Clinical trial NCT03724357
Conditions: Cholera Vaccination
Sponsor: University of Maryland, Baltimore
Start date: 2018-11-04; Completion date: 2021-10-28
Enrollment: 3 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
This is an open-label, non-randomized study. Volunteers will be vaccinated with the oral cholera vaccine, Vaxchora. Vaxchora has been licensed by the Food and Drug Administration (FDA) for travelers to developing countries. Volunteers will also be asked to provide blood specimens over a follow-up time period of up to eight years. The specimens obtained in this clinical research study will be used to further the investigator's understanding of the protective immunological mechanisms that can be elicited systemically and may be applicable to other enteric pathogens.

Detailed description:

Arms: arm 1: Volunteers receive immunization with Vaxchora cholera vaccine. Blood draws are performed at subsequent visits.
Interventions: intervention 1: Vaxchora
Intervention descriptions: intervention 1: One dose. Approximately 100 mL of cold or room temperature purified bottled water is placed into a clean disposable cup. The contents of the buffer sachet are added to the water and stirred. Then the contents of the active (lyophilized vaccine) sachet are added to the water and stirred for approximately 30 seconds. The reconstituted mixture should be completely ingested within 15 minutes.

Protocol documents:
Prot_SAP_001.pdf: 
Blood Donor 9000 
 
v1.0; 26 October 2018 
1 
 
Protocol Title: Collection of Blood for In Vitro Studies from Healthy Adults Who Have Received 
Oral Cholera Vaccine (CVD 103-HgR) 
Protocol Number: Blood Donor CVD 9000 
 
Sponsored by: 
Center for Vaccine Development 
University of Maryland School of Medicine 
Baltimore, MD 
 
Principal Investigator: 
Monica A. McArthur, MD, PhD 
 
 
 
 
 
 
 
 
 
 
Version Number: 1.0 
October 26, 2018 
 
 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
2 
 
Statement of Compliance 
This study will be carried out in accordance with the United States (US) Code of Federal 
Regulations (CFR), local regulations, and Good Clinical Practice (GCP) as required by the 
following  
US CFR applicable to clinical studies (45 CFR 46; and 21 CFR including part 50 and 56 
concerning informed consent and institutional review board (IRB) regulations and 21 CFR 11 
concerning electronic records,) 
• 
International Conference on Harmonisation (ICH) E6 (R1); 62 Federal Register 25691 (1997)  
• 
NIAID Clinical Terms of Award  
All individuals responsible for the design and conduct of this study have completed Human 
Subjects Protection Training and are qualified to be conducting this research prior to the 
enrollment of any volunteers. Curricula vitae for all investigators and sub-investigators 
participating in this study are on file in a central facility (21 CFR 312.23 [a] [6] [iii] [b] edition).  
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
3 
 
Signature Page 
The signature below constitutes approval of this protocol and the attachments and provides the 
required assurances that this study will be conducted according to all stipulations of the protocol, 
including all statements regarding confidentiality, and according to local legal and regulatory 
requirements, applicable US federal regulations, and (ICH E6 R1) guidelines.  
 
Clinical Principal Investigator:   
 
 
 
 
Signed: 
 
Date: 
 
Name: 
 
 
 
Title: 
 
 
 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
4 
 
Table of Contents 
STATEMENT OF COMPLIANCE ...................................................................................................................... 2 
SIGNATURE PAGE ............................................................................................................................................ 3 
TABLE OF CONTENTS ..................................................................................................................................... 4 
LIST OF ABBREVIATIONS ............................................................................................................................... 6 
PROTOCOL SUMMARY .................................................................................................................................... 9 
KEY ROLES ...................................................................................................................................................... 10 
1 
BACKGROUND INFORMATION AND SCIENTIFIC RATIONALE ........................................................................... 11 
1.1 
Background Information ..................................................................................................................... 11 
1.2 
Scientific Rationale ............................................................................................................................. 11 
2 
STUDY OBJECTIVES AND OUTCOME MEASURES ........................................................................................... 12 
2.1 
Objectives ............................................................................................................................................. 12 
2.2 
Outcome Measures............................................................................................................................. 12 
3 
STUDY DESIGN ............................................................................................................................................... 14 
3.1 
Screening ............................................................................................................................................. 14 
3.2 
Interventions ........................................................................................................................................ 14 
3.3 
Potential Risks ..................................................................................................................................... 15 
3.4 
Known Potential Benefits ................................................................................................................... 16 
3.5 
Risk/Benefit Ratio ................................................................................................................................ 16 
3.6 
Specimen Management ..................................................................................................................... 16 
4 
STUDY ENROLLMENT AND WITHDRAWAL ....................................................................................................... 17 
4.1 
Informed Consent ................................................................................................................................ 17 
4.2 
Inclusion Criteria .................................................................................................................................. 17 
4.3 
Exclusion Criteria ................................................................................................................................ 17 
4.4 
Delayed Vaccination ........................................................................................................................... 18 
4.5 
Withdrawal............................................................................................................................................ 18 
4.6 
Termination of the Study .................................................................................................................... 19 
5 
INVESTIGATIONAL PRODUCT(S) ..................................................................................................................... 20 
5.1 
Cholera Vaccine (CVD 103-HgR) ..................................................................................................... 20 
5.2 
Maintaining Blind for Investigational Product(s) ............................................................................. 20 
5.3 
Accountability/Final Disposition for the Investigational Product(s) .............................................. 20 
6 
MANAGEMENT OF SAFETY .......................................................................................................................... 21 
6.1 
Specification of Safety Parameters .................................................................................................. 21 
6.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety Parameters ................. 21 
6.3 
Reporting Procedures......................................................................................................................... 23 
6.4 
Other Adverse Events (if applicable) ................................................................................................ 23 
6.5 
Reporting of Pregnancy ..................................................................................................................... 23 
6.6 
Type and Duration of Follow-up of Volunteers after Adverse Events ......................................... 24 
6.7 
Independent Safety Monitor .............................................................................................................. 24 
6.8 
Oversight of Safety ............................................................................................................................. 24 
6.9 
Clinical Monitoring/Site Monitoring Plan .......................................................................................... 24 
7 
STATISTICAL CONSIDERATIONS ..................................................................................................................... 25 
7.1 
Study Hypothesis ................................................................................................................................ 25 


Blood Donor 9000 
 
v1.0; 26 October 2018 
5 
 
7.2 
Sample Size Considerations ............................................................................................................. 25 
7.3 
Final Analysis Plan .............................................................................................................................. 25 
8 
DATA HANDLING/RECORD KEEPING/SOURCE DOCUMENTS ......................................................................... 27 
8.1 
Data Capture Methods ....................................................................................................................... 27 
8.2 
Types of Data ...................................................................................................................................... 27 
8.3 
Study Records Retention ................................................................................................................... 28 
8.4 
Source Documents ............................................................................................................................. 28 
8.5 
Protocol Deviations ............................................................................................................................. 28 
8.6 
Quality Control and Quality Assurance ............................................................................................ 28 
9 
ETHICS/PROTECTION OF HUMAN SUBJECTS ................................................................................................. 30 
9.1 
Ethical Standard .................................................................................................................................. 30 
9.2 
Institutional Review Board ................................................................................................................. 30 
9.3 
Informed Consent Process ................................................................................................................ 30 
9.4 
Volunteer Confidentiality .................................................................................................................... 31 
9.5 
Principal Investigator Responsibility When Volunteer Withdraws or is Discontinued ............... 31 
9.6 
Future Use of Stored Specimens (if applicable) ............................................................................. 31 
10 
PUBLICATION POLICY ..................................................................................................................................... 33 
11 
LITERATURE REFERENCES............................................................................................................................. 34 
APPENDIX A. TYPICAL STUDY SCHEDULE: SERIAL BLOOD DONORS3 ............................................. 35 
APPENDIX B. STUDY SCHEDULE: LARGE VOLUME/UNIT BLOOD DONORS3 .................................... 36 
APPENDIX C. CLINICAL LABORATORY VALUES, FOR STUDY ELIGIBILITY ...................................... 37 
APPENDIX D. INSTRUCTIONS FOR VAXCHORA ....................................................................................... 38 
 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
6 
 
List of Abbreviations 
Ab 
Antibody 
AE 
Adverse Event 
AIDS 
Acquired Immunodeficiency Syndrome 
ALT 
Alanine Aminotransferase 
ASC 
Antibody Secreting Cell 
AST 
Aspartate Transaminase  
BM 
Memory B cells 
CBA 
Cytometry Bead Array 
CBC 
Complete Blood Count 
CCR9 
C-C Chemokine Receptor Type 9 
CD 
Cluster of Differentiation 
CD3 
Pan T-cell Marker 
CD4 
T-helper Cell 
CD8 
T-cytotoxic Cell 
CD14 
Pan-monocyte/macrophage Marker 
CD19 
Pan-B Cell Marker 
CD20 
Pan-B Cell Marker 
CD25 
Interleukin 2 Receptor Alpha Chain 
CD27 
Tumor Necrosis Factor Receptor Superfamily Member 7 
CD38 
ASC/plasmablasts/plasma cells Marker 
CD45RA 
Isoform A of CD45 (Leukocyte common antigen) 
CD62L 
L-selectin 
CD69 
Early T-cell Activation Antigen 1 
CD103 
Integrin Alpha E  
CD107 
Lysosome-associated Membrane Proteins (LAMP)-1 and LAMP-2 
CFR 
Code of Federal Regulations 
CICERO 
Comprehensive Institutional Collaborative Evaluation of Research 
Online 
CMI 
Cell-mediated Immunity 
CRF 
Case Report Form 
CTCAE 
Common Terminology Criteria for Adverse Events 


Blood Donor 9000 
 
v1.0; 26 October 2018 
7 
 
CTL 
Cytotoxic T Lymphocyte activity 
CVD 
Center for Vaccine Development 
CVD 103-HgR 
The strain designation of live oral cholera vaccine, Vaxchora 
DHHS 
Department of Health and Human Services 
DMEM 
Dulbecco’s Modiﬁed Eagle's Medium 
DNA 
Deoxyribonucleic Acid 
EBV 
Epstein-Barr virus 
e.g. 
For Example 
EU 
European Union 
FDA 
Food and Drug Administration, DHHS 
FOXP3 
Forkhead Box P3 
FWA 
Federalwide Assurance 
GAPDH 
Glyceraldehyde 3-phosphate dehydrogenase 
GCP 
Good Clinical Practice 
GI 
Gastrointestinal 
HIV 
Human Immunodeficiency Virus 
HLA-DR 
Human Leukocyte Antigen DR 
HRPO 
Human Research Protections Office 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IFN- 
Interferon- 
IgA 
Immunoglobulin A  
IgG 
Immunoglobulin G 
IL 
Interleukin 
IPC 
Internal Position Control 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
IV  
Intravenous 
LCL 
Lymphoblastoid Cell Lines 
MHC 
Major Histocompatibility Complex 
Ml 
Milliliter 


Blood Donor 9000 
 
v1.0; 26 October 2018 
8 
 
MOP  
Manual of Procedures 
mRNA 
Messenger Ribonucleic Acid 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, DHHS 
NIH 
National Institutes of Health, DHHS 
OCT 
Optimal Cutting Temperature 
OHRP 
Office for Human Research Protections, DHHS 
PBMC 
Peripheral Blood Mononuclear Cells 
PCR 
Polymerase Chain Reaction 
PI 
Principal Investigator 
PWM 
Pokeweed Mitogen 
QA 
Quality Assurance 
QC 
Quality Control 
QMP 
Quality Management Plan 
RPMI 
Roswell Park Memorial Institute medium 
rRNA 
Ribosomal Ribonucleic Acid 
SAC 
Staphylococcus aureus Cowan 
SAE 
Serious Adverse Event 
SEB 
Staphylococcal enterotoxin B 
SMC 
Safety Monitoring Committee 
SOP 
Standard Operating Procedure 
TCR 
T-cell Receptor 
TM 
Memory T cells                                                        
TEM 
Effector Memory T cells                                                        
TCM 
Central Memory T cells                                                        
TEMRA 
CD45RA+ Effector Memory T cells                                                     
TNF 
Tumor Necrosis Factor 
Tregs 
Regulatory T Cells 
UMB 
University of Maryland Baltimore 
US 
United States 
WHO 
World Health Organization 
Yrs 
Years 


Blood Donor 9000 
 
v1.0; 26 October 2018 
9 
 
Protocol Summary 
Title: 
Collection of Blood for In Vitro Studies from Healthy Adults Who Have 
Received Oral Cholera Vaccine (CVD 103-HgR) 
Protocol 
No.: 
Blood Donor CVD 9000 
 
Phase: 
Specimen and data collection study 
 
Population: 
Total of 200 healthy adults (ages 18 years or older) from the Baltimore-
Washington metropolitan area and from the UMB campus 
 
Number 
of 
Sites: 
1 
Study 
Duration: 
indefinite   
 
Volunteer 
Participation 
Duration: 
Up to 8 years 
 
Agent 
or 
Intervention: Licensed CVD 103-HgR oral V. cholerae O1 vaccine (Vaxchora®, PaxVax 
Inc., Redwood City, CA, USA) 
 
Objectives: 
1. To measure systemic and mucosal immune responses following 
immunization with the oral licensed cholera vaccine in consenting 
volunteers 
2. To measure epigenetic changes which occur in response to vaccination 
with oral cholera vaccines 
3. To integrate these multifaceted datasets into a systems biology approach 
to the understanding of the response to vaccination 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
10 
 
 
Key Roles 
 
 
Clinical PI 
 
Monica A. McArthur, MD, PhD 
Assistant Professor of Pediatrics 
Center for Vaccine Development 
University of Maryland School of 
Medicine 
685 W. Baltimore Street, Suite 
480 
Baltimore, MD 21201 
Office 410-706-0329 
mmcarthu@som.umaryland.edu 
Study 
Coordinator 
Robin Barnes, CRNP 
Center for Vaccine Development 
University of Maryland School of 
Medicine 
685 
W. 
Baltimore 
Street, 
Room422 
Baltimore, MD 21201 
Office 410-706-6192 
rbarnes@som.umaryland.edu  
 
 
 
 
 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
11 
 
1 
BACKGROUND INFORMATION AND SCIENTIFIC RATIONALE 
1.1 
Background Information 
Vibrio cholerae, the causative agent of cholera, has the ability to cause a rapidly dehydrating 
diarrheal disease and is also a public health problem of developing countries, travelers, and 
deployed military personnel to areas lacking safe water and sanitation.  A live oral cholera vaccine 
CVD 103-HgR has been licensed and a single oral dose demonstrated an efficacy of 80-90%.1  
Vaccination with CVD 103-HgR appears to confer sterilizing immunity in the gut.   
Despite the availability of such an efficacious vaccine, the development of new and improved 
vaccines is severely hampered by a lack of information concerning the “true” (i.e., operative) 
immunological mechanisms underlying the protection elicited by cholera, which does not invade 
the mucosal surface.  Specimens obtained from volunteers participating in this clinical study will 
be used to identify the immunological mechanisms of protection from V. cholerae.  These 
mechanisms may be broadly applied to other enteric pathogens for which vaccines are not yet 
available.   
1.2 
Scientific Rationale 
Immunological responses to V. cholerae 
The immunological responses that correlate with protection from V. cholerae remain largely 
undefined or poorly defined.  Thus, in depth studies of immune responses will be conducted with 
specimens from young adults and the elderly immunized with a single dose of CVD 103-HgR in 
this clinical research study.  These studies should provide key insights into the characteristics and 
longevity of the immune responses that best correlate with protection from infection with V. 
cholerae and contribute to further our knowledge of the mechanisms underlying 
immunosenescence.  Furthermore, these studies will enable a direct comparison of the magnitude 
and characteristics of the systemic and gut mucosa immune responses elicited by CVD 103-HgR 
immunization.  
Summary 
The specimens obtained in this clinical research study will be used to further our understanding 
of the protective immunological mechanisms that can be elicited systemically and may be 
applicable to other enteric pathogens.   
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
12 
 
2 
STUDY OBJECTIVES AND OUTCOME MEASURES 
2.1 
Objectives 
1. To measure the systemic immune response following immunization with oral licensed 
cholera vaccine in consenting volunteers 
2. To measure epigenetic change which occur in response to vaccination with oral cholera 
vaccine 
3. To integrate these multifaceted datasets into a systems biology approach to the 
understanding of the response to vaccination  
2.2 
Outcome Measures 
2.2.1 Primary outcome 
Percentage of responders by four-fold increase in vibriocidal antibody titers between pre- 
and post-immunization time-points. 
2.2.2 Exploratory outcomes 
The following brief descriptions provide some overview of the intended laboratory analyses to be 
performed with the specimens to be collected in this study. This is not meant to be an exhaustive 
list of all the possible laboratory studies to be performed. 
 1. Functional analysis of systemic (e.g., using serum, PBMC) anti-V. cholerae immune 
responses.  These studies will include the following measurements, depending on specimen 
availability:  
a. Ab levels to V. cholerae antigens in bodily fluids (e.g., serum, plasma) 
b. Ab-secreting cells (ASC) to V. cholerae antigens 
c. Cytokine production (e.g., intracellular IFN-γ, TNF-α, etc. by multichromatic flow 
cytometry or mass cytometry, IFN-γ production by Elispot and/or cytokine production 
in culture supernatants by multiplex technology.) 
d. Memory (TM) and BM responses 
e. CTL 
f. innate immune responses (macrophages and dendritic cells) 
2.  Evaluation of homing characteristics of the various V. cholerae-responsive CD4+ and CD8+ 
T cell subsets  
6. Study of TCRγδ T cells and Treg using multichromatic flow cytometry or mass cytometry 
using monoclonal antibodies to molecules that are characteristic of these cells (e.g., CD25 
and FOXP3) 
7.  Measure the expression of host response genes to vaccination using functional genomics, 


Blood Donor 9000 
 
v1.0; 26 October 2018 
13 
 
proteomics, and epigenetics 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
14 
 
3 
STUDY DESIGN 
This study is a single center, open label clinical research study to study the effect of oral 
immunization with the CVD 103-HgR cholera vaccine in younger adults and older adults (i.e., 
elderly).  Healthy adults, ages 18 years to 64 years, will be recruited from the Baltimore-
Washington area in response to campus flyers, advertising, and word-of-mouth. Study 
participants will receive the cholera vaccine.  Subsequent to vaccination, small volume (up to 200 
mL) or large volume (200 mL or greater) blood draws will be requested; the timing of these blood 
draws are not strictly set and the schedules in Appendix D and E are provided to give a general 
overview of the flow of research procedures.  The timing of the blood draws and the types (small 
or large volume blood) is dependent on the research laboratory’s needs. In general, more frequent 
scheduled blood draws will occur in the first year after vaccination and less often in the years after 
vaccination (up to 8 years of planned follow-up).  Therefore, we intend to regularly re-screen for 
anemia and have set limits on the total volume of blood to be collected over any 56 day period. 
3.1 
Screening 
After informed consent has been completed, a medical history, concomitant medications, and vital 
signs are collected.  Initial screening labs for eligibility include: 
• 
Complete blood count (CBC) with differential, to assess total white blood count (WBC), 
hemoglobin, and platelet count 
• 
Liver function tests to assess alanine aminotransferase (ALT) and aspartate 
aminotransferase (AST) 
• 
Serologies for HIV 
• 
Blood typing (this is not an eligibility criterion, but is used to guide the immunological 
analysis) 
3.2 
Interventions  
Administration of licensed Live Oral Cholera Vaccine CVD 103-HgR. Participants will be 
given the vaccine, as packaged, and are to administer the vaccine as per the package insert.  
Briefly, approximately 100 mL of cool or room temperature purified bottled water is to be placed 
into a clean disposable cup. The contents of the buffer sachet are to be placed into the water and 
stirred. Then the contents of the active (lyophilized vaccine) sachet are to be placed into the water, 
then stirred for ~30 seconds.  The reconstituted mixture should be completely ingested within 15 
minutes. Illustrated instructions for the administration of CVD 103-HgR is presented in Appendix 
E.  
Serial Blood Donor (small volume). After volunteers have been recruited, they may be 
contacted to have blood drawn, 1-20 times a year. No more than 2 ⅓ cups (550 ml) will be taken 
over a 2-month period of time.   The volunteer will be given a schedule of the dates when blood 
will be drawn and the amount of blood required each time they are contacted.  At any time a donor 
may decline a blood specimen but choose to remain in the study.  In most instances, 20 to 200 
mL (4 to 40 teaspoons) will be collected at one visit. Visits for one-time or serial blood donations 
will take approximately ten minutes each and are performed at the Center for Vaccine 


Blood Donor 9000 
 
v1.0; 26 October 2018 
15 
 
Development (CVD) Outpatient Research Clinic. 
Blood Unit Donor (large volume). When large volumes of blood are desired, participants may 
be directed to the Greenebaum Cancer Center (GCC) Cell Component Therapy (CCT) Lab.  Visits 
for the donation of a unit of blood/large volume (200- 500 mL) will take one hour. As part of the 
GCC CCT lab’s standard operating procedure, a CBC will be performed on the day of donation 
(prior to donation of a unit of blood) for verification of a hemoglobin 12.5 or greater.  If the 
hemoglobin is less than 12.5, the volunteer will be deferred and re-screened at a later date or 
they may have a smaller volume blood draw (≤ 200 mL) performed at the CVD Outpatient 
Research Clinic, if eligible by the hemoglobin criteria in Appendix C.   Additional blood draws will 
take place according to the schedules attached. 
Planned repeat screening labs. The screening blood tests (except for blood typing) will be 
repeated prior to each scheduled blood donation (small or large volume), but not more often than 
every 56 days. This is primarily to ensure that the participant is not significantly anemic, but also 
assesses for any interval abnormality in blood tests since the study duration may be over the 
course of 8 years. 
3.3 
Potential Risks 
CVD 103-HgR Vaccination 
The live oral cholera vaccine, known as Vaxchora (composed of strain CVD 103-HgR), was 
licensed by the U.S. FDA on June 10, 2016.  The adverse reactions to CVD 103-HgR vaccine are 
generally mild and may include tiredness (30%), headache (28%), abdominal pain (18%), 
nausea/vomiting (17%), lack of appetite (16%), and diarrhea (4%). The safety and 
immunogenicity of the live oral cholera vaccine has not been fully established in children (age <18 
years), pregnant women, or immunocompromised patients. However, a phase 2 study of the 
immune response to CVD 103-HgR was performed in HIV patients.2 
Risk with Venipuncture 
Venipuncture is associated with temporary local pain, some bruising, light-headedness or fainting, 
and a risk for infection. Infection risk is minimized by the use of alcohol swabs and sterile 
equipment.  Blood will be drawn by trained personnel using clean technique.  Volunteers who are 
found to experience lightheadedness will lie down during and after they have their blood drawn.  
Risk of Anemia 
Because serial blood draws will be performed in this protocol, there is a risk of anemia. The 
volumes of blood drawn from each volunteer will be limited to 550 mL every 56 days, and the 
hematocrit will be checked at regular intervals. Participants found to be anemic will have their 
scheduled blood draws postponed, be advised to increase iron intake, and have a hematocrit 
repeated and in the normal range in order to be eligible for the blood draw. 
Risk of Loss of Confidentiality 
There is always the risk for loss of confidentiality when participants are sharing their personal 
information.  As a standard procedure for all clinical studies, access to clinical information will be 
limited to the IRB-approved clinical investigators, study coordinator, and research staff.  Paper 
source documents and Case Report Forms (CRFs) will be stored in limited-access rooms and 


Blood Donor 9000 
 
v1.0; 26 October 2018 
16 
 
locked file cabinets.  Electronic CRFs will be password protected in a limited access database.  
IRB-approved research staff will be the only individuals with access to the personal identifiable 
information and personal health information of research participants. 
3.4 
Known Potential Benefits 
Vaccine against V. cholerae is intended to protect the immunized in endemic areas, as well as 
travelers (including military personnel) from industrialized countries to regions endemic for V. 
cholerae, and in the event of a threat or exposure to a biological weapon.  There is no known 
benefit to receipt of off-label use of the CVD 103-HgR vaccine. 
V. cholerae is a global pathogen that can disseminate rapidly in settings where there is crowding 
and inadequate sanitation.  The symptoms of cholera are predominantly a rapidly dehydrating 
diarrheal disease without significant systemic toxicity.  
This clinical research study is intended to greatly further our understanding of the protective 
immunological mechanisms that can be elicited systemically.  This information is intended to 
inform the development of newer generation cholera vaccines.    
3.5 
Risk/Benefit Ratio 
Alternative approaches are limited for these studies.  The risks of the study interventions are low.  
The CVD 103-HgR vaccine has been extensively used, and their side effect profiles are tolerable 
and well-understood.  Venipuncture also carries a low risk when conducted by a trained 
phlebotomist.  On the other hand, enteric infections are widespread throughout the globe and are 
associated with significant morbidity and mortality.  The complexity of the immune response after 
vaccination cannot be reproduced in other environments.  The risks associated with the 
procedures described in this protocol are small compared to the potential general benefit of better 
understanding the protective immunity elicited by these vaccines. 
3.6 
Specimen Management 
All specimens collected through this protocol will be labelled with the volunteer identification 
number and date, but not any volunteer identifiable information (such as date of birth). Specimens 
that are processed for storage will also be labelled in a manner which is de-identified and contains 
no identifiable information. Specimens are entered into specimen tracking form to link them with 
the volunteer identification number.  Only the clinical PIs and study coordinator(s) will have access 
to the linking information.  Specimen handling, processing, and storage will be according to 
internal CVD standard operating procedures.  
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
17 
 
4 
STUDY ENROLLMENT AND WITHDRAWAL 
The study enrollment will include adults from the local area.  The study will not exclude any race, 
ethnicity or gender from enrollment.  Women of child bearing potential will have a urine pregnancy 
test prior to vaccination.  Women testing positive will be excluded from the study.  Volunteers will 
have frequent contact with the study coordinator(s) to facilitate retention and complete data 
collection.  The inclusion/exclusion criteria will be reviewed with each volunteer during the consent 
process. The clinical principal investigator or designee is responsible for reviewing and signing 
off the inclusion and exclusion criteria for all enrollees. 
4.1 
Informed Consent 
All aspects of the protocol will be explained in depth to volunteers including the specific purpose 
of the blood draws.  Volunteers will be given a schedule of when, where, and how much blood 
will be drawn.  The consent form will be read by the volunteer and ample time for questions and 
answers will be provided.  Interested volunteers will provide written consent prior to any 
procedures being performed.  Volunteers will remain enrolled as a blood donor as long as the 
laboratory has need for the blood (which may continue for several years), or until the volunteer 
withdraws consent. The informed consent process and other health assessment questions will 
take place in a private or semi-private room.  Privacy and confidentiality of the volunteer will be 
maintained. 
 
4.2 
Inclusion Criteria  
Volunteers must meet all inclusion criteria in order to be eligible to participate. 
1. Age 18 years to 64 years at the time of enrollment 
2. Good general health as determined by a screening evaluation within 28 days before blood 
donation or vaccination 
3. Provides written informed consent prior to initiation of any study procedures 
4.3 
Exclusion Criteria 
Volunteers must meet no exclusion criteria in order to be eligible to participate  
1. History of any of the following medical conditions: 
• 
Diabetes 
• 
Cancer in past 5 years (except for basal cell carcinoma of the skin and cervical 
carcinoma in situ)  
• 
Heart disease (e.g., hospitalization for a heart attack, coronary artery bypass graft, 
arrhythmia, or syncope, within past 5 years. Current symptoms of heart disease, 
such as dyspnea, angina, or orthopnea) 
• 
Recurrent infections (e.g., more than 3 hospitalizations for invasive bacterial 


Blood Donor 9000 
 
v1.0; 26 October 2018 
18 
 
infections such as pneumonia or meningitis) 
• 
Current drug or alcohol abuse    
• 
Active ulcer disease or ongoing intestinal condition 
• 
Treatment for anemia in last 6 months 
• 
Treatment with anti-malarial drugs within ten days prior to study vaccination 
• 
Treatment with antibiotics within 14 days prior to study vaccination 
• 
Immunodeficiency or immunosuppression from illness or treatment 
2. Close contact within 7 days following study vaccination with a person who has an 
immunodeficiency or immunosuppression from illness or treatment 
3. History of cholera infection or cholera vaccination 
4. Any of the following CBC abnormalities during screening: 
• 
WBC <0.81 x LLN or > 1.09 x  ULN   
• 
Hemoglobin <0.91 x LLN  or >1.18 x ULN (women) or <0.92 x LLN or >1.18 x ULN  
(men)  
• 
Platelet count <0.8 x LLN or > 1.2 x ULN   
5. Any of the following laboratory abnormalities during screening: 
• 
SGOT or SGPT >1.5 times normal  
• 
Positive serology for HIV antibody 
 
       6.   Poor peripheral venous access for blood donation 
7.   Other condition that the opinion of the investigator would jeopardize the safety or rights of 
a volunteer participating in the trial or interfere with the scientific integrity of study 
8. Positive urine pregnancy test (HCG) on the day of vaccination  
4.4 
Delayed Vaccination 
Study participants that have an acute illness, fever, or gastrointestinal complaint (temporary) 
within the 3 days prior to scheduled vaccination should have their vaccination delayed until 
recovery from their temporary illness.  Study participants that are on a temporary course of 
concomitant antibiotics should have their vaccination delayed until at least 7 days after completion 
of antibiotics.  Study participants on certain potentially interfering malaria prophylaxis medications 
(chloroquine), should have their vaccination delayed—dosing may begin 2 weeks after completion 
of chloroquine. 
4.5 
Withdrawal 
A study volunteer will be discontinued from study participation for the following reasons: 
• 
Request by volunteer to terminate participation 
• 
Requirement for prohibited concomitant medication or treatment 
• 
Unable to comply with requirements of the protocol 
• 
Failure to comply with study procedures   
• 
At the request of the IRB or NIAID 
• 
The volunteer’s well-being, based on the opinion of the investigator 


Blood Donor 9000 
 
v1.0; 26 October 2018 
19 
 
The PI may remove volunteers from the study if they develop adverse events (AEs) that are 
believed to be related to the study or unrelated but may impact the safety of the volunteer or the 
integrity of the study.  These might include abnormal laboratory values or test procedure results, 
medical treatments that might interfere with immune responses or use of other investigational 
products or vaccines during the study period.  Development of exclusion criteria may also prompt 
exclusion from the study.  Volunteers may also be withdrawn for failure to comply with study 
procedures.  Volunteers may withdraw consent at any time upon request. 
4.6 
Termination of the Study 
This study may be closed by the NIH, the principal investigator, or the UMB IRB.  If the study is 
terminated prematurely the study participants will be notified in writing.  A final safety evaluation 
will be done for all volunteers enrolled at the termination of the study.  
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
20 
 
5 
INVESTIGATIONAL PRODUCT(S) 
5.1 
Cholera Vaccine (CVD 103-HgR)  
5.1.1 
Formulation, Packaging, and Labeling 
CVD 103-HgR, marketed as Vaxchora®, is provided as two powder sachets, containing buffer 
and lyophilized organisms, in a single package. 
5.1.2 
Acquisition  
Product will be obtained from the manufacturer PaxVax, Inc (Redwood City, CA, U.S.A.)  
5.1.3 
Product Storage, Stability, and Expiration  
Vaxchora (Cholera Vaccine, Live, Oral) is to be stored at -15° C or colder (5° F or colder).  Each 
package of vaccine shows an expiration date.  This expiration date is valid only if the product has 
been maintained at -15° C or colder. 
5.1.4 
Preparation/Handling/Administration 
Vaxchora (Cholera Vaccine, Live, Oral; CVD 103-HgR) is indicated for immunization of adults 
18-64 years of age against disease caused by V. cholerae. [NOTE: Vaxchora is currently not 
licensed for older adults (age 65 years or older) and thus older adults are not intended to be 
vaccinated with Vaxchora in this study.] Vaccine potency is dependent upon storage under 
freezing (-15° C or colder; 5° F or colder).  The vaccine should be reconstituted and ingested 
within 15 minutes of removing from the freezer. Recipients should avoid eating or drinking for 
60 minutes before and after ingestion of Vaxchora.  
Participant will take a single oral dose of the CVD 103-HgR vaccine.  A volume of ~100 mL of 
purified bottled water (at cold or room temperature) shall be placed into a clean disposable cup.  
The buffer component sachet is emptied into the cup and stirred. Then, the active vaccine 
(lyophilized V. cholerae CVD 103-HgR) component is emptied into the cup and stirred, for ~30 
seconds.  The mixed contents should be completely consumed within 15 minutes of removal from 
the freezer. 
5.2 
Maintaining Blind for Investigational Product(s) 
This study is not blinded.  
5.3 
Accountability/Final Disposition for the Investigational Product(s) 
Vaccine will be kept in a secure area and stored according to package directions.  The CVD will 
maintain an accurate record of shipping and dispensing of investigational products in an 
accountability ledger.  The ledger will include the date and amount of investigational product 
dispensed to each volunteer.   
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
21 
 
6 
MANAGEMENT OF SAFETY  
6.1 
  Specification of Safety Parameters 
The clinical investigators are responsible for reporting Adverse Events (AEs) that are observed 
or reported during the study. Serious Adverse Events (SAEs) must be reported as soon as the 
clinical-investigators become aware. See Section 7.2 for reporting windows.  Only AEs and SAEs 
occurring within 7 days post vaccination determined to be related to the study product or 1 day 
post blood donation will be collected.   No AE or SAE will be collected outside the window 
mentioned above.  
6.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety Parameters 
Adverse Event (AE) 
ICH E6 defines an AE as any untoward medical occurrence in a patient or clinical investigation 
volunteer 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 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.  
All AEs including local and systemic reactions not meeting the criteria for “serious adverse events” 
should be captured on the appropriate CRF.  Information to be collected includes event 
description, time of onset, clinician’s assessment of severity, relationship to study product 
(assessed only by those with the training and authority to make a diagnosis, which would include 
MD, PA, Nurse Practitioner, DO, or DDS), and time of resolution/stabilization of the event.  All 
AEs occurring while on study must be documented appropriately, AEs occurring within 7 days 
post vaccination determined to be related to the study product and/or 1 day post-blood donation 
and determined to be related to the procedure will be collected.  All AEs will be followed to 
adequate resolution.  
Any medical condition that is present at the time that the patient is screened should be considered 
as baseline and not reported as an AE.  However, if it deteriorates at any time during the study, it 
should be recorded as an AE. 
All AEs must be graded for severity and relationship to study product.  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:  The following guidelines will be used to quantify intensity.  
Mild: events require minimal or no treatment and do not interfere with the patient’s daily 
activities. 
Moderate: events result in a low level of inconvenience or concern with the therapeutic 
measures. Moderate events may cause some interference with functioning. 
Severe: events interrupt a patient’s usual daily activity and may require systemic drug therapy 


Blood Donor 9000 
 
v1.0; 26 October 2018 
22 
 
or other treatment.  Severe events are usually incapacitating. 
Life threatening: any adverse drug experience that places the patient or volunteer, in the view 
of the investigator, at immediate risk of death from the reaction as it occurred, ie, it does 
not include a reaction that had it occurred in a more severe form, might have caused 
death. 
Changes in the severity of an AE should be documented to allow an assessment of the duration 
of the event at each level of intensity to be performed.  Adverse events characterized as 
intermittent require documentation of onset and duration of each episode. 
Relationship to Study Products:  The clinician’s assessment of an AE's relationship to vaccine 
is part of the documentation process, but it is not a factor in determining what is or is not reported 
in the study.  If there is any doubt as to whether a clinical observation is an AE, the event should 
be reported.  All AEs must have their relationship to study product assessed using the terms: 
related or not related.  In a clinical study, the study product must always be suspect.  To help 
assess, the following guidelines are used. 
• 
Related – There is a reasonable possibility that the study product caused the adverse 
event. Reasonable possibility means that there is evidence to suggest a causal 
relationship between the study product and the adverse event. 
• 
Not Related – There is not a reasonable possibility that the administration of the study 
product caused the event. 
Serious Adverse Event (SAE) 
An adverse event or suspected adverse reaction is considered “serious” if, in the view of either 
the investigator or sponsor, it results in any of the following outcomes:  
• 
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 volunteer 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. 
* Life-threatening adverse event. An adverse event is considered “life-threatening” if, in the 
view of either the investigator or sponsor, its occurrence places the patient or volunteer at 
immediate risk of death.  It does not include an adverse event, had it occurred in a more 
severe form, might have caused death. 
All SAEs will be: 


Blood Donor 9000 
 
v1.0; 26 October 2018 
23 
 
• 
recorded on the appropriate SAE CRF 
• 
followed through resolution by a study clinician 
• 
reviewed and evaluated by a study clinician 
The SAEs must be reported as soon as the investigator becomes aware but within 5 working days 
for internal events, consistent with UMB policy   Investigators will report the SAE using the 
Adverse Event form in the Comprehensive Institutional Collaborative Evaluation of Research 
Online (CICERO) electronic system.  The investigator will provide a summary of the problem or 
event in the text box labeled “adverse event information”.  The summary should include type or 
nature of the problem, a full description of the activities leading to the problem, interventions 
and/or actions taken in response, temporal relationship of event to the use of the investigational 
product/procedure and the current status of the participant.  Additional information will be reported 
as it is received. 
In the event of any serious adverse experience which, in the clinical investigator’s opinion, justifies 
termination or modification of the study, it will be stopped and the IRB and ISM will be informed 
immediately.  The IRB will be notified via CICERO, the IRB electronic tracking system.   
Reactogenicity 
Reactogenicity will not be measured in this study. 
6.3 
Reporting Procedures 
AEs will be followed until resolution.  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. 
AEs and SAEs will be received, reviewed and managed by the ISM.  The investigator is 
responsible for reporting events to VAERS according to the VAERS reporting criteria provided in 
the federal regulations.  Regulatory Reporting of AEs that are determined to be vaccine related 
will be reported through the Vaccine Adverse Event Reporting System (VAERS).     
6.4 
Other Adverse Events (if applicable) 
An unexpected AE is an adverse reaction, the nature or severity of which is not consistent with 
the applicable product information (e.g., package insert for the respective vaccine).   
6.5 
Reporting of Pregnancy  
Although not AEs, pregnancies are reportable events (on the study-specific form) to the data 
management group.  The pregnancy outcome will be reported if known including the health status 
of the mother and child including date of delivery and infant’s gender and weight.  If a volunteer’s 
urine pregnancy test is positive prior to receiving the vaccination, they will be excluded from 
participating in the study. 
Volunteers who are pregnant prior to receiving the vaccination will be excluded from participation 
in this study.  Pregnancy will be reported as an unanticipated problem involving risk to research 
participants.  Pregnancy will be reported to the UMB IRB within 5 working days of the time the 
research staff become aware of the pregnancy.  In the event a volunteer becomes pregnant after 


Blood Donor 9000 
 
v1.0; 26 October 2018 
24 
 
taking the vaccine the volunteer will be encouraged to complete all visits for safety follow up.  
6.6 
Type and Duration of Follow-up of Volunteers after Adverse Events 
All 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.  The Clinical-PIs will follow all SAEs until resolution 
(return to pretreatment status or stabilization of the condition with the expectation that it will remain 
chronic) even if this extends beyond the study-reporting period. 
At any time after completion of the study, if the investigator becomes aware of an SAE that is 
suspected to be related to study product, the investigator will report the event to the IRB. 
6.7 
Independent Safety Monitor 
Dr. Monica A. McArthur will be the ISM for this clinical research study. 
6.8 
Oversight of Safety 
The clinical investigators reserve the right to terminate this study should serious AEs or any other 
safety issue occur during the study.  The UMB IRB upholds that all currently approved research 
that is not being conducted in accordance with the Human Research Protections Program and 
UMB IRB policies and procedures, Federal, State, and local requirements, or has been associated 
with unexpected serious harm to participants may be subject to suspension or termination. The 
IRB has the authority to suspend or terminate approval of research not being conducted in 
accordance with the IRB’s requirements or that had been associated with unexpected serious 
harm to participants.  
6.9 
Clinical Monitoring/Site Monitoring Plan  
The study will be monitored by the site clinical PIs.  On a regular basis the clinical PIs will review 
patient charts/clinical reports, laboratory tests, outcomes, enrollment, procedure reports, and all 
adverse events.  Site monitoring is conducted to ensure the human subject protection, study 
procedures, laboratory, study intervention administration and data collection processes are of 
high quality and meet sponsor, ICH E6, and other appropriate, regulatory guidelines and that the 
study is conducted in accordance with the protocol.  
  
 
 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
25 
 
7 
STATISTICAL CONSIDERATIONS 
7.1 
Study Hypothesis 
This is an exploratory study rather than a hypothesis-driven study. 
7.2 
Sample Size Considerations 
This study is exploratory in nature and is designed to identify associations among the presence 
and quantity of various intestinal bacteria, immune responses and clinical outcome.  Thus it is 
designed to generate hypotheses but not to test any specific hypothesis based on prior data since 
no such data is available in the literature.  The sample sizes planned for the study are therefore 
based on feasibility rather than power to reject any specific hypothesis.  For large differences 
between groups the study will have good power to find statistically significant associations.  
Data for withdrawn volunteers will be included in the intent to treat group.  We anticipate a dropout 
rate of 20%.  This is consistent with other studies of this nature performed at the University of 
Maryland.  
7.3 
Final Analysis Plan 
The statistical analysis will depend on the hypothesis being evaluated.  For example, if measuring 
cytokine levels, the level of cytokine, in pg/ml, will be obtained from extrapolation over standard 
curves. Seroconversion following challenge will be defined as a fourfold rise in serum Ab over 
pre-challenge levels.  The geometric mean Ab level will be calculated for each time point and the 
peak time determined. The proportion of volunteers in each group who seroconvert will be 
determined for each antigen and Ab class at the peak time post-challenge.  A positive ASC 
response will be defined for each immunoglobulin class and antigen as a post-vaccination count 
of at least 8 ASC/106 PBMC that is ≥ 3 standard deviations above the mean pre-vaccination count 
for all volunteers in all cohorts combined.  The proportion of volunteers in each group who develop 
each specific ASC response will be determined for each antigen and Ab class measured.  The 
geometric mean ASC level at each time point and the peak post-immunization geometric mean 
will be calculated.  Comparisons between systemic and local responses will be performed by 
using a two-sample t-test on log10 values or a Wilcoxon rank-sum test.   
Comparisons between pre- and post-immunization levels within a single group will use a one-
sample t-test on log10-transformed values or a Wilcoxon signed-rank test.  Comparisons between 
two groups will use a two-sample t-test on log10 values or a Wilcoxon rank-sum test or the 
Kolmogorov-Smirnov test.  Comparisons of proportions responding will use chi-square or exact 
tests.  For comparisons among three or more groups, an overall analysis of variance or Kruskal-
Wallis test will be done first; if that test indicates significant heterogeneity among groups, pairwise 
comparisons will then be made, with adjustment for the number of comparisons.  Continuous 
immunological responses will be compared in the young adults and elderly by analysis of variance 
or Kruskal-Wallis test and pairwise comparisons as appropriate.  Proportions of responders will 
be compared by chi-square or exact tests.  Relationships between immunological measurements 
will be assessed using correlation analyses (for two continuous responses), t-tests (for comparing 
a continuous and a dichotomous response), and chi-square tests or exact tests (for comparing 


Blood Donor 9000 
 
v1.0; 26 October 2018 
26 
 
two dichotomous responses).  Two-sided p-values will be reported whenever two groups, or pre- 
and post-immunization levels within a group, are compared. P < 0.05 will be considered 
statistically significant.  No imputation will be made for missing data. 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
27 
 
8 
DATA HANDLING/RECORD KEEPING/SOURCE DOCUMENTS 
The investigators are responsible to ensure the accuracy, completeness, legibility, and timeliness 
of the data reported.  Data will be recorded on source documents and CRFs in a neat, legible, 
and timely manner using black ink to ensure accurate interpretation of data and clarity of 
reproduced copies.  When making a change or correction, the original entry will be crossed out 
with a single line.  The initials of the person making the change and the date of the change will 
be recorded next to the change.  Corrections will NOT be made by erasing, overwriting, or using 
correction fluid or tape on the original.  Dates reported in the CRF derived from source documents 
will be consistent with the source documents or the discrepancies will be explained.  After data 
are quality controlled by the clinical or laboratory staff, they will be entered into a database and 
checked for accuracy, consistency and completeness, according to data entry standard operating 
procedures (SOPs). 
The Principal and Co-Principal investigators assume responsibility for the accuracy and security 
of study data obtained, managed, stored, and reported.  Data collection is the responsibility of 
trained study clinical and laboratory staff under the supervision of the project leader.  Data 
management core facilities such as the University of Maryland School of Medicine Bioinformatics 
Core Facility may be responsible for data management including data capture and quality review.  
Adverse events must be graded, assessed for severity and causality, and reviewed by the site PI 
or designee. 
All data collection and analysis will be performed using coded samples.  Coding will occur at 
sample collection.  Investigators will remain blinded to volunteer identifiers during data review and 
analysis. 
8.1 
Data Capture Methods 
Clinical data (including AEs, concomitant medications, and clinical laboratory data) will be entered 
into paper record clinical research forms.  The clinical research forms will remain the authoritative 
record. 
Data capture from these source documents will be stored in a password-protected electronic 
database managed by REDCap, a 21 CFR Part 11-compliant CRF and database. The data 
system includes password protection and internal quality checks, such as automatic range checks 
to identify data that appear inconsistent, incomplete, or inaccurate.  In some cases the CRF will 
serve as a source document. 
Source documents will be reviewed by study staff receiving the source document (as in clinical 
team receiving the screening laboratory report) or by study staff generating the data (as in and 
data entry staff, who will ensure that they are accurate and complete).  Adverse events will be 
graded, assessed for severity and causality, and reviewed by the Clinical Investigator or designee 
8.2 
Types of Data 
Clinical, immunological, microbiological, and other laboratory data may be generated as part of 
the projects.  Clinical data may be obtained from clinical source documents or directly from the 
participant as documented by trained clinical research staff.  Immunological, microbiological, and 


Blood Donor 9000 
 
v1.0; 26 October 2018 
28 
 
other data will be generated by laboratory staff.  Source documents are the documents on which 
data collected is first recorded. 
Safety data are collected on CRFs in the patient study record and recorded aggregately in a 
separate study log. 
8.3 
Study Records Retention  
Records for the study will be retained for 5 years as required by University of Maryland Human 
Research Protections Office policy.  
The site will maintain appropriate medical and research records for this study, in compliance with 
ICH E6 and regulatory and institutional requirements for the protection of confidentiality of 
volunteers.  The site participating in this study will permit authorized representatives of the 
sponsor  and regulatory agencies to examine (and when required by applicable law, copy) clinical 
records for the purposes of clinical site monitoring, quality assurance reviews, audits, and 
evaluation of the study safety and progress.  
8.4 
Source Documents 
The clinical history recorded in the volunteer’s medical records will provide source data for 
participants in this study.  This data is captured both electronically and in paper copy.  Clinical 
laboratory data are reported electronically. A paper printout of volunteer data will provide the 
source document for laboratory data that is generated electronically either in the clinical or 
research laboratory setting, whenever possible.  Data capture from source documents will be 
collected into a database managed by REDCap.  The data, as entered on the forms, constitutes 
the “official” study data and any computerized data is required to be in agreement with the 
information as recorded on the paper forms. 
8.5 
  Protocol Deviations  
A protocol deviation is any noncompliance with the clinical study protocol or GCP.  The 
noncompliance may be either on the part of the volunteer, the investigator, or the study site staff.  
As a result of deviations, corrective actions are to be developed by the site and implemented 
promptly. 
UMB IRB will be notified within 5 working days of all deviations and violations that result in an 
increase in risk or a decrease in benefit to participants and all unanticipated problems involving 
risk to research participants and others.  Deviations that do not alter the risk benefit analysis or 
integrity of the study can be submitted at the time of annual renewal or in an expedited fashion if 
the sponsor requires such reporting.  All deviations from the protocol must be addressed in study 
volunteer source documents.  Protocol deviation reporting will be according to CVD standard 
operating procedures.     
8.6 
Quality Control and Quality Assurance 
Internal audits serve to verify that the clinical study/research is conducted according to the IRB-


Blood Donor 9000 
 
v1.0; 26 October 2018 
29 
 
approved protocol and in compliance with the International Conference on Harmonization (ICH) 
and Good Clinical Practice Guideline (GCP), applicable U. S. Code of Federal Regulations (CFR) 
Titles 21, Parts, 50, 54, 56, 58 and 312, US FDA Guidances, Department of Health and Human 
Services (DHHS) Title 45 part 46, and University of Maryland Human Research Protections Office 
(HRPO) polices.   
A Quality Management Plan developed for this protocol will identify the protocol, state the 
frequency of quality control checks, quality assurance audits, priority of QA reviews, and the 
percentage of charts that will be selected for QA review.  The plan will describe how data will be 
evaluated for compliance with the protocol, completeness, and accuracy.  
Processes to be reviewed include, but are not limited to: eligibility, informed consent process and 
documentation, concomitant medication recordings, study product accountability and 
management, AE/SAE reporting, study/clinical endpoints, follow-up visits, missed visits, study 
discontinuation, and review of regulatory documents.  
The QMP and written quality assurance communication (e.g., audit reports, problem resolution 
worksheets, regulatory checklists, audit summaries, QC/QA trend reports, quarterly and annual 
reports, etc.) for this study will be kept in a separate quality management file.   
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
30 
 
9 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
9.1 
Ethical Standard 
The investigator(s) will ensure that this study is conducted in full conformity with the principles of 
The Belmont Report:  Ethical Principles and Guidelines for the Protection of Human Subjects of 
Research of The National Commission for the Protection of Human Subjects of Biomedical and 
Behavioral Research (April 18, 1979) and codified in 45 CFR 46, 21 CFR 312, and/or ICH E6; 62 
Federal Regulations 25691 (1997).  The PI/Institution will hold a current federal wide assurance 
(FWA) issued by Office of Human Research Protections, DHHS (OHRP) for federally funded 
research 
Volunteers will provide informed consent prior to any participation in the study.  Volunteers may 
withdraw consent at any point in the study.  Specimens may not be used for future research 
without the volunteer’s written permission.  Pregnant women will be excluded from the research 
because of the potential for increased risk from live vaccines.  No other special population will be 
excluded. 
9.2 
Institutional Review Board 
A copy of the protocol, informed consent forms (ICF), and other information to be completed by 
participants, such as survey instruments or questionnaires, and any proposed advertising / 
recruitment materials will be submitted to the IRB for written approval.  The investigator must 
submit and obtain approval from the IRB for all subsequent amendments to the protocol, informed 
consent documents and other study documentation referenced above.  The investigator will be 
responsible for obtaining IRB approval of the annual continuing review throughout the duration of 
the study.  The investigator will notify the IRB of violations from the protocol and serious adverse 
events. 
9.3 
Informed Consent Process  
The informed consent process will be initiated before a volunteer agrees to participate in the study 
and should continue throughout the individual’s study participation.  The consent will explain that 
volunteers may withdraw consent at any time throughout the course of the study.  Extensive 
explanation and discussion of risks and possible benefits of this investigation will be provided to 
the volunteers in understandable language.  Adequate time will be provided to ensure that the 
volunteer has time to consider and discuss participation in the protocol.  
The consent forms will describe in detail the study interventions/products/procedures and 
risks/benefits associated with participation in the study.  The rights and welfare of the volunteers 
will be protected by emphasizing that their access to and the quality of medical care will not be 
adversely affected if they decline to participate in this study. 
The volunteer will sign the informed consent document before any procedures are undertaken for 
the study.  A copy of the signed informed consent document will be given to the volunteer for 
his/her records.  


Blood Donor 9000 
 
v1.0; 26 October 2018 
31 
 
9.4 
Volunteer Confidentiality 
Volunteer confidentiality is strictly held in trust by the participating investigators, their staff, and 
the sponsor(s) and their agents.  This confidentiality is extended to cover testing of biological 
samples and genetic tests in addition to the clinical information relating to participating volunteers.  
All electronic records will be password protected in a limited access database.  Paper records will 
be stored in a locked cabinet with access limited to persons involved in the research.  Only the 
PIs and staff will have access to the data.  Specimens will be identified by barcode linked to 
volunteer ID.  The table linking the volunteer ID to the volunteer’s identifying information will be 
maintained in a password-protected network file, with access only by the clinical PIs and study 
coordinator(s).  Any published data will have all identifiers removed. 
The study protocol, documentation, data, and all other information generated will be held in strict 
confidence.  No information concerning the study or the data will be released to any unauthorized 
third party without prior written approval of the sponsor. 
The study monitor or other authorized representatives of the sponsor may inspect all documents 
and records required to be maintained by the investigator, including but not limited to, medical 
records (office, clinic, or hospital) and pharmacy records for the volunteers in this study.  The 
clinical study site will permit access to such records. 
Records for the study will be retained for 5 years as required by University of Maryland policy. 
Volunteer confidentiality will be protected to the fullest extent permitted by law.  Volunteer names 
will not be used in any published report.  Study records may be confidentially reviewed by federal 
agencies such as National Institutes of Health (NIH), and the IRB. 
9.5 
Principal Investigator Responsibility When Volunteer Withdraws or is 
Discontinued 
In the event that the study is discontinued volunteers will continue to receive standard clinical 
care. 
9.6 
Future Use of Stored Specimens (if applicable) 
Volunteers will indicate if specimens may be used for future research as part of the consent form.  
Specimens will be coded and banked in the CVD specimen bank.  Any future studies will be 
reviewed by the UMB IRB prior to specimen use.  Genetic testing may not be conducted on these 
specimens without the approval of the IRB.  Future genetic testing might include the presence of 
genes or particular haplotypes or variants that might impact either positively or negatively the host 
immune response to vaccination.  No genetic testing for hereditary diseases will be done.  We do 
not anticipate that all of the samples obtained as part of this research study will be used during 
the course of the study.  Remaining samples will not be discarded, but instead will be frozen and 
kept indefinitely for future CVD 103-HgR research.  Banked samples will not be made available 
to the research volunteer (or his/her medical doctor) for other testing.  If a participant withdraws 
from the study they will not have the option to get the remaining portion of their sample(s) back; 
however, the sample(s) and all data for that participant will be anonymized with a unique 
identification code.  This code will not be linked or identifiable to a specific participant.  Volunteers 


Blood Donor 9000 
 
v1.0; 26 October 2018 
32 
 
who withdraw from the study and decline in the consent form to have specimens used for future 
research will have specimens discarded. 
If specimens have to be shipped to other institutions for specialized measurements (e.g., 
functional genomics, proteomics), we will seek prior approval of the UMB IRB to enable the 
shipment of the specimens as well as approval of the recipient institution’s IRB to enable the 
receipt and processing of such specimens.  Only coded specimens will be shipped.  Under no 
circumstances will information linking the code in the specimens to the name of the volunteer be 
made available to investigators receiving these specimens at other institutions.   
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
33 
 
10 
PUBLICATION POLICY  
If the data merit, the investigators will prepare one or more manuscripts for publication in peer-
reviewed professional journals or abstracts for presentation, oral or written, to a learned society 
or symposium.  All manuscripts will contain appropriate acknowledgement of the funding 
mechanism.  
The PI is responsible for the timely submission of all abstracts, manuscripts and reviews 
(co)authored by investigators and supported in part or in total by this protocol.  The PI and project 
leaders are requested to submit manuscripts to the Program Officer within two weeks of 
acceptance for publication so that an up-to-date summary of program accomplishments can be 
maintained and joint press conferences and press releases prepared.  Publications or oral 
presentations of work performed under this protocol are the responsibility of the PI and 
appropriate project leaders and will require appropriate acknowledgement of NIAID support. 
All investigators will submit or have submitted for them to the National Library of Medicine’s 
PubMed Central 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.  
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
34 
 
11 
LITERATURE REFERENCES 
 
1. 
Chen WH, Cohen MB, Kirkpatrick BD, Brady RC, Galloway D, Gurwith M, Hall RH, Kessler 
RA, Lock M, Haney D, Lyon CE, Pasetti MF, Simon JK, Szabo F, Tennant S, Levine MM. 
Single-dose Live Oral Cholera Vaccine CVD 103-HgR Protects Against Human 
Experimental Infection With Vibrio cholerae O1 El Tor. Clin Infect Dis. 2016;62(11):1329-
35. doi: 10.1093/cid/ciw145. PubMed PMID: 27001804; PMCID: PMC4872293. 
2. 
Perry RT, Plowe CV, Koumare B, Bougoudogo F, Kotloff KL, Losonsky GA, Wasserman 
SS, Levine MM. A single dose of live oral cholera vaccine CVD 103-HgR is safe and 
immunogenic in HIV-infected and HIV-noninfected adults in Mali. Bull World Health Organ. 
1998;76(1):63-71. PubMed PMID: 9615498; PMCID: PMC2305629. 
 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
35 
 
Appendix A. Typical Study Schedule: Serial Blood Donors3 
Visit 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
Day -7 to -28 
1 
3 
±1 
6 
±1 
8 
±1 
29 
±2 
91 
±3 
181 
±5 
361 
±30 
721 
±3m 
4 Yr 
±3m 
5 Yr 
±3m 
6 Yr 
±3m 
Informed Consent obtained 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/exclusion Review 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Urine Pregnancy test 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Interim History 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Vaccinate with CVD 103-HgR 
 
C 
 
 
 
 
 
 
 
 
 
 
 
Screening laboratories1 
X 
15mL 
 
 
 
 
 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
CBC w/differential2 
 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
PBMC  
 
X 
100
mL 
X 
100
mL 
X 
85 
mL 
X 
85 
mL 
X 
85 
mL 
X 
100
mL 
X 
100
mL 
X 
100
mL 
X 
100
mL 
X 
100
mL 
X 
100
mL 
X 
100
mL 
Serum  
 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
Gene expression 
 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
 
1 – Screening laboratory can be performed up to 28 days prior to the scheduled clinic visit and includes: CBC with differential, AST, 
ALT, and HIV antibody. The screening laboratory results must be reviewed prior to the scheduled blood draw for the respective 
visit. 
2 – CBC w/differential is to be performed on the scheduled clinic visit, with the blood draw for PBMC. It is intended for the back-
calculation of the number and concentration of lymphocyte subsets. 
3 – Not all volunteers will follow the same study schedule. The schedule will vary according to laboratory needs.  No more than 550 
mL of blood will be drawn within a 56 day period. 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
36 
 
Appendix B. Typical Study Schedule: Large Volume/unit Blood Donors3 
Visit 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
Day -7 to -28 
1 
3 
±1 
8 
±1 
29 
±5 
91 
±7 
181 
±30 
361 
±30 
721 
±3m 
4 Yr 
±3m 
5 Yr 
±3m 
6 Yr 
±3m 
8 Yr 
±3m 
Informed Consent obtained 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/exclusion Review 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Urine Pregnancy test 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Interim History 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Vaccinate with CVD 103-HgR 
 
C 
 
 
 
 
 
 
 
 
 
 
 
Screening laboratories1 
X 
15mL 
 
 
 
 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
X 
15 
mL 
CBC w/differential2 
 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
X 
4mL 
PBMC  
 
X 
150
mL 
X 
150
mL 
X 
100 
mL 
X 
100 
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
X 
200
mL 
Serum  
 
X 
8.5 
mL 
 
 
 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
X 
8.5
mL 
Gene expression 
 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
X 
3mL 
 
1 – Screening laboratory can be performed up to 28 days prior to the scheduled clinic visit and includes: CBC with differential, AST, 
ALT, and HIV antibody. The screening laboratory results must be reviewed prior to the scheduled blood draw for the respective 
visit. 
2 – CBC w/differential is to be performed on the scheduled clinic visit, with the blood draw for PBMC. It is intended for the back-
calculation of the number and concentration of lymphocyte subsets. 
3 – Not all volunteers will follow the same study schedule. The schedule will vary according to laboratory needs.  No more than 550 
mL will be drawn within a 56 day period. 
 
 


Blood Donor 9000 
 
v1.0; 26 October 2018 
37 
 
Appendix C. Clinical Laboratory values, for study eligibility 
 
 
Parameter 
Values  Leading to Study Exclusion 
Low Outlier 
(or Change from Reference)* 
High Outlier 
(or Change from Reference)* 
ALT (SGPT) 
N/A 
> 1.5 x ULN 
AST (SGOT) 
N/A 
> 1.5 x ULN 
WBC† 
< 0.81 x LLN 
> 1.09 x ULN 
Hemoglobin† 
M: < 0.92 x LLN 
F: < 0.91 x LLN 
M: > 1.18 x ULN 
F: > 1.18 x ULN 
Platelet (x 103) † 
< 0.8 x LLN 
> 1.2 x ULN 
HIV 
Positive 
Blood Type§ 
Not evaluated for eligibility (this test does not need to be repeated) 
For blood unit donations, the following lab values are exclusionary:‡ 
WBC 
<3.5 x103 /mm3 
>11 x103 /mm3 
Hemoglobin 
<12.5 g/dL 
>18 g/dL 
Platelet Count 
<150 x103 /mm3 
>500 x103 /mm3 
ULN = Upper limit of normal range 
LLN = lower limit of normal range 
N/A = not applicable 
* Values that lead to exclusion are defined as either an absolute value based on the reference range provided by Garcia (the usual 
clinical laboratory used by CVD) or the deviation from the reference value of a particular clinical laboratory by the order of 
magnitude specified.   "N/A" means that any lower value is acceptable for inclusion 
† Note that different WBC, Hemoglobin, and Platelet Count criteria are defined, for those being evaluated for blood unit (large 
volume) blood donations 
§ Blood Type is performed only once, at initial screening only 
‡ to be performed by the Cell Component Therapy Facility at University of Maryland Greenebaum Cancer Center, as part of their 
standard operating procedures 


Blood Donor 9000 
 
v1.0; 26 October 2018 
38 
 
Appendix D. Instructions for Vaxchora 
 
 
Storage 
 
 
Vaxchora must be stored in a freezer between -13°F and 5°F. 
Protect from light and moisture. Packets do not require 
thawing prior to reconstitution.