study team via telephone or email and a study team member will provide information 
about the study and review eligibility criteria. Participants will receive a total of $130 over the 
course of the study to compensate for time and providing samples related to the study in addition 
to receiving the benefit of protection from invasive N. menigtidis infection as a result of receiving 
the FDA-approved 4CMenB vaccine over the course of the study. The payments for participation 
are $20 provided at screening visit, $20 at entry visit, and $30 each at week 5 visit, week 6 visit, 
and week 7 study completion visit. Participants will be paid at the end of each study visit.  
 
To promote a roughly even balance in the sex at birth of participants, enrollment of additional 
participants of one sex at birth will be discontinued when enrollment of participants of that sex at 
birth reaches 8 participants. 
 
Once a candidate has been identified, study details will be carefully discussed with the participant 
by the study coordinator or study investigator. The participant will be asked to read and sign the 
IRB approved informed consent form. For participants from whom a signed informed consent 
form has been obtained, a screening visit will be performed that will include obtaining 
information related to eligibility, completion of a targeted physical examination, and a point of 
care urine pregnancy test for female participants of child bearing potential.  For participants from 

IGHID # 11911 
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whom informed consent has been obtained, but who are deemed ineligible or who do not enroll 
into initial immunization step, a Screening Failure Results form must be completed.  
 
5.0 
STUDY TREATMENT  
Study treatment is defined as 4-component Neisseria menigitidis group B vaccine, 4CMenB 
(trade name: Bexsero™)  
 
5.1 
Regimens, Administration, and Duration 
All participants will receive the 4CMenB vaccine, 0.5 mL, at entry (Day 0) and at week 5. The 
study product will be administered as supplied, intramuscularly in the deltoid region of the upper 
arm or the higher anterolateral area of the thigh in the clinic by a licensed study nurse, nurse 
practitioner, or study physician with experience administering vaccines using aseptic technique 
during preparation and administration. All injections should preferably be given in the deltoid 
region and in the non-dominant arm; however, if this is not feasible, the dominant arm may be 
used. The vaccine must not be injected intravascularly, subcutaneously, or intradermally.  
Vaccination should be deferred at the discretion of the site investigator if the participant is febrile 
or acutely ill. Following each vaccination, a licensed study nurse, nurse practitioner, or study 
physician with experience administering vaccines will observe the participant for 15 minutes for 
any adverse reactions/events.  
 
5.2 
Study Product Formulation and Preparation 
4-Component Meningococcal Group B Vaccine  
Each 0.5 mL dose of 4CMenB contains approximately: 
• 
recombinant Neisserial adhesin A protein 
 
 
 
50 mcg 
• 
recombinant factor H binding protein 
 
 
 
50 mcg 
• 
recombinant Neisserial Heparin Binding Antigen protein 
 
50 mcg 
• 
Outer Membrane Vesicles  
 
 
 
 
25 mcg 
• 
aluminum hydroxide 
 
 
 
 
 
1.5 mg 
• 
Sodium chloride 
 
 
 
 
 
 
3.13 mg 
• 
L-histidine  
 
 
 
 
 
 
0.78 mg 
• 
sucrose 
 
 
 
 
 
 
 
10 mg 
• 
Water 
 
 
 
 
 
 
 
quantity sufficient 
 
5.3 
Pharmacy:  Study Product Acquisition/Distribution and Accountability 
5.3.1 
Study Product Acquisition/Distribution 
4CMenB Vaccine is an FDA-approved product.  Study vaccine will be purchased form the 
manufacturer through UNC Investigational Drug Service (IDS) Pharmacy. 
 
 
5.3.2 
Study Product Accountability 
The site pharmacist is required to maintain complete records of all study product received from 
the manufacturer and subsequently dispensed.  All unused study product will be distributed from 
the UNC Investigational Drug Service (IDS) Pharmacy to the primary investigator’s laboratory 
for use in in vitro experiments after the study is completed or terminated. 
 
5.4 
Concomitant Medications 
5.4.1 
Required Medications 
None 
 
 

IGHID # 11911 
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5.4.2 
Prohibited Medications 
• 
anticoagulants (aspirin or nonsteroidal anti-inflammatory drugs (NSAIDS) are acceptable) 
• 
systemic corticosteroids (inhaled or topical corticosteroids are acceptable) 
• 
systemic antineoplastic drugs 
• 
systemic immunomodulatory drugs (including but not limited to immune modulating 
monoclonal antibodies, calcineurin inhibitors, antiproliferative agents, mTOR inhibitors, 
interleukins, interferons, and growth factors) 
• 
investigational or approved vaccines (other than study product) 
• 
intravenous immunoglobulin (IVIG) 
• 
other investigational products 
 
5.4.3 
Precautionary Medications 
 
None 
 
1 
female participants only 
1a female participants of child bearing potential  
2   mucosal surface swabs, urine, serum, and PBMC preparations will be used for immunologic 
analysis that will be conducted on batched samples.   
3  male participants only 
 
6.2 
Timing of Evaluations 
6.2.1 
Screening Evaluations 
Screening evaluations include: medical history, medication history within the past 60 days, vital 
signs, targeted physical exam, and pregnancy test for female participants of child bearing 
potential). Screening visit must be conducted no greater than 30 days prior to enrollment visit.  
The screening evaluation and enrollment visit can all occur on the same day. Participants may be 
re-screened one time.  
 
6.0 
 CLINICAL AND LABORATORY EVALUATIONS 
6.1 
Schedule of Events  
Evaluation 
Screening 
 
Entry 
 
48-72 hr 
post 
injection 
Post-Entry Evaluations 
Week 5 
 
48-72 hr 
post 
injection 
Week 6 
 
Week 7 
 
Medical & medication history 
X 
X 
 
X 
 
X 
X 
Acute Illness Assessment 
 
X 
 
X 
 
 
 
Vital signs/targeted physical exam 
X 
X 
 
X 
 
X 
X 
Pregnancy test1a 
X 
X 
 
X 
 
 
 
Phlebotomy collected blood (up to 60 
mL per visit) 
 
X 
 
X 
 
X 
X 
Stored PBMCs2 
 
X 
 
X 
 
X 
X 
Stored serum2 
 
X 
 
X 
 
X 
X 
Pharyngeal swab collection2 
 
X 
 
X 
 
X 
X 
Self-collected vaginal swab1,2 
 
X 
 
X 
 
X 
X 
Urine collection2,3 
 
X 
 
X 
 
X 
X 
Vaccine injection  
 
X 
 
X 
 
 
 
Telephone contact  
 
 
X 
 
X 
 
 
 

IGHID # 11911 
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6.2.2 
Entry Evaluations 
Entry evaluations will be collected PRIOR to phlebotomy, specimen collection, and vaccine 
administration:  
 
Acute Illness Assessment will be completed before phlebotomy, specimen collection, and vaccine 
administration. Participants who have an acute illness may be rescheduled for their phlebotomy, 
specimen collection, and vaccine within their screening visit window.  
 
Update medical history and concomitant medications. Conduct a targeted physical exam and 
collect vital signs [heart rate (HR), blood pressure (BP), oral temperature (Temp), respiration rate 
(RR), and weight] prior to phlebotomy to collect up to 60 mL of blood, specimen collection, and 
vaccine administration.  
 
For females of reproductive potential, negative urine pregnancy testing results must be available 
before phlebotomy, specimen collection, and vaccine administration.   
 
Samples to be stored for immunologic testing will be batch run at the end of the study: 
pharyngeal swabs, urine sample (from male participants), self-collected vaginal swabs (from 
female participants), serum, and PBMC.  
 
6.2.3 
Post-Entry Evaluations 
Week 5 Evaluations:  
Acute Illness Assessment will be completed before phlebotomy, specimen collection, and vaccine 
administration. Participants who have an acute illness or who have taken exclusionary 
anticoagulant medications may be rescheduled for their vaccine within their screening visit 
window.  
 
Update medical history and concomitant medications before phlebotomy and vaccine 
administration. Conduct a targeted physical exam and collect vital signs (HR, BP, Temp, RR) 
before phlebotomy to collect up to 60 mL of blood, specimen collection, and vaccine 
administration. 
 
For females of reproductive potential, negative urine pregnancy testing results must be available 
before phlebotomy, specimen collection, and vaccine administration.   
 
Week 5 laboratory evaluations will be collected PRIOR to vaccine administration:  
 
Samples to be stored for immunologic testing will be batch run at the end of the study: 
pharyngeal swabs, urine sample (from male participants), self-collected vaginal swabs (from 
female participants), serum, and PBMC.  
 
Week 6 and Week 7 Evaluations: 
Update medical history and concomitant medications prior to phlebotomy to collect up to 60 mL 
of blood and specimen collection. 
 
Samples to be stored for immunologic testing will be batch run at the end of the study: 
pharyngeal swabs, urine sample (from male participants), self-collected vaginal swabs (from 
female participants), serum, and PBMC.  

IGHID # 11911 
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6.2.4 
Discontinuation Evaluations 
Participants who elect not to receive or cannot receive the initial immunization or second 
immunization within the appropriate study time window are permanently discontinued from the 
study and will have no further evaluations conducted upon discontinuation. Participants who 
remain eligible but elect to prematurely discontinue form the study will have no additional 
evaluations performed from the point at which they discontinue. Participants who experience a 
grade 3 or 4 Adverse Event related to the vaccine or study procedures prior to the second 
immunization will be discontinued from the study and will return for evaluation at week 6 and/or 
7 to ensure resolution but will not provide specimens for immunologic evaluation.  
 
6.3 Instructions for Evaluations 
6.3.1 
Medical & Medication History 
At screening, a standard medical history and medication history will be obtained and 
documented.  Specifically, information will be obtained regarding: 
• 
Known allergy/sensitivity or any hypersensitivity to latex or any of the components of the 
study product or its formulation (see section 5.2 for a list of components). 
• 
Serious illness or injury requiring hospitalization within 21 days prior to study entry. 
• 
A history of a medical condition resulting in impaired immunity (including complement 
deficiency, antibody deficiency, chronic granulomatous disease, or HIV infection).  
• 
Autoimmune disorders; mild autoimmune disorders such as eczema is not exclusionary 
after assessment by the investigator. 
• 
Known active infection with HIV, HCV, or HBV. 
• 
History of all cancers including leukemia or lymphoma  
• 
History of excessive alcohol consumption, drug abuse, psychiatric conditions, social 
conditions, or occupational conditions that in the opinion of the investigator would 
preclude compliance with the study. 
• 
Hemophilia or other bleeding diatheses. 
• 
Have donated blood or blood products within 30 days before study vaccination, plan to 
donate blood at any time during the duration of subject study participation, or plan to 
donate blood within 30 days after the last blood draw. 
• 
Pregnancy in women and nursing mothers or women who are planning to become 
pregnant within the study duration. 
• 
Prior history of gonorrhea, chlamydia, or syphillis 
The medication history will include inquiry and documentation of all medications used within the 
last 60 days and the participants will be asked specifically about the use of the following agents: 
• 
anticoagulants other than aspirin or nonsteroidal anti-inflammatory drugs (NSAIDS) 
within 14 days prior to study entry. 
• 
any systemic antineoplastic or immunomodulatory treatment, systemic corticosteroids, 
(inhaled or topical corticosteroids acceptable), investigational products, interleukins, 
interferons, growth factors, or intravenous immunoglobulin (IVIG) within 45 days prior 
to study entry. 
The medication history will also include inquiry and documentation of vaccine history and the 
participants will be asked specifically about the use of the following: 
• 
Previous immunization with a MenB vaccine 
• 
Have received any licensed vaccine within 30 days prior to study vaccination. 
• 
Have planned vaccination with any vaccine within  30 days of the anticipated date of 
second study immunization (approximately 65 days from study entry). 

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6.3.2 
Targeted Physical Exam & Vital Signs 
A physical exam performed at each visit will include at a minimum an examination of the skin, 
head, mouth, and neck; auscultation of the chest; cardiac exam; abdominal exam; and 
examination of the lower extremities for edema. A targeted physical exam and vital signs (oral 
temperature, pulse, respiration rate, and blood pressure) including weight will be collected at each 
visit.   
 
6.3.3 
Laboratory Evaluations 
 
Pregnancy Test for females of child bearing potential  
A point of care pregnancy test is performed and results need to be recorded only in source 
document at screening visit, at entry, and at week 5 visit prior to vaccine administration.  
 
6.3.4 
Immunologic Studies 
Stored PBMCs 
PBMCs for cell-surface marker phenotyping, N. gonorrhoeae-antigen stimulated cytokine 
production, and B-cell receptor cloning will be isolated and stored per the SOE (Schedule of 
Events). Cryopreserved PBMC will be sent to Duke Immune Profiling Core at Duke University 
for flow cytometry and cellular immune analysis; the shipped samples will have an identifying 
sample number (code) derived from the participant’s assigned PID and the visit number; no 
identifying information on the participant’s samples will be transmitted to the Duke Immune 
Profiling Core and leftover samples wil be destroyed or returned to UNC for storage if the 
participant has agreed to storage of their study specimens. The PI at UNC will keep a master log 
which links the codes on the stored specimens to the participants. Leftover PBMC from 
consenting participants may be used for future N. gonorrhoeae research.  
 
N. gonorrhoeae reactive antibody studies/Stored Serum 
Pharyngeal and vaginal swabs, urine, and serum for measures of N. gonorrhoeae-directed 
antibodies will be collected per the SOE. Antibodies eluted from swabs or present in urine and 
present in serum from samples collected at entry, week 5, week 6, and week 7 will be tested for 
N. gonorrhoeae antigen-specific antibodies using ELISA. Antibody containing samples will be 
sent to Regional Biocontainment Laboratory (RBL) Immunology Unit at Duke University for 
ELISA analysis. The shipped samples will have an identifying sample number (code) derived 
from the participant’s assigned PID and the visit number; no identifying information on the 
participant’s samples will be transmitted to the Duke RBL Immunology Unit and leftover 
samples will be destroyed or returned to UNC for storage if the participant has agreed to storage 
of their study specimens. The PI at UNC will keep a master log which links the codes on the 
stored specimens to the participants. Leftover sera from consenting participants may be used for 
future N. gonorrhoeae research. 
 
6.3.5 
Vaccine/ Injection 
Every effort should be made to administer vaccines within the protocol-defined visit window. 
Participants who do not receive their first vaccine within 30 days of screening or who do not 
receive the week 5 vaccine will be prematurely discontinued from the study.  
 
The week 5 visit should be conducted 35 days (± 7 days) after the first vaccination (enrollment 
visit). If the week 5 visit is not within the protocol-specified window, then the vaccine can be 
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The week 6 visit should occur 5-9 days after the second dose of vaccine; if week 6 visit samples 
are not collected, the participant may continue in the study and attend the week 7 visit 12-16 days 
after receiving the second dose of vaccine. If the week 7 visit is not collected between day 12-16 
after the second dose of vaccine, participant will be discontinued from the study.  
 
Vaccination should be deferred at the discretion of the site investigator if the participant is 
febrile with reported or observed temperature greater than or equal to 38 °C or acutely ill 
with symptoms (ongoing or present within 7 days of planned vaccination) consistent with 
infection of the respiratory tract, gastrointenstinal tract, urinary tract, or skin/soft tissues. 
 
Participants will remain at the clinic for a 15-minute post-injection observation period following 
each vaccination to observe for syncope or any immediate adverse reactions. Participants will be 
provided instructions on self-assessment of adverse reactions related to vaccination.  
 
6.3.6 
Telephone Contact  
The study staff will contact each participant by telephone 48 to 72 hours after each vaccination at 
the entry visit and at the week 5 visit to determine whether any side effects have occurred. 
Information from the telephone contact will be recorded on the electronic case report form 
(eCRF). If a participant experiences any side effects that are Grade 3 or 4, including edema 
(swelling), erythema (redness), or induration of the injection site, the participant should be 
scheduled to come into the research clinic for evaluation, including a targeted physical exam 
focused on the area involved, as soon as possible.  
 
NOTE: If telephone contact is not possible, then in person is permissible.   

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7.0 
CLINICAL MANAGEMENT ISSUES 
7.1 
Dose Modification 
No modification of the study vaccine dose is allowed. 
 
7.2 
Toxicity Management 
7.2.1 
Management of Injection Site and Allergic Reactions 
Local and systemic reactions will be graded according to the DAIDS Adverse Event (AE) 
Grading Table, Corrected Version 2.1, July 2017, found on the DAIDS RSC Web site: 
http://rsc.tech-res.com/safetyandpharmacovigilance/.  
 
Injection Site Reactions 
• 
Swelling, induration, or redness at the injection site 
• 
Pain or tenderness, or other reactions at the injection site 
 
Systemic reactions occurring within 48 hours after vaccination 
• 
Fever or chills  
• 
Malaise or fatigue  
• 
Headache or pain, e.g., myalgia or arthralgia  
• 
Nausea or vomiting 
• 
Allergic reactions, e.g., rash, hives, dyspnea, pruritis 
 
For any Grade 3 or 4 reactions thought definitely, possibly, or probably related to vaccination, 
further vaccines should not be given to that participant. The participant will be asked to return to 
the site for an evaluation and followed until AE has been resolved.  
 
7.2.2 
Other Adverse Events 
For toxicities not specifically addressed above, the following guidelines should be used for the 
management of AEs that are felt to be at least possibly related to vaccines: 
Grade 1 or 2 Toxicity/AE: 
Participants who develop a Grade 1 or 2 AE may continue in the study and receive the second 
dose of vaccine (which may be postponed as directed in section 6.3.5) and return for evaluation 
study visits as noted in section 6.2.3. For participants experiencing Grade 1 or 2 AEs who choose 
to discontinue study treatment or study participation prior to receiving the second vaccine dose, 
see section 6.2.4. 
Grade 3 or 4 Toxicity/AE: 
Participants who develop a Grade 3 or 4 AE that is at least possibly related to vaccine should be 
re-evaluated for that toxicity until the AE returns to Grade ≤2. Participants who experience Grade 
3 or 4 AE that is at least possibly related to vaccine should not receive the second immunization if 
the AE occurs prior to the week 5 study visit and their participation in the study should be 
discontinued. Participants who experience a grade 3 or 4 AE that is considered by the investigator 
to be unrelated to vaccine or that occurs after the second immunization may continue in the study 
and study specimens may be obtained for week 6 and 7 visits if the AE has returned to Grade ≤2. 
For instructions regarding participants experiencing Grade 3 or 4 AEs that are at least possibly 
related to vaccine that result in premature discontinuation of study vaccine and/or study 
participation, see section 6.2.4. These participants should be followed until resolution of the AE 
Toxicity. 
 
7.3 
Pregnancy 
Pregnant participants are discontinued from the study at the time pregnancy is discovered.   

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8.0 
ADVERSE EVENT REPORTING 
8.1 
Definitions 
 
An adverse event (AE) is any untoward medical occurrence in a patient or clinical investigation 
participant administered a pharmaceutical product and which does not necessarily have a causal 
relationship with this treatment.  An adverse event can therefore be any unfavorable and unintended 
sign (including an abnormal laboratory finding), symptom, or disease temporally associated with 
the use of a medicinal (investigational) product, whether or not related to the medicinal 
(investigational) product. 
 
An unexpected (unlisted) adverse event is an adverse reaction, the nature or severity of which is 
not consistent with the applicable product information (e.g., Investigator’s Brochure for an 
unapproved investigational product or package insert/summary of product characteristics for an 
approved product). 
 
A serious adverse event (SAE) is any untoward medical occurrence that: 
• 
results in death; 
• 
is life-threatening; 
• 
requires inpatient hospitalization or prolongation of existing hospitalization; 
• 
results in a persistent or significant disability/incapacity; 
 
or 
• 
 is a congenital anomaly/birth defect 
Other important medical events as assessed by medical and scientific judgment may also be 
considered SAEs by the investigator and should be reported in an expedited fashion.  
 
An Unanticipated Problem (UP)/New Safety Information (NSI) is defined by the UNC IRB as 
any incident, experience, or outcome that: 
• 
is unexpected (in terms of nature, severity, or frequency) given (a) the research 
procedures that are described in the protocol-related documents, such as the IRB-
approved research protocol and informed consent document; and (b) the characteristics of 
the participant population being studied;  
• 
is related or possibly related to a participant’s participation in the research; and  
• 
suggests that the research places participants or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) related to the research than was 
previously known or recognized.  
 
AE attribution will be assigned according to the following scale:  
• 
Not related:  The AE is clearly not related to the study procedures (i.e., another cause of 
the event is most plausible and/or a clinically plausible temporal sequence is inconsistent 
with the onset of the event).   
• 
Possibly related:  An event that follows a reasonable temporal sequence from the 
initiation of study procedures, but that could readily have been produced by a number of 
other factors. 
• 
Related:  The AE is clearly related to the study procedures.   
 

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8.2 
Reporting 
The protocol-defined expedited event reporting period for this protocol is the entire study 
duration for an individual participant (from study enrollment until study completion or 
discontinuation of the participant from study participation for any reason). 
 
All AEs defined below must be recorded on the eCRFs if any of the following criteria have been 
met: 
• 
All grade >3 AEs 
• 
All AEs that led to a change in study treatment/intervention regardless of grade 
• 
All AES meeting SAE definition 
 
For all of these adverse events reported or observed, the information should be recorded in the 
electronic Case Report Form (eCRF) for that participant and will include a full description of the 
event, its seriousness, its severity or toxicity grade (intensity), the relationship to the study drug, 
and the treatment, outcome, and sequelae of the event.  
 
Any serious or unexpected event, which occurs to any participant in the course of his/her treatment 
on this study or within 30 days following cessation of treatment, must be reported within 24 hours 
to the principal investigator.  The immediate reports should be followed promptly by detailed, 
written reports.  Copies of these reports should be sent to: 
• 
UNC IRB if it meets the UNC IRB UP/NSI reporting requirements 
• 
Events that are reportable to the UNC IRB will also be submitted to the appropriate 
representative of the study funding source/sponsor (NC TraCS): Marie Rape, RN, BSN, 
Associate Director of Regulatory, NC TraCS (marie_rape@med.unc.edu)   
  
Reporting is required of all unanticipated problems/new safety information to the UNC IRB, 
including those which may occur after the participant has completed or has withdrawn from the 
study, including after study closure.  
 
Clinically significant AEs and other problems related to vaccines will also be reported to the 
Vaccine Adverse Event Reporting System (VAERS), which is maintained by the FDA and CDC 
(http://vaers.hhs.gov/). 
 
All individual AE shall be maintained by the Principal Investigator.  For those events that are not 
reportable to the UNC IRB, a summary (i.e., not individual reports) of all adverse events that 
have occurred within the last approval period will be submitted to the UNC IRB at the time of 
continuing review. 
 
9.0 
STATISTICAL CONSIDERATIONS 
9.1 
General Design Issues 
This study is a single center, single arm interventional pilot study in which participants will 
receive two-doses of the US FDA-approved 4CMenB vaccine according to recommended 
administration schedule. Blood and mucosal surface specimens will be obtained at 4 study visits. 
The study population will include 15 individuals aged 18-25 years desiring immunization against 
group B N. meningitidis with no contraindication to vaccination and no known immune 
compromising medical condition or medication. Enrolled individuals who are discontinued will 
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9.2 
Endpoints 
9.2.1 
Primary Endpoints: 
• 
Geometric mean concentration of IgG binding to N. gonorrhoeae strain FA1090 surface 
antigens (Outer-membrane vesicles) in human serum at entry visit (prior to first 
vaccination), and week 7 visit (2 weeks after second vaccination) determined by ELISA 
• 
Geometric mean concentration of IgM binding to N. gonorrhoeae strain FA1090 surface 
antigens (Outer-membrane vesicles) in human serum at entry visit (prior to first 
vaccination), and week 7 visit (2 weeks after second vaccination) determined by ELISA 
• 
Geometric mean concentration of IgA binding to N. gonorrhoeae strain FA1090 surface 
antigens (Outer-membrane vesicles) in human serum at entry visit (prior to first 
vaccination), and week 7 visit (2 weeks after second vaccination) determined by ELISA 
• 
Frequency of CD4+  T cells expressing at least two different activation markers (IFN-g, 
TNF-a, IL-2, and CD107a) after in vitro stimulation with N. gonorrhoeae strain FA1090 
Outer-membrane vesicles in circulating PBMC obtained at entry visit (prior to first 
vaccination) and week 7 visit (2 weeks after second vaccination) determined by flow 
cytometry 
 
9.2.2  Exploratory Endpoints: 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (Outer-membrane vesicles) in human pharyngeal secretions at 
entry visit (prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), 
week 6 visit (1 week after second vaccination), and week 7 visit (2 weeks after second 
vaccination) determined by ELISA 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (Outer-membrane vesicles) in human vaginal secretions (female 
participants) at entry visit (prior to first vaccination), week 5 visit (4-6 weeks after first 
vaccination), week 6 visit (1 week after second vaccination), and week 7 visit (2 weeks 
after second vaccination) determined by ELISA 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (Outer-membrane vesicles)  in human urine (male participants) 
at entry visit (prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), 
week 6 visit (1 week after second vaccination), and week 7 visit (2 weeks after second 
vaccination) determined by ELISA 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (PorB, and MtrE) in human serum at entry visit (prior to first 
vaccination), week 5 visit (4-6 weeks after first vaccination), week 6 visit (1 week after 
second vaccination), and week 7 visit (2 weeks after second vaccination) determined by 
ELISA 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (PorB, and MtrE) in human pharyngeal secretions at entry visit 
(prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), week 6 visit (1 
week after second vaccination), and week 7 visit (2 weeks after second 
vaccination) determined by ELISA 
• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (PorB, and MtrE) in human vaginal secretions (female 
participants) at entry visit (prior to first vaccination), week 5 visit (4-6 weeks after first 
vaccination), week 6 visit (1 week after second vaccination), and week 7 visit (2 weeks 
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• 
Geometric mean concentration of IgM, IgG, and IgA binding to N. gonorrhoeae strain 
FA1090 surface antigens (PorB, and MtrE) in human urine (male participants) at entry 
visit (prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), week 6 
visit (1 week after second vaccination), and week 7 visit (2 weeks after second 
vaccination) determined by ELISA 
• 
Qualitative assessment of the presence of human IgG directed against N. gonorrhoeae 
strain FA1090 antigens (Outer-membrane vesicles, PorB, and MtrE) in human serum at 
entry visit (prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), 
week 6 visit (1 week after second vaccination), and week 7 visit (2 weeks after second 
vaccination) determined by immunoblot. 
• 
Frequencies of CD8+ T cells expressing functional IFN-g, TNF-a, IL-2, and CD107a 
after in vitro stimulation with N. gonorrhoeae strain FA1090 Outer-membrane vesicles in 
circulating PBMC obtained at entry visit (prior to first vaccination), week 5 visit (4-6 
weeks after first vaccination), week 6 visit (1 week after second vaccination),  and week 
7 visit (2 weeks after second vaccination) determined by flow cytometry 
• 
Frequencies of memory, activated, and proliferating CD4+ Tfh cells specific in 
circulating PBMC obtained at entry visit (prior to first vaccination), week 5 visit (4-6 
weeks after first vaccination), week 6 visit (1 week after second vaccination), and week 7 
visit (2 weeks after second vaccination) determined by flow cytometry 
• 
The quantity of IFN-g and IL-17 secreted after in vitro stimulation with N. gonorrhoeae 
strain FA1090 Outer-membrane vesicles from circulating PBMC obtained at entry visit 
(prior to first vaccination), week 5 visit (4-6 weeks after first vaccination), week 6 visit (1 
week after second vaccination), and week 7 visit (2 weeks after second vaccination) 
determined by Bio-Plex Multiplex immunoassay 
• 
The sequence of B-cell receptors found to associate with FA1090 Outer-membrane 
vesicles through flow cytometry-based cell sorting will be determined using sequencing 
of PCR amplicons generated using primers for non-variable regions of the B cell receptor  
 
 
9.3 
Sample Size and Accrual 
Due to the majority of the target population likely to be undergraduate students, we anticipate that 
enrollment will be limited to ~1 participant each month in July and August (months when student 
population residing in Chapel Hill is low) and that the remaining participants will be enrolled at 
~6/month in September and October. All Serum and PBMC will be stored until a complete 
specimen set is obtained from at least 8 subjects; the serum and PBMC specimens will then be 
transferred to Dr. Andrew MacIntyre’s laboratory at Duke University in batch for immunologic 
assays. We plan to enroll up to 15 subjects so we can obtain a complete specimen set at all 4 
study visits for at least 8 subjects.      
 
9.4 
Monitoring 
Oversight of the trial is provided by the Principal Investigator (PI).  The PI will be responsible for 
safety monitoring throughout the study. The PI will assure that informed consent is obtained prior 
to performing any research procedures, that all subjects meet eligibility criteria, and that the study 
is conducted according to the IRB-approved research plan.  Study data are accessible at all times 
for the PI to review. The PI will review study conduct   including: accrual, drop-outs, protocol 
deviations after 5 participants are enrolled and complete the study protocol and then on an annual 
basis until study is complete.  The PI reviews AEs individually in real-time and in aggregate after 
5 individuals have completed the protocol and then annually until study is complete.  The PI 
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section 8.0 in real-time. The PI ensures all protocol deviations, AEs, and SAEs are reported to the 
NC TraCS and the IRB according to the applicable reporting requirements. 
 
9.5. Analyses 
9.5.1 
Statistical Methods: 
Descriptive statistics such as age and gender will be reported in means, standard deviations, 
frequencies, and proportions when appropriate. As recently recommended for pilot studies, ours 
will focus on point estimates of immuse responses  and their 80% confidence intervals (CIs) in 
place of 95% CI used for traditional statistical inferences26 .The geometric mean concentration 
(anti-log of the arithmetic mean of log transformed concentrations) of antibodies while the 
arithmetic mean of T cell population frequencies and secreted cytokine levels from in vitro 
antigen-stimulated PBMC will be identified.  Comparisons between measurements of immune 
response in pre- and post-immunization specimens (as specified for primary outcome 
measurements) will be completed using paired Student's t-test for normally distributed data or 
Wilcoxon rank signed test for non-normally distributed data.  For repeated outcome 
measurements at 4 time points (as specified in exploratory outcomes), we will use generalized 
estimating equations (GEE) with unstructured covariance matrix to estimate the time difference. 
The link function for GEE will be chosen depending on the data type.  
 
9.5.2 
Sample Size and Power: 
There is only one published report describing the anti-gonococcal antibody titers in 10 humans 
who received 3 immunizations with 4CMenB. The geometric mean titer of serum antibodies 
directed against whole cell N. gonorrhoeae (strain 1291) demonstrated a statistically significant 
increase between pre (95% CI : 25,906–90,811) and post (95% CI : 49,228–126,115) 
immunization samples15. We do not have preliminary data and there is no published data 
regarding cross- reactive antibodies in 4CMenB-immunized humans against the N. gonorrhoeae 
strain FA1090 (the strain used by our center in both human challenge model and preclinical 
laboratory studies) or specific FA1090 outer-membrane antigens. Further, there is no published 
data on anti-gonococcal antibody subtype or levels of these antibodies in mucosal surface 
secretions in any 4CMenB-immunized humans. Finally, there is no published data regarding 
cellular immune responses to gonococcal antigens in 4CMenB-immunized individuals. Thus, the 
proposed study is a pilot study and the power to detect significant immune responses towards N. 
gonorrhoeae antigens is unknown. The study will provide baseline values and variability in 
measurements for non-immune individuals as well as post immunization measurements for the 
studied immunologic parameters. We will enroll up to 15 individuals, including similar numbers 
of men and women; this sample size is proposed based on size of potential participant pool and 
available funding for the study. The data generated will provide valuable information for how to 
deploy these assays in human populations receiving 4CMenB immunization for testing efficacy in 
preventing N. gonorrhoeae infection in the future.  
 
  
10.0 
STUDY MANAGEMENT 
10.1 
Institutional Review Board (IRB) Review and Informed Consent 
This protocol and the informed consent document and any subsequent modifications 
(amendments) will be reviewed and approved by the IRB or ethics committee responsible for 
oversight of the study.  
 
It is expected that the IRB will have the proper representation and function in accordance with 
federally mandated regulations.  The IRB should approve the consent form(s) and protocol. 

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In obtaining and documenting informed consent, the principal investigator should comply with the 
applicable regulatory requirement(s), and should adhere to Good Clinical Practice (GCP) and to 
ethical principles that have their origin in the Declaration of Helsinki. 
 
Before recruitment and enrollment onto this study, the participant will be given a full explanation 
of the study and will be given the opportunity to review the consent form(s). Each consent form 
must include all the relevant elements currently required by the FDA Regulations and local or 
state regulations, which include elements such as the purpose of the study, the procedures to be 
followed, and the risks and benefits of participation.  Once this essential information has been 
provided to the participant and the investigator is assured that the participant understands the 
implications of participating in the study, the participant will be asked to give consent to 
participate in the study by signing an IRB-approved consent form. 
 
Prior to a participant’s participation in the trial, the written informed consent form(s) should be 
signed and personally dated by the participant or the participant’s legally authorized representative 
(legal guardian or person with power of attorney for participants who cannot consent for 
themselves), and by the person who conducted the informed consent discussion. 
 
A copy of the consent form will be given to the participant or legal guardian, and this fact will be 
documented in the participant’s record. 
 
10.2  
Registration Procedures 
All participants must be registered with the Global HIV Prevention and Treatment Clinical Trials 
Unit Office at the University of North Carolina before enrollment onto study.  Prior to registration, 
eligibility criteria must be confirmed with the Study Coordinator.   
 
Each participant will be assigned a participant identification (PID) number prior to study entry.  
Therefore, all participants are identified by a PID number and not by name on all case report forms.  
 
10.3 
Criteria for Discontinuation 
Removal of participants from study can occur for the following reasons: 
 
 
 
10.3.1 Premature Treatment Discontinuation 
• 
Treatment-related AEs as discussed in section 7.2.  
• 
Pregnancy or breastfeeding. 
• 
Clinical reasons believed life-threatening by the site principal investigator, even if not 
addressed in section 7.2. 
 
10.3.2 Premature Study Discontinuation 
• 
Request by the participant to discontinue treatment prior to receiving second immunization 
or to withdraw from study. 
• 
Request of the primary care provider if s/he thinks the study is no longer in the best interest 
of the participant. 
• 
Failure to start study vaccination within 30 days after screening or to receive second 
vaccination within 8 weeks of initial dose. 
• 
Failure to attend week 6 or week 7 study visits within the indicated time window. 

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• 
At the discretion of the IRB, NC TraCS, industry supporter, FDA, Office for Human 
Research Protections (OHRP), investigator, or other country-specific government agencies 
as part of their duties to ensure that research participants are protected.  
 
10.4 
Deviations from the Protocol 
The investigator may implement a deviation from, or a change of, the protocol to eliminate an 
immediate hazard(s) to study participants without prior IRB approval.  As soon as possible, the 
implemented deviation or change, the reasons for it, and, if appropriate, the proposed protocol 
amendment(s) should be submitted to: 
 
a) IRB for review and approval 
b) to the sponsor for agreement and, if required 
c) to the appropriate regulatory authority(ies) 
 
10.5 
Clinical Site Monitoring and Record Availability 
Personnel from the Global HIV Prevention and Treatment Clinical Trials Unit at the University of 
North Carolina (UNC) will monitor the trial for quality assurance.  These monitors will review all 
source documents used in the preparation of the case report forms to assure proper conduct of the 
trial and proper collection of the data.   
 
10.6 
Required Documentation 
Before the study can be initiated, the following documentation must be provided to the IGHID 
Regulatory Office at the University of North Carolina. 
 
A copy of the official IRB approval letter for the protocol and informed consent(s) 
 
IRB membership list 
 
CVs and medical licensure for the principal investigator and any associate investigators who 
will be involved in the study 
 
A copy of the IRB-approved consent form(s) 
 
10.7 
Adherence to the Protocol 
Except for an emergency situation in which proper care for the protection, safety, and well-being 
of the study participant requires alternative treatment, the study shall be conducted exactly as 
described in the approved protocol.  Any deviation from the protocol must have prior approval by 
the Principal Investigator and must be recorded and explained. 
 
10.8 
Amendments to the Protocol 
Should amendments to the protocol be required, the amendments will be originated and 
documented by the Principal Investigator.  The written amendment will be approved by the 
principal investigator and must be submitted to the IRB at the principal investigator's site for  
approval.  It should also be noted that when an amendment to the protocol substantially alters the 
study design or the potential risk to the participant, a revised consent form might be required. 
 
10.9 
Data Management  
Data to be obtained during the study includes directed medical history provided by participant self-
report, personally identifying information (including name, contact information, results of directed 
physical examination, and point of care pregnancy tests administered to female participants, as well 
as results of research tests to monitor immunologic response to immunization.  Medical history and 
personal identification information will be recorded with identifiers and kept in participants’ 
research charts. After written informed consent is provided, consecutive study code numbers will 

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be assigned to the participants to ensure confidentiality. This study code number will be used to 
identify research specimens and the results of research tests.  At the end of each individual’s 
participation in the trial, research charts will be reviewed by the study coordinator and stored in 
locked cabinets in the PI’s office. At the completion of the study, the research charts and all 
identifying information related to participants will be archived in an off-site storage facility. 
 
De-identified data entered into computerized files will be stored in a password-protected, encrypted 
file on a UNC-owned server with access provided only to authorized personnel directly involved 
with the study.  Only authorized personnel directly involved with the study will have access to 
individually identifiable private information about participants.  All data entered into the study 
database will be coded and unlinked to personal identifiers.  Data from the trial to be stored 
electronically or shared will be unlinked to all identifiable information. 
 
10.10 Record Retention 
Study documentation includes all Case Report Forms, data correction forms or queries, source 
documents, Sponsor-Investigator correspondence, monitoring logs/letters, and regulatory 
documents (e.g., protocol and amendments, IRB correspondence and approval, signed participant 
consent forms). 
 
Source documents include all recordings of observations or notations of clinical activities and all 
reports and records necessary for the evaluation and reconstruction of the clinical research study. 
 
Government agency regulations and directives require that all study documentation pertaining to 
the conduct of a clinical trial must be retained by the study principal investigator.  Study documents 
will be kept on file until three years after the completion and final study report of this investigational 
study. 
 
10.11   Obligations of Investigators 
The Principal Investigator is responsible for the conduct of the clinical trial at the site in accordance 
with Title 21 of the Code of Federal Regulations and/or the Declaration of Helsinki.  The Principal 
Investigator is responsible for personally overseeing the treatment of all study participants.  The 
Principal Investigator must assure that all study site personnel, including co-investigators and other 
study staff members, adhere to the study protocol and all FDA/GCP regulations and guidelines 
regarding clinical trials both during and after study completion. 
 
The Principal Investigator will be responsible for assuring that all the required data will be 
collected and entered onto the Case Report Forms (CRFs). At the completion of the study, all 
case report forms will be reviewed by the Principal Investigator and will require his/her final 
signature to verify the accuracy of the data. 
 
The site principal investigator will make study documents (e.g., consent forms, drug distribution 
forms, CRFs) and pertinent hospital or clinic records readily available for inspection by the local 
IRB, the site monitors, the FDA, the NCATS, the OHRP, or NC TraCS for confirmation of the 
study data.  
 
10.12 Participant Confidentiality 
All laboratory specimens and accompanying evaluation forms, reports, and other records that are 
sent to Duke University for immunologic analysis will be de-identified to maintain participant 
confidentiality.  All records will be kept locked in secure areas/secure offices at UNC.  All 

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computer entry and networking programs at UNC will be done with coded numbers only.  
Clinical information will not be released without written permission of the participant, except as 
necessary for monitoring by the IRB, the NCATS, the OHRP, or NC TraCS. 
 
10.13 Study Discontinuation 
The study may be discontinued at any time by the IRB, NCATS, the OHRP, NC TraCS or other 
government agencies as part of their duties to ensure that research participants are protected.  
 
11.0 
BIOHAZARD CONTAINMENT 
As the transmission of HIV and other blood-borne pathogens can occur through contact with 
contaminated needles, blood, and blood products, appropriate blood and secretion precautions 
will be employed by all personnel in the drawing of blood and shipping and handling of all 
specimens for this study, as currently recommended by the Centers for Disease Control and 
Prevention and the NIH. 
 
All dangerous goods materials, including diagnostic specimens and infectious substances, must 
be transported using packaging mandated by CFR 42 Part 72. Please refer to instructions detailed 
in the International Air Transport Association (IATA) Dangerous Goods Regulations.  
 
 
12.0 
REFERENCES 
 
1 
Tapsall, J. W. Antibiotic resistance in Neisseria gonorrhoeae. Clin Infect Dis 41 Suppl 4, S263-268, 
doi:10.1086/430787 (2005). 
2 
Unemo, M. et al. High-level cefixime- and ceftriaxone-resistant Neisseria gonorrhoeae in France: 
novel penA mosaic allele in a successful international clone causes treatment failure. Antimicrob 
Agents Chemother 56, 1273-1280, doi:10.1128/AAC.05760-11 (2012). 
3 
European Centre for Disease Prevention and Control. Extensively drug-resistant (XDR) Neisseria 
gonorrhoeae in the United Kingdom and Australia. (2018). 
4 
Fox, K. K. et al. Longitudinal evaluation of serovar-specific immunity to Neisseria gonorrhoeae. 
Am J Epidemiol 149, 353-358 (1999). 
5 
Schmidt, K. A. et al. Experimental gonococcal urethritis and reinfection with homologous 
gonococci in male volunteers. Sex Transm Dis 28, 555-564 (2001). 
6 
Finne, J., Leinonen, M. & Makela, P. H. Antigenic similarities between brain components and 
bacteria causing meningitis. Implications for vaccine development and pathogenesis. Lancet 2, 
355-357 (1983). 
7 
Naess, L. M. et al. Human IgG subclass responses in relation to serum bactericidal and opsonic 
activities after immunization with three doses of the Norwegian serogroup B meningococcal 
outer membrane vesicle vaccine. Vaccine 17, 754-764 (1999). 
8 
Peeters, C. C. et al. Phase I clinical trial with a hexavalent PorA containing meningococcal outer 
membrane vesicle vaccine. Vaccine 14, 1009-1015 (1996). 
9 
de Moraes, J. C. et al. Protective efficacy of a serogroup B meningococcal vaccine in Sao Paulo, 
Brazil. Lancet 340, 1074-1078 (1992). 
10 
Holst, J. et al. Properties and clinical performance of vaccines containing outer membrane 
vesicles from Neisseria meningitidis. Vaccine 27 Suppl 2, B3-12, 
doi:10.1016/j.vaccine.2009.04.071 (2009). 

IGHID # 11911 
PI:  Joseph A. Duncan, MD, PhD 
Version 1.2/June 24, 2020 
 
Page 27 
 
 
 
11 
Nokleby, H. et al. Safety review: two outer membrane vesicle (OMV) vaccines against systemic 
Neisseria meningitidis serogroup B disease. Vaccine 25, 3080-3084, 
doi:10.1016/j.vaccine.2007.01.022 (2007). 
12 
Pérez, O. et al. Mucosal approaches in Neisseria Vaccinology.  (2009). 
13 
Whelan, J., Klovstad, H., Haugen, I. L., Holle, M. R. & Storsaeter, J. Ecologic Study of 
Meningococcal B Vaccine and Neisseria gonorrhoeae Infection, Norway. Emerg Infect Dis 22, 
1137-1139, doi:10.3201/eid2206.151093 (2016). 
14 
Petousis-Harris, H. et al. Effectiveness of a group B outer membrane vesicle meningococcal 
vaccine against gonorrhoea in New Zealand: a retrospective case-control study. Lancet 390, 
1603-1610, doi:10.1016/S0140-6736(17)31449-6 (2017). 
15 
Semchenko, E. A., Tan, A., Borrow, R. & Seib, K. L. The serogroup B meningococcal vaccine 
Bexsero elicits antibodies to Neisseria gonorrhoeae. Clin Infect Dis, doi:10.1093/cid/ciy1061 
(2018). 
16 
Goldschneider, I., Gotschlich, E. C. & Artenstein, M. S. Human immunity to the meningococcus. I. 
The role of humoral antibodies. J Exp Med 129, 1307-1326 (1969). 
17 
Goldschneider, I., Gotschlich, E. C. & Artenstein, M. S. Human immunity to the meningococcus. 
II. Development of natural immunity. J Exp Med 129, 1327-1348 (1969). 
18 
Gotschlich, E. C., Rey, M., Triau, R. & Sparks, K. J. Quantitative determination of the human 
immune response to immunization with meningococcal vaccines. J Clin Invest 51, 89-96, 
doi:10.1172/JCI106801 (1972). 
19 
Hedges, S. R., Mayo, M. S., Mestecky, J., Hook, E. W., 3rd & Russell, M. W. Limited local and 
systemic antibody responses to Neisseria gonorrhoeae during uncomplicated genital infections. 
Infect Immun 67, 3937-3946 (1999). 
20 
Simpson, S. D., Ho, Y., Rice, P. A. & Wetzler, L. M. T lymphocyte response to Neisseria 
gonorrhoeae porin in individuals with mucosal gonococcal infections. J Infect Dis 180, 762-773, 
doi:10.1086/314969 (1999). 
21 
Feinen, B., Jerse, A. E., Gaffen, S. L. & Russell, M. W. Critical role of Th17 responses in a murine 
model of Neisseria gonorrhoeae genital infection. Mucosal Immunol 3, 312-321, 
doi:10.1038/mi.2009.139 (2010). 
22 
Liu, Y. & Russell, M. W. Diversion of the Immune Response to Neisseria gonorrhoeae from Th17 
to Th1/Th2 by Treatment with Anti-Transforming Growth Factor {beta} Antibody Generates 
Immunological Memory and Protective Immunity. MBio 2, doi:10.1128/mBio.00095-11 (2011). 
23 
Gagliardi, M. C. et al. Circulating levels of interleukin-17A and interleukin-23 are increased in 
patients with gonococcal infection. FEMS Immunol Med Microbiol 61, 129-132, 
doi:10.1111/j.1574-695X.2010.00759.x (2011). 
24 
Walker, T. Y. et al. National, Regional, State, and Selected Local Area Vaccination Coverage 
Among Adolescents Aged 13-17 Years - United States, 2017. MMWR Morb Mortal Wkly Rep 67, 
909-917, doi:10.15585/mmwr.mm6733a1 (2018). 
25 
Soeters, H. M. et al. University-Based Outbreaks of Meningococcal Disease Caused by Serogroup 
B, United States, 2013-2018. Emerg Infect Dis 25, 434-440, doi:10.3201/eid2503.181574 (2019). 
26 
Lee, E. C., Whitehead, A. L., Jacques, R. M. & Julious, S. A. The statistical interpretation of pilot 
trials: should significance thresholds be reconsidered? BMC Med Res Methodol 14, 41, 
doi:10.1186/1471-2288-14-41 (2014).