A Staged Phase I/II Study, to Assess Safety, Efficacy and Immunogenicity of a New Hepatitis C Prophylactic Vaccine Based on Sequential Use of AdCh3NSmut1 and MVA-NSmut
Phase: PHASE1
Conditions: Hepatitis C
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

DMID Protocol 10-0069  
Version 9.0 
 
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i 
REDACTED VERSION: MAY 14, 2019 FOR NLM 
 
 
A Staged Phase I/II Study, to Assess Safety, Efficacy and 
Immunogenicity of a New Hepatitis C Prophylactic Vaccine 
Based on Sequential Use of AdCh3NSmut1 and MVA-NSmut 
 
DMID Protocol Number:  10-0069 
 
DMID Funding Mechanism:  HHSN 26620040073C 
 
Pharmaceutical Support Provided by:  Okairos AG 
 
IND Sponsor:  Division of Microbiology and Infectious Diseases (DMID), 
National Institute of Allergy and Infectious Diseases, National Institutes of 
Health 
 
Co-Principal Investigators:  Kimberly Page, Ph.D., M.P.H. and  
Andrea Cox, M.D., Ph.D. 
 
DMID Clinical Project Manager:  Peter Wolff, M.H.A. 
 
Version Number:  9.0 
 
 6 October 2015 

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STATEMENT OF COMPLIANCE 
The study will be carried out in accordance with Good Clinical Practice (GCP) as 
required by the following:  
• United States (US) Code of Federal Regulations (CFR) applicable to clinical 
studies (45 CFR Part 46; 21 CFR Part 50, 21 CFR Part 56, and 21 CFR Part 
312) 
• ICH E6; 62 Federal Register 25691 (1997)  
• NIH Clinical Terms of Award 
All key personnel (all individuals responsible for the design and conduct of this study) 
have completed Human Subjects Protection Training. 
 
 

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SIGNATURE PAGE 
The signatures below constitute the approval of this protocol and appendices, and 
provide the necessary assurances that this trial will be conducted according to all 
stipulations of the protocol, including all statements regarding confidentiality, and 
according to local legal and regulatory requirements and applicable US federal 
regulations and ICH guidelines. 
 
 
 
 
 
Signed:  
Date: 
 
 
Andrea L. Cox, M.D., Ph.D. 
Co-Principal Investigator 
The Johns Hopkins University, Baltimore 
 
 
 
 
 
Signed:  
Date: 
 
 
Kimberly Page, Ph.D., M.P.H. 
Co-Principal Investigator 
University of New Mexico, Albuquerque 
 
 
 
Signed:  
Date: 
 
 
Paula Lum, M.D., M.P.H. 
Site Investigator 
University of California, San Francisco 
 

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TABLE OF CONTENTS 
Statement of Compliance ................................................................................................. ii 
Signature Page ............................................................................................................... iii 
Table of Contents ............................................................................................................ iv 
List of Abbreviations ...................................................................................................... viii 
List of Tables .................................................................................................................. xii 
Protocol Summary ......................................................................................................... xiii 
Schematic of Study Design ............................................................................................ xv 
1 
Key Roles ................................................................................................................ 1 
2 
Background Information and Scientific Rationale .................................................... 4 
2.1 
Background Information ............................................................................... 5 
2.2 
Previous Human Experience ...................................................................... 13 
2.2.1 Chimpanzee Adenovirus Vaccines .................................................. 13 
2.2.2 MVA Derived Vaccine Vectors ......................................................... 13 
2.2.3 HCV Vaccine Using NSmut Antigen by Okairos .............................. 14 
2.2.4 Prime/boost with Chimpanzee Derived Adenovirus and MVA 
Vectored Vaccines ........................................................................... 16 
2.3 
Scientific Rationale (including dose selection) ............................................ 17 
2.4 
Potential Risks and Benefits ....................................................................... 18 
2.4.1 Potential Risks ................................................................................. 18 
2.4.2 Unknown Risks ................................................................................ 19 
2.4.3 Potential Benefits ............................................................................. 20 
3 
Study Objectives ................................................................................................... 21 
3.1 
Primary Objectives: Safety and Efficacy ..................................................... 21 
3.1.1 Primary Safety Endpoints ................................................................ 21 
3.1.2 Primary Efficacy Endpoint ................................................................ 21 
3.2 
Secondary Objective: Immunogenicity ....................................................... 22 
3.2.1 Secondary Immunogenicity Endpoint .............................................. 22 
3.3 
Exploratory endpoints in the evaluation of efficacy ..................................... 22 
4 
Study Design ......................................................................................................... 25 
5 
Study Population ................................................................................................... 27 
5.1 
Inclusion Criteria ......................................................................................... 27 
5.2 
Exclusion Criteria ........................................................................................ 28 
5.3 
Randomization and Code Breaking Procedures ......................................... 30 
5.4 
Withdrawal .................................................................................................. 31 
5.4.1 Reasons for Withdrawal ................................................................... 31 
5.4.2 Handling of Discontinuation of Vaccine Regimen ............................ 32 
5.5 
Termination of the Study ............................................................................ 32 
6 
Investigational Products ........................................................................................ 33 
6.1 
Study Products Description ........................................................................ 33 
6.1.1 Acquisition ....................................................................................... 35 

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6.1.2 Product Storage, Stability, and Expiration........................................ 35 
6.1.3 Preparation & Administration ........................................................... 36 
6.1.4 Maintaining Blind for Investigational Product ................................... 37 
6.1.5 Accountability/Final Disposition for the Investigational Product(s) ... 37 
7 
Study Procedures/Evaluations .............................................................................. 38 
7.1 
Clinical Evaluations .................................................................................... 38 
7.2 
Concomitant Medications/Treatments ........................................................ 38 
7.3 
Prevention: Counseling and Risk Reduction .............................................. 39 
7.4 
Contraception Status and Pregnancy ......................................................... 40 
7.5 
Behavioral Risk Assessment Questionnaire ............................................... 40 
7.6 
Laboratory Evaluations ............................................................................... 40 
7.6.1 Clinical Laboratory Evaluations ........................................................ 41 
7.6.2 Endpoint Assays:  HCV serology and virology ................................. 41 
7.6.3 Immunogenicity Evaluations ............................................................ 43 
7.6.4 Exploratory Assays .......................................................................... 43 
7.6.5 Ancillary Assays ............................................................................... 44 
7.6.6 Future use of stored specimens ....................................................... 44 
7.7 
Specimen Preparation, Handling, and Shipping ......................................... 44 
7.7.1 Instructions for Specimen Preparation, Handling, and Storage ....... 44 
7.7.2 Specimen Shipment ......................................................................... 45 
8 
Study Schedule ..................................................................................................... 46 
8.1 
Screening and Enrollment .......................................................................... 46 
8.1.1 Anti-HCV Pre-Screening Day -30 to -1 ............................................ 46 
8.1.2 Day -30 to -1, Visit 00, Study Screening .......................................... 46 
8.1.3 Day 0, Visit 1, Dose 1 ...................................................................... 47 
8.2 
Subject Follow-Up ...................................................................................... 48 
8.2.1 Day 3, Visit 2 .................................................................................... 49 
8.2.2 Day 7, Visit 3 .................................................................................... 49 
8.2.3 Day 14, Visit 4 .................................................................................. 49 
8.2.4 Day 30, Visit 5 .................................................................................. 50 
8.2.5 Day 56, Visit 6, Dose 2 .................................................................... 50 
8.2.6 Day 59 [Target 3 Days Post Second Vaccination], Visit 7 ............... 52 
8.2.7 Day 63 [Target 7 Days Post Second Vaccination], Visit 8 ............... 52 
8.2.8 Day 90, Visit 9 .................................................................................. 53 
8.2.9 Day 120, Visit 10 .............................................................................. 53 
8.2.10 Days 150–270, Visits 11–15 ............................................................ 54 
8.2.11 Day 300, Visit 16 .............................................................................. 54 
8.2.12 Day 330, Visit 17 .............................................................................. 55 
8.2.13 Day 360, Visit 18 .............................................................................. 55 
8.2.14 Days 390–570, Visits 19–25 ............................................................ 56 
8.2.15 Day 600, Visit 26 .............................................................................. 56 
8.3 
Follow-up for Subjects with HCV Infection .................................................. 57 

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8.3.1 HCV Day 30, Visit H01 .................................................................... 58 
8.3.2 HCV Days 60–210, Visits H02–H07 ................................................ 58 
8.3.3 HCV Day 240, Visit H08 .................................................................. 59 
8.3.4 HCV Day 270, Visit H09 .................................................................. 60 
8.4 
Unscheduled Visit(s) ................................................................................... 60 
8.5 
Early Termination ....................................................................................... 61 
8.5.1 For HCV-uninfected subjects ........................................................... 61 
8.5.2 For HCV-infected subjects ............................................................... 61 
8.6 
Referral for Acute HCV Infection ................................................................ 62 
8.7 
Subsequent Positivity for Anti-HCV: Distinguishing Intercurrent HCV 
Infection from Vaccine Induced Positive HCV Serology ............................. 62 
9 
DMID Safety Reporting and Safety Assessment Monitoring ................................. 63 
9.1 
Responsibilities........................................................................................... 63 
9.2 
Definitions ................................................................................................... 63 
9.3 
Safety Reporting Requirements.................................................................. 64 
9.3.1 Reporting Interval ............................................................................ 64 
9.3.2 Notification of the Sponsor of Serious Adverse Events .................... 65 
9.3.3 Regulatory Reporting for Studies Conducted Under 
DMID-Sponsored IND ...................................................................... 65 
9.3.4 Reporting of Pregnancy ................................................................... 66 
9.4 
Investigator’s Assessment of Adverse Events ............................................ 66 
9.4.1 Assessment of Seriousness ............................................................. 66 
9.4.2 Assessment of Severity ................................................................... 66 
9.4.3 Assessment of Association .............................................................. 67 
9.4.4 Assessment of Reactogenicity ......................................................... 67 
9.5 
Safety Monitoring by the DMID Safety Oversight Mechanism .................... 68 
9.5.1 Data Safety and Monitoring Board (DSMB) ..................................... 68 
9.5.2 Independent Safety Monitor (ISM) ................................................... 68 
9.5.3 Interruption or Discontinuation of Study Enrollment and Study 
Product Administration for all Subjects in the Study ........................ 68 
9.6 
Halting Criteria/Rules .................................................................................. 69 
10 
Clinical Monitoring ................................................................................................. 70 
10.1 
Site Monitoring Plan ................................................................................... 70 
11 
Statistical Considerations ...................................................................................... 71 
11.1 
Study Hypotheses ...................................................................................... 71 
11.2 
Sample Size Considerations ...................................................................... 71 
11.3 
Planned Interim Analyses ........................................................................... 71 
11.4 
Final Analysis Plan ..................................................................................... 73 
11.4.1 Definition of Analysis Cohorts .......................................................... 73 
11.4.2 Safety Analyses ............................................................................... 73 
11.4.3 Evaluation of the Primary Efficacy Endpoint .................................... 74 
11.4.4 Immunogenicity Analyses ................................................................ 75 

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11.4.5 Evaluation of Exploratory Efficacy Endpoints ................................... 76 
12 
Data Handling/Record Keeping/Source Documents ............................................. 78 
12.1 
Source Documents and Access to Source Documents .............................. 78 
12.2 
Data Management Responsibilities ............................................................ 78 
12.3 
Data Capture Methods ............................................................................... 79 
12.4 
Types of Data ............................................................................................. 79 
12.5 
Timing/Reports ........................................................................................... 79 
12.6 
Study Records Retention ............................................................................ 79 
13 
Quality Control and Quality Assurance.................................................................. 80 
14 
Ethics/Protection of Human Subjects .................................................................... 81 
14.1 
Ethical Standard/Declaration of Helsinki ..................................................... 81 
14.2 
Institutional Review Board .......................................................................... 81 
14.3 
Informed Consent Process ......................................................................... 81 
14.4 
Subject Confidentiality ................................................................................ 83 
14.5 
PI Responsibility When Subject Withdraws or Is Discontinued .................. 84 
14.6 
Referrals for HCV Treatment and Care ...................................................... 84 
15 
Protocol Conduct ................................................................................................... 85 
15.1 
Subject Confidentiality ................................................................................ 85 
15.2 
Data Collection, Handling, Source Documents ........................................... 85 
15.3 
Protocol Deviations ..................................................................................... 86 
15.4 
Recruitment and Retention of Study Subjects ............................................ 86 
15.5 
Study Subject Reimbursement ................................................................... 88 
16 
Publication Policy .................................................................................................. 90 
17 
References ............................................................................................................ 91 
Supplements/Appendices .............................................................................................. 99 
Appendix A:  Acceptable Ranges for Clinical Laboratory Evaluations at Screening .... 100 
Appendix B:  Toxicity Table ......................................................................................... 101 
Appendix C:  Study Visit Chart .................................................................................... 106 
Appendix D:  Study Visit Chart – Confirmed HCV Infection ......................................... 108 
Appendix E:  Alcohol Use Disorders Identification Test (AUDIT) C test ...................... 109 
 

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LIST OF ABBREVIATIONS 
Ab 
Antibody 
AdCh 
Chimpanzee Derived Adenovirus 
AE 
Adverse Event/Adverse Experience 
AIDS 
Acquired Immune Deficiency Syndrome 
ALT 
Alanine Aminotransferase 
ATP 
According To Protocol 
AUDIT 
Alcohol Use Disorders Identification Test 
BP 
Blood Pressure 
BSRI 
Blood Systems Research Institute 
C 
Celsius 
CBC 
Complete Blood Count 
CD4 
Cluster of Differentiation 4 
CD8 
Cluster of Differentiation 8 
CDC 
Centers for Disease Control and Prevention 
CEF 
Chicken Embryo Fibroblast 
CIN 
Cervical Intraepithelial Neoplasia 
CFR 
Code of Federal Regulations 
cGMP 
Current Good Manufacturing Practices 
Cl 
Chlorine 
CLIA 
Clinical Laboratory Improvement Amendments 
CPM  
Clinical Project Manager 
CRF 
Case Report Form 
CROMS 
Clinical Research Operations and Management Support, DMID 
CRO 
Contract Research Organization 
CTL 
Cytotoxic T Lymphocyte 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH, DHHS 
DMSO 
Dimethyl Sulfoxide 
DSMB 
Data and Safety Monitoring Board 
DUIT 
Drug Users Intervention Trial 
eCRF 
Electronic Case Report Form 
EDTA 
Ethylenediaminetetraacetic Acid 
EIA or ELISA 
Enzyme Immunoassay or Enzyme-Linked Immunosorbant Assay 
ELISpot 
Enzyme-Linked Immunospot Assay 
F 
Fahrenheit 
FDA 
Food and Drug Administration, DHHS 
FDAAA 
Food and Drug Administration Amendments Act 
FWA 
Federalwide Assurance 

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GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
GMT 
Geometric Mean Titer 
GP 
General Practitioner 
HBsAg 
Hepatitis B Surface Antigen 
HBV 
Hepatitis B Virus 
HCC 
Hepatocellular Carcinoma 
HCV 
Hepatitis C Virus 
HEENT 
Head, Ears, Eyes, Nose, Throat 
HgB 
Hemoglobin 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HIVNET 
HIV Network for Prevention Trials 
HLA 
Human Leukocyte Antigen 
IATA 
International Air Transport Association 
IB 
Investigator’s Brochure 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
IDES 
Internet Data Entry System 
IDU or IDUs 
Injection Drug Use or Injection Drug Users 
IEC 
Independent or Institutional Ethics Committee 
IgG 
Immunoglobulin G 
IFN 
Interferon 
IL28B 
Interleukin-28B 
IND 
Investigational New Drug  
INR 
International Normalized Ratio 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
IU 
International Units 
JHU 
The Johns Hopkins University 
mcg 
Microgram 
MedDRA 
Medical Dictionary for Regulatory Activities 
ME-TRAP 
Multiple Epitope - Thrombospondin-Related Adhesion Protein 
Mg 
Magnesium 
mg 
Milligram 
MHC 
Major Histocompatibility Complex 
mITT 
Modified Intention to Treat 
mL 
Milliliter 
mm 
Millimeter 
mM 
Millimole 

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MM 
Medical Monitor 
mmHg 
Millimeter of Mercury 
MOP 
Manual of Procedures 
MVA 
Modified Vaccinia Ankara 
N 
Number (Typically Refers to Subjects) 
Na 
Sodium 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, DHHS 
NIH 
National Institutes of Health, DHHS 
NLM 
National Library of Medicine 
NS 
Nonstructural Region 
OCRA 
Office of Clinical Research Affairs, DMID, NIAID, NIH, DHHS 
OHRP 
Office for Human Research Protections, DHHS 
OHSR 
Office for Human Subjects Research, NIH, DHHS 
ORA 
Office of Regulatory Affairs, DMID, NIAID, NIH, DHHS 
PBMC 
Peripheral Blood Mononuclear Cell 
PBS 
Phosphate Buffered Saline 
PCR 
Polymerase Chain Reaction 
PD 
Protocol Deviation 
pfu 
Plaque-Forming Units 
PI 
Principal Investigator 
POWs 
Peer Outreach Workers/Trained Recruiters 
PTT 
Partial Thromboplastin Time 
PVG 
Pharmacovigilance Group 
PY 
Person-Years 
PYO 
Person-Years of Observation 
QA 
Quality Assurance 
QC 
Quality Control 
QM 
Quality Management 
RBV 
Ribavirin 
RCA 
Replication Competent Adenovirus 
RIBA 
Recombinant Immunoblot Assay 
RNA 
Ribonucleic Acid 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SDCC 
Statistical and Data Coordinating Center 
SGPT 
Serum Glutamic Pyruvic Transaminase 
SIV 
Simian Immunodeficiency Virus 
SFC 
Spot-Forming Cell 
SMC 
Safety Monitoring Committee 
SMS 
Short Message Service 
SOP 
Standard Operating Procedure 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 

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TRAP 
Thrombospondin-Related Adhesion Protein 
UCSF 
University of California San Francisco 
UK 
United Kingdom 
UNM 
University of New Mexico 
US 
United States 
vp 
Virus Particles 
WBC 
White Blood Cell 
WHO 
World Health Organization 
 

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LIST OF TABLES 
Table 1. Okairos’ on-going clinical trials with NSmut Antigen HCV Vaccine……………13 
 
LIST OF FIGURES 
Figure 1. The nonstructural region of the hepatitis C virus encoded by the vaccine. ...... 7 
Figure 2. The phylogenetic tree of adenovirus’ hexons. .................................................. 9 
Figure 3. Kinetics of IFN-γ ELISpot responses over time. ............................................. 12 
 
 

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PROTOCOL SUMMARY 
 
Title: 
A Staged Phase I/II Study, to Assess Safety, Efficacy and 
Immunogenicity of a New Hepatitis C Prophylactic 
Vaccine Based on Sequential Use of AdCh3NSmut1 and 
MVA-NSmut 
 
Phase: 
 
Design: 
 
I/II 
 
Double-blinded, randomized, placebo-controlled two 
stage study. 
 
Stage 1:  68±4 evaluable subjects 
Arm A:  34±2 evaluable subjects; 1 dose AdCh3NSmut1 
2.5 x 1010 vp at Day 0, and 1 dose MVA-NSmut 
1.8 x 108 pfu at Day 56 
Arm B:  34±2 evaluable subjects; 1 placebo dose at Day 
0, and 1 placebo dose at Day 56 
 
Stage 2:  472±4 subjects  
Enrollment will be initiated after an interim analysis of the 
Stage 1 safety and immunogenicity data.  
Arm A:  236±2 subjects; 1 dose AdCh3NSmut1 2.5 x 1010 
vp at Day 0, and 1 dose MVA-NSmut 1.8 X 108 
pfu at Day 56 
Arm B:  236±2 subjects; 1 placebo dose at Day 0, and 1 
placebo dose at Day 56 
 
Total number of subjects: 540±8 subjects 
  
Population: 
 
 
 
Number of Sites: 
Males and females negative for HCV antibodies and HCV 
RNA at screening, active injection drug users at high risk 
for hepatitis C, and 18 to 45 years old. 
 
Three Sites (University of California San Francisco; The 
Johns Hopkins University, Baltimore; University of New 
Mexico, Albuquerque) 
 
Study Duration: 
Approximately 63 months 
 

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Subject Participation 
Duration: 
12 months to 29 months. Non-viremic subjects will be 
followed for 20 months; subjects who are viremic will be 
followed for 9 months after detection of HCV (up to month 
29). 
 
Description of Agent or 
Intervention: 
Two investigational vaccines administered sequentially 
as a prime and boost: 
 
AdCh3Nsmut1:  2.5 x 1010 viral particles and  
MVA-NSmut:  1.8 x 108 pfu 
 
As intramuscular injections in the deltoid region of the 
arm. 
 
Objectives: 
Primary: 
• To assess the safety of the new candidate hepatitis C 
virus vaccines, AdCh3NSmut1 and MVA-NSmut, 
compared to placebo when administered to HCV-
uninfected injection drug users (IDUs). 
• To determine if AdCh3NSmut1 and MVA-NSmut HCV 
vaccines will reduce incidence of chronic HCV 
infection compared to placebo among HCV-uninfected 
IDUs. 
 
Secondary: 
• To evaluate the immunogenicity of the new candidate 
hepatitis C virus vaccines, AdCh3NSmut1 and MVA-
NSmut, compared to placebo when administered to 
HCV-uninfected IDUs. 
Estimated Date of 
Completion: 
 
2016 
 

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SCHEMATIC OF STUDY DESIGN
FOLLOW UP VISITS AT DAYS 3, 7, 
14, AND 30 
TOTAL N: 540±8 
STAGE 1 (n=68±4) 
STAGE 2 (n=472±4) 
SCREENING VISIT 
(Consent and screen) 
1st Vaccination visit at Day 0 
ARM A 
(n=34±2) 
1st vaccination 
with 
AdCh3NSmut1 
ARM B 
(n=34±2) 
1st Placebo 
2nd Vaccination visit at Day 56 
ARM A  
2nd 
vaccination 
with MVA-
ARM B  
2nd Placebo 
FOLLOW UP VISITS AT DAYS 59 
AND 63  
INTERIM ANALYSIS OF SAFETY 
AND IMMUNOGENICITY DATA 
18 MONTHLY FOLLOW UP VISITS 
AFTER THE 2ND VACCINATION, 
WITH 9 MONTHLY EVALUATIONS 
FOLLOWING FIRST DETECTED 
HCV INFECTION  
SCREENING VISIT 
(Consent and screen) 
1st Vaccination visit at Day 0 
ARM B 
(n=236±2) 
1st Placebo 
ARM A 
(N=236±2) 
1st vaccination 
with 
AdCh3NSmut1 
FOLLOW UP VISITS AT DAYS 3, 7, 
14, AND 30 
2nd Vaccination visit at Day 56 
ARM B  
2nd Placebo 
ARM A  
2nd 
vaccination 
with MVA-
FOLLOW UP VISITS AT DAYS 59 
AND 63 

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1 Key Roles 
 
 
University of New Mexico (UNM) 
Kimberly Page, Ph.D., 
M.P.H. 
Co-Principal Investigator 
Department of Internal Medicine 
University of New Mexico Health Sciences Center 
MSC10 5550 
1 University of New Mexico 
Albuquerque, NM 87131-0001 USA 
Phone: +1 (505) 272-2520 (office) 
Mobile: XXXXXXXXXX 
Email: Pagek@salud.unm.edu 
 
 
Karla Thornton, M.D., 
M.P.H. 
 
Physician Investigator  
University of New Mexico Health Sciences Center 
Albuquerque, NM 87131-0001 USA 
Phone: +1 (505) 272-6335 (office) 
Mobile: XXXXXXXXXX 
Email: kthornton@salud.unm.edu  
 
 
University of California, San Francisco (UCSF) 
Paula Lum, M.D., M.P.H. Site Investigator 
University of California, San Francisco 
995 Potrero Ave, SFGH, Bldg 80, Ward 84 
San Francisco, CA 94110 USA 
Phone: +1 (415) 476-4082, ext 411 
Pager: XXXXXXXXXX 
Email: plum@php.ucsf.edu   
 
 
The Johns Hopkins University, Baltimore (JHU) 
Andrea L. Cox, M.D., 
Ph.D. 
Co-Principal Investigator  
The Johns Hopkins University School of Medicine 
Rangos Building, Suite 536 
855 N. Wolfe St. 
Baltimore, MD 21205 USA 
Phone: +1 (410) 502-2715 (office)  
Mobile: XXXXXXXXXX 
Pager: XXXXXXXXXX   
Fax: XXXXXXXXXX    
Email: acox@jhmi.edu 
 

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Okairos AG, Basel, Switzerland and ReiThera SRL, formerly Okairos SRL, 
Rome, Italy 
Alfredo Nicosia, Ph.D. 
Chief Scientific Officer 
Viale Città d’Europa, 679 
00144 Rome – Italy 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX 
 
 
Antonella Folgori, Ph.D. Director of Immunology 
Viale Città d’Europa, 679 
00144 Rome – Italy 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX 
 
 
Dario Ruscica, M.D. 
Clinical Research Physician 
Viale Città d’Europa, 679 
00144 Rome – Italy 
Phone: XXXXXXXXXX 
Email: XXXXXXXXXX     
 
 
NIH – Division of Microbiology and Infectious Disease (DMID) 
Clinical Project 
Manager 
Peter A. Wolff, M.H.A. 
DMID/NIAID/NIH/DHHS 
5601 Fishers Lane, Rm 8E47, MSC 9825 
Bethesda, MD 20892 USA 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
Scientific Lead 
Rajen Koshy, Ph.D. 
DMID/NIAID/NIH/DHHS 
5601 Fishers Lane, Rm 8F49, MSC 9826 
Rockville, MD 20852 USA 
Phone: XXXXXXXXXX  
Email: XXXXXXXXXX  
 
 
 
 

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The Emmes Corporation 
Statistical and Data 
Coordinating Center 
(SDCC) 
The Emmes Corporation 
401 N. Washington St., Suite 700 
Rockville, MD 20850 USA 
Phone: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
Statistician 
Michael R. Wierzbicki, Ph.D. 
The Emmes Corporation 
401 N. Washington St., Suite 700 
Rockville, MD 20850 USA 
Phone: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
FHI 360 
 
Protocol Specialist 
Anne Rinaldi 
FHI 360 
359 Blackwell Street, Suite 200 
Durham, NC  27701 
Phone: XXXXXXXXXX  
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
Fisher BioServices 
 
Study Agent Repository DMID-Clinical Agents Repository Contract 
Fisher BioServices 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 USA 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
 
 
Clinical Research Operations and Management Support (CROMS) 
Safety and 
Pharmacovigilance 
Organization/Contact 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management 
Support (CROMS) 
6500 Rock Spring Dr., Suite 650 
Bethesda, MD 20814 USA 
SAE line: XXXXXXXXXX 
SAE Fax line: XXXXXXXXXX 
SAE Email: XXXXXXXXXX  
 
 

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Institutional Review Boards (IRBs) 
University of California 
San Francisco 
The Committee on Human Research (CHR) 
Office of Research 
3333 California Street, Suite 315 
University of California 
San Francisco, CA 94118 USA 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
Johns Hopkins 
University 
Johns Hopkins Bloomberg School of Public Health 
Institutional Review Board 
615 N. Wolfe Street 
Suite E1100 
Baltimore, Maryland 21205 USA 
Phone: XXXXXXXXXX 
Toll-Free: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX  
 
 
University of New 
Mexico  
UNM Health Sciences Center Institutional Review 
Board 
B71, MSC08 4560 
1 University of New Mexico 
Albuquerque, NM 87106 USA 
Phone: XXXXXXXXXX 
Fax: XXXXXXXXXX 
Email: XXXXXXXXXX  
       

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2 Background Information and Scientific Rationale 
2.1 Background Information 
Hepatitis C: the need for a vaccine 
HCV is the world’s most common blood borne viral infection for which there is no vaccine.  
The WHO estimates that about 180 million people, some 3% of the world's population, 
have been infected, 130 million of whom are chronic HCV carriers.1,2  They estimate that 
three to four million persons are newly infected each year.3  HCV is a major etiological 
agent of liver disease including cirrhosis and hepatocellular carcinoma (HCC) in the U.S. 
and worldwide and the principal cause of liver transplantation.4,5  Population estimates of 
prevalence of HCV infection range from 1-2% in Australia, Canada, US and most of 
Europe, to between 5% and 10% in many countries of Africa, Latin America, Central and 
South-Eastern Asia.  In the U.S., it is estimated that over four million have been infected, 
and 3.2 million have chronic infection.6  In the U.S. in 2008, the CDC estimated that 
18,000 new HCV infections occurred,7 the majority attributed to IDU.8 
 
Acute HCV infection is typically asymptomatic and spontaneously resolves in about 
20% of cases.  Jaundice and fulminant hepatitis occur but are not common.  In 70–80% 
of cases, the immune system is nevertheless unable to eliminate the virus and the 
disease becomes chronic.  Patients chronically infected with HCV typically show some 
liver function test abnormalities (elevated transaminase levels) and in some cases 
develop mixed cryoglobulinemia.  Chronic infection is associated with progressive liver 
fibrosis.  Although the clinical course of liver disease upon HCV infection is very 
variable, about 20% of chronically infected individuals will eventually develop cirrhosis, 
and some of them hepatocellular carcinoma.4  It has been estimated that only about 
50% of HCV-infected persons are diagnosed in most developed countries and that two-
thirds of them need to undergo antiviral treatment.9  However, the proportion of patients 
who access and complete treatment remains low.10-13 
 
The standard of care for patients with chronic HCV infection consists of the combination 
of pegylated interferon α (IFN) and ribavirin (RBV).14,15  Therapy achieves sustained 
viral clearance in no more than ~40-50% of patients infected with the most common 
genotype of HCV.  More than 20% of the IFN treated patients develop significant 
adverse effects and RBV frequently causes haemolytic anemia.  Current treatment 
regimens are considered inadequate due to limited efficacy and toxicity resulting in poor 
tolerability.  The various HCV genotypes respond differently to the treatment.  Genotype 
1 infections are more resistant to IFN therapy than infections with virus of genotypes 2 
and 3.  Novel therapies based on antiviral drugs that specifically target HCV protease 
are currently approved for clinical use in combination with IFN and RBV.  Recent clinical 
trials showed an increase in the number of genotype 1 infected individuals responding 
to the triple antiviral treatment (IFN/RBV and protease inhibitor).16-18  These new 

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treatments, however, are associated with the rapid emergence of viral escape 
variants.19  Furthermore, many patients are not eligible for therapy because of co-
morbid medical or psychiatric conditions or severe liver disease.10,20  For these reasons, 
as well as for the cost of the combined standard therapy, it is widely believed that HCV 
therapy will not have a significant impact on the disease in many parts of the world and 
will have a minimal impact in blocking the spread of infection within the human 
population.   
 
A significant epidemiologic impact on the spread of HCV infection is expected to occur 
only subsequent to the introduction of an effective vaccine.  Extensive work in the 
chimpanzee model suggests that immunity against HCV can be generated by initial 
infection 21-23 or vaccination, and protects chimpanzees from HCV disease after 
challenge with homologous or heterologous virus.22-25  Moreover, prospective studies in 
acutely infected humans have shown that clearance of the infection is correlated with 
the type, strength and breadth of the immune T-cell response.26-28  Recently, Osburn et 
al. have demonstrated that prior HCV clearance is associated with control of 
subsequent HCV infections in injection drug users.29  Reinfection is associated with 
broadening of the cellular immune response and shorter peak viremia with decreased 
duration of viremia, suggestive of protective immunity against HCV infection.  All these 
observations provide a rational basis for vaccine design and suggest that an HCV 
vaccine will be efficacious in prevention of chronic infection if capable of eliciting a T-cell 
response equivalent or superior to that observed in acute/resolving individuals. 
 
A vaccine is needed to reduce HCV disease especially for groups at high risk of infection.  
Injection drug users (IDUs) have the highest prevalence of any population, ranging from 
25% to 80%, and an incidence rate of new infections ranging from 10% to 30% 
(annualized rate).30,31  IDUs are also not considered good candidates for standard therapy 
due to numerous behavioral and clinical challenges.  Few effective prevention modalities 
are available or implemented in this population.32  The rates of infection remain highest in 
this group.  Given the asymptomatic nature of infection, it is also likely that the number of 
infections and the proportion attributed to IDU, are significantly underestimated.33  
Moreover, as many as 30% of the estimated one million HIV-infected people in US 
(approximately 300,000 subjects in all) are co-infected with HCV; a similar percentage is 
reported for HIV patients co-infected with HCV in Europe.9  The course of HCV-related 
disease is accelerated by HIV co-infection; the US Public Health Service has recognized 
HCV as an opportunistic infection of HIV.  The death rate from HCV is seven times higher 
among HIV-infected individuals than among patients infected with HCV alone.  Despite 
enormous research efforts, a successful HCV vaccine has not yet been developed.  
Importantly, a recent study evaluating the potential impact of HCV vaccination predicted 
that the introduction of a prophylactic HCV vaccine in an IDU population can have 
substantial impact on the overall incidence of chronic HCV infection.34  A vaccination 
program, optimized through targeting strategies and high vaccination rates, is expected to 

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reduce HCV prevalence of chronic infection to a state where, after several years, other 
interventions may drive HCV to extinction.34  The Co-Principal Investigators believe the 
IDU population represents one group that will benefit greatly if a hepatitis C vaccine is 
licensed; therefore, we propose to conduct a Phase I/II clinical trial in this population. 
 
The rationale for using HCV NS polyprotein as antigen for HCV prophylactic 
vaccine  
HCV belongs to the genus Hepacivirus in the family Flaviviridae.  It is an enveloped virus 
with an icosahedral capsid that contains a 9.6 kb-long, single-stranded, positive sense 
genomic RNA.  Its envelope contains two glycoproteins, E1 and E2, which form 
heterodimers at the surface of the virion.  The genomic RNA is translated into a viral 
polyprotein which is cleaved by cellular and viral-encoded proteases to generate the 
capsid protein (C), the two glycoproteins E1 and E2, p7, the viral proteases NS2 and 
NS3, and nonstructural proteins NS4a, NS4b, NS5a and NS5b, which are required for 
viral RNA replication.  HCV can be classified into a number of distinct genotypes and 
subtypes, whose distribution varies both geographically and between risk groups.35,36  
Genotype 1 represents the majority of known isolates (88.5% in US, 83.3% in Europe and 
62.2% in Asia).  In Europe, subtypes 1a and 1b are widely distributed, particularly in older 
age groups, with those infected through drug use also having significant prevalence of 
subtype 3a infections.  The 1b subtype is also the most frequently found in Japan, while in 
the United States (US), the closely related 1a subtype is slightly more prevalent.  
Moreover, HCV displays very high inter- and intra-individual genetic variability, especially 
in the E1 and E2 envelope glycoproteins where neutralizing determinants have been 
found.  In fact, neutralizing antibody responses have been shown to be effective against 
only a handful of viral variants.  Consistent with this scenario, the role of antibodies in viral 
clearance is still uncertain.  For instance, vaccination protocols in chimpanzees aimed at 
eliciting neutralizing antibodies have not been able to induce a sterilizing immunity, 
especially against heterologous viral inocula.37 
 
The prophylactic HCV vaccines proposed for this study are based on adenoviral and 
Modified Vaccinia virus Ankara (MVA) vectors.  The vaccines vectors encode the 1985 
amino acid-long HCV nonstructural region (NS), with genetically inactivated RNA-
dependent RNA polymerase activity (NS5b).  The mutated NS region is called NSmut 
(Figure 1). 
 
 

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Figure 1. The nonstructural region of the hepatitis C virus encoded by the vaccine. 
 
The HCV nonstructural region (NS) is commonly conserved between all six HCV 
genotypes and several of the major subtypes, making this region of HCV a good 
vaccine candidate.  The NS region encompasses about two thirds of the HCV genome 
and encodes five different proteins.  Upon transduction of mammalian cells with the 
vaccine, the encoded NS fragment is translated into a single polyprotein which is then 
proteolytically cleaved by the encoded NS3 protease into five mature products (NS3, 
NS4a, NS4b, NS5a and NS5b).  This process recapitulates the same cascade of events 
occurring in vivo upon viral infection and therefore provides for a physiological way of 
presenting the viral antigens to the host immune system.  An effective HCV vaccine will 
need to induce a broad cellular immune response due to the genetic diversity of human 
MHC alleles and of the virus.  The NS region from a 1b genotype BK strain isolate was 
chosen as vaccine candidate because its amino acid sequence is well conserved 
across several genomes covering 14 different subtypes belonging to the 6 major types 
(between 74 and 79% sequence identity).  The HCV database group has mapped a 
large number of T-cell epitopes in the NS region: more than 90 T helper and more than 
70 cytotoxic T lymphocyte (CTL ) epitopes.38  Recent published data in chronically and 
acutely infected subjects suggest a predominant role for epitopes mapped in the NS 
region in inducing T-cell immunity.26,39,40  Poly-specific T-cell response could be 
detected in human subjects infected by different HCV genotypes by using peptides 
reproducing the NS 1b/Bk vaccine sequence as antigens in the assay.26,41  Because of 
the multiple antigens encoded in this region, it is reasonable to speculate that NS 
proteins will induce T-cell responses specific for several viral determinants capable of 
binding to a large panel of different Human Leukocyte Antigen (HLA) molecules. 
 
Okairos researchers have performed a number of immunogenicity studies in non-human 
primates with adenovirus and DNA-based vectors encoding for HCV NS 1b/Bk 
sequence, and have shown that a high and broad T-cell response could be induced not 
only against a set of antigens spanning the 1b sequence,42,43 but also against antigens 
derived from the highly divergent genotype 3a which has 23% amino acid diversity.44  
Importantly, genotype 3 is the second most prevalent genotype in IDUs, the specific 
population we intend to enroll in the Phase I/II trial.  Furthermore, Okairos researchers 
NS5bmut 
AAG 
NS4 
NS5 
a 
b 
a 
b 
NSNS
NS3 protease 
Me

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have demonstrated that genetic vaccination with the 1b/BK NS antigen, inducing broad 
and long-lasting T-cell responses, protected chimpanzees from HCV disease after 
challenge with a heterologous virus belonging to the 1a genotype and differing from the 
vaccine sequence at more than 13% of the amino acid positions.25 
 
Chimpanzee Adenovirus as vaccine vectors 
Potent immunogenicity and lack of prolonged transgene expression have made 
adenovirus attractive viral vectors for vaccine development.  Replication defective 
adenovirus can be engineered by deletion of genes from the E1 locus, which is required 
for viral replication, and these viruses can be propagated easily with good yields in cell 
lines such as HEK 293 and PER.C6 (Crucell).45  They possess a stable virion allowing 
inserts of foreign genes without deletion.  They can infect many different cell types and 
the transferred information remains epichromosomal thus avoiding the risk of insertional 
mutagenesis.  Preclinical and clinical results conclusively showed superiority of 
adenovirus-vectored genetic vaccines based on the most common human adenovirus 
serotype 5 (Ad5) for the induction of T-cell responses in preclinical animal models and 
in Phase I safety and immunogenicity studies in humans.46-49  However, recent studies 
have shown that pre-existing immunity to Ad5 is capable of significantly blunting the 
immunological response induced by Ad5 vectored vaccines in rodents, in non-human 
primates and in Phase I clinical trials in humans.42,46  Various attempts have been made 
to overcome the problem of pre-existing immunity to Ad5, and thus exploit the full 
potential of the adenovirus vectors for the development of vaccines.  However, 
alternative adenovirus serotypes to be developed as vaccine vectors have to fulfill a 
number of requisites including efficient growth in cell lines suitable for GMP production 
and immunological potency comparable to that of Ad5. 
 
Simian adenoviruses are not known to cause pathological illness in humans and the 
prevalence of antibodies to chimpanzee derived adenoviruses (AdCh) have low/no 
seroprevalence (0%-18%) in the human population.50  In Equatorial Africa, the natural 
habitat for chimpanzees, seroprevalence is higher, but still significantly below that of 
Ad5.  Okairos performed a large screening of several adenoviruses isolated from 
chimpanzees and identified several chimpanzee adenoviruses that meet the necessary 
requirements for vaccine development.51  In chimpanzee adenoviruses the E1 locus can 
be deleted to render virus replication deficient and to allow trans-complementation in 
Ad5 E1 complementing cell line.51  Moreover, because of lack of sequence homology 
between the E1 flanking regions of Ad5 and AdCh adenoviruses, no recombination and 
production of replication competent virus occurs if the AdCh vectors are propagated in 
HEK 293 cells and PER.C6® (Crucell)52 eliminating the problem of replication 
competent adenovirus (RCA) generation during vector manufacturing.  Chimpanzee 
derived adenoviruses exhibit sequence similarity to human adenoviruses.  Hexon is the 
major capsid protein in adenoviruses; adenoviruses can be classified in subgroups 
based on homology in the hexon amino acid sequence.  Phylogenetic analysis of the 

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hexons of simian and human adenoviruses (Figure 2) shows substantial overlap 
indicating that there is no clear sequence feature that distinguished a simian from a 
human adenovirus.  In fact, these sequences suggest one large family of higher primate 
adenoviruses.  The potency of chimpanzee derived Ad vectors were assessed in mice, 
macaques and, recently, in humans.  The T-cell immunogenicity of some of these 
vectors matched or even exceeded the immunogenicity of the standard Ad5 vector used 
as a comparator.  The safety of these vectors has been similar to that of human 
adenovirus vectors suggesting that these might be suitable for widespread use. 
 
The simian virus AdCh3 has been chosen for the HCV vaccine development.  AdCh3 
can be classified based on the Hexon protein in subgroup C as human Ad5 and Ad6.  
The sequences of Ad5 and AdCh3 are clearly divergent in the hypervariable regions of 
hexons considered the target of neutralizing antibodies.53 
 
 
Figure 2. The phylogenetic tree of adenovirus’ hexons. 
 
Amino Acid Substitutions 
Bootstrap Trials = 1000, seed 
 
0 
21.7 
5 
10 
15 
20 
99.0 
62.7 
69.0 
100.0 
100.0 
97.3 
99.4 
99.3 
96.8 
100.0 
90.4 
98.6 
57.9 
76.6 
59.1 
NA 
51.2 
99.1 
91.9 
37.7 
59.6 
94.4 
100.0 
100.0 
99.3 
97.5 
Ad11 
Ad35 
Ad21 
Ad34 
AdCh1 
Ad7 
Ad3 
AdCh63  
Ad4 
Ad16 
AdCh7 
AdCh5 
AdCh9 
Ad33 
Ad24 
Ad44 
Ad32 
Ad30 
Ad25 
AdCh6 
Ad1 
Ad6 
Ad2 
AdCh3 
Ad5 
Ad40 
Ad41 
Ad12 

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Modified Vaccinia virus Ankara–based vaccines 
Modified Vaccinia virus Ankara (MVA), an attenuated strain of vaccinia virus that was 
originally developed as a smallpox vaccine, was obtained following extensive serial 
passage on primary chicken embryo fibroblasts (CEFs).  During this process of 
attenuation, MVA underwent deletion of 31 kb (~15%) of its genome, as compared to its 
parental strain, including a number of genes that contribute to viral evasion from host 
immune responses and that determine virus host range.54,55  As a result, MVA is unable 
to replicate productively in most mammalian cell types, including primary human cells.  
The resultant inability of MVA to undergo more than one infection cycle in a human host 
has imbued this virus with inherent safety that was demonstrated historically through the 
immunization of ~120,000 individuals during the smallpox eradication campaign.  More 
recently, the safety of MVA has been demonstrated in pre-clinical studies of immune-
deficient mice and immune-suppressed macaques,56,57 and in Phase I clinical trial 
evaluations of MVA as a next-generation smallpox vaccine.58  The desirable safety 
profile exhibited by MVA, in concert with its ability to express high levels (and large 
numbers) of foreign genes, has rendered MVA a leading candidate for evaluation as a 
vaccine vector against an array of infectious diseases and human cancers.  MVA-based 
vaccines against HIV/AIDS,59 malaria,60,61 tuberculosis,62 Human Papilloma Virus-
induced cervical intraepithelial neoplasia63 and melanoma64 are being evaluated in 
human clinical trials. 
 
Heterologous prime-boost vaccination strategy 
The main limitation of vaccination approaches based on viral vectors is linked to the 
induction of anti-vector immunity after the first immunization.  In fact, repeated 
administration of both recombinant adenoviruses and MVA vaccine vectors typically 
results in an increasingly diminished efficacy of such booster immunizations due to the 
elicitation of vector-specific neutralizing antibody responses.47,65  Several studies have 
shown that priming/boosting with different viral vector elicits higher immune response 
than repeated vaccination with an individual viral vector.  The combined use of these 
vectors in a heterologous prime-boost regimen is the best way to overcome the antiviral 
immunity induced by the first vaccination.  The utility of MVA-based vaccines to prime 
immune responses against foreign antigens appears to be limited due to unfavorable 
competition for immunodominance between the relatively large number of vector-
specific gene products55 and the much smaller number of intended vaccine antigens.66  
Therefore, an optimal regimen would use adenovirus first to prime and MVA later to 
boost the previously vaccine induced immune response.  An immunization protocol 
based on adenovirus as prime followed by MVA has demonstrated to be a powerful 
strategy to induce potent and durable T-cell responses.  This strategy enabled induction 
of protective immune response against mouse malaria67 and simian immunodeficiency 
virus (SIV) challenge in rhesus monkeys.68  Moreover, this approach is now in Phase I 
and IIa trials in the UK for a malaria vaccine with very promising results.69 
 

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Monkeys’ pre-clinical data in support of the choice of the vaccination strategy 
Here we summarize the assessments of the immunogenic potency of the heterogonous 
prime-boost regimen based on AdCh3NSmut1 followed by MVA-NSmut conducted in 
primates.  A group of 3 male macaques were primed at Week 0 with AdCh3NSmut1 
(1x1010 vp intramuscularly) and boosted at Week 8 with MVA-NSmut (2x108 pfu 
intramuscularly).  A second group of 3 male macaques was vaccinated with 2 injections 
of MVA-NSmut 8 weeks apart.  Analysis of HCV specific T-cell responses was 
performed at various time points following vaccination using an ELISpot assay.  Briefly, 
freshly isolated peripheral blood mononuclear cells (PBMC) were stimulated with 6 
pools of peptides encompassing the NS3-NS5B sequence and IFN-γ secreting cells 
were measured.   
 
T-cell responses to the vaccine insert were induced in all 3 animals of the first group 
after adenovirus vaccination, peaking at Week 4 post prime (Figure 3, below).  The sum 
of responses to the 6 NS pools at peak post-prime, ranged from 598 to 1908 spot 
forming cells (SFC)/106 PBMC.  The immune response was contracted at the time of 
boosting and was readily and efficiently expanded by MVA-NSmut injection, with a fold 
increase >30 compared to pre-boost residual responses.  Moreover, the peak values 
obtained post-boost were 3-fold higher than those at peak post-prime (sum range: 
3372-5021 SFC/106 PBMC).  Four weeks after MVA-NSmut boost (T=12 weeks), the 
effector T-cell response is again contracted, as expected.  For the MVA-primed/MVA-
boosted control group of animals data collected up to four weeks after boost show that 
no or very weak (58 and 61 SFC/million PBMC against a single peptide pool after boost 
in two animals) T-cell responses were mounted.  We expected, based on previous data 
performed with MVA vectors carrying other antigens, that the homologous prime–boost 
strategy would induce a much weaker immune response compared to the heterologous 
strategy.  Results in this study confirm previous findings and highlights the crucial role of 
AdCh3NSmut1 as priming agent in order to achieve a strong and broad T-cell response 
to the vaccine. 
 
 

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T=0
T=1
T=2
T=4
T=8
T=9
T=12
0
2000
4000
6000
3065
3071
2155
3053
6023
6147
weeks
IFNγ SFC/106 PBMC
T=0
T=1
T=2
T=4
T=8
T=9
T=12
0
1000
2000
3000
4000
5000
AdCh3/MVA
MVA/MVA
weeks
IFNγ SFC/10 6 PBMC
 
Figure 3. Kinetics of IFN-γ ELISpot responses over time. 
Data are expressed as IFN-γ Spot Forming Cells (SFC)/106 PBMC.  Individual data points (left panel), 
represent total NS response in each animal, obtained by summing reactivity to each of the 6 NS pools 
and subtracting 6 times the (Dimethyl sulfoxide) DMSO background.  Right panel shows each group’s 
median.  Arrows indicate vaccinations. 
2.2 Previous Human Experience 
2.2.1 Chimpanzee Adenovirus Vaccines 
The initial clinical experience with Adenoviral vectors as vaccine was based on the use 
Human Ad5 derived vectors.50  Ad5 based vectors are safe but unfortunately, pre-
existing immunity to Ad5 has been shown to blunt the induction of an immunological 
response.42,46  Therefore, novel Adenoviral vectors have been identified and tested for 
clinical use. 
2.2.2 MVA Derived Vaccine Vectors 
MVA is widely considered as the vaccinia virus strain of choice for clinical investigation 
because of its high safety profile.  The MVA virus can be administered in 
intracutaneous, subcutaneous, or intramuscular injections.  Over 120,000 humans have 
been inoculated and successfully vaccinated against smallpox with MVA during the 
campaign for the eradication of smallpox in Germany in the 1970s, including elderly and 
immunocompromised persons.  More recently, the use of MVA in recombinant vaccines 
is being tested in a number of clinical studies, including those for vaccines against HIV.  
59,70-72  An HCV vaccine program is in progress in France (Transgene vaccine TG4040), 
using MVAHCV vaccine that is similar to the MVA-NSmut vaccine proposed here.73-75  
The safety data obtained from previous trials using other MVA based vaccines have 
been excellent and no serious adverse events or suspected unexpected serious 
adverse reactions (SUSARs) have been reported. 

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2.2.3 HCV Vaccine Using NSmut Antigen by Okairos 
The HCV NSmut antigen, vectored by either adenoviral vectors or by MVA, is currently 
being used in three trials (HCV001 HCV002TV and HCV003) (Table 1). 
 
Table 1. Okairos’ on-going clinical trials with NSmut Antigen HCV Vaccine 
Study 
code(s) 
Current status 
Phase 
Administered 
products  
Subjects Population 
and Sample Size 
HCV001 
Completed 
I 
AdCh3NSmut 
Ad6NSmut 
Healthy subjects (41) 
 
HCV002TV In progress 
(Estimated end date: 
3rd Qtr, 2012) 
I 
AdCh3NSmut 
Ad6NSmut 
HCV-infected subjects 
(36) 
 
HCV003 
In progress 
(Estimated end date: 
4th Qtr, 2011) 
I 
AdCh3NSmut 
MVA-NSmut 
Healthy subjects (9) 
and HCV subjects (14) 
 
HCV001 is a Phase I study with the candidate HCV vaccines AdCh3NSmut and 
Ad6NSmut to address safety and immunogenicity of heterologous prime/boost regimen 
with two different adenovirus vectors in healthy subjects.  The same vectors are also 
under investigation in HCV-infected subjects (HCV002TV).  The HCV003 study mimics 
the immunization schedule proposed in this trial using the AdCh3NSmut to prime, and 
the MVA-NSmut to boost the immune response. 
 
The AdCh3NSmut vector used in HCV001 is essentially identical to AdCh3NSmut1 
proposed here for the Phase I/II clinical trial in IDU.  Minimal changes have been 
introduced in the manufacturing process to improve production.  These changes have 
not been shown to affect the safety profile and immunogenicity as determined by the 
pre-clinical toxicology assessment as described in more detail in the AdCh3NSmut1 
Investigator’s Brochure (AdCh3NSmut1 and MVA-NSmut repeated dose toxicity study 
in rat RTC Study No.: 82930).  Immunogenicity data in monkeys for the AdCh3NSmut1 
product are shown in Section 2.1 (Monkeys pre-clinical data in support of the choice of 
the vaccination strategy). 
 
AdCh3NSmut safety and immunogenicity data in healthy subjects 
Thirty-eight volunteers received AdCh3NSmut at least once during the HCV001 trial.  
Priming vaccinations of AdCh3NSmut were administered to 22 volunteers at doses of 5 
x 108vp (4 volunteers, 8 injections), 5 x 109 vp (4 volunteers, 8 injections), 2.5 x 1010 vp 
(10 volunteers, 15 injections) and 7.5 x 1010 vp (4 volunteers). 
 
A further 16 individuals received a single boosting vaccination of AdCh3NSmut at 2.5 x 
1010 vp following priming vaccinations with Ad6NSmut at varying doses.  The adverse 

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events described below have been deemed definitely, probably or possibly related to 
vaccination.  The complete final safety report of the study is shown in the appendix 2 of 
the Investigator’s Brochure (IB).  The following safety data pertain to the priming 
vaccinations with AdCh3NSmut in the HCV001 study. 
 
The observed frequency of local reactions to AdCh3NSmut was 66% overall and 
appeared to increase with dose; occurring in 25% of volunteers after 5x108vp, 75% after 
5x109vp, 80% after 2.5x1010vp, and 75% after 7.5x1010vp.  Local pain (51%), erythema 
(26%), warmth (14%) and swelling (11%) were the most commonly reported reactions.  
The frequency of local pain and erythema increased from 25% and 13% in low dose to 
75% and 50% in highest dose recipients.  Ninety-three percent of the reported local 
reactions were mild in severity.  All were self-limiting, with 86% resolved within 72 hours 
and 98% within 1 week.  
 
Systemic adverse events were observed in approximately 74% of recipients of 
AdCh3NSmut, with an observable increase in frequency at highest dose.  Fatigue 
(43%), headache (29%), and malaise (23%) were the most commonly reported 
systemic adverse events overall; however, in those receiving the highest dose (n=4) the 
frequencies were myalgia (100%), headache (75%), fever (50%), malaise (50%), and 
fatigue (50%).  
 
Ninety-six percent of observed systemic adverse events were mild in severity, with the 
remaining 4% moderate.  Eighty-nine percent of the observed systemic adverse events 
had resolved by 72 hours post-vaccination and 93% by 1 week.  No distinct difference in 
local or systemic reactogenicity following first and second vaccinations with 
AdCh3NSmut was observed.  Immunogenicity was assessed by ex-vivo IFN-γ ELISpot 
using 6 peptide pools spanning the HCV immunogen.  Following priming vaccinations 
with AdCh3NSmut the average peak HCV specific ELISpot responses were 142.5 
(5x108vp), 897.8 (5x109vp) and 1863 (2.5 x1010vp) spot forming units/106 PBMC.  No 
further increase of induced immune response was observed at the 7.5X1010 vp dose.  
All vaccinees responded to the 2.5 x1010vp vaccine dose.  HCV specific responses were 
generally multi-specific; at the highest vaccine doses all individuals responded to at 
least 2 peptide pools and 7/10 individuals responded to 4 or more peptide pools (data 
not shown).  
 
Moreover, in the HCV003 study, an additional 5 volunteers received one priming 
injection with AdCh3NSmut at 2.5x1010 doses and showed a safety profile comparable 
to that observed in the previous HCV001 trial. 
 

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MVA-NSmut safety and immunogenicity data in healthy subjects 
In the HCV003 study, 9 volunteers in total received MVA-NSmut intramuscularly at the 
dosage of 2 x 108 plaque forming units (pfu).  Four subjects received only a single 
injection of MVA (group A1), while 5 subjects received the injection of MVA 8 weeks 
after the priming injection with AdCh3NSmut at 2.5x1010 (group A2), ipsilateral to the 
priming injection.  No vaccine related serious adverse events occurred.  The overall 
observed percentage of volunteers with one or more local reactions after MVA injection 
was 100%.  Local pain (100%), erythema (67%), warmth (67%) and swelling/induration 
(56%) were the most commonly reported reactions.  74% of the reported local reactions 
were mild in severity and 4% were classified as severe.  Local reactions persisted for no 
longer than 7 days.  Systemic AEs deemed definitely, probably or possibly related to 
vaccination were observed in approximately 90% of recipients.  Most commonly 
reported systemic AEs were: malaise (67%), fatigue (56%), myalgia (56%), feverish 
(56%), nausea (56%) headache (44%), and arthralgia (44%).  66% of observed 
systemic AEs were classified as mild and 17% as severe.  The majority of systemic AEs 
resolved within 48 hours.  Immunogenicity data have been collected up to 22 weeks 
post MVA vaccination for group A1 and up to 10 weeks post MVA boost for group A2.  A 
single MVA-NSmut administration was unable to prime T-cell responses to NS antigens 
in any of the 4 volunteers in group A1 during the 22 weeks of follow up.  Conversely, 
and corresponding to previous results, priming with ChAd3NSmut promptly induced T-
cell responses 2 weeks after first vaccination in all 5 volunteers.  Boosting with MVA-
NSmut resulted in great expansion of T-cell responses in all 5 volunteers ranging from 
1,503 to 6,147 sum NS SFC/million PBMC.  
2.2.4 Prime/boost with Chimpanzee Derived Adenovirus and MVA Vectored 
Vaccines 
A series of clinical trials have recently been conducted or are in progress at the 
University of Oxford against malaria using a prime/boost regimen with a chimpanzee 
adenoviral vector followed by MVA using antigens other than NSmut.  For the VAC033 
and MAL 034 trials, the candidate vaccines multiple epitope thrombospondin-related 
adhesion protein, AdCh63 ME-TRAP and MVA ME-TRAP were given to healthy 
subjects.76  These vectors encode for ME-TRAP, a surface protein from the sporozoite 
stage of P. falciparum malaria with a selection of T-cell epitopes.  47 subjects have 
received AdCh63 ME-TRAP at doses from 1 x 108 vp to 2 x 1011 vp, followed by MVA 
ME-TRAP boosting 2 x 108 plaque forming units (pfu) administered by intradermal and 
intramuscular routes.  The simian adenovirus/MVA-vectored heterologous prime-boost 
regimen generates unprecedented cellular and humoral immune responses to TRAP 
and has recently demonstrated sterile protection against heterologous sporozoite 
challenge in 2/8 vaccinated subjects.76  There have been no SAEs following AdCh63 
ME-TRAP or MVA ME-TRAP administration to date.  These studies also showed that 
AdCh63 ME-TRAP administered intramuscularly is better tolerated and more 

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immunogenic than the same dose administered by the intradermal route76.  Moreover, a 
significant reduction in the frequency of local injection site reactions and systemic 
reactions was demonstrated when MVA ME-TRAP is administered intramuscularly 
compared to intradermally.76 
2.3 Scientific Rationale (including dose selection) 
Route of injection:  The choice of the intramuscular route for AdCh3 is based on the 
assumption that no co-infection of natural human Adenovirus could occur at this site.  
Furthermore, there is a large body of data from clinical trials in humans using replication 
defective Ad5- and Ad6-based HIV vaccines injected intramuscularly showing an 
excellent safety profile, no viral shedding, and high levels of immunogenicity.  MVA-
NSmut will also be given by the intramuscular route because it is better tolerated than 
when administered by subcutaneous route (see Section 2.2.4: Prime/boost with 
chimpanzee derived Adenovirus and MVA vectored vaccines). 
 
Vaccine Doses:  In this study AdCh3NSmut1 vaccine will be administered at 2.5x1010 
vp, and MVA-NSmut at 1.8 x 108 pfu. 
 
AdCh3NSmut1:  Clinical data are not available for the AdCh3NSmut1 vector.  
AdCh3NSmut1 has minimal changes from AdCh3NSmut vector used in the 3 clinical 
studies shown in Table 1.  Pre-clinical assessment has shown that these changes do 
not affect the safety or immunogenicity profile.  Safety data from the study HCV001 and 
the ongoing HCV003 have shown that AdCh3NSmut is safe and well tolerated in 
healthy subjects at the dosage (2.5x1010 vp) selected for this study.   
 
Phase I studies assessing safety and immunogenicity in healthy individuals of Ad5- and 
Ad6-based HIV vaccine candidates were shown to induce higher rates of responses 
and more potent responses when they were administered at 1010 vp/dose versus 109 
vp/dose.77  Similarly, there was a greater frequency of detectable cellular responses 
after 6 months from the first administration of the vector in the higher dose groups 
(between 1010 and 1011 vp/dose).77  These studies have shown that adenoviral vectors 
are safe and well tolerated up to 1011 vp/dose with some more reactogenicity associated 
with the highest dose.78-80 
 
MVA-NSmut:  In this study, MVA-NSmut will be administered at 1.8 x 108 pfu.  This 
dose has been selected based on the use of MVA vectors in primate and human 
studies.  In Oxford, MVA malaria vaccines have been safely administered in multiple 
Phase I and Phase II studies at 2 x 108.  MVA ME-TRAP and MVA-vectored P. 
falciparum CS protein has been well tolerated with good immunogenicity at 5 x 108 pfu 
and no severe adverse events.  In study VAC037, MVA MSP1 was associated with 
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MVA vectors at a dose of 2 x 108pfu.  In the HCV003 study the MVA-NSmut 
administered at 2 x 108 has shown a good safety profile and the induction of very potent 
immune responses.  A slightly lower dosage (1.8 x 108 pfu) has been chosen for study, 
aiming to inject a volume of 0.2mL. 
 
Number of injections and interval between injections:  The vaccine schedule will 
consist of a single priming injection with AdCh3NSmut1, followed by a single boost with 
MVA-NSmut 56 days later in a heterologous prime/boost regimen.  This is based on 
promising data from ongoing malaria clinical trials in Oxford, where optimal 
immunogenicity is achieved with an 8 week interval between vectors and from pre-
clinical data shown in the previous section.  Subjects randomized to placebo will receive 
2 doses of placebo at Day 0 and Day 56 (approximately week 8).  As placebo, a volume 
of the normal saline equal to the volume of the investigational vaccines will be used. 
2.4 Potential Risks and Benefits 
2.4.1 Potential Risks 
Potential risks 
The general risks to subjects in this study are associated with phlebotomy and with 
vaccination.  The volume of blood drawn over the study period, should not compromise 
health status.  Potential risks include local and systemic reactions as vaccine-related 
side effects, which are described below.  Safety data from the ongoing studies 
described above predicts for mild to moderate reactogenicity of vaccination. 
 
Local reactions 
Mild tenderness, bruising, light-headedness or, rarely, syncope, may result from 
venipuncture.  Vaccination usually precipitates a local inflammatory reaction.  This may 
include redness, swelling, tenderness, or itching. 
 
Systemic Reactions  
The potential systemic reactions to immunization with vaccines include a flu-like illness 
with low-grade fever, chills and malaise.  As with any vaccine, serious allergic reactions 
may occur.  Monitoring will be ongoing for all of these potential rare events. 
 
Subsequent positivity for hepatitis C antibody (anti-HCV) 
It is expected that after vaccination subjects may become anti-HCV positive.  In the 
previous clinical trial, HCV001, anti-HCV positivity in the absence of documented 
infection occurred in 10% of vaccinated subjects.  Anti-HCV seroconversion may occur 
as a result of exposure to antigens derived from the hepatitis C virus that are included in 
the vaccine.  In order to reassure subjects and maintain coherent health records, 
subjects will have blood taken for HCV RNA testing [using polymerase chain reaction 

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(PCR) methods] prior to the trial to exclude the presence of HCV.  No anti-HCV testing 
will occur systematically during the trial. Only HCV RNA results will be used for 
determination of HCV infection status during study visits.  Anti-HCV testing will be 
conducted at the end of the trial to examine whether subjects seroconverted (became 
anti-HCV antibody positive) during the trial. Among subjects who did not have 
detectable HCV infection, testing will be done to best ascertain whether subjects 
seroconverted: (1) in association with undetected (HCV RNA) infection, or (2) as a 
result of vaccination. This testing will be done at the end of the trial, so as not to unblind 
subjects or investigators during the trial.  If the determination is made that subjects 
became anti-HCV positive as a result of vaccination rather than infection, they will be 
provided with documentation explaining the reasons for their anti-HCV positivity.  These 
cards will have contact information for study investigators so that providers can 
communicate as necessary regarding subject’s anti-HCV status (with the subject’s 
consent).  This will be discussed in detail with subjects at screening and in informed 
consent forms (ICFs). 
 
Confidentiality 
The risk of study participation includes possible loss of confidentiality.  Risks to privacy 
and confidentiality exist with all human research.  Subjects will be assigned a unique 
study identification number which will be used on data collection forms, case report 
forms (CRFs), and specimens collected for laboratory analyses.  No samples or data 
will be labeled with the subject’s name.  The key to subjects’ names will be accessible 
only to the study co-principal investigators, and hard-copies and electronic copies of 
data will be stored in secured locations. 
2.4.2 Unknown Risks 
The experimental vaccine may have side effects that are not yet known or identified.   
 
For example, a series of experimental HIV vaccine studies (STEP Study - HVTN 502; 
Phambili Trial - HVTN 503 and HVTN 505) which utilized a human adenovirus serotype 
5 (Ad5) vector did not work to prevent HIV infections.  The STEP Study data showed an 
increased risk of HIV infection associated with the active vaccine  (adjusted HR, 1.40; 
95% CI 1.03-1.92).81  The HVTN 505 Study was stopped for meeting futility criteria and 
while not statistically different, there were more infections in the active vaccine arm 
compared to the placebo arm.  The researchers who conducted these HIV vaccine 
studies have not yet determined the explanation for these findings.   
 
In this prime/boost strategy HCV vaccine study a simian adenovirus serotype 3 (Ad3) 
vector administered at baseline and a Modified Vaccinia virus Ankara (MVA) vector 
given at week 8 are utilized.  There are significant differences between the HIV studies 
and this study in the risk of HIV infection that make an increased risk of HIV acquisition 

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in this trial unlikely.  Those differences include exclusion from enrollment in this trial of 
subjects who are at high risk for HIV acquisition such as men having sex with men and 
an HIV prevalence of <0.5% among the cohorts from which subjects are drawn.  
Regarding the potential to increase the risk of HCV infection, for which the subjects in 
this trial are at risk, there are key differences in HIV and HCV biology that make an 
increase the risk of HCV infection less likely.  Hepatitis C virus infects hepatocytes and 
not activated T cells.  The leading explanation for the increased risk of HIV infection 
following vaccination is that the vaccine activates T cells, the target of HIV infection.  
Activation of T cells is not likely to affect HCV transmission given that they are not 
targets of HCV infection.  However, there may be other unknown risks of adenoviral 
vaccination.  
 
The researchers will inform subjects of any as-yet-unknown risks if they occur.  Subjects 
have the option of not participating in future study activities at any time. 
2.4.3 Potential Benefits 
There is no previous clinical evidence showing the effectiveness of the AdCh3NSmut1 
and MVA-NSmut vaccine for prevention of HCV infection.  Subjects in both arms will 
receive no direct benefit from participating in the trial other than receiving information 
about their general health status and counseling, education and tools to prevent HCV 
infection. 
 
Society may benefit from the potential development of a new prophylactic hepatitis C 
vaccine. 
 

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3 Study Objectives 
3.1 Primary Objectives: Safety and Efficacy  
To assess the safety of new candidate hepatitis C virus vaccines, AdCh3NSmut1 and 
MVA-NSmut, compared to placebo when administered sequentially to IDUs.  To 
determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce incidence of 
chronic HCV infection compared to placebo among HCV-uninfected IDUs. 
3.1.1 Primary Safety Endpoints  
The primary endpoints in the evaluation of safety are as follows:  (1) occurrence of 
vaccine-related serious adverse events (SAEs) from the time of first vaccination through 
the entire study period; (2) occurrence of severe local and/or systemic solicited 
reactogenicity signs and symptoms in the 8 days (Day 0-7) after each vaccination; and 
(3) occurrence of clinical safety laboratory adverse events (AEs) assessed at baseline 
and 1 month following each vaccination.  The occurrence of these safety parameters 
will be assessed by treatment arm (vaccine and placebo). 
 
Signs and symptoms will be categorized as either local (pain, tenderness, erythema, 
induration and warmth) or systemic (fever, malaise/fatigue, myalgia, headache, nausea, 
vomiting, chills, arthralgia and abdominal pain).  Clinical laboratory measures will 
include measures of white blood cell (WBC), hemoglobin (HgB), platelets, ALT, and 
creatinine. 
 
The protocol includes safety evaluations at multiple time points in order to better 
characterize the impact of the vaccine on the at-risk population.  Although the principal 
goal is safety signal detection in association with the experimental vaccine, these safety 
data will be an important resource for future clinical trials in this population.     
3.1.2 Primary Efficacy Endpoint 
The primary endpoint for evaluation of efficacy is chronic HCV infection at 6 months 
defined by persistent viremia over a period of 6 months after initial detection of primary 
infection.  Persistent viremia will be determined by:  (1) the presence of the same virus 
(as confirmed by HCV core-E1 phylogenetic analysis testing) in blood samples collected 
at the first visit where HCV RNA is detected (initial infection) provided sufficient sample 
is available, and a subsequent sample collected at month 6 (not less than 159 or more 
than 201 days following initial infection); and (2) third HCV RNA positive sample taken 
at a time point in between these two samples (initial and month 6).  If there are two or 
more samples within the 159 to 201 day window for the month 6 collection, the test 
result from the collection date closest to 180 days after initial infection will be used. 

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3.2 Secondary Objective: Immunogenicity 
3.2.1 Secondary Immunogenicity Endpoint  
The secondary immunogenicity endpoint in the evaluation of immunogenicity is the 
frequency of positive cell mediated immune response within 14 days after the last 
vaccination by treatment group (vaccine vs. placebo) measured by interferon gamma 
(IFN-γ) production by T-cells in response to at least one out of the six HCV genotype 1b 
(Bk strain) peptide pools representing NS region present in the vaccines, as determined 
by ELISpot assays. 
3.3 Exploratory endpoints in the evaluation of efficacy 
Exploratory objective 1:  
To determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce incidence of 
primary HCV infection compared to placebo among HCV-uninfected IDU. 
 
Exploratory endpoint 1:  
An incident HCV infection will be defined as a new confirmed positive HCV RNA test 
after a previous negative HCV RNA test.  The interval during which incident HCV 
infection occurs will be defined by the date of the last documented negative HCV RNA 
results, and the date of the first positive HCV RNA result. 
 
In the event that seroconversion is detected in the absence of detectable viremia (no 
positive HCV RNA results), incident HCV infection date will be defined by results from 
anti-HCV testing conducted on stored blood samples tested retrospectively.  
Seroconversion is defined as a subject developing a positive anti-HCV antibody test 
after a previous negative anti-HCV antibody test.  Detection of infection-induced 
antibodies to HCV core antigens not present in the vaccine construct will be used to 
differentiate infection- from vaccine-induced seroconversion.  The seroconversion date 
will be defined by the interval between the date of the last negative anti-HCV and the 
date of the first positive anti-HCV test result.   
 
Exploratory objective 2: 
To determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce 9-month 
incidence of chronic HCV infection compared to placebo among HCV-uninfected IDU. 
 
Exploratory endpoint 2: 
The exploratory endpoint for evaluation of efficacy is chronic HCV infection at 9 months 
defined by persistent viremia over a period of 9 months after initial detection of primary 
infection. Persistent viremia will be determined by:  (1) the presence of the same virus 
(as confirmed by HCV core-E1 phylogenetic analysis testing) in blood samples collected 
at the first visit where HCV RNA is detected (initial infection) provided sufficient sample 

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is available, and a subsequent sample collected at month 9 (not less than 249 or more 
than 291 days following initial infection); and (2) third HCV RNA positive sample taken 
at a time point in between these two samples (initial and month 9).  If there are two or 
more samples within the 249 to 291 day window for the month 9 collection, the test 
result from the collection date closest to 270 days after initial infection will be used.  
Cases classified as not-chronically infected at 9 months will be considered resolved 
(cleared of infection). 
 
Exploratory objective 3:  
To determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce peak 
concentration (magnitude) of HCV RNA compared to placebo, in blood samples of 
persons with incident HCV infection. 
 
Exploratory endpoint 3: 
Quantitative viremia measures, including the initial peak and total HCV RNA levels 
detected, from the date of first HCV RNA detection and at study visits in the first 6 
months of viremia will be compared, between vaccinated and placebo arms, using 
logarithmic IU/mL values. 
 
Exploratory objective 4: 
To determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce duration of 
HCV viremia (shorter time to spontaneous resolution) during acute infection (6-month 
period following incident HCV infection) compared to placebo in blood samples of 
persons with incident HCV infection. 
 
Exploratory endpoint 4: 
The duration of acute infection will be defined as the interval between incident HCV 
infection and viral clearance.  For subjects who do not clear HCV viremia, the duration 
will be measured as six months following incident infection.  The interval during which 
incident HCV infection occurs will be defined by the date of the last documented anti-
HCV and HCV RNA negative results, and the date of the first positive HCV RNA result.  
The interval during which viral clearance occurs will be defined by the date of the last 
test with detectable HCV RNA and the first of two consecutive samples with 
undetectable HCV RNA. 
 
Exploratory objective 5: 
To determine if AdCh3NSmut1 and MVA-NSmut HCV vaccines will reduce incidence of 
chronic HCV infection with genotype 1 compared to non-genotype 1 among HCV-
uninfected IDU. 
 
 
 

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Exploratory endpoint 5: 
A chronic HCV infection genotype 1 will be defined as a chronic HCV infection at 6 
months, as defined in Section 3.1.2, with the additional requirement that the infection is 
genotype 1 as determined from blood samples obtained at the first HCV viremic visit, 
provided sufficient sample is available.  
 
 

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4 Study Design 
Overall trial design 
This is a Phase I/II double-blind (subject, investigator, and clinical personnel monitoring 
safety and laboratory assay results, and laboratory personnel), randomized, and 
placebo-controlled study in approximately 540 individuals at high risk for HCV infection 
due to active injection drug use (IDU) between the ages of 18 and 45.  HCV-uninfected 
(negative for both anti-HCV and HCV RNA at screening) male and female adults who 
are current IDUs will be randomized at a 1:1 ratio, stratified by gender and IL28B status, 
to receive AdCh3NSmut1 and MVA-NSmut HCV vaccines or placebo at Day 0 and Day 
56.  The study is designed to be implemented in two stages and enroll a total of 270 
subjects/arm. 
 
Stage 1:  Blinded, randomized, controlled assessment of AdCh3NSmut1 and MVA-
NSmut HCV vaccine compared to placebo in 68±4 (34±2/arm) evaluable subjects.  This 
study stage will evaluate safety of the combined use of both vectors in a prime/boost 
regimen in the target population.  Each individual will be followed with extensive safety 
and immunogenicity measures.  Interim analysis (safety and immune responses) is 
planned immediately following the end of stage 1 (at Day 63 follow-up (34±2/arm) for all 
evaluable subjects). 
 
Stage 2:  After the safety and immunogenicity interim analyses, the study will be 
extended into a 2nd stage, wherein an additional (n=472±4) HCV-uninfected subjects will 
be enrolled.  A 2nd interim analysis to evaluate progress (statistical and enrollment) 
toward the efficacy endpoint will occur when roughly one-third of enrolled and currently 
per-protocol subjects (N = 98) have completed six months of post-vaccination follow-up, 
but no later than one month prior to the planned completion of enrollment to the trial.  
Immune responses will be evaluated following each vaccination and HCV viremia status 
will be monitored monthly for 18 months following the 2nd vaccination.  This stage is 
designed as a blinded, randomized placebo-controlled study, with safety and efficacy 
assessments. 
 
Study duration 
The planned duration of the study is approximately 63 months total including accrual 
time for subjects (assuming 31 months of screening/enrollments, plus 3 months of 
halted enrollment for the first interim analysis), 2 months vaccination, 18 months follow-
up of each enrolled subject, and 9 months extended observation (monthly), from the 
time of infection, for subjects becoming viremic in the last month of follow-up. 
 
The follow-up period for detection of HCV infection will start after the second injection of 
vaccine and will last 18 months.  Subjects who become viremic will be followed for 9 
months after detection of HCV (up to month 29).  The trial will be considered closed 

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when the observation period for the last subject that became viremic during the 18 
months follow-up is completed. 

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5 Study Population 
All subjects in this study will be adults between the ages of 18 and 45, who are at high 
risk for HCV infection due to active injection drug use (IDU). 
5.1 Inclusion Criteria 
Inclusion criteria that must met prior to the first vaccination: 
1. 
Comprehension of informed consent. 
2. 
18-45 year old men or women with acknowledged active IDU in the past 90 days 
and have no travel plans that would interfere with ability to meet the study visit 
schedule.  
3. 
In good general health as determined by a participating study physician and 
results within acceptable ranges for clinical laboratory evaluations as detailed in 
Appendix A.   
4. 
Negative for antibodies to hepatitis C virus (anti-HCV). 
5. 
Negative for HCV RNA. 
6. 
Negative antibodies to HIV. 
7. 
Negative for HBsAg. 
8. 
Able and willing (in the Investigator’s opinion) to comply with all study 
requirements. 
9. 
Willing to allow the investigators access to their medical records. 
10. 
Willingness to practice continuous effective contraception from the screening visit 
through 90 days after the last vaccination (males and females). 
11. 
Among females, a negative pregnancy test within 24 hours prior to vaccination. 
12. 
Agreement to refrain from blood donation during the course of the study or after 
the study. 
13. 
Provide written informed consent prior to initiation of any study procedures. 
14. 
Willing to provide contact information for study follow-up activities, including the 
address, name and contact information of three people who can be contacted to 
facilitate follow-up compliance. 
 
 

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5.2 Exclusion Criteria 
Exclusion criteria that apply prior to the first vaccination: 
1. 
The subject is currently participating in a study that involves an experimental 
agent (vaccine, drug, biologic device, blood product, or medication) or has 
received an experimental agent within 30 days prior to enrollment in this study, or 
expects to receive another experimental agent during participation in this study. 
2. 
Prior receipt of a recombinant simian or human adenoviral vaccine or MVA 
vaccine. 
3. 
Administration of immunoglobulins and/or any blood products within the 90 days 
preceding the planned administration of the vaccine candidate. 
4. 
Any confirmed or suspected immunosuppressive or immunodeficient state, 
including: HIV infection; asplenia; recurrent, severe infections. 
5. 
History of allergic disease or reactions likely to be exacerbated by any 
component of the vaccine (i.e., known hypersensitivity to aminoglycoside 
antibiotics or to egg proteins). 
6. 
History of clinically significant contact dermatitis or other significant 
dermatological conditions such as psoriasis. 
7. 
Any history of anaphylaxis in reaction to vaccination. 
8. 
Pregnancy, lactation or willingness/intention to become pregnant during the 
study. 
9. 
History of cancer (except for successfully treated basal cell carcinoma of the skin 
and cervical carcinoma in situ). 
10. 
History of severe psychiatric illness, including severe depression, history of 
suicidal ideation, suicidal attempts, or psychosis requiring medication.  The 
subject has a diagnosis of schizophrenia, bi-polar disease, or other severe 
(disabling) chronic psychiatric diagnosis that is uncontrolled and would interfere 
with the ability to adhere to the protocol.   
11. 
Any other serious chronic illness requiring hospital specialist supervision. 
12. 
Suspected or known current alcohol abuse as defined by a score of 10 or more 
on the Alcohol Use Disorders Identification Test (AUDIT) C test (a standardized 
screening tool used to identify hazardous drinkers or those with active alcohol 
use disorders, including abuse or dependence).82 
13. 
At high risk of HIV infection by the following criteria (adapted from HIV Network 
for Prevention Trials (HIVNET) behavioral criteria for high risk of HIV):  (1) 
sexually active male who has sex with men (MSM), defined as (i) male who has 
had anal sex with male sexual partner or partners in the past year or (ii) a male 

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who exchanged sex with male partner(s) for money or drugs in the past year; and 
(2) female and in a current relationship with a high risk male (active MSM, HIV 
positive male).83  
14. 
Any other significant disease, disorder or finding, which, in the opinion of the 
Investigator, may put the subject at risk because of participation in the study, 
may influence the result of the study, or may influence the subject’s ability to 
participate in the study. 
15. 
History of or current diagnosis of Diabetes mellitus. 
16. 
History of or current diagnosis of autoimmune disease. 
17. 
History of or current cardiac disease including history of myocardial infarction or 
arrhythmia. 
18. 
Current diagnosis of active liver disease. 
19. 
History of seizure disorder or currently taking anti-convulsant therapy that would 
interfere with safety evaluation. 
20. 
Uncontrolled hypertension (defined as systolic blood pressure being greater than 
140mm Hg or diastolic blood pressure being greater than 90mm Hg).   
21. 
History of splenectomy. 
22. 
Long term immunosuppressive use (defined as taken for 14 days or more in total 
at any time during the past 180 days) of high dose oral or parenteral 
glucocorticoids (high dose defined as prednisone ≥ 20 mg total daily dose, or 
equivalent dose of other glucocorticoids); or high-dose inhaled steroids (high 
dose defined as >800 mcg/day of beclomethasone dipropionate or equivalent); or 
any use of hepatotoxic or non-FDA approved medication. 
23. 
Have an acute illness, including an oral temperature greater than or equal to 
100.4°F, within 7 days prior to the first vaccination. 
24. 
Immunization against another pathogen within 14 days of planned injection. 
Second vaccination exclusion criteria: 
The following events associated with vaccine immunization constitute absolute 
contraindications to further administration of vaccine.  The subject will not receive 
additional vaccination, but will continue with scheduled follow-up procedures except 
vaccination. 
1. Anaphylactic reaction following administration of vaccine. 
2. Pregnancy.  If a woman reports having a positive home urine pregnancy test or a 
positive in clinic urine pregnancy test prior to a scheduled second vaccination, she is 
not eligible to receive the study vaccine.  If she later returns to clinic and reports 

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having a negative home urine pregnancy test and this is confirmed by a negative in 
clinic urine pregnancy test and a negative blood pregnancy test she may be eligible 
for the second vaccination if she is still within the vaccination window and no other 
exclusion criteria are met. 
The following adverse events constitute contraindications to administration of vaccine at 
that point in time.  If any one of these adverse events occurs at the time scheduled for 
vaccination, the subject may be vaccinated at a later date (within 28 days of scheduled 
administration) or withdrawn at the discretion of the investigator.  The subject will not 
receive additional vaccination, but will continue with all scheduled follow-up procedures 
except vaccination. 
3. Acute disease at the time of vaccination (acute disease is defined as the presence of 
a moderate or severe illness with or without fever).  The vaccine dose can be 
administered to persons with a minor illness such as diarrhea, mild upper respiratory 
infection with or without low-grade febrile illness, i.e., temperature of <38°C 
(100.4°F). 
4. Temperature of >38°C (100.4°F) at the time of vaccination. 
5. Immunization against another pathogen within 14 days of vaccination. 
5.3 Randomization and Code Breaking Procedures 
The list of randomized treatment assignments stratified by gender and IL28B status will 
be prepared by statisticians at The Emmes Corporation and included in the enrollment 
module of The Emmes Corporation’s Internet Data Entry System (IDES).  IDES will 
assign each subject a treatment code from the list after demographic and eligibility data 
have been entered into the system.  A designated individual at each site will be 
provided with a treatment key, which links the treatment code to the actual treatment 
assignment, which will be kept in a secure place. 
 
Instructions for use of the enrollment module are included in the IDES User’s Guide.  
Manual back-up randomization procedures are provided for use in the event that the 
site temporarily loses access to the Internet or the online enrollment system is 
unavailable. 
 
Unblinding 
Unblinding is unlikely to be necessary for the provision of medical treatment or to 
otherwise protect the safety of study subjects.  In the event that an investigator is 
concerned that a subject might be put at undue risk by continuing product use, the 
investigator may discontinue product use by this subject; however, knowledge of the 
specific product (vaccine or placebo) to which the subject was assigned should not be 
necessary to guide further follow-up and/or treatment.   

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The co-principal investigators and/or Independent Safety Monitor (ISM) should be 
available to review and evaluate any notable reactions.  In the case of a medical 
emergency, the co-principal investigators or ISM may deem it medically necessary to 
unblind the subject's treatment assignment.  If the co-principal investigators or ISM 
believe that unblinding would benefit the medical care of the subject and time permits, 
DMID will be consulted prior to unblinding, and concurrence will be obtained. 
 
Otherwise, subjects will be unblinded to their drug only at the close of the trial.  Subjects 
who develop HCV infections or drop out will not be unblinded early.  The Data Safety 
and Monitoring Board (DSMB) may receive data in aggregate and presented by 
treatment group, but without the treatment group identified.  The DSMB may be 
unblinded to individual study treatment assignments, as needed, to adequately assess 
safety issues. 
5.4 Withdrawal 
5.4.1 Reasons for Withdrawal 
Subjects may withdraw or be withdrawn for any of the reasons given below.  The reason 
for withdrawal will be recorded in the data collection form.  The DSMB may recommend 
withdrawal of subjects. 
 
In accordance with the current revision of the Declaration of Helsinki (amended October 
2000, with additional footnotes added 2002 and 2004) and any other applicable 
regulations, a subject has the right to withdraw from the study at any time and for any 
reason and is not obliged to give his or her reasons for doing so.  The Investigator may 
withdraw the subject at any time in the interests of the subject’s health and well-being.  
In addition the subject may withdraw/be withdrawn for any of the following reasons: 
• Administrative decision by the Investigator. 
• Ineligibility (either arising during the study or retrospective, having been 
overlooked at screening). 
• Significant protocol deviation. 
• Subject non-compliance with treatment regime or study requirements. 
• An adverse event which requires discontinuation of the vaccination regimen or 
results in inability to continue to comply with study procedures (see below). 
If a subject is withdrawn from the study, all study procedures (except for another 
vaccination) including safety follow-up will be continued, if possible.  If voluntary 
withdrawal occurs, the subject will be asked to continue scheduled study procedures 
(except for another vaccination) including safety evaluations, if possible, and be given 

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appropriate care under medical supervision if symptoms of any AE related to 
participation in the study are continuing.  The subject will be followed until the AE is 
resolved or until the subject’s condition becomes stable.  (Note:  These plans regarding 
follow-up only apply if the subject received at least one study dose.) 
5.4.2 Handling of Discontinuation of Vaccine Regimen 
Any subject whose vaccinations have been discontinued will be seen for appropriate 
follow-up visits or medical care will be arranged until the adverse event has resolved or 
stabilized. 
5.5 Termination of the Study 
The NIAID/DMID and/or the FDA have the right to terminate this study at any time.  
Reasons for terminating the study may include, but are not limited to, the following: 
• Incidence or severity of AEs indicating a potential health hazard. 
• Data recording is inaccurate or incomplete. 
• The Investigator(s) has not been adhering to the protocol or applicable regulatory 
guidelines in conducting the study. 

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6 Investigational Products 
6.1 Study Products Description 
Two investigational vaccines will be used in this study, namely AdCh3NSmut1 and 
MVA-NSmut, along with the 0.9% sodium chloride as placebo. 
 
AdCh3NSmut1 formulation and dose 
The vaccine, AdCh3NSmut1, consists of the attenuated virus, simian adenovirus 3, 
(AdCh3), encoding the Nonstructural region (NS), from the Hepatitis C virus 1b 
genotype (BK strain isolate), with a mutation (mut) introduced in the active site of the 
HCV polymerase. 
 
AdCh3NSmut1 is manufactured under Good Manufacturing Practice (GMP) conditions.  
The AdCh3NSmut1 vaccine is formulated in A195 buffer (10 mM Tris, 75 mM NaCl, 1 
mM MgCl2, 0.02% PS80, 5% sucrose, 0.1 mM EDTA, 10 mM Histidine, 0.5% ethanol, 
pH 7.4).  AdCh3NSmut1 has been vialed in 2 mL nominal Type I borosilicate glass vial 
with a siliconized 13 mm butyl rubber stopper at concentration of 5X1010 vp/mL at 0.5 
mL extractable volume /vial.  The dose of AdCh3NSmut1 to be used in this study is 
2.5x1010 vp.  This dose is obtained by injecting 0.5 mL of the vaccine without 
dilution. 
 
MVA-NSmut formulation and dose 
The vaccine, MVA-NSmut, consists of the attenuated replication-defective 
orthopoxvirus, modified vaccinia virus Ankara, (MVA), encoding the Nonstructural 
region (NS) from the Hepatitis C virus 1b genotype (BK strain isolate), with a mutation 
(mut) introduced in the active site of the HCV polymerase.  The investigational 
medicinal product MVA-NSmut is manufactured under GMP conditions.  The 
investigational medicinal product is supplied as liquid presented in Tris buffer (10 mM 
Tris, 140 mM NaCl, pH 7.7), at a concentration of 8.9 x 108 plaque-forming units (pfu) 
per mL.  The virus suspension is supplied as sterile 0.5mL/vial extractable volume in 2 
mL injection vials, made of clear borosilicate glass, FIOLAX® -klar, WBK 1, DIN ISO 
8362/1.  The vials were sealed with 13 mm bromobutyl rubber stoppers, grey, FM 
457/0, V 9128, SAF 1, DIN ISO 8362-2, aluminium injection closures with a 6 mm hole.  
The dose of MVA-NSmut to be used in this study will be 1.8 x 108 pfu delivered 
intramuscularly.  The volume administered will be 0.2 mL without dilution. 
 

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0.9% Sodium Chloride for Injection and Dose 
The placebo agent to be utilized will be 0.9% sodium chloride injection, USP which is a 
sterile, nonpyrogenic, isotonic solution of sodium chloride and water for injection.  Each 
mL contains sodium chloride 9 mg.  It is supplied as single-dose containers.  The 
volume to be administered will be 0.5 mL and 0.2 mL without dilution delivered 
intramuscularly. 
 
A pharmacy facility will be located at the study site and staffed by pharmacists who are 
licensed to practice in the US.  The pharmacy will have adequate space to store 
sufficient quantities of study placebo product to assure continuous access to all study 
subjects.   
 
Illustrations of the Test Article outer box labels are provided below: 
 
Study Drug for DMID Protocol 10-0069  
Lot # XXXX 
AdCh3NSmut1 Vaccine 
 
Directions: administer 0.5 mL by intramuscular injection 
 
Store at or below -70°C  
 
Caution: New Drug – limited by United States Law to 
investigational use. 
 
 
 
Study Drug for DMID Protocol 10-0069  
Lot # XXXX 
MVA-NSmut Vaccine 
 
Directions: administer 0.2 mL by intramuscular injection 
 
Store at or below -70°C  
 
Caution: New Drug – limited by United States Law to 
investigational use. 
 
 
 

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Study Drug for DMID Protocol 10-0069  
Lot # XXXX 
0.9% Sodium Chloride 
 
Directions: administer 0.5 mL by intramuscular injection 
 
Store at 20 to 25°C  
Expires: MM/DD/YYYY 
  
Caution: New Drug – limited by United States Law to 
investigational use. 
 
 
 
Study Drug for DMID Protocol 10-0069  
Lot # XXXX 
0.9% Sodium Chloride 
 
Directions: administer 0.2 mL by intramuscular injection 
 
Store at 20 to 25°C  
Expires: MM/DD/YYYY 
  
Caution: New Drug – limited by United States Law to 
investigational use. 
 
6.1.1 Acquisition 
The AdCh3NSmut1 and MVA-NSmut vaccines will be provided by Okairos.  The normal 
saline placebo will be provided by DMID. 
 
Clinical sites will obtain vaccines from the DMID Clinical Agent Repository at Fisher 
Bioservices. 
6.1.2 Product Storage, Stability, and Expiration 
Vials of both recombinant viruses are stored frozen at -70°C or below in a clearly 
marked secure, limited-access storage area with temperature-controlled freezers.  
Access to the area will be controlled and temperature records will be maintained.   
 
A pharmacy facility will be located at the study site and staffed by pharmacists who are 
licensed to practice in the US.  The pharmacy will have adequate space to store 
sufficient quantities of study product to assure continuous access to all study subjects.  
 
AdCh3NSmut1 Stability 
Research grade purified virus has been stored for 12 months in buffer A195 at -70°C 
with no significant loss of biological activity.  Genetic stability has also been assessed 

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on a research batch of AdCh3NSmut1, subjected to ten passages in PER.C6 cells and 
no evidence of genetic instability has been observed. 
 
AdCh3NSmut1shelf life extension plan 
The AdCh3NSmut1 clinical lot will have an initial expiry date of 12 months from 
manufacture.  A shelf-life extension program is in place to assign new expiry date to the 
product.  Samples of the clinical lot will be tested by a potency assay, based on in vitro 
infectivity in HEK (human embryonic kidney) 293 cells by the AdCh3 hexon immune 
staining method.  Testing will be carried out for a period of 36 months. 
 
MVA-NSmut Stability 
The clinical batch of MVA-NSmut has undergone testing to demonstrate genetic stability 
of the virus.  An initial expiration date of 2 years from date of manufacture was assigned 
based on the vast stability data for MVA based vectors stored at or below -70°C.  
Titrations on chicken embryo fibroblast (CEF) cells will be performed in order to check 
that the titer does not decrease over time.  Long term stability testing for MVA-NSmut 
stored in a freezer (-80°C ±10°C) will be carried out for a period of 36 months to allow 
the expiration date to be lengthened. 
 
0.9% Sodium chloride for injection 
The 0.9% sodium chloride should be stored at a controlled room temperature of 20°C to 
25°C (68 to 77°F).  The solution should be inspected visually for particulate matter and 
discoloration prior to administration. 
6.1.3 Preparation & Administration 
Vaccine administration 
On vaccination day, both for priming and boosting administration, vaccines will be 
allowed to thaw to room temperature.  The pharmacist or nurse trained to prepare 
vaccine will wear gloves.  Using aseptic technique, the pharmacist or trained nurse will 
withdraw the volume specified in Section 6.1 into a 1 mL syringe and add a blinding 
sleeve.  The syringe containing the product must be held at room temperature.  Once 
drawn into the syringe, the vaccine should be administered as soon as possible after 
preparation as intramuscular injection in the deltoid region of the arm.  During 
administration of the vaccines, medicines and resuscitation equipment will be 
immediately available for the management of anaphylaxis.  All subjects will remain on 
the unit for at least 30 minutes for observation. 

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6.1.4 Maintaining Blind for Investigational Product 
Vaccine will be prepared by a pharmacist or trained nurse and administered by a 
blinded vaccine administrator.  All study assessments will be performed by blinded 
study personnel.  All other staff, as well as all subjects, will be blinded to treatment 
assignment. 
6.1.5 Accountability/Final Disposition for the Investigational Product(s) 
A pharmacist at the study site will receive the study products and store them in a 
designated secure area within the pharmacy.  Access will be restricted to the 
pharmacist, nurse trained to prepare vaccine and the site monitor.  The pharmacist will 
be responsible for keeping accurate records of the material received.  S/he will keep a 
daily log of the amount of material received, the amount dispensed, and the amount 
remaining at the end of the study.  At the end of the study, the pharmacist will perform 
the final drug accounting of unused study material on the proper log documents.  At the 
completion of the study, a clinical monitor will review final accountability records.  All 
unused vials should be returned to the DMID Clinical Repository at Fisher BioServices. 
 
The pharmacist or trained nurse will be responsible for dispensing the drug to the 
blinded vaccine administrator.  The pharmacist will be responsible for storing, 
managing, and dispensing all drug supplies for the study.  The pharmacist will be 
responsible for tracking the flow of all study product in a log book, including managing 
and logging all supplies in storage, and logging all study product dispensed to subjects.  
The on-site investigator will oversee the duties of the pharmacist and will be responsible 
for ensuring that the pharmacist is accountable for these responsibilities. 

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7 Study Procedures/Evaluations 
7.1 Clinical Evaluations 
Medical History:  Will be obtained by interview of the subjects.  Subjects will be queried 
regarding a history of significant medical disorders of the head, eyes, ears, nose, throat 
(HEENT), mouth, cardiovascular system, lungs, gastrointestinal tract, liver, pancreas, 
kidney, urologic, nervous system, blood, lymph glands, endocrine system, 
musculoskeletal system, skin, and genital/reproductive tract.  A history of any allergies, 
cancer, immunodeficiency, psychiatric illness, and autoimmune disease will be solicited. 
 
Targeted Physical Examination:  At screening or Day 0 visit, a targeted physical exam 
will be performed based on subject’s clinical history and height and weight will be 
collected.  Vital signs (temperature, heart rate, respiration and blood pressure) will be 
collected at screening and prior to each vaccination.  The exam will be performed by 
appropriate study personnel to assess general physical condition and will include the 
following areas/systems:  oral cavity, supraclavicular and axillary lymph nodes, 
cardiovascular, pulmonary, abdomen, skin. 
 
At visits following the first vaccination a targeted physical examination may be 
conducted based on interim medical history.   
 
Reactogenicity Assessments:  Will include brief history for assessment of AE/SAEs just 
prior to and following vaccination, which includes an assessment of pain, tenderness, 
erythema, induration and warmth at the injection site; fever, chills, arthralgia/joint pain, 
malaise/fatigue, myalgia/body aches, headache, nausea, vomiting and abdominal pain.  
The vaccination site will be examined at the end of the 30 minute observation period 
following each vaccination as well as at the Day 3/7 and Day 59/63 clinic visits following 
each vaccination. 
 
Memory Aids:  All subjects will complete a subject memory aid for 8 days following each 
vaccination (Days 0-7).  Subject memory aids will be reviewed with the subject in 
person or by telephone for AE/SAEs at the Day 1-3 follow-up visit or phone call and at 
the Day 5-9 clinic visit following each vaccination.  Following review, subject memory 
aids will be discarded. 
7.2 Concomitant Medications/Treatments 
Administration of any concomitant medications, therapies or vaccines will be 
documented in the subject’s data collection form and reported on the subject’s data 
acquisition screen.  Concomitant medications will include all current medications and 
medications taken within the 30 days prior to enrollment (prescription and over-the-

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counter drugs, vitamins and supplements, topical products, vaccinations, and allergy 
shots) through 34 days after the second vaccination (approximately Day 90 for subjects 
who receive both doses of vaccine or approximately Day 30 for subjects who receive 
only one dose of vaccine) or early termination (if prior to Day 90), whichever occurs first.  
Assessment of eligibility also will include a review of permitted and prohibited 
medications (per the exclusion criteria). 
 
Immunosuppressant medications, as defined by exclusion criterion #22, and 
medications for the treatment of HCV infection will be collected for the duration of the 
study.   
 
Medications that might interfere with the evaluation of the investigational product should 
not be used unless absolutely necessary.  Medications in this category include, but are 
not limited to, glucocorticoids, i.e., oral, parenteral and high-dose inhaled steroids, and 
immunosuppressive or cytotoxic drugs.  Other than from participation in this study, 
subjects should not receive experimental agents including vaccines during the 20-month 
study period.  The administration of licensed vaccines should be delayed until 14 days 
after the last study vaccination.  Subjects should also not receive licensed products from 
participation in another clinical trial. 
7.3 Prevention: Counseling and Risk Reduction  
Prevention counseling for risk avoidance and harm prevention for HCV and other blood 
borne infections (HIV, HBV) will be provided to all study subjects (in both study arms) at 
each study visit where HCV testing occurs.  Counseling will emphasize the unknown 
efficacy of the study product in preventing HCV infection (and or other blood borne 
infections).  Subjects will be counseled that stopping injecting will significantly reduce 
risk of HCV, and that reducing injecting, consistent use of clean needles and drug 
preparation equipment may also prevent parenteral transmission and acquisition of 
HCV, if injecting.  As well, at each visit study subjects will be provided with free sterile 
needles and syringes (supplied by local Public Health Departments and/or local syringe 
exchange programs) and condoms.  Members of the Protocol Team will monitor self-
reported injection rates, including use of clean and used needles and drug preparation 
equipment.  Risk counseling methods will be updated if needed to address higher than 
expected rates of reported unsafe behavior and to promote reduction of parenteral risk 
of HCV by study subjects.  In addition to risk reduction information, referral lists of 
educational resources, community and social service providers, and clinics will be made 
available.  Individuals who are identified as anti-HCV positive at eligibility screening will 
be referred to established services in each locale.  Where possible, medical and social 
service providers who are experienced in working with the trial population will receive 
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7.4 Contraception Status and Pregnancy 
Women of childbearing potential (not surgically sterile via tubal ligation, bilateral 
oophorectomy or hysterectomy or who have not been postmenopausal for ≥1 year) 
must agree to practice adequate contraception from the day of the screening visit 
through 90 days after the second vaccination.  Acceptable birth control methods for the 
purposes of this study may include, but are not limited to:  abstinence; monogamous 
relationship with vasectomized partner; barrier methods such as condoms, diaphragms, 
spermicides, and intrauterine devices; and licensed hormonal methods. 
Contraceptive status is assessed and documented at every scheduled clinic visit for 
subjects who were born female and who are sexually active in a way that could lead 
them to becoming pregnant.  Prior to enrollment and throughout the study, staff will ask 
subjects to verbally confirm their use of adequate contraceptive methods.  A subject 
who was born female and is sexually active in a way that could cause that subject to 
become pregnant should be reminded at all scheduled clinic visits of the importance of 
using contraception and should be referred to specific counseling, information, and 
advice as needed.  This reminder should be documented in the subject’s study record.  
Self-reported infertility—including having reached menopause (no menses for 1 year) or 
having undergone hysterectomy, bilateral oophorectomy, or tubal ligation—must be 
documented in the subject’s study record. 
Vaccinations will be discontinued for subjects who become pregnant during the course 
of the study.  Pregnant subjects will attend the remaining visits and study procedures 
should be completed unless medically contraindicated.  If the subject terminates from 
the study prior to the pregnancy outcome, the site must keep in touch with the subject in 
order to ascertain the pregnancy outcome. 
7.5 Behavioral Risk Assessment Questionnaire 
A clinic interviewer/clinician will administer a Behavioral Risk Assessment Questionnaire 
as specified in the study schedule (Section 8).  The Behavioral Risk Assessment will 
query subjects’ self-reported HCV risk including injection frequency, use of clean and 
used needles and drug preparation equipment, drugs used, and behaviors injecting 
partners.  All subjects will receive risk reduction counseling as detailed in Section 7.3.  
7.6 Laboratory Evaluations 
All laboratory testing will be conducted at CLIA certified laboratories at each site.  Blood 
specimens and volumes will be obtained in accordance with the schedule (Appendix C).  
Blood specimens that will be collected as part of the standard specimen collection 
protocol will include serum and anticoagulated plasma.  These specimens may be used 
for (1) repeat analysis of safety and efficacy parameters; (2) analyses of anti-HCV; (3) 
HCV viral nucleic acids; and (4) antibodies to adenovirus (anti-ChAd3).  Blood samples 

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for isolation and cryopreservation of peripheral blood mononuclear cells (PBMCs) are 
collected for use in immunology studies.  Blood samples collected for immunology 
analyses at the San Francisco site will be processed for PBMC at Blood Systems 
Research Institute (BSRI) using a standardized research operating protocol, and 
shipped to the HCV Immunology Laboratory of Dr. Andrea Cox at Johns Hopkins 
University in Baltimore for laboratory analyses of host antiviral immune responses. 
PBMC for the UNM site will be processed at the UNM Division of Epi/Biostat Molecular 
Epidemiology Lab and shipped to JHU. 
7.6.1 Clinical Laboratory Evaluations 
Urine or serum pregnancy tests will be performed within 24 hours prior to each 
vaccination on all female subjects of childbearing potential.  Results must be negative 
and known prior to vaccination. 
 
Subjects will be tested for HIV antibodies and Hepatitis B surface antigen at screening 
and the last visit. 
 
Clinical laboratory evaluations for safety will be performed by local (clinical) laboratories.  
Venous blood samples (approximately 23 mL) will be collected from each subject prior 
to each vaccination and according to the Study Schedule in Appendix C.  Clinical safety 
lab parameters to be followed include WBC, HgB, platelets, ALT, and creatinine. Urine 
will be collected to evaluate glucose and protein, which will be performed at screening 
only. 
 
Human Leukocyte Antigen (HLA) and IL28B typing will be performed for all subjects.  
IL28B testing and typing will be done at screening/enrollment to inform the 
randomization process.  HLA typing will be done on collected blood samples from 
vaccinated subjects to inform immunogenicity assessments.   
7.6.2 Endpoint Assays:  HCV serology and virology 
HCV RNA 
HCV RNA will be monitored using both qualitative and quantitative assays. 
 
Qualitative HCV RNA testing: 
A standardized HCV RNA qualitative assay (Procleix® HIV-1/HCV assay, by Gen-Probe 
Inc., San Diego, CA) with 50% limit of detection of 12.1 copies/mL of HCV RNA when 
using the recommended 0.5 mL specimen input (95% CI, 11.1 to 13.2) will be used to 
screen for HCV RNA at monitoring visits.  HCV reactive samples will be confirmed using 
a quantitative HCV RNA assay based on real-time PCR (Procleix HCV assay; Gene-
Probe).  (This assay will not be used for HIV screening). 
 

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Quantitative HCV RNA testing: 
Quantitative HCV RNA testing (based on real-time PCR (Procleix HCV assay; Gene-
Probe) will be used to confirm HCV infection. Monitoring of HCV infection relies on 
sensitive and accurate HCV RNA detection by quantitative assays.  Recently developed 
tests based on real time PCR have a high sensitivity (based on the lower limit for 
detecting HCV RNA) and a great dynamic range.  The principle of real-time PCR 
techniques is to detect amplicon synthesis and to deduce the number of viral genomes 
in the start of the clinical sample during the PCR rather than at the end.  The main 
advantage of tests based on real time PCR is that each has been standardized, which 
makes results by different laboratories comparable through reporting in international 
units (IU). 
 
HCV subtypes 
HCV genotype will be determined by reverse transcription/nested PCR (nt 85--1326 
core-E1 region amplicon) following nucleic acid extraction on automated 
instrumentation.  Genotype is determined by sequence comparison with GenBank using 
BLAST.  Viral sequencing of HCV Core-E1 sequences of initial infections will be 
conducted to determine HCV subtype on the first quantitative measurement of HCV 
RNA, dependent on specimen availability and RNA level.  In patients who demonstrate 
alternating HCV RNA positive and negative results or who have missed more than two 
follow up study visits, viral sequencing will be used to confirm reinfection with a different 
virus or intercalating viremia with the same virus.29,84  
 
HCV antibodies 
Anti-HCV testing (counseling will be given prior to testing blood for these blood-borne 
viruses), will be performed at screening and at indicated time points during the study.  
Anti-HCV antibody testing will be done using a commercial enzyme immunoassay (EIA). 
Following a positive EIA test in individuals who are not documented to have evidence of 
an HCV infection (positive HCV RNA), additional testing for HCV antibodies will be done 
at the end of the study to differentiate infection-induced antibodies from vaccine-induced 
HCV antibodies (anti-HCV).  Differentiation will be based on detection or not of 
antibodies to antigens not present in the vaccine.  
 
Anti ChAd3 antibodies 
Antibodies specific for the ChAd3 virus will be monitored at indicated time points 
(Appendix C) at Okairos laboratories by using an experimental assay. 
 
 

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7.6.3 Immunogenicity Evaluations 
Interferon-gamma (IFN-γ) ELISpot will be used to examine HCV specific T-cell 
responses following vaccination. 
 
ELISpot Assay 
Samples for assessment of vaccine immunogenicity will be collected at the indicated 
time points (Appendix C).  The primary immunogenicity assay is the ex vivo ELISpot 
assay for IFN-γ with peptide pools derived from the HCV genotype1b BK virus used in 
the vaccine as antigen.  IFN-γ is a cytokine critical in mediating the adaptive T-cell 
response to viral infections.  It is secreted by activated antigen-specific CD8 T-cells and 
by the CD4 helper T-cells required to sustain effective CD8 T-cell responses.  The 
ELISpot assay is an efficient, simple, and sensitive method of assessing the total T-cell 
(sum of CD8+ and CD4+) response to viral antigens.  Peripheral blood mononuclear 
cells (PBMCs) will be isolated from whole blood and frozen according to validated 
SOPs. PBMC will be thawed according to Okairos SOP IU008 (Human PBMCs triage) 
and assessed for HCV specific immune responses via a human IFN-у ELISpot assay 
based on Okairos’ standard operating procedure. This Okairos SOP has been already 
used to measure anti-HCV immune responses in vaccinated volunteers in previous 
trials: HCV001 (EudraCT number 2007-004259-12), HCV002TV (EudraCT number 
2008-006127-32), HCV003 (EudraCT number 2009-018260-10) and HCV004 (EudraCT 
number 2010-022700-49).  Thawed PBMCs will be incubated for 16 hours with HCV 
antigens in 15-mer peptide pools (overlapping by 11 amino acids) covering the entire 
NS3-NS5b sequence contained in the vaccine.  Peptides 15 amino acid-long have been 
shown to optimally allow stimulation, and consequently detection, of both CD8+ and 
CD4+ T-cell responses.85  To facilitate the analysis the 494 nonstructural HCV peptides, 
the peptides are dissolved in DMSO, and divided into 6 mixtures, or pools for screening: 
pool NS3p (NS3 protease, aa 1027-1349), pool NS3h (NS3 helicase, aa 1339-1661), 
pool NS4 (NS4, aa 1655-1977), pool NS5a (NS5a, aa 1971-2425), pool NS5bI (NS5b 
aa 2415-2725) and pool NS5bII (NS5b aa 2715-3011) 
 
Depending on availability of PBMCs, additional assays will be performed on selected 
samples to further characterize cross-reactivity, function, and phenotype of the induced 
response as detailed in Sections 7.6.4-5. 
7.6.4 Exploratory Assays 
Exploratory studies may include measurements of serum or plasma cytokine and 
chemokine profiles, intracellular cytokine staining to determine if HCV specific T-cells 
can secrete multiple cytokines in addition to IFN-γ and if they degranulate, or additional 
T-cell assays for function based on specimen availability. 
 
 

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7.6.5 Ancillary Assays 
Ancillary studies may include but are not limited to epitope mapping, tetramer analysis 
by flow cytometry, and vector-specific immunogenicity.  Vector-specific studies may be 
performed to better understand the immune responses to the vector and vaccine insert.  
This information may help further product development.  Cryopreserved samples may 
also be used to perform assays to support standardization and validation.  Additional 
cell surface markers, cytokines or functional markers may also be analyzed. 
7.6.6 Future use of stored specimens 
HCV vaccine researchers aim not only to test vaccine candidates but also to continue to 
explore the correlates of immunity to HCV.  In order to do so, the research team intends 
to store plasma, serum, and PBMC samples from subjects at UCSF and JHU.  These 
stored samples will be used for future testing and research related to furthering the 
understanding of HCV or vaccines, such as immunologic analyses in addition to those 
outlined in sections 7.6.3-7.6.5 and future potential bridging studies.  
 
The protocol ICF is written so that the subject is informed that by agreeing to participate 
in the study they are also agreeing to have samples stored for future additional research 
studies.  Some of these samples will be sent to a DMID central storage facility.  
Subjects will not be contacted with the results of these future research studies.  Future 
testing on specimens will only occur to the extent authorized in each study site’s ICF or 
as otherwise authorized under applicable law and after review and approval by the 
DMID and the IRB of the researcher requesting the specimens. 
7.7 Specimen Preparation, Handling, and Shipping 
Biological specimens are to be collected in accordance with protocol specifications, as 
outlined in the Protocol Schedule of Activities.  Study staff will follow all safety 
guidelines of local safety committee and/or institutional policies for handling biological 
specimens.  Blood samples will be labeled and processed according to the protocol-
specific MOP.  Samples will be barcoded and entered into the GlobalTrace Inventory 
system before shipping, as indicated in the protocol-specific MOP. 
 
The blood test for detection of anti-HCV, urine test for glucose and protein, and urine 
pregnancy test will be performed on site.  All other specimens will be collected and 
shipped as indicated in the protocol-specific MOP. 
7.7.1 Instructions for Specimen Preparation, Handling, and Storage 
Instructions for specimen preparation, handling, and storage are included in the 
protocol-specific MOP. 

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7.7.2 Specimen Shipment 
Instructions for specimen shipment are included in the protocol-specific MOP. 
 

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8 Study Schedule 
8.1 Screening and Enrollment  
8.1.1  Anti-HCV Pre-Screening Day -30 to -1  
• Potential subjects will be provided with a description of the anti-HCV test and will 
provide consent for anti-HCV testing.  Anti-HCV screening will be conducted for 
detection of anti-HCV obtained using finger stick or venipuncture whole blood 
sample.   
• Potential subjects who test positive for anti-HCV at this test will not proceed to 
the next screening step.  Those who test negative for anti-HCV will be asked if 
they wish to continue to the Screening Visit Procedures. 
8.1.2 Day -30 to -1, Visit 00, Study Screening  
• Potential subjects with previous negative anti-HCV test may be screened for 
eligibility. All pre-screening and screening activities outlined in Sections 8.1.1 and 
8.1.2 must be completed within 30 days prior to enrollment. The following 
activities will be performed at the screening visit: 
o Potential subjects will be provided with a description of the study (purpose 
and study procedures).  Subjects will be asked to read and sign the informed 
consent form (ICF).  The ICF will be signed prior to performing any other 
screening procedures. 
o Eligibility criteria for study entry and receipt of Dose 1 will be reviewed with 
the subject – see Section 5.1 and 5.2. 
o Vital signs will be obtained, including oral temperature, pulse, and blood 
pressure. 
o Height and weight will be obtained. 
o Medical history will be reviewed. 
o All concomitant medications will be recorded. 
o A targeted physical examination will be performed by appropriate study 
personnel to assess general physical condition.  Additional physical 
assessments may be performed as indicated. 
o Urine sample will be obtained to evaluate glucose and protein. 
o Review of acceptable contraceptive methods (male and female subjects) and 
recent menstrual history (female subjects only). 

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o Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
o Referral to drug treatment and needle exchange programs will be provided. 
o A urine or serum pregnancy test must be performed within 24 hours prior to 
vaccination for all female subjects of childbearing potential.  Results must be 
negative and known prior to vaccination.  If the screening visit is Day -1, a 
negative pregnancy test obtained at Day -1 may be used for eligibility at Day 
0.  If the pregnancy test is > 24 hours prior to vaccination, it must be repeated 
to determine eligibility. 
o Approximately 5 mL of venous blood will be collected for IL28B, as described 
in Section 7.6.1. 
o Approximately 23 mL of venous blood will be collected for serological studies 
and chemistry (see hematology and biochemistry parameters, as described in 
Section 7.6.1). 
o Approximately 20 mL of venous blood will be collected for anti-HCV, HbsAg, 
anti-HIV, and HCV viremia. 
8.1.3 Day 0, Visit 1, Dose 1  
• Eligibility criteria for study entry and receipt of Dose 1 will be reviewed with the 
subject – see Section 5.1 and 5.2. 
• Vital signs will be obtained, including oral temperature, pulse, and blood 
pressure. 
• Medical history will be reviewed to assure continued eligibility. 
• All concomitant medications will be reviewed for accuracy and completeness.  
Any new medications will be recorded and assessed for continuing eligibility. 
• A targeted physical examination may be performed, as indicated, based on 
subject’s recent clinical history since the screening visit. 
• Behavioral risk will be assessed using an interviewer administered questionnaire. 
• A urine or serum pregnancy test must be performed within 24 hours prior to 
vaccination for all female subjects of childbearing potential.  Results must be 
negative and known prior to vaccination.  If the screening visit is Day -1, a 
negative pregnancy test obtained at Day -1 may be used for eligibility at Day 0.  If 
the pregnancy test is > 24 hours prior to vaccination, it must be repeated to 
determine eligibility. 
• Approximately 8 mL of venous blood will be collected for HLA typing/genetic 
studies, as described in Section 7.6.1. 

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• Approximately 23 mL of venous blood will be collected prior to vaccination for 
hematology, coagulation and chemistry/metabolic biochemistry parameters, as 
described in Section 7.6.1.  Clinical safety laboratory assessments will be 
performed by the local laboratory.  The results from this blood draw will not be 
available or reviewed prior to vaccination, and will serve as a baseline safety 
assessment only. 
• Approximately 160 mL of venous blood will be collected prior to vaccination for 
baseline immunology assessments. 
• Plasma saved from the blood collected for immunology assessments at this visit 
will be used for anti-adenovirus Ab testing. 
• Subjects will be enrolled in AdvantageEDC (IDES) and randomly assigned to a 
treatment arm. 
• Subjects will receive a single dose of AdCh3NSmut1 vaccine or placebo per 
randomized assignment via intramuscular injection in the deltoid muscle of the 
preferred arm.  Following vaccination the injection site will be covered with a 
sterile dressing and subjects will be observed in the clinic for at least 30 minutes.  
The vaccination site will be examined, and any AE/SAEs will be assessed prior to 
discharge from the clinic. 
• Subjects will be provided with a memory aid and other study related materials to 
record daily oral temperature and systemic and local AE/SAEs.  Subjects will be 
encouraged to take their temperature around the same time each day.  Subjects 
will be instructed on how to use the memory aid and how to measure and record 
AE/SAEs prior to discharge from the clinic.  Subjects will be instructed to notify 
the study center if they develop any severe reactions following vaccination.  If the 
site principal investigator or appropriate sub-investigator deems the reaction 
severe enough, s/he will give further instructions on the proper course of action, 
including a return to the clinic for immediate evaluation if appropriate. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2 Subject Follow-Up 
Follow-up visits are scheduled in reference to dosing dates as indicated for each visit 
window. 
 
 

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8.2.1 Day 3, Visit 2  
(Window: Day 1 to 5) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review the memory aid information with the subject and 
record all AE/SAEs and concomitant medications on the appropriate data 
collection form. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.2 Day 7, Visit 3 
(Window: Day 5 to 9) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review the memory aid information with the subject and 
record all AE/SAEs and concomitant medications on the appropriate data 
collection form. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.3 Day 14, Visit 4 
(Window: Day 7 to 21)  
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined if ongoing Memory Aid symptoms are 
reported after Visit 3. 

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• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review and record all AE/SAEs and concomitant 
medications on the appropriate data collection form  
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.4 Day 30, Visit 5 
(Window: Day 23 to 37) 
• Current health status will be reviewed and any changes since the last visit will be 
recorded. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review and record all AE/SAEs and concomitant 
medications on the appropriate data collection form. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Approximately 160 mL of venous blood will be collected for immunologic assays. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.5 Day 56, Visit 6, Dose 2 
(Window: Day 49 to 63) 
• Eligibility criteria for receipt of Dose 2 will be reviewed with the subject – see 
Section 5.1 and 5.2. 
• Current health status will be reviewed and any changes since the last visit will be 
recorded. 
• The vaccination site will be examined. 

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• Vital signs will be obtained, including oral temperature, pulse, and blood 
pressure. 
• All concomitant medications will be reviewed for accuracy and completeness. 
Any new medications will be recorded and assessed for continuing eligibility. 
• A targeted physical examination may be performed, as indicated, based on 
subject’s recent clinical history since the screening visit. 
• A urine or serum pregnancy test must be performed within 24 hours prior to 
vaccination for all female subjects of childbearing potential.  Results must be 
negative and known prior to vaccination.  If the screening visit is Day -1, a 
negative pregnancy test obtained at Day -1 may be used for eligibility at Day 0.  If 
the pregnancy test is > 24 hours prior to vaccination, it must be repeated to 
determine eligibility. 
• Approximately 23 mL of venous blood will be collected prior to vaccination for 
hematology, coagulation and chemistry/metabolic biochemistry parameters, as 
described in Section 7.6.1, and performed by the local laboratory. 
• Approximately 10 mL of venous blood will be collected prior to vaccination for 
HCV viremia.  
• Approximately 100 mL of venous blood will be collected prior to vaccination for 
baseline immunology assessments. 
• Subjects will receive a single dose of MVA-NSmut vaccine or placebo per 
randomized assignment via intramuscular injection in the deltoid muscle of the 
preferred arm.  Following vaccination the injection site will be covered with a 
sterile dressing and subjects will be observed in the clinic for at least 30 minutes.  
The vaccination site will be examined, and any AE/SAEs will be assessed prior to 
discharge from the clinic. 
• Subjects will be provided with a memory aid and other study related materials to 
record daily oral temperature and systemic and local AE/SAEs.  Subjects will be 
encouraged to take their temperature around the same time each day.  Subjects 
will be instructed on how to use the memory aid and how to measure and record 
AE/SAEs prior to discharge from the clinic.  Subjects will be instructed to notify 
the study center if they develop any severe reactions following vaccination.  If the 
site principal investigator or appropriate sub-investigator deems the reaction 
severe enough, s/he will give further instructions on the proper course of action, 
including a return to the clinic for immediate evaluation if appropriate. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 

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• Subjects who become HCV PCR+ after the 1st vaccine may receive a 2nd 
vaccine dose. These subjects will return for their Visit 7 and 8 assessments and 
follow the Confirmed HCV Infection visit schedule outlined in Appendix D. 
8.2.6 Day 59 [Target 3 Days Post Second Vaccination], Visit 7 
(Window: 1-5 days after receipt of second dose of vaccine) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review the memory aid information with the subject and 
record all AE/SAEs and concomitant medications on the appropriate data 
collection form. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.7 Day 63 [Target 7 Days Post Second Vaccination], Visit 8 
(Window: 7-21 days after receipt of second dose of vaccine) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Study personnel will review the memory aid information with the subject and 
record all AE/SAEs and concomitant medications on the appropriate data 
collection form. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 
• Approximately 100 mL of venous blood will be collected for immunologic assays. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 

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8.2.8 Day 90, Visit 9 
(Window: Day 83 to 97) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire.  
• Study personnel will review and record all AE/SAEs and concomitant 
medications on the appropriate data collection form. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Approximately 160 mL of venous blood will be collected for immunologic assays. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.9 Day 120, Visit 10 
(Window: Day 113 to 127) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 

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8.2.10 Days 150–270, Visits 11–15 
Visit 
Target (Day) 
Window (Days) 
11 
150 
+ 7 
12 
180 
+ 7 
13 
210 
+ 7 
14 
240 
+ 7 
15 
270 
+ 7 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire 
at visits 12 and 15.  
• Approximately 100 mL of venous blood will be collected for immunologic assays 
at Visit 14. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.11 Day 300, Visit 16 
(Window: Day 293 to 307) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status.  
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 
local laboratory. 

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• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.12 Day 330, Visit 17 
(Window: Day 323 to 337) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.13 Day 360, Visit 18 
(Window: Day 353 to 367) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAES that have occurred since the last contact will be collected. 
• Approximately 20 mL of venous blood will be collected for anti-adenovirus Ab 
and HCV viremia testing. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 

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8.2.14 Days 390–570, Visits 19–25 
Visit 
Target (Day) 
Window (Days) 
19 
390 
+ 7 
20 
420 
+ 7 
21 
450 
+ 7 
22 
480 
+ 7 
23 
510 
+ 7 
24 
540 
+ 7 
25 
570 
+ 7 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status.  
• Behavioral risk will be assessed using an interviewer administered questionnaire 
at visits 21 and 24. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 10 mL of venous blood will be collected for HCV viremia. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
8.2.15 Day 600, Visit 26 
(Window: Day 593 to 607) 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire. 
• Immunosuppressant concomitant medications and medications for the treatment 
of HCV infection will be recorded. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1, and performed by the 

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local laboratory. 
• Approximately 20 mL of venous blood will be collected for anti-HCV, HbsAg, anti-
HIV and HCV viremia. 
• Approximately 100 mL of venous blood will be collected for immunologic assays. 
• Counseling on avoidance of HCV and HIV infection, and pregnancy will be 
conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
• Subjects will be thanked for participating in the study and contact information for 
end of study results will be confirmed. 
8.3 Follow-up for Subjects with HCV Infection 
Subjects who are confirmed HCV RNA positive will be followed for approximately 9 
months after their first HCV-positive quantitative RNA result.  The schedule of 
assessments for HCV RNA positive subjects is outlined in Appendix D:  Study Visit 
Chart – Confirmed HCV Infection.  Subjects will exit the study after visit H09, 
approximately Day 270 after initial HCV detection.  
 
Subjects who are confirmed HCV RNA positive prior to the 2nd dose of vaccine will be 
followed for safety as scheduled from the date of their first HCV-positive quantitative 
RNA result.  Subjects will exit the study after visit H09, approximately Day 270 after 
initial HCV detection. 
 
Subjects who are confirmed HCV RNA positive prior to the 2nd dose of vaccine and 
choose to terminate early will do the full schedule of assessments for HCV RNA positive 
subjects for visit H09 (exit visit). 
 
Trial subjects who test positive for HCV RNA will be provided with a referral to providers 
external to the study for evaluation for appropriate care and treatment of HCV infection.  
Treatment may include interferon alpha with or without ribavirin.  Referrals will be 
provided at every visit at which a participant tests positive for HCV RNA.  The providers’ 
lists will include primary care and specialist resources with experience working with and 
providing care to those at high risk for HCV infection, including injecting populations.  
This approach is based upon the NIH Consensus Statement on Management of 
Hepatitis C: 2002 regarding acute hepatitis C treatment which states, “it is 
recommended that treatment of active injection drug use be considered on a case-by-
case basis, and that active injection drug use in and of itself not be used to exclude 
such patients from antiviral therapy.”86   
 

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Follow-up visits are scheduled in reference to the first positive HCV RNA quantitative 
result. 
8.3.1 HCV Day 30, Visit H01 
(Window: HCV Day 23 to 37) 
• Study personnel will inform subjects that they have tested positive for the 
hepatitis C virus.  
• Counseling and written information on acute hepatitis C treatment options will be 
provided along with a list of healthcare providers who are separate from this 
study; a referral to a community-based healthcare provider will be offered. 
• Counseling on avoidance of HCV transmission, HIV infection, and pregnancy will 
be conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• The vaccination site will be examined. 
• A targeted physical examination may be performed, if indicated, based on review 
of the current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire, 
if applicable. 
• SAEs that have occurred since the last contact will be collected 
• Approximately 138 mL of venous blood for hematology, biochemistry, HCV 
genotyping, HBsAg, anti-HIV, HCV quantitative viremia, and immunology will be 
collected. 
8.3.2 HCV Days 60–210, Visits H02–H07 
Visit 
Target (Day) 
Window (Days) 
H02 
60 
± 7 
H03 
90 
± 7 
H04 
120 
± 7 
H05 
150 
± 7 
H06 
180 
± 21 
H07 
210 
± 7 
• Counseling and written information on acute hepatitis C treatment options will be 
provided along with a list of healthcare providers who are separate from this 
study; a referral to a community-based healthcare provider will be offered. 

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• Counseling on avoidance of HCV transmission, HIV infection, and pregnancy will 
be conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of the current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire, 
if applicable. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 133 mL of venous blood for hematology, biochemistry, HCV 
genotyping, HCV quantitative viremia, and immunology will be collected. 
8.3.3 HCV Day 240, Visit H08 
(Window: HCV Day 233 to 247) 
• Counseling and written information on acute hepatitis C treatment options will be 
provided along with a list of healthcare providers who are separate from this 
study; a referral to a community-based healthcare provider will be offered. 
• Counseling on avoidance of HCV transmission, HIV infection, and pregnancy will 
be conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of the current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire, 
if applicable. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 33 mL of venous blood for hematology, biochemistry, HCV 
genotyping, and HCV quantitative viremia will be collected. 
 
 

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8.3.4 HCV Day 270, Visit H09  
(Window: HCV Day 263 to 277) 
• Counseling and written information on acute hepatitis C treatment options will be 
provided along with a list of healthcare providers who are separate from this 
study; a referral to a community-based healthcare provider will be offered. 
• Counseling on avoidance of HCV transmission, HIV infection, and pregnancy will 
be conducted. 
• Referral to drug treatment and needle exchange programs will be provided. 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of the current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 143 mL of venous blood for hematology, biochemistry, HCV 
genotyping, anti-HCV, HbsAg, anti-HIV, HCV quantitative viremia, and 
immunology will be collected. 
• Subjects will be thanked for participating in the study and contact information for 
end of study results will be confirmed. 
8.4 Unscheduled Visit(s) 
Unscheduled visits may occur at any time during the study.  They may occur for the 
following reasons:   
(1) For operational reasons, e.g., a subject may request to reschedule, or to ask 
questions;  
(2) For AE related reasons.  When interim contacts or visits are completed in response 
to subject reports of AEs, study staff will assess the reported event clinically and 
provide or refer the subject to appropriate medical care.  All AEs will be evaluated 
and reported as required (Section 9.3); 
(3) If a subject presents to the study site after having missed a scheduled visit (e.g., in 
response to locator/tracing efforts) on a day that does not fall within a scheduled visit 
window; 
(4) For other reasons at subject request.  All interim contacts and visits will be 
documented in subjects' study records and on applicable data collection forms. 

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8.5 Early Termination  
Subjects may voluntarily withdraw their consent for participation in the study at any time 
and for any reason, without penalty.  Subjects may also withdraw voluntarily from 
receiving the study intervention for any reason.  A site principal investigator may also 
withdraw a subject from receiving further study intervention.  Follow-up safety 
assessments and immunogenicity evaluations will be completed, if possible, whether 
the subject withdraws from the study or is withdrawn from receiving further study 
product.  Subjects will be encouraged to permit continued follow-up of AE/SAEs and to 
donate scheduled venous blood samples for clinical safety laboratory and 
immunogenicity evaluations, if possible – see the protocol-specific MOP for alternate 
follow-up requirements.  For subjects who withdraw within 180 days of their last dose of 
vaccine, a 6-month follow-up phone call should be conducted to collect any new SAEs. 
8.5.1 For HCV-uninfected subjects 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• All concomitant medications will be recorded (if prior to Day 90 visit).  
Immunosuppressant medications and medications for the treatment of HCV 
infection will be collected throughout the study as applicable. 
• Approximately 23 mL of venous blood will be collected for hematology and 
biochemistry parameters, as described in Section 7.6.1. 
• Approximately 100 mL of venous blood will be collected for immunologic assays. 
• Approximately 20 mL of venous blood will be collected for anti-HCV, HbsAg, anti-
HIV, and HCV viremia. 
• A targeted physical examination may be performed, if indicated based on review 
of current health status. 
• Information regarding adverse events will be collected (if before Day 90).  All 
SAEs that have occurred since the last contact will be collected. 
• Subjects will be thanked for participating in the study and contact information for 
end of study results will be confirmed.  
8.5.2 For HCV-infected subjects 
• Counseling and written information on acute hepatitis C treatment options will be 
provided along with a list of health care providers who are separate from this 
study; a referral to a community-based healthcare provider will be offered. 
• Counseling on avoidance of HCV transmission, HIV infection, and pregnancy will 
be conducted. 

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• Referral to drug treatment and needle exchange programs will be provided. 
• Current health status will be reviewed and any changes since the last visit will be 
noted. 
• A targeted physical examination may be performed, if indicated, based on review 
of the current health status. 
• Behavioral risk will be assessed using an interviewer administered questionnaire. 
• SAEs that have occurred since the last contact will be collected. 
• Approximately 138 mL of venous blood for hematology, biochemistry, HCV 
genotyping, anti-HCV, HCV quantitative viremia, and immunology will be 
collected. 
• Approximately 5 mL of venous blood will be collected for HbsAg and anti-HIV for 
subjects who become HCV-infected prior to the 2nd dose of vaccine and choose 
to terminate early. 
• Subjects will be thanked for participating in the study and contact information for 
end of study results will be confirmed. 
8.6 Referral for Acute HCV Infection 
All trial subjects with positive HCV RNA test results will be provided a referral to 
providers external to the study for evaluation for appropriate care and treatment of HCV 
infection.  Treatment may include interferon alpha with or without ribavirin.  Referrals will 
be provided at every visit at which a participant tests HCV RNA positive.   
8.7 Subsequent Positivity for Anti-HCV: Distinguishing Intercurrent 
HCV Infection from Vaccine Induced Positive HCV Serology 
It is expected that after vaccination some subjects may become positive for anti-HCV in 
the absence of infection as a result of exposure to antigens derived from HCV that are 
included in the vaccine.  In the previous clinical trial, HCV001, seroconversion to HCV in 
absence of documented infection occurred only in 10% of vaccinated subjects.  Anti-
HCV testing will be done using a commercial enzyme immunoassay (EIA) on subjects 
who do not develop HCV infection during the trial. Following a positive EIA test, 
additional testing will be done to differentiate infection- from vaccine-induced antibodies 
(See Section 7.6). Testing will be conducted on stored blood samples obtained from 
monthly visits. Differentiation will be based on detection or not of antibodies to antigens 
not present in the vaccine.  Subjects who develop vaccine induced anti-HCV will be 
informed at the end of the study and be provided with documentation explaining the 
reasons for their anti-HCV positivity for communication with their medical providers (with 
the subjects’ consents).  This will be discussed in detail at screening and in ICFs. 

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9 DMID Safety Reporting and Safety Assessment Monitoring 
Regulatory requirements, including the FDA regulations and the ICH Guidelines for 
Good Clinical Practice, define safety monitoring and reporting responsibilities of 
sponsors and investigators to ensure the safety and protection of human subjects 
participating in clinical trials. 
9.1 Responsibilities 
Investigators participating in this clinical trial are responsible for and will: 
• Evaluate subject safety including assessment of adverse events (AEs) for 
seriousness, severity, and causality 
• Notify the sponsor (DMID) through their Pharmacovigilance contractor of SAEs 
immediately 
• Provide detailed written reports promptly following immediate initial reports, 
including necessary documentation requested by the sponsor or IRB  
• Inform the IRB of AEs as required by applicable regulatory requirements 
9.2 Definitions 
Adverse Event (AE)  
An AE is any untoward medical occurrence in a clinical investigation subject who has 
received a study intervention and that does not necessarily have to have a causal 
relationship with the study product.  An AE can, therefore, be any unfavorable and 
unintended sign (including an abnormal laboratory finding, for example), symptom, or 
disease temporally associated with the use of the study product, whether or not 
considered related to the study product. 
 
Unexpected 
An unexpected AE is an adverse reaction, the nature or severity of which is not 
consistent with the applicable product information (e.g., investigator’s brochure [IB] for 
an unapproved investigational medicinal product).  Unexpected refers to an experience 
that has not been previously observed. This includes events that are more serious than 
expected or occur more frequently than expected. 
 
Expected 
Any adverse experience, the nature, severity or frequency of which is consistent with 
the current IB; or with the risk information described in the investigational plan or 
protocol or consent form. 
 
 
 

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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 subject and may require 
medical or surgical intervention to prevent one of the outcomes listed in this 
definition.  Examples of such medical events include allergic bronchospasm 
requiring intensive treatment in an emergency room or at home, blood dyscrasias 
or convulsions that do not result in subject 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 subject 
or subject at immediate risk of death.  It does not include an adverse event, had it 
occurred in a more severe form, might have caused death. 
9.3 Safety Reporting Requirements 
9.3.1 Reporting Interval 
All SAEs and new onset chronic illnesses will be documented from randomization 
through subject’s last visit (Visit 26 or H09)/subject completion of all study assessments.   
• New-onset chronic illnesses will be collected from randomization through a 
subject’s last visit (Visit 26 or Visit H09).  New-onset chronic illnesses are defined 
as any new ICD-10 diagnosis that is applied to the subject during the duration of 
the study, after receipt of the study agent, that is expected to continue for at least 
6 months and requires continued health care intervention.  To reference the 
International Statistical Classification of Diseases and Related Health Problems, 
10th revision (ICD-10): http://www.cdc.gov/nchs/about/otheract/icd9/abticd10.htm      
• Non-serious Adverse Events will be collected from randomization through 34 
days post-vaccination 2 (approximately Day 90).  All SAEs and AEs will be 
followed until they are resolved or determined by the PI to be medically stable, 

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even if this extends beyond the study-reporting period.  Resolution of an adverse 
event is defined as the return to pre-treatment status or stabilization of the 
condition with the expectation that it will remain chronic. 
 
At any time after completion of the study, if the investigator becomes aware of a serious 
adverse event that is suspected to be related to study product, the investigator will 
report the event to the DMID Pharmacovigilance contractor, DMID CROMS 
Pharmacovigilance Group. 
9.3.2 Notification of the Sponsor of Serious Adverse Events 
Any AE that meets a protocol-defined serious criterion must be submitted on an SAE 
form to the DMID CROMS Pharmacovigilance Group (PVG), the DMID 
Pharmacovigilance contractor immediately (within 24 hours of site awareness), at the 
following address:  
 
DMID CROMS Pharmacovigilance Group (PVG) 
Technical Resources International, Inc  
6500 Rock Spring Dr., Suite 650 
Bethesda, MD 20817 USA  
Email: XXXXXXXXXX  
Fax:  XXXXXXXXXX 
SAE line: XXXXXXXXXX 
 
Other supporting documentation of the event may be requested by the DMID 
Pharmacovigilance contractor and should be provided as soon as possible. 
 
The DMID Medical Monitor (MM) and Clinical Project Manager (CPM) will be notified of 
the SAE.  The DMID MM will review and assess the SAE for regulatory reporting and 
potential impact on study subject safety and protocol conduct. 
 
Questions about SAE reporting can be referred to the SAE line  
(available 24 hours a day/7 days a week). 
9.3.3 Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND 
Following notification from the investigator, DMID, the IND sponsor, will report any 
suspected adverse reaction that is both serious and unexpected.  DMID will report an 
adverse event as a suspected adverse reaction only if there is evidence to suggest a 
causal relationship between the drug and the adverse event.  DMID will notify FDA and 
all participating investigators (i.e., all investigators to whom the sponsor is providing 
drug under its INDs or under any investigator’s IND) in an IND safety report of potential 
serious risks from clinical trials or any other source, as soon as possible, but in no case 
later than 15 calendar days after the sponsor determines that the information qualifies 

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for reporting as specified in 21 CFR Part 312.32.  DMID will also notify FDA of any 
unexpected fatal or life-threatening suspected adverse reaction as soon as possible but 
in no case later than 7 calendar days after the sponsor’s initial receipt of the information.  
Relevant follow-up information to an IND safety report will be submitted as soon as the 
information is available.  Upon request from FDA, DMID will submit to FDA any 
additional data or information that the agency deems necessary, as soon as possible, 
but in no case later than 15 calendar days after receiving the request. 
 
All serious events designated as “not related” to study product(s), will be reported to the 
FDA at least annually in a summary format. 
9.3.4 Reporting of Pregnancy 
Although not AEs, pregnancies are reportable events.  Any pregnancies that occur 
during the study period in women who have received any study product will be reported 
via the Emmes Internet Data Entry System (IDES) on the Pregnancy Report form within 
5 days of site awareness.  The report will include pregnancy outcome (e.g., any 
premature terminations, elective or therapeutic, and any spontaneous abortions or 
stillbirths), as well as the health status of the mother and child, including date of delivery 
and infant’s gender and weight.  Pregnancies will be followed until birth.  All 
pregnancies will be reported to the clinical data management group.  If the database is 
locked at time of pregnancy, a supplemental report will be generated and completed 
after birth and 3 month follow up, which will be appended to the database. 
9.4 Investigator’s Assessment of Adverse Events 
The determination of seriousness, severity, and causality will be made by an onsite 
investigator who is qualified (licensed in the state) to diagnose adverse event 
information, provide a medical evaluation of adverse events, and classify adverse 
events based upon medical judgment.  This includes but is not limited to physicians, 
physician assistants, and nurse practitioners.  Laboratory abnormalities will be reported 
as AEs based on the normal range of values for the laboratory.  The grading of the 
laboratory AEs will be based on the DMID toxicity table (Appendix B). 
9.4.1 Assessment of Seriousness 
Event seriousness will be determined according to the protocol definition of an SAE 
(Section 9.2). 
9.4.2 Assessment of Severity 
Event severity will be graded according to parameters shown in Appendix B or if not, 
noted in the tables using the definitions below: 

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1 = Mild (awareness of a symptom but the symptom is easily tolerated; the symptom 
requires minimal or no treatment) 
2 = Moderate (discomfort enough to cause interference with usual activity) 
3 = Severe (incapacitating; unable to perform usual activities; requires 
absenteeism/bed rest) 
4 = Life-threatening  
5 = Death 
9.4.3 Assessment of Association 
Relationship to Study Products:  The clinician’s assessment of an AE's relationship to 
test article (vaccine or study drug) 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 trial, 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. 
9.4.4 Assessment of Reactogenicity 
Subjects will be asked to record events daily on an 8-day subject memory aid.  The 
subject will record temperature and the presence and intensity of post-vaccination 
reactogenicity events, including pain, tenderness, erythema, induration and warmth at 
the at injection site; fever, chills, arthralgia/joint pain, malaise/fatigue, myalgia/body 
aches, headache, nausea, vomiting, abdominal pain and any other symptoms they 
might be experiencing after vaccination.  Erythema and induration at the vaccination site 
will be measured in millimeters.  Any symptoms still present at Day 8 will continue to be 
followed until symptom resolution.  Subjects will also be asked to record any 
medications taken and any emergency room or physician visits (other than routine 
check-ups).  The subject memory aid will be reviewed with the subject at subsequent 
clinic visits and phone calls as needed. 
 
Symptoms will be graded using the following scale: 
• Grade 0:  Not present 
• Grade 1:  Mild (present but easily tolerated) 

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• Grade 2:  Moderate (able to tolerate routine activity with effort) 
• Grade 3:  Severe (unable to continue routine activity) 
 
This information will be reviewed by study staff during follow-up subject contacts. 
 
In the event a subject experiences an adverse event or injection site reaction that is still 
present at the end of the subject’s participation in the study, the subject must be 
followed until resolution or until the event or abnormality resolves to the investigator's 
and/or sponsor's satisfaction.  Follow-up procedures, evaluations, and outcomes will be 
recorded on the subject's data collection forms. 
9.5 Safety Monitoring by the DMID Safety Oversight Mechanism 
9.5.1 Data Safety and Monitoring Board (DSMB) 
Safety oversight will be under the direction of a DSMB.  The DSMB will meet at least 
annually (to be determined by the DSMB) to assess safety and enrollment data in 
addition to the planned interim analysis.  If halting rules are initiated or the DSMB 
requests additional meetings, more frequent meetings may be held.  The DSMB will 
operate under the rules of a DMID-approved charter that will be written at the 
organizational meeting of the DSMB.  The DSMB will advise DMID of its findings.  The 
DSMB will review blinded aggregate safety data for increased rate of occurrence of 
serious suspected adverse reactions 
9.5.2 Independent Safety Monitor (ISM) 
Safety oversight for this study will include an ISM in close proximity to each of the 
clinical research sites.  The ISM will receive and evaluate SAEs blinded and provide an 
independent written summary to the DMID Clinical Project Manager and DMID Medical 
Monitor.  The ISM may be asked by the DMID Medical Monitor to review additional 
safety events (i.e. significantly increased frequency of non-serious adverse events). 
9.5.3 Interruption or Discontinuation of Study Enrollment and Study Product 
Administration for all Subjects in the Study 
DMID, as the study sponsor, may interrupt study dosing and/or study entry at any time if 
medically indicated.  To minimize risk, cumulative safety data will be reviewed by the 
Data and Safety Monitoring Board (DSMB), as expeditiously as is reasonable.  The 
DSMB may recommend terminating the study if deemed necessary following a vaccine-
related SAE. 

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9.6 Halting Criteria/Rules 
If the trial is halted, study drug will not be administered to any additional subjects, and 
enrollment will stop until the DSMB has reviewed the event. 
 
After their review, the DSMB will make a recommendation to DMID whether the study 
should continue per protocol, proceed with caution, be further investigated, be 
discontinued, or be modified and then proceed.  Suspension of enrollment (for a 
particular group or for the entire study) is another potential outcome of a safety review. 
 
Subsequent review of serious, unexpected, and related AEs by the medical monitor, 
DSMB, IRB, the Sponsor, the FDA, or relevant local regulatory authorities may also 
result in suspension of further trial interventions/administration of study product at a site.  
The FDA and study Sponsor retain the authority to suspend additional enrollment and 
study interventions/administration of study product for the entire study, as applicable. 
The following events will trigger a safety review: 
• If two subjects experience a systemic adverse event that is judged to be severe 
(Grade 3) and related to vaccination.  Number of events contributing to halting 
rule is the sum of a and b: 
a. Solicited severe (Grade 3) systemic reaction that is judged to be related to the 
vaccination.  Solicited reactions are assumed to be associated with 
vaccination. 
b. Severe (Grade 3) adverse event that is judged to be related to the 
vaccination. 
• If 1 subject experiences an SAE judged by an investigator to be related to 
vaccination. 
• If 1 subject develops a laboratory value that is considered Severe (Grade 3) and 
that is judged to be related to vaccination. Includes all lab values that are 
considered severe.  Relatedness must be determined by the medical monitor. 

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10 Clinical Monitoring 
10.1  Site Monitoring Plan 
Clinical monitoring at the site will be conducted to verify that the study is conducted 
according to the approved protocol, human subjects are protected, and data collection 
processes are of high quality and meet sponsor SOPs, GCP/ICH guidelines, and federal 
regulations.  DMID, the sponsoring agency, or its designee will conduct site monitoring 
visits as detailed in the monitoring plan or in the Manual of Procedures (MOP). 
 
Site visits will be made at standard intervals or more frequently as defined by DMID.  
Monitoring visits will include, but are not limited to, review of regulatory files, study 
product accountability records, data collection forms, case report forms, informed 
consent forms, medical and laboratory reports, and protocol compliance.  Study 
monitors will meet with investigators to discuss any problems and actions to be taken 
and document visit findings and discussions.  Clinical monitoring reports will be 
submitted to DMID. 
 
 

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11 Statistical Considerations 
11.1 Study Hypotheses 
The primary objective of the trial is to assess the safety and evaluate the efficacy of new 
candidate hepatitis C virus vaccines, AdCh3NSmut1 and MVA-NSmut, compared to 
placebo when administered sequentially to IDU.  This Phase I/II study is intended 
primarily to uncover any safety issues that occur at a sufficiently high rate that they 
might be observed in a study of this size, and to obtain meaningful estimates of the 
efficacy of the vaccine against chronic HCV infection.  While the limited sample 
precludes reaching definitive conclusions, descriptive analyses will be supplemented by 
hypothesis testing and estimation of vaccination effects when feasible.  For the primary 
safety and efficacy endpoints, the null hypothesis of no difference in vaccine versus 
placebo arms will be tested against a two-sided alternative at the p = 0.05 level. 
11.2 Sample Size Considerations 
Sample size calculations are based on the assumption of detecting a 60% reduction in 
incidence of chronic infection over 18 months of post-vaccination follow-up among 
vaccinated subjects compared to unvaccinated controls in an according to protocol 
(ATP) analysis.  A total of 43 observed events of chronic infection in the ATP cohort 
would provide power of 85% for a two-sided log rank test conducted at the significance 
level (alpha) of 0.05 to detect a 60% reduction in hazard rate.  The incidence of chronic 
infection among controls is assumed to be 14% annually.30  Assuming 14 cases/100 
person-years (PY) in the control arm, and 5.6 cases/100 PY in the vaccine arm, a total 
of 292.5 subjects in the ATP cohort followed for 1.5 years would provide on average 43 
events.  We further assume that 65% of enrolled subjects will be retained in the ATP 
cohort.  Therefore, we inflate the required number of subjects by a factor of 1/0.65 and 
obtain a target enrollment of 540 subjects, or 270 subjects per vaccination arm. 
 
As described in Section 11.3, the DSMB will conduct a blinded examination of progress 
towards the goal of 43 observed events of chronic infection in the ATP cohort, and 
decide whether to recommend an extension of the enrollment period.  At the September 
2, 2015 DSMB meeting, the DSMB reviewed the progress towards this goal and 
recommended increasing the target enrollment from 450 to 540 subjects, or 270 
subjects per vaccination arm. 
11.3 Planned Interim Analyses 
The DSMB will meet and review safety data and enrollment data at least annually as 
outlined in the DSMB charter.  The study will be monitored to determine if any of the 

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safety halting rules described above in Section 9.6 are met.  Additionally, there will be 
two planned interim analyses.   
 
The first interim analysis is planned when study Day 63 (approximately week 9) follow-
up data is available on the last of approximately 68 subjects enrolled in Stage 1 of the 
trial.  The second interim analysis is planned to occur when roughly one-third of enrolled 
and currently per-protocol subjects (N = 98) have completed six months of post-
vaccination follow-up, but no later than one month prior to the planned completion of 
enrollment to the trial. Both the first and second interim analysis will be fully blinded and 
will report on trial data aggregated across both treatment arms. 
 
The first stage of the study is designed to establish the safety and immunogenicity of 
the sequential administration of the AdCh3NSmut1 and MVA-NSmut vaccines in a 
prime/boost regimen in the target population.  The first interim analysis will focus on 
presenting descriptive tabulations of the safety parameters to the DSMB along with the 
ELISpot assay immunological data. The immunological data analysis will not require 
study un-blinding.  A positive immune response is defined by two criteria:  i) more than 
48 spot forming cells per million PBMC; ii) at least three times the mean background 
spots per million PBMC found in ELISpot wells containing cells and peptide diluent 
(DMSO).39,87,88  A subject will be considered to be a responder if a positive immune 
response is detected in at least one of six different pools of peptides utilized from at 
least one of the blood samples drawn after the first or second vaccination.  Please note 
that results obtained from blood samples drawn after the onset of HCV infection will not 
be used in this first interim analysis.  A subject will be considered evaluable for the first 
interim immunological analysis if ELISpot assay data is available from a sample drawn 
7-21 days after receipt of the second vaccination, the anticipated timeframe in which a 
positive response to vaccine will be detected if present.  Note that this requirement does 
not alter the definition of a responder provided above, and in particular, a positive 
ELISpot result after the first vaccination still suffices to define a responder.  The study 
will proceed to Stage 2 in the absence of any safety concerns and if at least 30% of the 
evaluable subjects in Stage 1 of the study (n = 68 ± 4) have a positive immune 
response to HCV.  If this threshold is not met, then a more complete unblinded analysis 
of immunogenicity responses will be undertaken by an independent assessor appointed 
by DMID.     
 
As noted in the section 11.2 (Sample Size Considerations), the statistical information 
goal for the efficacy endpoint of prevention of chronic HCV is 43 cases of chronic HCV 
in the per protocol analysis cohort.  At the second interim analysis, the DSMB will do a 
blinded examination of progress towards this goal, and decide whether to recommend 
an extension of the enrollment period, while it is still possible to do so without halting 
enrollment and losing momentum in accrual. 

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11.4 Final Analysis Plan 
11.4.1 Definition of Analysis Cohorts 
The safety analysis cohort will consist of all randomized subjects who receive at least 
one vaccination. 
 
The mITT efficacy analysis cohort will consist of all randomized subjects who receive at 
least one vaccination, are not HCV infected at the first vaccination, and have sufficient 
follow-up to be evaluable for efficacy.  For the primary efficacy endpoint of chronic HCV 
infection, sufficient follow-up will consist of at least three clinic visits following the 
second vaccination.  If the second dose of vaccine was not administered, the target 
date for the second dose will be used in this calculation.  Subjects who receive 
treatment for acute HCV infection will be censored from time to event analyses of 
chronic infection at the date of first treatment. 
 
The according to protocol (ATP) efficacy analysis cohort is a subset of the mITT 
analysis cohort, excluding subjects with major protocol deviations that would 
compromise the assessment of vaccine efficacy.  Subjects who meet any of the criteria 
listed below will be censored from time to event analyses according to the date of first 
occurrence of the criteria, and will be excluded from all other types of analyses.  The 
ATP cohort will exclude subjects who:  a) receive treatment for acute HCV infection; b) 
were enrolled but subsequently found to have been ineligible at enrollment based on a 
criterion that could reasonably be expected to affect vaccine efficacy; c) did not receive 
both doses of vaccine or control; d) acquired HCV infection prior to receipt of the 
second vaccination; e) received the wrong (non-randomized) product at either dose; f) 
received the second dose fewer than 42 days or more than 70 days after the first dose; 
g) received an immunosuppressant other than inhaled or topical steroids; h) were 
immunized against another pathogen or received immunoglobulins or other blood 
products within 14 days of either dose of study vaccine; i) have autoimmune disease; 
and j) have a confirmed or suspected immunosuppressive or immunodeficient state.  
Membership in the ATP efficacy cohort will be assessed by a centralized clinical review 
prior to unblinding of the study data. 
11.4.2 Safety Analyses 
In safety analyses, if a subject receives the wrong (non-randomized) study product at 
either dose, he/she will be included in the vaccine group if they received one or more 
doses of vaccine and in the placebo group otherwise. 
 
Solicited AEs will be analyzed by taking the most severe response over the follow-up 
period, dichotomizing into a binary variable (none or mild versus moderate or severe) 
and using standard techniques, such as exact confidence intervals, to summarize the 

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reactogenicity rates.  Analyses will be conducted separately for each dose, and both 
doses will be compared to see if a reaction to the first dose is predictive of a reaction to 
the second dose. 
 
Results of safety laboratories assessed prior to and following each vaccination will be 
summarized by treatment group and grade.  Changes in laboratory parameters pre- and 
post-vaccination will also be examined.  Standard summary statistics, including 95% 
confidence intervals will be computed.  Additionally, graded results will either be 
analyzed as a binary variable (normal, abnormal) or multinomial variable (Grade 0, 
Grade 1, Grade 2, Grade 3 and above) using either logistic regression or ordinal logistic 
regression respectively to make treatment group comparisons. 
 
Unsolicited AEs will be coded by MedDRA® for preferred term and system organ class.  
The rate and exact 95% confidence intervals of AEs in aggregate, as well as by 
MedDRA® categories, will be computed.  The number of SAEs is likely to be small in 
this study and will be reported by a detailed listing showing the type, MedDRA® coding, 
relevant dates (vaccinations and AEs), severity, relatedness, and outcome for each 
event. 
 
Local Laboratory Values 
Analyses of local laboratory values may include boxplots or scatterplots.  These 
analyses will be conducted for baseline values and for values measured during the 
course of the study. 
11.4.3 Evaluation of the Primary Efficacy Endpoint 
Reduction in the incidence of chronic HCV infection associated with AdCh3NSmut1 and 
MVA-NSmut HCV vaccines. 
HCV RNA assays will be used to classify chronic vs. cleared infection, using the 
definition described in Section 3.1.2 (page 19).  Based on previous analysis performed 
in IDU cohorts in Baltimore and San Francisco, we expect very few cases of re-infection 
to occur during the trial time.29,30  Blood samples from individuals who demonstrate 
alternating positive and negative HCV RNA results, suggestive of reinfection will be 
analyzed to determine infection status using genotyping tests and viral sequencing 
where necessary to confirm infection outcome.  A small number of cases may occur in 
which individuals will clear HCV viremia after 6 months of infection.  All possible cases, 
including cases with uncertain classification for chronicity, will be reviewed by a blinded 
independent case review committee to make a determination of infection outcome. 
 
The primary efficacy analysis will be performed in the ATP cohort, and repeated in the 
mITT cohort, as defined in Section 11.4.1 above.  A time to event analysis will be 
performed as follows.  For subjects in the mITT (ATP) cohort without acute HCV 

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infection, the 6-month chronic infection endpoint will be coded as censored, with time to 
event defined as the elapsed time from the first (second) vaccination, to six months post 
completion of follow-up for acute HCV infection (26 months), or to the first occurrence of 
loss to follow-up or of a criterion listed under the censoring criteria for the mITT (ATP) 
cohort.  For subjects in the mITT (ATP) cohort with acute HCV infection, the 6-month 
chronic infection endpoint will be coded as an observed event if the 6-month chronic 
infection endpoint definition is met, and as censored otherwise, with time to event 
defined as the elapsed time from the first (second) vaccination to the date of the sample 
collection with available test result nearest to the target assessment date of six months 
after primary acute infection.  Time to event and censoring status for 9-month chronic 
infection are defined analogously.  The reduction in chronic HCV incidence will be 
calculated from the hazard ratio of a Cox model stratified by gender and IL28B status fit 
to the time to event data which includes a binary indicator for membership in the 
vaccine as compared to the placebo arm.  A sensitivity analysis will be performed, in 
which subjects with acute infection who do not go on to develop chronic infection will be 
treated as experiencing a competing risk in the Fine-Gray proportional hazards 
regression model. 
11.4.4 Immunogenicity Analyses 
The primary assay to monitor vaccine immunogenicity is the IFN-γ ELISpot, which will 
be performed at time points indicated in the study visit flow chart in Appendix C.  A 
positive immune response is defined by two criteria: i) more than 48 spot forming cells 
per million PBMC; ii) at least three times the mean background spots per million PBMC 
found in ELISpot wells containing cells and peptide diluent (DMSO).39,87,88  An individual 
will be considered a responder if a positive response to at least one in 6 mixtures 
(pools) of peptides is detected.  Additional immunological parameters will be evaluated.  
These parameters include:  (1) the strength of the total anti-NS immune response 
defined as the sum of positive responses (total SFC/million PBMC) against pools; and 
(2) the breadth of the immune response defined as the number of individual pools 
recognized by each positive subjects.  The peptides recognized in the matrix will be 
then identified and tested individually in duplicate to confirm recognition and to measure 
the number of SFC produced.  If additional cells remain, T-cells specific for antigen will 
be assessed for the ability to secrete other cytokines via intracellular cytokine staining 
and for the ability to respond to peptides spanning the NS region from other HCV 
genotypes by IFN-γ ELISpot and potentially other tests if cells permit.  All immunological 
parameters listed will be correlated with the presence of HCV infection and HCV 
persistent infection.  Both cross-sectional and longitudinal analyses of this data will be 
performed.  Analyses will consider the geometric mean level of response, and the 
proportion of responders.  The analysis will be primarily descriptive, with tables and 
graphs displaying the results by treatment arm.  Additionally, the null hypotheses of no 
difference in peak immune response, and of no difference in proportion responding, in 

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the vaccine versus placebo arms will be tested against two-sided alternatives at the p = 
0.05 level.  Analysis of immunogenicity data collected will be performed at the first 
interim analysis and the end of the study. 
11.4.5 Evaluation of Exploratory Efficacy Endpoints 
1. Reduction in incidence of primary HCV infection associated with AdCh3NSmut1 and 
MVA-NSmut HCV vaccines 
HCV RNA and and/or anti-HCV will be used to classify incident infection, using the 
definition described in Section 3.3 (page 20).  The observed cumulative incidence of 
primary HCV infection in the IDU populations participating to the study has been 
observed at 27%/100 PYO30.  Analyses will be performed in both ATP and mITT 
populations.  The analyses will be based on comparisons of incidence of HCV infection 
between study groups using a log-rank test for time to midpoint in both ATP and mITT 
populations.  A sensitivity analysis will also be performed using interval censoring 
methods. 
2. Reduction of HCV chronic infection with a 9 month chronic cut-off associated with 
AdCh3NSmut1 and MVA-NSmut HCV vaccines 
Previous data have shown that acute HCV infection in individuals followed for two years 
is cleared in 86% of cases by 6 months, but in a small number, clearance occurs after 6 
months (95% at 12 months30).  We will assess the reduction in incidence of chronic 
infection based on 9 months follow up after incident infection.  HCV RNA assays will be 
used to classify chronic infections at 9 months using the definition described in Section 
3.3 (page 20).  Analysis will be performed in both ATP and mITT populations. 
3. Reduction of peak concentration (magnitude) of HCV RNA associated with 
AdCh3NSmut1 and MVA-NSmut HCV vaccines 
Quantitative measures of HCV RNA concentration will be compared between 
vaccinated and controls.  We expect to observe a reduction of 3 log values of the 
maximum concentration of RNA detected in the blood in vaccinated subjects who 
develop HCV viremia (similar to that observed in subjects with HCV reinfection29), 
compared to the placebo group.  Quantitative viremia measures, including the initial 
peak and total HCV RNA levels (described in Section 3.3 page 20) be compared, 
between vaccinated and placebo arms, using logarithmic IU/mL values, by Kruskal-
Wallis test, and a non-parametric equality of median tests.  Analysis will performed in 
both on ATP, and mITT populations. 
4. Reduction in the duration of HCV viremia following incidence HCV infection 
associated with AdCh3NSmut and MVA-NSmut HCV vaccines 
The duration of acute infection is expected to decrease 4-fold in vaccinated subjects who 
develop viremia (similar to that observed in subjects with HCV reinfection29), compared 
to the placebo group, after 6 months of observation following initial infection.  The midpoint 

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between sample collection dates will be used to determine duration as previously 
described (Section 3.3 page 21).  Time to event analyses using interval based censoring 
will be performed in both ATP and mITT populations. 
5. Reduction in the incidence of chronic HCV infection with genotype 1, compared to 
other HCV genotypes associated with AdCh3NSmut and MVA-NSmut HCV vaccines 
Because the vaccine is developed from HCV genotype 1b sequences, we hypothesize 
that we may see greater reduction in chronic infection with HCV genotype 1, compared 
to other genotypes as a result of stronger genotype specific immune responses:  75% 
reduction in chronic infection with genotype 1 compared to 45% reduction in chronic 
infection with non-1 genotypes.  These exploratory analyses will be based on 
comparisons of incidence of HCV genotype 1 infection and non-1 genotypes between 
study groups using methods for censored survival regression (Cox proportional 
hazards). 

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12 Data Handling/Record Keeping/Source Documents 
The principal investigator at each site is responsible for ensuring the accuracy, 
completeness, legibility, and timeliness of the data reported. 
 
Forms for use as source documents will be derived from the electronic case report 
forms (eCRFs) and provided by the SDCC to the sites to record and maintain data for 
each subject enrolled in the study.  All source documents should be completed in a 
neat, legible manner to ensure accurate interpretation of data.  Black or blue ink is 
required to ensure clarity of reproduced copies.  When making a change or correction, 
the original entry should be crossed out with a single line, and the change should be 
initialed and dated.  Do not erase, overwrite, or use correction fluid or tape on the 
original. 
 
Data reported in the eCRF should be consistent with the source documents or the 
discrepancies should be documented. 
 
The sponsor will provide guidance to investigators on making corrections to the source 
documents and eCRFs. 
12.1 Source Documents and Access to Source Documents 
Each participating site will maintain appropriate medical and research records for this 
trial, in compliance with ICH E6, Section 4.9 and regulatory and institutional 
requirements for the protection of confidentiality of subjects.  Each site will permit 
authorized representatives of the DMID, its designees, and appropriate regulatory 
agencies to examine (and when required by applicable law, to copy) clinical records for 
the purposes of quality assurance reviews, audits, and evaluation of the study safety 
and progress.  These representatives will be permitted access to all source data, which 
include, but are not limited to, hospital records, clinical and office charts, laboratory 
notes, memoranda, evaluation checklists, pharmacy dispensing records, recorded data 
from automated instruments, copies or transcriptions certified after verification as being 
accurate and complete, microfiches, photographic negatives, microfilm or magnetic 
media, x-rays, and subject files and records kept at the pharmacy, at the laboratories, 
and medico-technical departments involved in the clinical trial.  Forms for use as source 
documents will be derived from the eCRFs and be provided by the Statistical and Data 
Coordinating Center (SDCC). 
12.2 Data Management Responsibilities 
Data collection is the responsibility of the clinical trial staff at the site under the 
supervision of the site principal investigator.  During the study, the investigator must 

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maintain complete and accurate documentation for the study.  All source documents 
and laboratory reports must be reviewed by the clinical team and data entry staff, who 
will ensure that they are accurate and complete.  Adverse events must be graded, 
assessed for severity and causality, and reviewed by the site principal investigator or 
designee.   
 
The Emmes Corporation will serve as the Statistical and Data Coordinating Center for 
this study, and will be responsible for data management, quality review, analysis, and 
reporting of the study data. 
12.3 Data Capture Methods 
Clinical data (including AEs, concomitant medications, and reactogenicity data) will be 
entered into a 21 CFR Part 11-compliant internet data entry system provided by The 
Emmes Corporation.  The data system includes password protection and internal quality 
checks, such as automatic range checks, to identify data that appear inconsistent, 
incomplete, or inaccurate.  Clinical data will be entered directly from the source 
documents. 
12.4 Types of Data 
Data for this study will include safety, laboratory, and outcome measures (e.g., 
reactogenicity, immunogenicity, infection status). 
12.5 Timing/Reports 
Safety data will be provided to the DSMB and investigators as per the DSMB charter.  
Immunogenicity reports will be provided to the investigators following preliminary 
analysis by the SDCC. 
 
A final clinical study report will be prepared following the last subject visit and upon 
completion of assays related to the immunogenicity endpoints. 
12.6 Study Records Retention 
Records and documents pertaining to the conduct of this study, including CRFs, source 
documents, ICFs, laboratory test results, and medication inventory records, must be 
retained in a secure storage location by the investigator for at least 2 years after a 
marketing application is approved for the drug; or, if an application is not approved for 
the drug, until 2 years after shipment and delivery of the drug for investigational use is 
discontinued and FDA has been so notified.   No study records will be destroyed without 
prior authorization from NIAID. 
 

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13 Quality Control and Quality Assurance 
Following a written DMID-accepted site quality management plan, the investigational 
site is responsible for conducting routine, quality assurance (QA) and quality control 
(QC) activities to internally monitor study progress and protocol compliance.  The 
Principal Investigator(s) will provide direct access to all trial-related sites, source 
data/documents and reports for the purpose of monitoring and auditing by the sponsor, 
and inspection by local and regulatory authorities.  The Principal Investigator will ensure 
all study personnel are appropriately trained and applicable documentations are 
maintained on site. 
 
The Statistical and Data Coordinating Center (SDCC) will implement quality control 
procedures beginning with the data entry system and generate data quality control 
checks that will be run on the database.  Any missing data or data anomalies will be 
communicated to the site(s) for clarification and resolution. 
 
 

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14 Ethics/Protection of Human Subjects 
14.1 Ethical Standard/Declaration of Helsinki 
The investigator(s) will ensure that this study is conducted in full conformity with 
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 50 and 56, and ICH E6; 62 Federal Regulations 25691 (1997), if 
applicable.  The investigator’s Institution will hold a current Federalwide Assurance 
(FWA) issued by OHRP for federally funded research. 
14.2 Institutional Review Board 
Each participating institution will provide for the review and approval of this protocol and 
the associated informed consent documents, by an appropriate ethics review committee 
or Institutional Review Board (IRB) listed on their Federal Wide Assurance.  Any 
amendments to the protocol or informed consent materials must also be approved 
before they are placed into use unless change is for the safety of the subject.  Only 
those IRB members who are independent of the investigators and the sponsor should 
provide an opinion on study related matters.  Verification of IRB approval of the protocol 
and the written informed consent will be transmitted by the investigator or designee prior 
to the shipment of vaccine.  No deviations from or changes to the protocol will be 
initiated without prior approval of an appropriate amendment unless change is for the 
safety of the subject. 
14.3 Informed Consent Process  
The written informed consent document will embody the elements of informed consent 
as described in the Declaration of Helsinki and will adhere to the ICH Harmonised 
Tripartite Guideline for Good Clinical Practice.  Informed consent should be 
implemented before any protocol-specified procedures or interventions are carried out.  
Informed consent will be obtained in accordance with 21 CFR 50.25 and 45 CFR 46.  
Information should be presented both orally and in written form. 
 
All subjects will sign and date the informed consent form before any study specific 
procedures are performed.  An information sheet will also be made available to all 
potential subjects prior to screening.  At the screening visit, the subject will be fully 
informed of all aspects of the trial, the potential risks and their obligations.  The following 
general principles will be emphasized: 
 
• Participation in the study is entirely voluntary 

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• Refusal to participate involves no penalty or loss of medical benefits 
• The subject may withdraw from the study at any time 
• The subject is free to ask questions at any time to allow him or her to understand 
the purpose of the study and the procedures involved 
• The study involves research of an investigational vaccine 
• There is no direct benefit for participating 
• The subject’s general practitioner (GP) will be contacted to corroborate their 
medical history 
 
The aims of the study and all tests to be carried out will be explained.  The subject will 
be given the opportunity to ask about details of the trial, and will then have time to 
consider whether or not to participate.  If they do decide to participate, they will sign and 
date two copies of the ICF, one for them to take away and keep, and one to be kept by 
the investigator.  These forms will also be signed and dated by the study staff person 
obtaining the informed consent. 
 
An investigator or designee will describe the protocol to potential subjects face-to-face.  
The ICF may be read to the subjects, but, in any event, the investigator shall give the 
subjects ample opportunity to inquire about details of the study and ask any questions 
before the signing and dating the ICF. 
 
Study staff must inform subjects that the trial involves research, and explain the purpose 
of the trial, those aspects of the trial that are experimental, any expected benefits, all 
possible risks (including a statement that the particular treatment or procedure may 
involve risks to the subject or to the embryo or fetus, if the subject is or may become 
pregnant, that are currently unforeseeable), the expected duration of the subject’s 
participation in the trial, the procedures of the research study, including all invasive 
procedures, and the probability for random assignment to treatment groups.  Subjects 
will be informed that they will be notified in a timely manner if information becomes 
available that may be relevant to their willingness to continue participation in the trial.  
They must also be informed of alternative procedures that may be available, and the 
important potential benefits and risks of these available alternative procedures.  
Subjects must receive an explanation as to whether any compensation and any medical 
treatments are available if injury occurs, and, if so, what they consist of, or where further 
information may be obtained.  Subjects must be informed of the anticipated financial 
expenses, if any, to the subject for participating in the trial, as well as any anticipated 
prorated payments, if any, to the subject for participating in the trial.  They must be 
informed of whom to contact (e.g., the investigator) for answers to any questions 
relating to the research project.  Information will also include the foreseeable 

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circumstances and/or reasons under which the subject’s participation in the trial may be 
terminated.  The subjects must be informed that participation is voluntary and that they 
are free to withdraw from the study for any reason at any time without penalty or loss of 
benefits to which the subject is otherwise entitled. 
 
Neither the investigator, nor the trial staff, should coerce or unduly influence a subject to 
participate or continue to participate in the trial.  The extent of the confidentiality of the 
subjects’ records must be defined, and subjects must be informed that applicable data 
protection legislation will be followed.  Subjects must be informed that the monitor(s), 
auditors(s), IRB, NIAID, and regulatory authority(ies) will be granted direct access to the 
subject’s original medical records for verification of clinical trial procedures and/or data 
without violating the confidentiality of the subject, to the extent permitted by the 
applicable laws and regulations, and that, by signing a written ICF, the subject is 
authorizing such access.  Subjects must be informed that records identifying the subject 
will be kept confidential, and, to the extent permitted by the applicable laws and/or 
regulations, will not be made publicly available and, if the results of the trial are 
published, the subject’s identity will remain confidential. 
 
Informed consent forms must be in a language fully comprehensible to the prospective 
subjects.  Comprehension of informed consent will be verified using a brief 
comprehension tool.  Informed consent shall be documented by the use of a written ICF 
approved by the IRB and signed and dated by the subject and the person who 
conducted the informed consent discussion.  The signature confirms that the consent is 
based on information that has been provided and all questions have been answered to 
the prospective subject’s satisfaction.  Each subject’s signed ICF must be kept on file by 
the investigator for possible inspection by Regulatory Authorities and/or the sponsor and 
Regulatory Compliance persons.  The subject should receive a copy of the signed and 
dated written ICF and any other written information provided to the subjects, and should 
receive copies of any signed and dated ICF updates and any amendments to the written 
information provided to subjects. 
14.4 Subject Confidentiality 
Subject confidentiality is strictly held in trust by the participating investigators, their staff, 
and the sponsor(s) and their agents.  This confidentiality includes documentation, 
investigation data, subject’s clinical information, and all other information generated 
during participation in the study.  This material may be shown to other professional staff, 
used for educational purposes, or included in a scientific publication. 
 
No information concerning the study or the data generated from the study will be 
released to any unauthorized third party without prior written approval of the sponsor 
and the subject. 

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The study monitor or other authorized representatives of the sponsor or governmental 
regulatory agencies 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 subjects in this study.  The clinical study site will 
permit access to such records. 
14.5 PI Responsibility When Subject Withdraws or Is Discontinued 
See Section 5.4 
14.6 Referrals for HCV Treatment and Care 
See Section 8.6. 

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15 Protocol Conduct 
The protocol will be conducted in compliance with federal regulations, the principles of 
GCP and DMID standards, policies and procedures, including the following processes: 
• Protocol registration, activation, and implementation; 
• Informed consent, screening, and enrollment; 
• Clinical and safety assessments; 
• Safety monitoring and reporting; 
• Data collection and documentation; 
• Study follow-up and close-out; 
• Unblinding of staff and subjects; 
• Quality management; 
• Protocol monitoring and compliance; 
• Risk reduction counseling; and 
• Specimen collection, processing, and analysis. 
 
Any policies or procedures that vary from DMID standards or require additional 
instructions will be described in the study Manual of Procedures. 
15.1 Subject Confidentiality  
The investigators will maintain appropriate medical and research records for this trial, in 
compliance with ICH E6 GCP and regulatory and institutional requirements for the 
protection of confidentiality of subjects.  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 principal investigators will obtain a Certificate of 
Confidentiality from NIH to further protect subjects’ privacy. 
15.2 Data Collection, Handling, Source Documents  
Clinical research data will be collected as described in detail in Section 12. 
 
Standard GCP will be followed to ensure accurate, reliable, and consistent data 
collection.  Each participating site will maintain appropriate medical and research 
records for this trial, in compliance with ICH, GCP, regulatory, and institutional 
requirements for the protection of confidentiality of subjects. 

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15.3 Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, Good Clinical 
Practice (GCP), or Manual of Procedures requirements.  The noncompliance may be on 
the part of the subject, the investigator, or the study site staff.  As a result of deviations, 
corrective actions are to be developed by the site and implemented promptly.   
 
These practices are consistent with Good Clinical Practice: 
4.5  
Compliance with Protocol, sections 4.5.1, 4.5.2, and 4.5.3 
5.1  
Quality Assurance and Quality Control, section 5.1.1 
5.20 
Noncompliance, sections 5.20.1 and 5.20.2d 
 
It is the responsibility of the site to use continuous vigilance to identify and report 
deviations within 5 working days of identification of the protocol deviation, or within 5 
working days of the scheduled protocol-required activity.  All deviations must be 
promptly reported to DMID, via The Emmes Corporation’s IDES. 
 
All deviations from the protocol must be addressed in study source documents.  A 
completed copy of the DMID Protocol Deviation (PD) Form must be maintained in the 
Regulatory File.  Protocol deviations must be submitted to the local IRB/IEC per their 
guidelines.  The site PI/study staff is responsible for knowing and adhering to their IRB 
requirements. 
15.4 Recruitment and Retention of Study Subjects 
All subjects in this study will be adult subjects who are part of the cohort maintained by 
the study sites.   
 
Recruitment and retention of young adult IDUs is challenging and requires an 
experienced culturally sensitive operational structure.  The two original sites, Baltimore 
(MD) and San Francisco (CA) have been studying HCV in IDUs for over 10 years each 
and have developed successful accrual and follow up methods.  Both established study 
sites have experience recruiting and retaining IDUs in prospective studies.  An urban 
IDU cohort has been established in Baltimore under the supervision of Dr. Cox to create 
and maintain a cohort of persons at risk for acute HCV infection.  This cohort will be 
continued and expanded.  The infrastructure was originally established during parallel 
research on injection drug use that was funded by CDC and NIDA under the leadership 
of several principal investigators in the Bloomberg School of Public Health.89-93  At the 
San Francisco study site, prospective studies (under the collective name of The UFO 
Study) of young IDUs at risk of HCV and with acute HCV have been ongoing for 10 
years.30,94  The study is directed and supervised by Dr. Page; and currently has over 

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100 high risk HCV negative subjects in active follow up.  The infrastructure is in 
existence, including trained personnel and a community-clinic based research site. Dr. 
Page has recently initiated studies of HCV in young adult users in New Mexico where 
there is a significant HCV epidemic.  The clinical study site and supporting laboratory 
infrastructure for conducting the study is located in Albuquerque, NM, proximal to the 
University of New Mexico Health Sciences Center. 
 
The Co-Principal Investigators are leaders in the field of acute HCV research in the US 
and both have been studying HCV infection in injection drug users and other at-risk 
populations for over 10 years.  They have extensive professional networks to which they 
refer research participants for care, including HCV treatment, vaccinations and primary 
care, drug treatment, mental health, housing, and other health resources.  These 
investigators have extensive experience working with, and in referral and treatment of, 
HCV in injection drug users.  They have working relationships with local providers who 
can evaluate, assess available resources for care, and treat HCV, including acute HCV, 
on a case-by-case basis.  Both investigators are leaders in advocacy for access to HCV 
testing and treatment in injection drug users and other at-risk populations.  In addition, 
Dr. Cox is an expert in the treatment of acute HCV infections and clinically manages 
therapy of HCV in IDUs. 
 
We estimate that 2160 persons will need to be screened to enroll the target study 
population size of 540 (screening: enrollment ratio, 4:1); corresponding to screening 70 
subjects/month and enrolling 17.5/month, corresponding to per site screening of 23 
subjects/month and enrollment of 6/month.  The San Francisco study had an average 
screening of 45/month and enrollment of 25/month during peak operations and 
operating study site three nights per week (screening for and enrolling only anti-HCV 
negative subjects).  Baltimore has had similar accrual numbers.  These estimates are 
based on a previous intervention study conducted with HCV negative IDUs (the DUIT 
Study95).  The vaccine trial will include resources for enhanced outreach to achieve the 
required 90% follow-up rate including: monthly visits, expanded clinic hours, and subject 
reimbursement, facilitated by a subject tracking database to ensure targeted rates. 
 
All procedures for retention are described during informed consent.  Retention is a 
coordinated activity involving several groups within the study team, including POWs, 
subject advocates, physicians, pharmacists, and counselors.  Once a subject enrolls in 
this study, the study site will make every effort to retain him/her in follow-up to minimize 
possible bias associated with loss-to-follow-up.  This study aims to achieve a minimum 
of 90% retention.  Retention procedures and methods will include: 
(1) Thorough explanation of the study visit schedule and procedural requirements 
during the informed consent process and re-emphasis at each study visit, and at 
study termination. 

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(2) Compilation of detailed contact tracing and locator information at the study screening 
visits, and active review and updating of this information at each subsequent visit. 
(3) Use of mapping techniques to establish the location of subject residences and other 
locator venues. 
(4) Use of appropriate and timely visit reminder mechanisms. 
(5) Immediate and multi-targeted follow-up on missed visits. 
(6) Use of trained outreach workers to complete in-person contact with subjects at their 
homes and/or other community locations. 
(7) The study site will have a subject tracking database to facilitate visit scheduling and 
timely identification and follow-up on missed visits.  The study team will generate 
weekly reports on the number and percentage of subjects completing the follow-up 
visits throughout the course of the study.  Members of the Protocol Team will track 
retention rates closely and work with the study site as needed to take any required 
action to address below-target retention rates. 
 
The key role for POWs in retention:  Retention of IDU in prospective studies is 
extremely challenging and at all three sites, significant effort will be made to maximize 
retention.  Every enrolled subject will be assigned to a POW, in accordance to a POW’s 
geographical coverage during fieldwork.  It is estimated that there will be approximately 
50 subjects assigned to each POW.  During enrollment, the counselor will notify the 
subject that he will be assigned a POW and the counselor will give the subject the 
POW’s name.  POWs will verify all the addresses of the subjects.  Throughout the 
study, the recruitment coordinator or POW or their designee will remind his/her subjects 
by e-mail, text short message service (SMS) messaging, or by telephone of their 
upcoming appointments and invite them to any activities hosted at the study sites.  
Retention and follow-up rate will be assessed and reported periodically.  Study 
coordinating meetings will include review and discussion of recruitment and retention, 
with case reviews and problem solving aimed at maximizing both. 
15.5 Study Subject Reimbursement 
Reimbursement of study subjects for attendance at study visits is at the discretion of 
each study site.  Reimbursement should be comparable to the reimbursement offered 
for similar research in the local community, if possible.  The study site is encouraged to 
confer with its local resources (including Community Advisory Boards, and other 
researchers) in deciding appropriate reimbursement. 
 
The study informed consent submitted to the site IRB/IEC will state the plan for 
reimbursement (if any).  The DMID relies upon local IRBs to determine whether the 
proposed plan for reimbursement meets ethical requirements in the local context.  The 

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exact amounts may be modified during the course of the study in consideration of 
changes in costs such as bus fares, exchange rates, child care, or other factors that 
affect the ability of a subject to comply with study visit requirements.  Reviewing IRBs 
must be made aware of the changes in reimbursement before they occur.  Study 
subjects will not be charged for study injections, research clinic visits, research-related 
examinations, or research-related laboratory tests. 
 

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16 Publication Policy  
All manuscripts resulting from this trial will be reviewed by representatives from the 
site(s), DMID and the product owner(s) (Okairos).  Each institution will have thirty days 
to review the publication prior to submission. 
 
Following completion of the study, the co-principal investigators will publish the results 
of this research in a scientific journal.  The NIH Public Access Policy ensures the public 
has access to the published results of NIH funded research.  It requires that all 
investigators submit or have submitted for them an electronic version of their final peer-
reviewed journal manuscripts that arise from NIH funds to the digital archive of the 
National Library of Medicine’s 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 the official date of publication. 
 
Refer to: 
http://publicaccess.nih.gov 
http://grants.nih.gov/grants /guide/notice-files/NOT-OD-08-033.html  
 
The International Committee of Medical Journal Editors (ICMJE) member journals have 
adopted a trials-registration policy as a condition for publication.96  This policy requires 
that all clinical trials be registered in a public trials registry such as ClinicalTrials.gov, 
which is sponsored by the National Library of Medicine (NLM).  Other biomedical 
journals are considering adopting similar policies.  This trial will be registered in NLM in 
accordance with the new NLM requirements under the FDAAA. 
 
 

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17 References 
1. Global surveillance and control of hepatitis C. Report of a WHO Consultation 
organized in collaboration with the Viral Hepatitis Prevention Board, Antwerp, 
Belgium. J Viral Hepat 1999;6:35-47. 
2. Shepard CW, Finelli L, Alter MJ. Global epidemiology of hepatitis C virus infection. 
Lancet Infect Dis 2005;5:558-67. 
3. An international comparative study of HIV prevalence and risk behaviour among 
drug injectors in 13 cities. WHO Collaborative Study Group. Bull Narc 1993;45:19-
46. 
4. Perz JF, Armstrong GL, Farrington LA, Hutin YJ, Bell BP. The contributions of 
hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer 
worldwide. J Hepatol 2006;45:529-38. 
5. Perz JF, Alter MJ. The coming wave of HCV-related liver disease: dilemmas and 
challenges. J Hepatol 2006;44:441-3. 
6. Armstrong GL, Wasley A, Simard EP, McQuillan GM, Kuhnert WL, Alter MJ. The 
prevalence of hepatitis C virus infection in the United States, 1999 through 2002. 
Ann Intern Med 2006;144:705-14. 
7. Centers for Disease Control and Prevention (CDC), Division of Viral Hepatitis - 
Statistics and Surveillance, 2010. Disease burden from hepatitis A, B, and C in the 
United States. U.S. Department of Health and Human Services,   
http://www.cdc.gov/hepatitis/PDFs/disease_burden.pdf;Accessed December 29, 
2010. 
8. Wasley A, Grytdal S, Gallagher K, Surveillance for acute viral hepatitis--United 
States, 2006. MMWR Surveill Summ 2008;57:1-24. 
9. World Health Organization (WHO), Global Alert and Response (GAR), 2002. 
Hepatitis C.  http://www.who.int/csr/disease/hepatitis/Hepc.pdf; Accessed February 
21, 2011. 
10. Bini EJ, Brau N, Currie S, Shen H, Anand BS, Hu KQ, et al.  Prospective multicenter 
study of eligibility for antiviral therapy among 4,084 U.S. veterans with chronic 
hepatitis C virus infection. Am J Gastroenterol 2005;100:1772-9. 
11. Tsui JI, Currie S, Shen H, Bini EJ, Brau N, Wright TL.  Treatment eligibility and 
outcomes in elderly patients with chronic hepatitis C: results from the VA HCV-001 
Study. Dig Dis Sci 2008;53:809-14. 
12. Rifai MA, Moles JK, Lehman LP, Van der Linden BJ. Hepatitis C screening and 
treatment outcomes in patients with substance use/dependence disorders. 
Psychosomatics 2006;47:112-21. 

DMID Protocol 10-0069  
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13. Hagan H, Latka MH, Campbell JV, Golub ET, Garfein RS, Thomas DA, et al. 
Eligibility for treatment of hepatitis C virus infection among young injection drug 
users in 3 US cities. Clin Infect Dis 2006;42:669-72. 
14. Feld JJ, Hoofnagle JH. Mechanism of action of interferon and ribavirin in treatment 
of hepatitis C. Nature 2005;436:967-72. 
15. Chevaliez S, Pawlotsky JM. Hepatitis C virus: virology, diagnosis and management 
of antiviral therapy. World J Gastroenterol 2007;13:2461-6. 
16. Poordad F, McCone J, Jr., Bacon BR, Bruno S, Manns MP, Sulkowski MS, et al. 
Boceprevir for untreated chronic HCV genotype 1 infection. N Engl J Med 
2011;364:1195-206. 
17. McHutchison JG, Manns MP, Muir AJ, Terrault NA, Jacobson IM, Afdhal NH, et al. 
Telaprevir for previously treated chronic HCV infection. N Engl J Med 
2010;362:1292-303. 
18. Bacon BR, Gordon SC, Lawitz E, Marcellin P, Vierling JM, Zeuzem S, et al. 
Boceprevir for previously treated chronic HCV genotype 1 infection. N Engl J Med 
2011;364:1207-17. 
19. Pawlotsky JM, Chevaliez S, McHutchison JG. The hepatitis C virus life cycle as a 
target for new antiviral therapies. Gastroenterology 2007;132:1979-98. 
20. Falck-Ytter Y, Kale H, Mullen KD, Sarbah SA, Sorescu L, McCullough AJ. 
Surprisingly small effect of antiviral treatment in patients with hepatitis C. Ann Intern 
Med 2002;136:288-92. 
21. Lanford RE, Bigger C, Bassett S, Klimpel G. The chimpanzee model of hepatitis C 
virus infections. ILAR J 2001;42:117-26. 
22. Bassett SE, Guerra B, Brasky K, Miskovsky E, Houghton M, Klimpel GR, et al. 
Protective immune response to hepatitis C virus in chimpanzees rechallenged 
following clearance of primary infection. Hepatology 2001;33:1479-87. 
23. Prince AM, Brotman B, Lee DH, Pfahler W, Tricoche N, Andrus L, et al. Protection 
against chronic hepatitis C virus infection after rechallenge with homologous, but not 
heterologous, genotypes in a chimpanzee model. J Infect Dis 2005;192:1701-9. 
24. Lanford RE, Guerra B, Chavez D, Bigger C, Brasky KM, Wang XH, et al. Cross-
genotype immunity to hepatitis C virus. J Virol 2004;78:1575-81. 
25. Folgori A, Capone S, Ruggeri L, Meola A, Sporeno E, Ercole BB, et al. A T-cell HCV 
vaccine eliciting effective immunity against heterologous virus challenge in 
chimpanzees. Nat Med 2006;12:190-7. 
26. Spada E, Mele A, Berton A, Ruggeri L, Ferrigno L, Garbuglia AR, et al. Multispecific 
T cell response and negative HCV RNA tests during acute HCV infection are early 
prognostic factors of spontaneous clearance. Gut 2004;53:1673-81. 
27. Elliot LN, Lloyd AR, Ziegler JB, Ffrench RA. Protective immunity against hepatitis C 
virus infection. Immunol Cell Biol 2006;84:239-49. 

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 6 October 2015 
 
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28. Folgori A, Spada E, Pezzanera M, Ruggeri L, Mele A, Garbuglia AR, et al. Early 
impairment of hepatitis C virus specific T cell proliferation during acute infection 
leads to failure of viral clearance. Gut 2006;55:1012-9. 
29. Osburn WO, Fisher BE, Dowd KA, Urban G, Liu L, Ray SC, et al. Spontaneous 
Control of Primary Hepatitis C Virus Infection and Immunity Against Persistent 
Reinfection. Gastroenterology 2010;138:315-24. 
30. Page K, Hahn JA, Evans J, Shiboski S, Lum P, Delwart E, et al. Acute hepatitis C 
virus infection in young adult injection drug users: a prospective study of incident 
infection, resolution, and reinfection. J Infect Dis 2009;200:1216-26. 
31. Hagan H, Pouget ER, Des Jarlais DC, Lelutiu-Weinberger C. Meta-regression of 
hepatitis C virus infection in relation to time since onset of illicit drug injection: the 
influence of time and place. Am J Epidemiol 2008;168:1099-109. 
32. Page-Shafer K, Hahn JA, Lum PJ. Preventing hepatitis C virus infection in injection 
drug users: risk reduction is not enough. AIDS 2007;21:1967-9. 
33. Edlin BR. Five million Americans infected with the hepatitis C virus: a corrected 
estimate. Oral presentation at the 56th Annual Meeting of the American Association 
for the Study of Liver Diseases, (AASLD). San Francisco: Hepatology 2005;42(4 
Suppl 1):213. 
34. Hahn JA, Wylie D, Dill J, Sanchez MS, Lloyd-Smith JO, Page-Shafer K, et al. 
Potential impact of vaccination on the hepatitis C virus epidemic in injecting drug 
users. Epidemics 2009;1:47-57. 
35. Simmonds P. The origin and evolution of hepatitis viruses in humans. J Gen Virol 
2001;82:693-712. 
36. Simmonds P, Bukh J, Combet C, Deleage G, Enomoto N, Feinstone S, et al. 
Consensus proposals for a unified system of nomenclature of hepatitis C virus 
genotypes. Hepatology 2005;42:962-73. 
37. Houghton M, Abrignani S. Prospects for a vaccine against the hepatitis C virus. 
Nature 2005;436:961-6. 
38. Yusim K, Richardson R, Tao N, Dalwani A,  Agrawal A, Szinger J, et al. Los alamos 
hepatitis C immunology database. Appl Bioinformatics 2005;4:217-25. 
39. Lauer GM, Ouchi K, Chung RT, Nguyen TN, Day CL, Purkis DR, et al. 
Comprehensive analysis of CD8(+)-T-cell responses against hepatitis C virus 
reveals multiple unpredicted specificities. J Virol 2002;76:6104-13. 
40. Bowen DG, Walker CM. Adaptive immune responses in acute and chronic hepatitis 
C virus infection. Nature 2005;436:946-52. 
41. Al-Sherbiny M, Osman A, Mohamed N, Shata MT, Abdel-Aziz F, Abdel-Hamid M, et 
al. Exposure to hepatitis C virus induces cellular immune responses without 
detectable viremia or seroconversion. Am J Trop Med Hyg 2005;73:44-9. 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
94 
42. Capone S, Meola A, Ercole BB, Vitelli A, Pezzanera M, Ruggeri L, et al. A novel 
adenovirus type 6 (Ad6)-based hepatitis C virus vector that overcomes preexisting 
anti-ad5 immunity and induces potent and broad cellular immune responses in 
rhesus macaques. J Virol 2006;80:1688-99. 
43. Capone S, Zampaglione I, Vitelli A, Pezzanera M, Kierstead L, Burns J, et al. 
Modulation of the immune response induced by gene electrotransfer of a hepatitis C 
virus DNA vaccine in nonhuman primates. J Immunol 2006;177:7462-71. 
44. Fattori E, Zampaglione I, Arcuri M, Meola A, Ercole BB, Cirillo A, et al. Efficient 
immunization of rhesus macaques with an HCV candidate vaccine by heterologous 
priming-boosting with novel adenoviral vectors based on different serotypes. Gene 
Ther 2006;13:1088-96. 
45. Bett AJ, Haddara W, Prevec L, Graham FL. An efficient and flexible system for 
construction of adenovirus vectors with insertions or deletions in early regions 1 and 
3. Proc Natl Acad Sci U S A 1994;91:8802-6. 
46. Casimiro DR, Chen L, Fu TM, Evans RK, Caulfield MJ, Davies ME, et al. 
Comparative immunogenicity in rhesus monkeys of DNA plasmid, recombinant 
vaccinia virus, and replication-defective adenovirus vectors expressing a human 
immunodeficiency virus type 1 gag gene. J Virol 2003;77:6305-13. 
47. Casimiro DR, Bett AJ, Fu TM, Davies ME, Tang A, Wilson KA, et al. Heterologous 
human immunodeficiency virus type 1 priming-boosting immunization strategies 
involving replication-defective adenovirus and poxvirus vaccine vectors. J Virol 
2004;78:11434-8. 
48. Duerr A, Wasserheit JN, Corey L. HIV vaccines: new frontiers in vaccine 
development. Clin Infect Dis 2006;43:500-11. 
49. Ali L. Safety and immunogenicity of the MRK Adenovirus type 5 gag/pol/nef HIV-1 
trivalent vaccine in healthy adults, Merck protocol 016. 
http://www.hvtn.org/ppt/oct06/RobertsonMerckPhaseITrivalent.ppt. October 2006. 
50. Tatsis N, Ertl HC. Adenoviruses as vaccine vectors. Mol Ther 2004;10:616-29. 
51. Farina SF, Gao GP, Xiang ZQ, Rux JJ, Burnett RM, Alvira MR, et al. Replication-
defective vector based on a chimpanzee adenovirus. J Virol 2001;75:11603-13. 
52. Tatsis N, Tesema L, Robinson ER, Giles-Davis W, McCoy K, Gao GP, et al. 
Chimpanzee-origin adenovirus vectors as vaccine carriers. Gene Ther 2006;13:421-
9. 
53. Xue F, Burnett RM. Capsid-like arrays in crystals of chimpanzee adenovirus hexon. 
J Struct Biol 2006;154:217-21. 
54. Meyer H, Sutter G, Mayr A. Mapping of deletions in the genome of the highly 
attenuated vaccinia virus MVA and their influence on virulence. J Gen Virol 1991;72 
(Pt 5):1031-8. 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
95 
55. Antoine G, Scheiflinger F, Dorner F, Falkner FG. The complete genomic sequence 
of the modified vaccinia Ankara strain: comparison with other orthopoxviruses. 
Virology 1998;244:365-96. 
56. Wyatt LS, Earl PL, Eller LA, Moss B. Highly attenuated smallpox vaccine protects 
mice with and without immune deficiencies against pathogenic vaccinia virus 
challenge. Proc Natl Acad Sci U S A 2004;101:4590-5. 
57. Stittelaar KJ, Kuiken T, de Swart RL, van Amerongen G, Vos HW, Niesters HG, et 
al. Safety of modified vaccinia virus Ankara (MVA) in immune-suppressed 
macaques. Vaccine 2001;19:3700-9. 
58. Parrino J, McCurdy LH, Larkin BD, Gordon IJ, Rucker SE, Enama ME, et al. Safety, 
immunogenicity and efficacy of modified vaccinia Ankara (MVA) against Dryvax 
challenge in vaccinia-naive and vaccinia-immune individuals. Vaccine 2007;25:1513-
25. 
59. Vasan S, Schlesinger SJ, Chen Z, Hurley A, Lombardo A, Than S, et al. Phase 1 
safety and immunogenicity evaluation of ADMVA, a multigenic, modified vaccinia 
Ankara-HIV-1 B'/C candidate vaccine. PLoS One 2010;5:e8816. 
60. Moorthy VS, Imoukhuede EB, Keating S, Pinder M, Webster D, Skinner MA, et al. 
Phase 1 evaluation of 3 highly immunogenic prime-boost regimens, including a 12-
month reboosting vaccination, for malaria vaccination in Gambian men. J Infect Dis 
2004;189:2213-9. 
61. Moorthy VS, Imoukhuede EB, Milligan P, Bojang K, Keating S, Kaye P, et al. A 
randomised, double-blind, controlled vaccine efficacy trial of DNA/MVA ME-TRAP 
against malaria infection in Gambian adults. PLoS Med 2004;1:e33. 
62. McShane H, Pathan AA, Sander CR, Keating SM, Gilbert SC, Huygen K, et al. 
Recombinant modified vaccinia virus Ankara expressing antigen 85A boosts BCG-
primed and naturally acquired antimycobacterial immunity in humans. Nat Med 
2004;10:1240-4. 
63. Corona Gutierrez CM, Tinoco A, Navarro T, Contreras ML, Cortes RR, Calzado P, et 
al. Therapeutic vaccination with MVA E2 can eliminate precancerous lesions (CIN 1, 
CIN 2, and CIN 3) associated with infection by oncogenic human papillomavirus. 
Hum Gene Ther 2004;15:421-31. 
64. Smith CL, Dunbar PR, Mirza F, Palmowski MJ, Shepherd D, Gilbert SC, et al. 
Recombinant modified vaccinia Ankara primes functionally activated CTL specific for 
a melanoma tumor antigen epitope in melanoma patients with a high risk of disease 
recurrence. Int J Cancer 2005;113:259-66. 
65. Hirsch VM, Fuerst TR, Sutter G, Carroll MW, Yang LC, Goldstein S, et al. Patterns of 
viral replication correlate with outcome in simian immunodeficiency virus (SIV)-
infected macaques: effect of prior immunization with a trivalent SIV vaccine in 
modified vaccinia virus Ankara. J Virol 1996;70:3741-52. 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
96 
66. Smith CL, Mirza F, Pasquetto V, Tscharke DC, Palmowski MJ, Dunbar PR, et al. 
Immunodominance of poxviral-specific CTL in a human trial of recombinant-modified 
vaccinia Ankara. J Immunol 2005;175:8431-7. 
67. Reyes-Sandoval A, Berthoud T, Alder N, Siani L, Gilbert SC, Nicosia A, et al. Prime-
boost immunization with adenoviral and modified vaccinia virus Ankara vectors 
enhances the durability and polyfunctionality of protective malaria CD8+ T-cell 
responses. Infect Immun 2010;78:145-53. 
68. Wang HB, Kondo A, Yoshida A, Yoshizaki S, Abe S, Bao LL, et al. Partial protection 
against SIV challenge by vaccination of adenovirus and MVA vectors in rhesus 
monkeys. Gene Ther 2010;17:4-13. 
69. Hill AV, Reyes-Sandoval A, O'Hara G, Ewer K, Lawrie A, Goodman A, et al. Prime-
boost vectored malaria vaccines: Progress and prospects. Hum Vaccin 2010;6:78-
83. 
70. Guerra S, Gonzalez JM, Climent N, Reyburn H, Lopez-Fernandez LA, Najera JL, et 
al. Selective induction of host genes by MVA-B, a candidate vaccine against 
HIV/AIDS. J Virol 2010;84:8141-52. 
71. Currier JR, Ngauy V, de Souza MS, Ratto-Kim S, Cox JH, Polonis VR, et al. Phase I 
safety and immunogenicity evaluation of MVA-CMDR, a multigenic, recombinant 
modified vaccinia Ankara-HIV-1 vaccine candidate. PloS One 2010;5:e13983. 
72. Earl PL, Cotter C, Moss B, VanCott T, Currier J, Eller LA  et al. Design and 
evaluation of multi-gene, multi-clade HIV-1 MVA vaccines. Vaccine 2009;27:5885-
95. 
73. Habersetzer F, Zarsky JP, Leroy V, Maynard-Muet M, Bronowicki JP, Feray C, et al. 
A novel vectorized HCV therapeutic vaccine (TG4040): results of a phase 1 study in 
naive patients chronically infected by HCV. J Hepatol 2009;Supplement No. 1; 
50:S18. 
74. Bain C, Honnet G, Veron L. Biological activity of TG4040, a novel HCV therapeutic 
vaccine, in treatment-naive HCV chronic patients: interim results of biomarker and 
immunomonitoring analyses. 16th International Symposium on Hepatitis C Virus & 
Related Viruses. Nice, France; 2009. 
75. Fournillier A, Gerossier E, Evlashev A, Schmitt D, Simon B, Chatel L, et al. An 
accelerated vaccine schedule with a poly-antigenic hepatitis C virus MVA-based 
candidate vaccine induces potent, long lasting and in vivo cross-reactive T cell 
responses. Vaccine 2007;25:7339-53. 
76. O’Hara GA, Duncan CJA, Ewer KJ, Collins KA, Elias SC, Halstead FD, et al. Clinical 
assessment of a recombinant simian adenovirus ChAd63: a potent new vaccine 
vector. J Infect Dis 2012,205:772-781. 
77. Casimiro DR, for HIV Vaccine R&D Team Merck Research Laboratories, 2007. 
Next-Generation HIV Vaccine Candidates: Improved Antigen Coverage and New 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
97 
Adenovirus Vectors, V526-001. HVTN May 2007 Meeting: 
http://www.hvtn.org/ppt/may07/1/HVTN_dcasimiro_01May07.pdf 
78. Catanzaro AT, Koup RA, Roederer M, Bailer RT, Enama ME, Moodie Z, et al. Phase 
1 safety and immunogenicity evaluation of a multiclade HIV-1 candidate vaccine 
delivered by a replication-defective recombinant adenovirus vector. J Infect Dis 
2006;194:1638-49. 
79. Harro CD, Robertson MN, Lally MA, O'Neill LD, Edupuganti S, Goepfert PA, et al. 
Safety and immunogenicity of adenovirus-vectored near-consensus HIV type 1 clade 
B gag vaccines in healthy adults. AIDS Res Hum Retroviruses 2009;25:103-14. 
80. Priddy FH, Brown D, Kublin J, Monahan K, Wright DP, Lalezari J, et al. Safety and 
immunogenicity of a replication-incompetent adenovirus type 5 HIV-1 clade B 
gag/pol/nef vaccine in healthy adults. Clin Infect Dis 2008;46:1769-81. 
81. Duerr A, Huang Y, Buchbinder S, Coombs RW, Sanchez J, del Rio C, et al.  
Extended follow-up confirms early vaccine-enhanced risk of HIV acquisition and 
demonstrates waning effect over time among participants in a randomized trial of 
recombinant adenovirus HIV vaccine (Step Study). J Infect Dis 2012;206:258-66. 
82. Rubinsky AD, Kivlahan DR, Volk RJ, Maynard C, Bradley KA. Estimating risk of 
alcohol dependence using alcohol screening scores. Drug Alcohol Depend 
2010;108:29-36. 
83. Buchbinder SP, Metch B, Holte SE, Scheer S, Coletti A, Vittinghoff E. Determinants 
of enrollment in a preventive HIV vaccine trial: hypothetical versus actual willingness 
and barriers to participation. J Acquir Immune Defic Syndr 2004;36:604-12. 
84. Page K, Osburn W, Evans J, Hahn J, Cox A, Busch MP. Controlling Hepatitis C 
Virus (HCV): Reinfection and Intercalation and Clearance of Viremia in Highly 
Exposed Young Injectors. Proceedings of the 46th Annual Meeting of the European 
Association for the Study of the Liver (EASL). Berlin, Germany; 2011. 
85. Draenert R, Altfeld M, Brander C, Basgoz N, Corcoran C, Wurcel AG, et al. 
Comparison of overlapping peptide sets for detection of antiviral CD8 and CD4 T cell 
responses. J Immunol Methods 2003;275:19-29. 
86. Management of hepatitis C: 2002. NIH Consens State Sci Statements 2002 Jun 10-
12;19:1-46. 
87. Cox AL, Mosbruger T, Lauer GM, Pardoll D, Thomas DL, Ray SC. Comprehensive 
analyses of CD8+ T cell responses during longitudinal study of acute human 
hepatitis C. Hepatology 2005;42:104-12. 
88. Barnes E, Folgori A, Capone S, Swadling L, Aston S, Kurioka A, et al. Novel 
adenovirus-based vaccines induce broad and sustained T cell responses to HCV in 
man. Sci Transl Med 2012;4:115ra1. 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
98 
89. Cox AL, Netski DM, Mosbruger T, Sherman SG, Strathdee S, Ompad D, et al. 
Prospective evaluation of community-acquired acute-phase hepatitis C virus 
infection. Clin Infect Dis 2005;40:951-8. 
90. Galai N, Vlahov D, Bareta JC, Wang C, Cohn S, Sterling TR. Prognostic factors for 
survival differ according to CD4+ cell count among HIV-infected injection drug users: 
pre-HAART and HAART eras. J Acquir Immune Defic Syndr 2005;38:74-81. 
91. Mehta S, Galai N, Astemborski J, Celentano DD, Strathdee SA, Vlahov D, et al. HIV 
incidence among injection drug users in Baltimore, Maryland (1988-2004). J Acquir 
Immune Defic Syndr 2006;43(3):368-72. 
92. Mehta S, Genberg B, Astemborski J, Kavasery R, Kirk GD, Vlahov D, et al. Limited 
uptake of hepatitis C treatment among injection drug users. J Comm Health 
2008;33(3):126-33. 
93. Shah N, Galai N, Celentano D, Vlahov D, Strathdee S. Longitudinal predictors of 
injection cessation and subsequent relapse among a cohort of injection drug users in 
Baltimore, MD, 1988-2000. Drug Alcohol Depend 2005;83(2):147-56. 
94. Hahn JA, Page-Shafer K, Lum PJ, Bourgois P, Stein E, Evans JL, et al. Hepatitis C 
virus seroconversion among young injection drug users: relationships and risks. The 
J Infect Dis 2002;186:1558-64. 
95. Garfein RS, Golub ET, Greenberg AE, Hagan H, Hanson DL, Hudson SM, et al. A 
peer-education intervention to reduce injection risk behaviors for HIV and hepatitis C 
virus infection in young injection drug users. AIDS 2007;21:1923-32. 
96. De Angelis C, Drazen JM, Frizelle FA, Haug C, Hoey J, Horton R, et al. Clinical trial 
registration: a statement from the International Committee of Medical Journal 
Editors. N Engl J Med 2004;351:1250-1. 
 
 

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SUPPLEMENTS/APPENDICES 

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Appendix A:  Acceptable Ranges for Clinical Laboratory Evaluations 
at Screening 
 
NOTE:  When the site-specific laboratory normal range is available, if there is a 
discrepancy between the laboratory value in the Appendix and the site specific 
laboratory normal range, the site normal range will take precedence in eligibility 
determination. 
 
HEMATOLOGY 
 
Acceptable Range 
Hemoglobin F (g/dl) 
11.5-15.0 
Hemoglobin M (g/dl) 
12.5-17.0 
Platelet (K) 
120-499  
WBCs (thou/mcl) 
3.0-10.8 
 
CHEMISTRIES 
 
Acceptable Range 
CreatinineA 
Within or below the normal range 
A  Values lower than the normal range are not exclusionary. 
 
ENZYMES 
 
Acceptable Range 
ALT (SGPT) 
< 1.25 x ULN 
 
URINALYSIS 
 
Acceptable Range 
Glucose (mg/dl) 
≤ 1+ 
Protein (mg/dl) 
≤ 1+ 
 

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Appendix B:  Toxicity Table 
 
ABBREVIATIONS:  Abbreviations utilized in the Table: 
ULN = Upper Limit of Normal 
LLN = Lower Limit of Normal 
Rx = Therapy 
Req = Required 
Mod = Moderate 
IV = Intravenous 
ADL = Activities of Daily Living 
Dec = Decreased 
 
NOTE:  For any of the lab values listed below, if the Grade 1 value conflicts with the 
LLN value, the LLN value takes precedence and the event will not be 
considered a Grade 1 Adverse Event.  
When the site-specific laboratory normal range is available, if there is a 
discrepancy between the laboratory value in the Appendix and the site specific 
laboratory normal range, the site normal range will take precedence in 
determining Grade 1 Adverse Events. 
ESTIMATING SEVERITY GRADE 
For abnormalities NOT found elsewhere in the Toxicity Tables use the scale below to 
estimate grade of severity: 
 
GRADE 1:   
Mild (events require minimal or no treatment and do not interfere with the 
subject’s daily activities.) 
GRADE 2:   
Moderate (events result in a low level of inconvenience or concern with the 
therapeutic measures.  Moderate events may cause some interference with 
functioning) 
GRADE 3:   
Severe (events interrupt a subject’s usual daily activity and may require systemic 
drug therapy or other treatment.  Severe events are usually incapacitating.) 
GRADE 4:   
Life-threatening (Any adverse drug experience that places the subject or 
subject, in the view of the investigator, at immediate risk of death from the 
reaction as it occurred, i.e., it does not include a reaction that had it occurred in a 
more severe form, might have caused death). 
 
SERIOUS OR LIFE-THREATENING AEs 
ANY clinical event deemed by the clinician to be serious or life-threatening should be 
considered a grade 4 event.  Clinical events considered to be serious or life-threatening 
include, but are not limited to:  seizures, coma, tetany, diabetic ketoacidosis, 
disseminated intravascular coagulation, diffuse petechiae, paralysis, acute psychosis, 
severe depression. 
 

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HEMATOLOGY 
 
Grade 1  
Grade 2  
Grade 3  
Grade 4  
Hemoglobin (g/dl) 
Male: 
Female: 
 
11.0-12.4 
9.6-10.8 
 
9.6-10.9 
8.4-9.5 
 
8.3-9.5 
7.2-8.3 
 
<8.3  
<7.2 
Platelets (per cumm) 
84,500-117,000 
65,000-84,499 
25,000-64,999 
< 25,000 
WBCs (thou/mcl) 
Lower Limit 
Upper Limit 
 
2.5-2.9 
11.9-15.1 
 
1.9-2.4 
15.2-21.6 
 
1.0-1.8 
21.7-25.0 
 
<1.0 
>25 
 
CHEMISTRIES 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Hypokalemia 
3.0 - 3.4 mEq/L 
2.5 - 2.9 mEq/L 
2.0 - 2.4 mEq/L  
or intensive 
replacement 
therapy  
or hospitalization 
required 
<2.0 mEq/L  
or abnormal 
potassium with 
paresis, ileus  
or life-threatening 
arrhythmia 
Hyperkalemia 
5.6 - 6.0 mEq/L 
6.1 - 6.5 mEq/L 
6.6 - 7.0 mEq/l 
>7.0 mEq/L  
or abnormal 
potassium  with 
life-threatening 
arrhythmia 
Hyperglycemia 
(nonfasting and no 
prior diabetes) 
NOTE: (normal range for 
Glucose, Random, Serum is 
60-199 mg/dL for one site 
laboratory) 
200 - 274 mg/dL 
275 - 349 mg/dL 
350 - 499 mg/dL 
>500 mg/dL  
or abnormal 
glucose with 
ketoacidosis 
or seizures 
Hypophosphatemia 
2.0 - 2.4 mg/dL 
1.5 -1.9 mg/dL  
or replacement 
Rx required 
1.0 -1.4 mg/dL 
intensive therapy  
or hospitalization 
required 
< 1.0 mg/dL  
or abnormal 
phosphate with 
life-threatening 
arrhythmia 
Hyperbilirubinemia 
(when accompanied by 
any increase in other 
liver function test) 
1.1 - <1.25 x 
ULN 
1.25 - <1.5 x ULN 
1.5 – 1.75 x ULN 
>1.75 x ULN 
Hyperbilirubinemia 
(when other liver 
function are in the 
normal range) 
1.1 - <1.5 x ULN 
1.5 - <2.0 x ULN 
2.0 – 3.0 x ULN 
>3.0 x ULN 
 
 

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CHEMISTRIES cont. 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
BUN 
1.25 - 2.5 x ULN 
2.6 - 5 x ULN 
5.1 - 10 x ULN 
>10 x ULN 
Creatinine* 
1.2 - 1.5 x ULN  
>1.5- 2 x ULN 
>2 x ULN 
Dialysis required 
CPK 
2.0-3.4 x ULN 
3-5.9 x ULN 
6-9.9 x ULN 
>10 x ULN 
* Use age and gender appropriate values 
 
ENZYMES 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
AST (SGOT) 
>1.25 -2.5 x ULN 
>2.5 – 4 x ULN 
>4 -8 x ULN 
>8 x ULN 
ALT (SGPT) 
>1.25 - 2.5 x 
ULN 
>2.5 – 4 x ULN 
>4 -8 x ULN 
> 8 xULN 
LDH 
>1.25 - 2.5 x 
ULN 
>2.5 – 4 x ULN 
>4-8xULN 
> 8 x ULN 
GGT 
1.1 - <2.0 x ULN 
2.0 – <3.0 x ULN 
3.0 – 8.0 x ULN 
>8 x ULN 
Alkaline Phosphatase 
1.1 - <2.0 x ULN 
2.0 – <3.0 x ULN 
3.0 – 8.0 x ULN 
>8 x ULN 
Amylase 
1.1 - 1.5 x ULN 
1.6 - 2.0 x ULN 
2.1 - 5.0 x ULN 
>5.0 x ULN 
Lipase  
1.1 - 1.5 x ULN 
1.6 - 2.0 x ULN 
2.1 - 5.0 x ULN 
>5.0 x ULN 
 
URINALYSIS 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Proteinuria 
1+ or 200 mg – 1 
gm loss/day 
2-3 + or 1-2 gm 
loss/day 
4+ or 2-3.5 gm 
loss/day 
Nephritic 
syndrome or >3.5 
gm loss/day 
Hematuria 
Microscopic only 
<10 rbc/hpf 
Gross, no clots 
>10rbc/hpf 
Gross, with or 
without clots, OR 
red blood cell 
casts 
Obstructive or 
required 
transfusion 
Glucose 
1+ 
2+ 
3+ 
4+ 
 
CARDIOVASCULAR 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Cardiac Rhythm 
Not defined 
asymptomatic, 
transient signs, no 
Rx required 
recurrent/ 
persistent; 
symptomatic Rx 
required 
unstable 
dysrhythmia; 
hospitalization and 
treatment required 
 
 

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CARDIOVASCULAR cont. 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Hypertension 
Asymptomatic 
transient (<24 hrs) 
increase >20 
mmHg in diastolic 
or to >140/90 and 
≤150/95 mmHg; no 
treatment 
recurrent or 
persistent (>24 hrs) 
or symptomatic 
increase >20 mmHg 
in diastolic or to 
>151/96 and 
≤155/100 mmHg; 
monotherapy may be 
indicated 
requiring more 
than one drug or 
more intensive 
therapy than 
previously or to 
>155/100 mmHg 
Life-threatening 
consequences 
(e.g. hypertensive 
crisis) 
Hypotension 
non applicable 
Symptomatic 
corrected with oral 
fluids 
Symptomatic IV 
fluids indicated 
Shock–requiring 
use of 
vasopressors or 
mechanical 
assistance to 
maintain blood 
pressure 
Pericarditis 
minimal effusion 
mild/moderate 
asymptomatic 
effusion, no 
treatment 
symptomatic 
effusion; pain; 
EKG changes 
tamponade; 
pericardiocentesis 
or surgery required 
Hemorrhage, 
Blood Loss 
microscopic/occult 
mild, no transfusion 
gross blood loss; 
1-2 units 
transfused 
massive blood 
loss; >3 units 
transfused 
 
SKIN 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Mucocutaneous 
erythema; 
pruritus 
diffuse, maculo 
papular rash, 
dry 
desquamation 
vesiculation or moist 
desquamation 
or ulceration 
exfoliative dermatitis, 
mucous membrane 
involvement  
or erythema, multiforme 
or suspected Stevens-
Johnson  
or necrosis requiring 
surgery 
Induration 
<25mm  
25 - 50 mm 
>50mm 
 
Erythema 
<25mm  
25 - 50 mm 
>50mm 
 
Edema 
<25mm  
25 - 50 mm 
>50mm 
 
Rash at Injection 
Site 
<25mm  
25 - 50 mm 
>50mm 
 
Regional 
lymphadenopathy 
Soft/spongy 
Firm 
Hard 
 
 
 

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105 
SKIN cont. 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Pruritus 
slight itching at 
injection site 
moderate itching 
at injection 
extremity 
itching over entire 
body 
 
 
SYSTEMIC 
 
Grade 1 
Grade 2  
Grade 3  
Grade 4  
Allergic Reaction 
pruritus without rash 
localized urticaria 
generalized 
urticaria; 
angioedema 
Anaphylaxis 
Fever: oral 
38.0 – 39.0 oC 
100.4 F - 102.2 oF  
>39.0 – 40.0 oC 
102.3F - 104.0 oF 
>40.0 oC 
>104.0 oF 
 
 
MUSCULOSKELETAL 
 
Grade 1  
Grade 2  
Grade 3  
Grade 4  
Arthritis 
mild pain with 
inflammation, 
erythema or joint 
swelling – but not 
interfering with 
function 
moderate pain with 
inflammation, 
erythema  
or joint swelling – 
interfering with 
function, but not 
with activities of 
daily living 
severe pain with 
inflammation, 
erythema  
or joint swelling –
and interfering with 
activities of daily 
living 
permanent 
and/or disabling 
joint destruction 
 
 

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Appendix C:  Study Visit Chart 
Arms A and B 
S 
V 
 
 
 
 
V 
 
 
 
 
 
 
 
Timeline from enrollment 
(months) 
<-1 
0 
0 
0 
0 
1 
2 
2 
2 
3 
4 
5 
6 
7 
Timeline from enrollment 
(days) 
<-30 
0 
3 
7 
14 
30 
56 
59 
63 
90 
120 
150 
180 
210 
Window (days) 
-30 
0 
±2 
±2 
±7 
±7 
±7 
±2C 
+14C 
±7 
±7 
±7 
±7 
±7 
Visit Number 
00 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
Review Inclusion/Exclusion 
criteria 
X 
X 
 
 
 
 
X 
 
 
 
 
 
 
 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Behavioral Risk Assessment 
Questionnaire 
 
X 
 
 
 
 
 
 
 
X 
 
 
X 
 
Medical history 
X 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Examine vaccination site 
 
X 
X 
X 
(X) 
 
X 
X 
X 
 
 
 
 
 
Targeted Physical Examination 
X 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Vital Signs (Temp, Pulse, BP) 
X 
X 
 
 
 
 
X 
 
 
 
 
 
 
 
Height/Weight 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy Test* 
X 
X 
 
 
 
 
X 
 
 
 
 
 
 
 
Counseling on avoidance of HIV, 
HCV, and pregnancy 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Concomitant Medications 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
XB 
XB 
XB 
XB 
Vaccination 
 
X 
 
 
 
 
X 
 
 
 
 
 
 
 
Distribute Memory Aid 
 
X 
 
 
 
 
X 
 
 
 
 
 
 
 
Review Memory Aid 
 
 
X 
X 
 
 
 
X 
X 
 
 
 
 
 
AE/SAE Assessment 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
XA 
XA 
XA 
XA 
Urinalysis 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Hematology (mL) 
9 
9 
 
 
9 
9 
9 
 
9 
9 
9 
 
 
 
Biochemistry (mL) 
14 
14 
 
 
14 
14 
14 
 
14 
14 
14 
 
 
 
IL28B (mL) 
5 
 
 
 
 
 
 
 
 
 
 
 
 
 
Anti-HCV (mL) 
5 
 
 
 
 
 
 
 
 
 
 
 
 
 
HBsAg, anti-HIV (mL) 
5 
 
 
 
 
 
 
 
 
 
 
 
 
 
HCV qualitative viremia  (mL) 
5 
 
 
 
 
5 
5 
 
 
5 
5 
5 
5 
5 
HCV quantitative viremia (mL) 
5 
 
 
 
 
5 
5 
 
 
5 
5 
5 
5 
5 
Anti-adenovirus Ab (mL) 
 
XD 
 
 
 
 
 
 
 
 
 
 
 
 
HLA typing/genetic studies (mL) 
 
8 
 
 
 
 
 
 
 
 
 
 
 
 
Immunology (mL) 
 
160 
 
 
 
160 
100 
 
100 
160 
 
 
 
 
Blood per visit (mL) 
48 
191 
0 
0 
23 
193 
133 
0 
123 
193 
33 
10 
10 
10 
Cumulative blood volume (mL) 
48 
239 
239 
239 
262 
455 
588 
588 
711 
904 
937 
947 
957 
967 
 
 
 

DMID Protocol 10-0069  
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 6 October 2015 
 
 
 
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Arms A and B (continued) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Timeline from enrollment 
(months) 
8 
9 
10 
11 
12 
13 
14 
15 
16 
17 
18 
19 
20 
 
Timeline from enrollment 
(days) 
240 
270 
300 
330 
360 
390 
420 
450 
480 
510 
540 
570 
600 
 
Window (days) 
-±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
±7 
 
Visit Number 
14 
15 
16 
17 
18 
19 
20 
21 
22 
23 
24 
25 
26 
Early 
Termi-
nation 
Review Inclusion/Exclusion 
criteria 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Examine vaccination site 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Targeted Physical 
Examination 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Behavioral Risk 
Assessment Questionnaire 
 
X 
 
 
X 
 
 
X 
 
 
X 
 
X 
X 
Counseling on avoidance of 
HIV, HCV, and pregnancy 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Concomitant Medications 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
XB 
X or 
XB 
AE/SAE Assessment 
XA 
XA  
XA 
XA 
XA 
XA 
XA 
XA 
XA 
XA 
XA 
XA 
XA 
X 
Hematology (mL) 
 
 
9 
 
 
 
 
 
 
 
 
 
9 
9 
Biochemistry (mL) 
 
 
14 
 
 
 
 
 
 
 
 
 
14 
14 
Anti-HCV (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
5 
5 
HBsAg, anti-HIV (mL) 
 
 
 
 
 
 
 
 
 
 
 
 
5 
5 
HCV Qualitative viremia  
(mL) 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
HCV Quantitative viremia 
(mL) 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
Anti-adenovirus Ab (mL) 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
Immunology (mL) 
100 
 
 
 
 
 
 
 
 
 
 
 
100 
100 
Blood per visit (mL) 
110 
10 
33 
10 
20 
10 
10 
10 
10 
10 
10 
10 
143 
143 
Cumulative blood volume 
(mL) 
1077 
1087 
1120 
1130 
1150 
1160 
1170 
1180 
1190 
1200 
1210 
1220 
1363 
 
S, screening visit;  
V, vaccination visit;  
(X), if considered necessary, emphasizing any acute complaint; 
A, only SAE’s collected;  
B, All concomitant medications will be recorded through Day 90 (Day 30 if only receiving 1 vaccination). 
At visits after Day 90 (Day 30 if only receiving 1 vaccination), only immunosuppressants and 
medications for the treatment of HCV infection will be recorded;  
C, windows dependent on Visit 6, Dose 2 
D, Plasma saved from the blood collected for immunology assessments at this visit will be used for anti-
adenovirus Ab testing. 
* Pregnancy test must be performed and documented as negative within 24 hours prior to each 
vaccination.   
 

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
 
 
108 
Appendix D:  Study Visit Chart – Confirmed HCV Infection 
 
HCV
+ test 
 
 
 
 
 
 
 
 
 
 
Timeline from HCV infection 
(months) 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
 
Timeline from first positive 
HCV Quantitative RNA 
result (days) 
0 
30 
60 
90 
120 
150 
180 
210 
240 
270 
 
Window (days) 
 
±7 
±7 
±7 
±7 
±7 
±21 
±7 
±7 
±21 
 
Visit Number 
 
H01 
H02 
H03 
H04 
H05 
H06 
H07 
H08 
H09 
Early 
Termi-
nationG 
Review confirmed HCV 
infection 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Medical history 
 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Examine vaccination site 
 
(X)C 
 
 
 
 
 
 
 
 
 
Targeted Physical 
Examination 
 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
Behavioral Risk Assessment 
Questionnaire 
 
XD 
XD 
XD 
XD 
XD 
XD 
XD 
XD 
XD 
XD 
Concomitant Medications 
 
XE 
XE 
XE 
XE 
XE 
XE 
XE 
XE 
XE 
XE 
AE/SAE Assessment 
 
XF 
XF 
XF 
XF 
XF 
XF 
XF 
XF 
XF 
XF 
Counseling on avoidance of 
HIV, HCV transmission and 
reinfection 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Hematology (mL) 
 
9 
9 
9 
9 
9 
9 
9 
9 
9 
9 
Biochemistry (mL) 
 
14 
14 
14 
14 
14 
14 
14 
14 
14 
14 
HCV genotyping (mL) 
 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
Anti-HCV (mL) 
 
 
 
 
 
 
 
 
 
5 
5 
HBsAg, anti-HIV (mL) 
 
5 
 
 
 
 
 
 
 
5 
 
HCV Quantitative viremia 
(mL) 
 
5 
5 
5 
5 
5 
5 
5 
5 
5 
5 
Immunology (mL) 
 
100 
100 
100 
100 
100 
100 
100 
 
100 
100 
Blood per visit (mL) 
 
138 
133 
133 
133 
133 
133 
133 
33 
143 
138 
Cumulative blood volume 
(mL) post HCV Infection 
 
138 
271 
404 
537 
670 
803 
936 
969 
1112 
 
(X), if considered necessary, emphasizing any acute complaint;  
C, Vaccination site is only to be examined at Day 59/63;  
D, For confirmed HCV infected subjects Behavioral Risk Assessment Questionnaire is to be administered at Day 90, 180, 
270, 360, 450, and 540 (and Early Termination Visit, if conducted) if applicable, as the follow-up period allows; if the 
subject’s follow-up period is extended as a result of confirmed HCV infection, the Behavioral Risk Assessment 
Questionnaire will be administered at Day 630, 720, 810, and the final study visit, as the follow-up period allows. 
E, All concomitant medications will be recorded through Day 90 (Day 30 if only receiving 1 vaccination). At visits after Day 
90 (Day 30 if only receiving 1 vaccination), only immunosuppressants and medications for the treatment of HCV infection 
will be recorded;  
F, AE’s and SAE’s recorded through Day 90. At visits following Day 90, only SAE’s recorded. 
G, Subjects who become HCV-infected prior to the 2nd dose of vaccine and choose to terminate early will follow the full 
schedule of tests for visit H09. 
.

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
 
 
109 
Appendix E:  Alcohol Use Disorders Identification Test (AUDIT) C test 
 
AUDIT-C- Overview 
 
The AUDIT-C is a 3-item alcohol screen that can help identify persons who are hazardous 
drinkers or have active alcohol use disorders (including alcohol abuse or dependence). The 
AUDIT-C is a modified version of the 10 question AUDIT instrument. 
 
Clinical Utility 
The AUDIT-C is a brief alcohol screen that reliably identifies patients who are hazardous 
drinkers or have active alcohol use disorders. 
 
Scoring 
The AUDIT-C is scored on a scale of 0-12. 
Each AUDIT-C question has 5 answer choices. Points allotted are: 
A = 0 points, b = 1 point, c = 2 points, d = 3 points, e = 4 points 
 In men, a score of 4 or more is considered positive, optimal for identifying hazardous 
drinking or active alcohol use disorders. 
 In women, a score of 3 or more is considered positive (same as above). 
 However, when the points are all from Question #1 alone (#2 & #3 are zero), it can be 
assumed that the patient is drinking below recommended limits and it is suggested that the 
provider review the patient’s alcohol intake over the past few months to confirm accuracy.1 
 Generally, the higher the score, the more likely it is that the patient’s drinking is affecting his 
or her safety. 
 
Psychometric Properties 
For identifying patients with heavy/hazardous drinking and/or Active-DSM alcohol abuse or 
dependence: 
 
Men2 
Women3 
>3 
Sens: 0.95 / Spec. 0.60 
Sens: 0.66 / Spec. 0.94 
>4 
Sens: 0.86 / Spec. 0.72 
Sens: 0.48 / Spec. 0.99 
 
For identifying patients with active alcohol abuse or dependence: 
>3 
Sens: 0.90 / Spec. 0.45 
Sens: 0.80 / Spec. 0.87 
>4 
Sens: 0.79 / Spec. 0.56 
Sens: 0.67 / Spec. 0.94 
 
                                                     
1 Frequently Asked Questions guide to using the AUDIT-C can be found via the website: 
http://www.queri.research.va.gov/tools/alcohol-misuse/alcohol-faqs.cfm  
2 Bush K, Kivlahan DR, McDonell MB, et al. The AUDIT Alcohol Consumption Questions (AUDIT-C): An effective brief 
screening test for problem drinking. Arch Internal Med. 1998 (3): 1789-1795. 
3 Bradley KA, Bush KR, Epler AJ, et al. Two brief alcohol-screening tests from the Alcohol Use Disorders Identification Test 
(AUDIT): Validation in a female veterans affairs patient population. Arch Internal Med Vol 163, April 2003: 821-829.  

DMID Protocol 10-0069  
Version 9.0 
 
 6 October 2015 
 
 
 
110 
AUDIT-C Questionnaire 
 
Patient Name_______________________________________ Date of Visit________________ 
Now I’m going to ask you about your use of alcohol beverages during this past year. 
1. How often did you have a drink containing alcohol?   
0 
1 
2 
3 
4 
-8 
-9 
never 
monthly or 
less 
2 to 4 
times/month 
2 to 3 
times/week 
4 or more 
times/week 
Don’t 
Know 
Decline 
2. How many drinks containing alcohol do you have on a typical day when you are 
drinking? 
0 
0 
2 
3 
4 
5 
-8 
-9 
none 
1 or 2 
3 or 4 
5 or 6 
7 to 9 
10 or 
more 
Don’t 
Know 
Decline 
3. How often do you have six or more drinks on one occasion? 
0 
1 
2 
3 
4 
-8 
-9 
never 
less than 
monthly 
monthly 
weekly 
daily or 
almost daily 
Don’t 
Know 
Decline