Dose-Proportionality and Intra-Individual Variability of Intracellular Tenofovir Diphosphate and Emtricitabine Triphosphate In Healthy Volunteers
Phase: PHASE1
Conditions: HIV
Sponsor: HIV Prevention Trials Network

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
DAIDS ID: 10851
A Study of the HIV Prevention Trials Network
Sponsored by:
Division of AIDS, U.S. National Institute of Allergy and Infectious Diseases
U.S. National Institutes of Health
Pharmaceutical Support Provided by:
Gilead Sciences
Protocol Chair(s):
Craig W. Hendrix
Johns Hopkins University
Baltimore, MD, U.S.A
Protocol Co-Chair:
Ken Mayer
Brown University
Boston, MA, U.S.A
Kristine Patterson
University of North Carolina, Chapel Hill 
Chapel Hill, NC, U.S.A
Version 2.0
29 November 2010
Non-IND study

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
TABLE OF CONTENTS
LIST OF ABBREVIATIONS AND ACRONYMS .................................................................... 5
LIST OF TABLES AND FIGURES............................................................................................ 7
TERMINOLOGY FOR TENOFOVIR, EMTRICITABINE, AND THEIR DERIVATIVES
......................................................................................................................................................... 8
PROTOCOL TEAM ROSTER................................................................................................... 9
PROTOCOL SIGNATURE PAGE........................................................................................... 11
SCHEMA..................................................................................................................................... 12
OVERVIEW OF PARTICIPANT DISTRIBUTION AND STUDY DESIGN ..................... 14
1.0
INTRODUCTION........................................................................................................... 16
1.1
Background and Prior Research.................................................................................... 16
1.2
Rationale ....................................................................................................................... 22
2.0
STUDY OBJECTIVES AND DESIGN......................................................................... 25
2.1
Primary Objectives........................................................................................................ 25
2.2
Secondary Objectives.................................................................................................... 25
2.3
Study Design................................................................................................................. 25
3.0
STUDY POPULATION ................................................................................................. 27
3.1
Inclusion Criteria .......................................................................................................... 28
3.2
Exclusion Criteria ......................................................................................................... 29
3.3
Recruitment Process...................................................................................................... 30
3.4
Participant Retention..................................................................................................... 30
3.5
Participant Withdrawal ................................................................................................. 31
4.0
STUDY TREATMENT/PRODUCT/INTERVENTION............................................. 31
4.1
Study Product Formulation/Content/Storage................................................................ 31
4.2
Study Product Regimen(s)............................................................................................ 32
4.3
Study Product Administration....................................................................................... 32
4.4
Product Supply and Accountability .............................................................................. 34
4.5
Toxicity Management................................................................................................... 34
4.6
Management of Specific Toxicities .............................................................................. 36
4.7
Clinical Management of Pregnancy.............................................................................. 41
4.8
Criteria for Early Termination of Study Participation.................................................. 41
4.9
Concomitant Medications ............................................................................................. 42
5.0
STUDY PROCEDURES ................................................................................................ 43
5.1
Screening Visit.............................................................................................................. 44
5.2
Enrollment Visit............................................................................................................ 46

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5.3
Non-Intensive Sampling Visits..................................................................................... 47
5.4
Intensive Sampling Visits ............................................................................................. 48
5.5
Follow up Procedures for Participants who Discontinue Study Product and for 
Participants who Receive a Biopsy on the Last Study Visit..................................................... 48
5.6
Interim Contacts and Visits........................................................................................... 49
5.7
Clinical Evaluations and Procedures ............................................................................ 49
5.8
HIV and STI Counseling .............................................................................................. 49
5.9
Participant Preparation for Sampling............................................................................ 49
6.0
SAFETY MONITORING AND ADVERSE EVENT REPORTING ........................ 50
6.1
Safety Monitoring......................................................................................................... 50
6.2
Clinical Data Safety Review......................................................................................... 51
6.3
Adverse Event Definition and Reporting Requirements .............................................. 52
6.4
Social Harms................................................................................................................. 54
7.0
STATISTICAL CONSIDERATIONS .......................................................................... 55
7.1
Review of Study Design ............................................................................................... 55
7.2
Endpoints ...................................................................................................................... 55
7.3
Accrual, Follow-up, Retention, and Sample Size......................................................... 56
7.4
Study Arm Assignment................................................................................................. 60
7.5
Blinding......................................................................................................................... 60
7.6
Data Analysis................................................................................................................ 60
8.0
HUMAN SUBJECTS CONSIDERATIONS ................................................................ 62
8.1
Ethical Review.............................................................................................................. 62
8.2
Informed Consent.......................................................................................................... 63
8.3
Risks.............................................................................................................................. 63
8.4
Benefits ......................................................................................................................... 66
8.5
Incentives...................................................................................................................... 66
8.6
Confidentiality .............................................................................................................. 66
8.7
Communicable Disease Reporting Requirements ........................................................ 67
8.8
Study Discontinuation................................................................................................... 67
9.0
LABORATORY SPECIMENS AND BIOHAZARD CONTAINMENT .................. 67
9.1
Local Laboratory Specimens ........................................................................................ 67
9.2
Network Laboratory Specimens ................................................................................... 69
9.3
Quality Control and Quality Assurance Procedures..................................................... 71
9.4
Specimen Storage and Possible Future Research Testing ............................................ 72
9.5
Biohazard Containment ................................................................................................ 72
10.0
ADMINISTRATIVE PROCEDURES.......................................................................... 72
10.1
Study Activation ........................................................................................................... 72
10.2
Study Coordination ....................................................................................................... 73
10.3
Study Monitoring.......................................................................................................... 74
10.4
Protocol Compliance..................................................................................................... 74
10.5
Investigator's Records................................................................................................... 75
10.6
Use of Information and Publications ............................................................................ 75

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11.0
REFERENCES................................................................................................................ 76
APPENDICES............................................................................................................................. 78
APPENDIX I: SCHEDULE OF EVALUATIONS AND PROCEDURES............................ 79
APPENDIX IA: Schedule of Evaluations and Procedures - General Procedures................ 80
APPENDIX IB: Schedule of Evaluations and Procedures - Clinical Procedures and 
Laboratory Tests......................................................................................................................... 81
APPENDIX IC: Schedule of Evaluations and Procedures - Sample Collection for PK 
Evaluation (Drug Testing).......................................................................................................... 84
APPENDIX 1D: Additional Procedures from Study Participants with Confirmed HIV 
Infection ....................................................................................................................................... 86
APPENDIX II: SCREENING/ENROLLMENT SAMPLE INFORMED CONSENT FORM 
(Intensive Sampling Cohort)...................................................................................................... 87
APPENDIX III: SCREENING/ENROLLMENT SAMPLE INFORMED CONSENT 
FORM (Non-Intensive Sampling Cohort) .............................................................................. 103

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LIST OF ABBREVIATIONS AND ACRONYMS
AE
adverse event
AIDS
Acquired Immunodeficiency Syndrome
aPTT  
activated partial thromboplastin time
ALT
alanine aminotransferase
AST
aspartate aminotransferase
BV
bacterial vaginosis
Ca
calcium
CBC
complete blood count
CDC
Centers for Disease Control and Prevention
CORE
(HPTN) Coordinating and Operations Center
Cl
chloride
CRPMC
(NIAID) Clinical Research Products Management Center
CT
Chlamydia (Chlamydia trachomatis)
DAIDS
Division of AIDS
DOT
Directly observed therapy
EAE
expedited adverse event (reporting) 
EC
ethics committee
FDA
(United States) Food and Drug Administration
FTC
emtricitabine
FTC-TP
emtricitabine triphosphate
GC
gonorrhea (Neisseria Gonorrhoeae)
GCLP
good clinical laboratory practice
GCP
good clinical practice
HBsAg
hepatitis B surface antigen
HC03
bicarbonate
HIV
Human Immunodeficiency Virus
HPTN
HIV Prevention Trials Network
IDS
investigational drug service
IRB
institutional review board
K
potassium
LDMS
Laboratory Data Management System
LL
local laboratory
LLN
lower limit of normal 
Na
sodium
NAAT
nucleic acid amplification test
NIAID
(United States) National Institute of Allergy and Infectious Diseases
NIH
(United States) National Institutes of Health

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NL
(HPTN) Network Laboratory
NSAID
Non-steroidal anti-inflammatory drug
OHRP
Office for Human Research Protections
PD
pharmacodynamics
PK
pharmacokinetics
PoR
pharmacist of record 
PrEP
pre-exposure prophylaxis
PSRT
Protocol Safety Review Team
PT
prothrombin test
QA
quality assurance
QC
quality control
RPR
rapid plasma reagin test for syphilis 
RTI
reproductive tract infection 
RSC
Regulatory Support Center (formerly RCC)
RUQ
right upper quadrant
SAE
serious adverse event
SDMC
(HPTN) Statistical and Data Management Center
SMC
Study Monitoring Committee
SOP
standard operating procedures
SSP
study specific procedures (manual)
STI
sexually transmitted infection
TDF
tenofovir disoproxil fumarate
TDF/FTC
Truvada® (tenofovir disoproxil fumarate and emtricitabine in fixed dose 
combination) 
TFV
tenofovir
TFV-DP
tenofovir diphosphate
Trich
trichomoniasis (Trichomonas vaginalis)
ULN
upper limit of normal
UTI
urinary tract infection

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LIST OF TABLES AND FIGURES
Figure 1-1.  Simulation of tenofovir diphosphate (TFV-DP) concentration ................................ 18
Figure 1-2. Simulation of tenofovir concentration in blood serum .............................................. 19
Table 1-1. Accumulation index for parent and phosphorylated moieties of TDF and FTC dosed 
either weekly (168 hours) or daily (24 hours). ........................................................... 21
Table 4-1. Study Product Dosing Arms........................................................................................ 32
Table 5-1. Screening Visit (up to 30 days prior to Enrollment Visit)........................................... 45
Table 5-2. Enrollment Visit (Day 0)............................................................................................. 46
Table 5-3. Non-Intensive Sampling Visits (Days 1‡, 3 ‡, 7, 14, 21, 28, 35, and 49) .................... 47
Table 5-4. Intensive Sampling Visit (Days 35 and 49)................................................................. 48
Table 7-1. Power for dose proportionality assessment, 
05
.0
=
................................................ 58
Table 7-2. Power to detect the minimal detectable difference (in fold) for various sample sizes 
and coefficients of variance (CV)............................................................................... 59
Table 7-3. Biological matrices for analyses.................................................................................. 61

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
TERMINOLOGY FOR TENOFOVIR, EMTRICITABINE, AND THEIR 
DERIVATIVES
Abbreviation
Compound 
name
Comments
TDF
Tenofovir 
disoproxil 
fumarate
This is the inactive, oral formulation of tenofovir (trade 
name: Viread®). The ester form enhances oral absorption 
and bioavailability. TDF is rapidly metabolized after dosing 
to the de-esterified pro-drug, tenofovir (TFV), which is also 
inactive. 
TFV
Tenofovir
This is the inactive, de-esterified form of TDF. This form of 
the drug that is measured in serum, blood, other body fluids, 
and tissue samples. 
TFV-DP
Tenofovir 
diphosphate
This is the active, phosphorylated form of tenofovir that is 
generated in cells. This is the form of the drug that is 
measured in cells (e.g., PBMCs). It is rapidly 
dephosphorylated to the inactive form outside of cells, and 
has a very short half-life outside of cells in tissue.
FTC
Emtricitabine
This antiretroviral drug (trade name: Emtriva®) is co-
formulated with TDF in Truvada® (TDF/FTC). FTC is an 
inactive pro-drug that is activated in cells by 
phosphorylation. This is the form of emtricitabine that is 
measured in serum, blood, other body fluids, and tissue 
samples.
FTC-TP
Emtricitabine 
triphosphate
This is the active form of FTC that is generated in cells. 
This is the form measured in cells (e.g., PBMCs). 
Truvada®
Tenofovir plus 
emtricitabine
This is the co-formulated drug produced by Gilead Sciences, 
Inc. Each pill contains 300 mg of TDF and 200 mg of FTC.

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
PROTOCOL TEAM ROSTER
David Borasky
Office of Research Protection 
RTI International 
3040 Cornwallis Rd
RTP, NC 27709 , U.S.A.
Phone:  919-316-3903
Fax:
 919-316-3897
E-mail:  dborasky@rti.org
Ying Chen
SCHARP/FHCRC
1100 Fairview Avenue N
M2-C200
PO Box 19024
Seattle, WA 98109, U.S.A
Email: yqchen@scharp.org  
Phone: 206-667-7051 
Fax: 206-667-4812
Myron Cohen
University of North Carolina 
School of Medicine, 
Department of Medicine, 
Division of Infectious 
Diseases
130 Mason Farm Road
Bioinformatics Bldg.
Chapel Hill, NC 27599, 
U.S.A
Email:
myron_cohen@med.unc.edu
Phone: 919-966-2536 
Fax: 919-966-6714
Vanessa Elharrar
Medical Officer 
Prevention Science 
Program/DAIDS/NIAID/NIH 
6700-B Rockledge Drive, 5th 
Fl. Rm. 5125 
Bethesda, MD 20892, U.S.A. 
Email: 
elharrarva@niaid.nih.gov
Phone: 301-827-0845
Lynda Emel
Senior Project Manager -
Perinatal and Prevention
SCHARP/FHCRC
1100 Fairview Avenue North
LE-400
PO Box 19024
Seattle, WA 09109 U.S.A
Email: lemel@scharp.org
Phone: 206-667-5803
Fax: 206-667-4812
Susan Eshleman
Johns Hopkins University
720 Rutland Avenue
Ross Bldg., Room 646
Baltimore, MD 21205, U.S.A
Email: seshlem@jhmi.edu 
Phone: 410-614-4734 
Fax: 410-502-9244
Alexia Exarchos
Pharmaceutical 
Representative
Gilead Sciences 
333 Lakeside Drive
Foster City, California 94404
U.S.A
Email: 
Alexia.Exarchos@gilead.com 
Phone: 650-522-1201
Fax: 650-522-5418
Fang Gai
Statistical Research Associate
SCHARP/FHCRC
1100 Fairview Ave. N. 
LE-400
P.O. Box 19024
Seattle, WA 98109-1024 
U.S.A
Email: fang@scharp.org
Phone: 206-667-7524
Fax: 206-667-4812
Craig Hendrix
Protocol Chair
Johns Hopkins University
600 North Wolfe Street
Harvey 502
Baltimore, MD 21287, U.S.A
Email: cwhendrix@jhmi.edu 
Phone: 410-955-9707 
Fax: 410-955-9708

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Angela Kashuba
3318 Kerr Hall, 
School of Pharmacy
UNC-Chapel Hill
Chapel Hill NC, 27599, 
U.S.A
Email: akashuba@unc.edu 
Phone: 919-966-9998 
Fax: 919-962-0644
Cheryl Cokley
FHI
2224 E. NC Hwy 54
Durham, NC 27713, U.S.A
Email: ccokley@fhi.org 
Phone: 919-544-7040 ext. 
11359
Fax: 919-544-0207
Ken Mayer
Protocol Co-Chair
Fenway Health
1340 Boylston Street
Boston, Ma, U.S.A
Email:
kenneth_mayer@brown.edu 
Phone: 401-793-4711 
Fax: 401-793 4709 
Fax: 617-536-8602
Laura McKinstry
Project Manager
SCHARP/FHCRC
1100 Fairview Ave. N. 
LE-400
P.O. Box 19024
Seattle, WA 98109-1024 
U.S.A
Email: 
lmckinst@scharp.org
Phone: 206-667-7322
Fax: 206-667-4812
Ayana Moore
FHI
2224 E. NC Hwy 54
Durham, NC 27713, U.S.A
Email: amoore@fhi.org 
Phone: 919-544-7040 ext. 
11244 
Fax: 919-544-0207
Kristine Patterson
Protocol Co-Chair
Division of Infectious 
Diseases 
CB# 7030, Bioinformatics 
Building 
130 Mason Farm Road, 2nd 
Floor 
Chapel Hill, NC 27599, 
U.S.A
Email:
kristine_patterson@med.unc.
edu 
Phone: 919-966-2536 
Fax: 919-966-6714
Paul Richardson
HPTN NL QC 
Representative
Johns Hopkins Univ. Dept. of 
Pathology
Pathology Building 306
600 N. Wolfe St.
Baltimore, MD 21287, 
U.S.A.
Email: pricha18@jhmi.edu
Phone: 410-502-0435
Fax: 410-614-0430
James F. Rooney
Pharmaceutical 
Representative
Gilead Sciences 
Vice President Medical 
Affairs
333 Lakeside Drive
Foster City, California 94404
U.S.A
Email: 
jim.rooney@gilead.com
Phone: (650) 522-5708
Fax: (650) 522-5854
Katherine Shin
Pharmacist
Pharmaceutical Affairs 
Branch
Division of AIDS
6700-B Rockledge Drive, 
Room 4227
Bethesda, MD 20892
Phone: 301-594-1517
Fax: 301-402-1506
Email:
KaShin@niaid.nih.gov
Nirupama Sista
FHI
2224 E. NC Hwy 54
Durham, NC 27713, U.S.A
Email: nsista@fhi.org 
Phone: 919-544-7040 ext. 
11590 
Fax: 919-544-0207
Lei Wang
SCHARP/FHCRC
1100 Fairview Avenue N
M2-C200
PO Box 19024 
Seattle, WA 98109, U.S.A
Email: lwang@scharp.org 
Phone: 206-667-7049 
Fax: 206-667-4812

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
PROTOCOL SIGNATURE PAGE
Version 2.0
29 November 2010
A Study of the HIV Prevention Trials Network (HPTN)
Sponsored by:
Division of AIDS, U.S. National Institute of Allergy and Infectious Diseases
U.S. National Institutes of Health
I, the Investigator of Record, agree to conduct this study in full accordance with the provisions of 
this protocol. I agree to maintain all study documentation for a minimum of three years after 
submission of the site’s final Financial Status Report to the Division of AIDS (DAIDS), unless 
otherwise specified by DAIDS or the HIV Prevention Trials Network (HPTN) Coordinating and 
Operations Center.  Publication of the results of this study will be governed by HPTN policies. 
Any presentation, abstract, or manuscript will be made available by the investigators to the 
HPTN Manuscript Review Committee and DAIDS for review prior to submission.
I have read and understand the information in this protocol and will ensure that all associates, 
colleagues, and employees assisting in the conduct of the study are informed about the 
obligations incurred by their contribution to the study.
__________________________________       
Name of Investigator of Record
__________________________________
_________________________________
Signature of Investigator of Record
Date

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TRIPHOSPHATE IN HEALTHY VOLUNTEERS
Version 2.0
29 November 2010
SCHEMA
Purpose:
Describe the dose-proportionality and intra-individual variability of 
tenofovir diphosphate (TFV-DP) and emtricitabine triphosphate (FTC-TP) 
at steady-state in healthy human participants taking Truvada® (FTC 
200mg/TDF 300 mg) under direct observation. 
Design: 
Phase 1, multi-site, open label, randomized, 4-arm, dose-ranging, 
pharmacokinetic (PK) study.
Study Population:
HIV-uninfected, healthy, sexually-active volunteers, including both men 
and women recruited from two sites in the United States (U.S.).
Study Size:
32 participants distributed into four 8-person cohorts with 4 men and 4 
women per cohort. 16 of the original 32 participants (2 men, 2 women per 
dosing cohort), will undergo intensive tissue sampling.
Treatment Regimen:Research participants will receive doses under the direct observation of 
study personnel for 5 weeks. Research participants will be enrolled 
randomly into one of four study arms: 
1. FTC 200 mg/TDF 300 mg, one tablet orally once weekly for 5 weeks
2. FTC 200 mg/TDF 300 mg, one tablet orally twice weekly for 5 weeks
3. FTC 200 mg/TDF 300 mg, two tablets orally twice weekly for 5 weeks
4. FTC 200 mg/TDF 300 mg, one tablet once daily for 5 weeks
Study Duration:
Approximately 12 months total. Accrual will require approximately 6 months. 
Each participant will be followed for approximately 2 months (screening, 
treatment, and follow-up).  

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Primary Objectives:
1. Assess dose-proportionality of intracellular TFV-DP and FTC-TP 
from weekly to daily dosing.
2. Describe intra-individual variability in intracellular TFV-DP and FTC-
TP concentrations at steady-state (comparison of Day 28 and Day 35)
Secondary Objectives: 
1. Describe the relationship between pre-dose (Ct) and decaying 
concentrations of TFV, FTC, and their phosphorylated derivatives 
(TFV-DP and FTC-TP) in blood serum, peripheral blood mononuclear 
cells (PBMCs), CD4+ blood cells, total tissue cells, CD4+ tissue cells, 
tissue homogenate, semen, and luminal fluid at steady-state (Day 35 
[pre-dose] and Day 49 [decaying, greater than one half-life for TFV-
DP]).
2. Describe differences in intracellular TFV-DP and FTC-TP steady-state 
Ct between men and women.
3. Characterize the safety profiles of four different TDF/FTC regimens 
for pre-exposure chemoprophylaxis (PrEP).
Study Sites:
·
Johns Hopkins University, Baltimore MD; USA
·
University of North Carolina at Chapel Hill, Chapel Hill NC; USA

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Version 2.0
29 November 2010
OVERVIEW OF PARTICIPANT DISTRIBUTION AND STUDY DESIGN 
A) Distribution Scheme 
2
men
Arm 1: One(1) Tablet 
Weekly (1x); N=8
Arm 2: One(1) Tablet 
Twice Weekly (2x); N=8
2
women
2
men
2
women
Arm 4: One(1) Tablet 
Daily (7x); N=8
2
men
2
women
Arm 3: Two(2) Tablets
Twice Weekly (4x); N=8
2
men
2
women
FTC 200mg/TDF 300 mg 
Tablet (Truvada®) N=32
2
men
2
women
2
men
2
women
2
men
2
women
2
men
2
women
Non-Intensive Cohort
(Blood Only)
Intensive Cohort
(Blood + Tissue, Lumen, 
Semen)

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OVERVIEW OF PARTICIPANT DISTRIBUTION AND STUDY DESIGN (cont.)
B) Study Design
Screening
Accumulation
Steady
-State
Decay
Study day
Dosing (Arm 1-4)
PK Blood Sampling
(Both Cohorts)
PK Tissue/Lumen/Semen 
Sampling
(Intensive Cohort only)
0
7
14
21
28
35
49
42

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1.0
INTRODUCTION
1.1
Background and Prior Research
1.1.1
Using PrEP for HIV Prevention 
With 2.5 million new HIV infections expected in the coming year, there is an urgent need to find 
effective ways to reduce HIV transmission (UNAIDS, 2008). Pre-exposure antiretroviral 
chemoprophylaxis (PrEP) has been shown to decrease viral transmission in animal models 
(Garcia-Lerma et al., 2008; Subbarao et al., 2007; Veazey et al., 2005), and post-exposure 
prophylaxis has decreased HIV transmission in health care workers (Cardo et al., 1997) and men 
who have sex with men (MSM) (Schechter et al., 2004). The first human efficacy trials of daily 
PrEP are likely to be completed by late 2010. These trials are only evaluating daily dosing. For 
the majority of the “at-risk” populations, HIV risk-taking is unlikely to occur on a daily basis. 
Therefore, less frequent drug dosing (i.e., intermittent PrEP) will result in decreased drug costs 
and lower risk of toxicity. However, any benefit from intermittent PrEP as compared to daily 
PrEP could be mitigated by an increased risk of HIV infection if intermittent PrEP were to 
provide insufficient drug concentrations in relevant tissues. The most encouraging data 
suggesting that intermittent PrEP may be a plausible prevention strategy comes from a study by 
Garcia-Lerma and colleagues (Garcia-Lerma et al., 2009), in which monkeys were well-
protected from SIV infection after receiving one dose of tenofovir disoproxil fumarate (TDF) 
plus emtricitabine (FTC) at either 7 days, 3 days, or 2 hours pre-SIV exposure followed by a 
second dose 2 hours post-exposure. While encouraging, these data cannot be directly 
extrapolated to humans due to the differences in simian anatomy and pharmacokinetics as well as 
differences between HIV and the SIV challenge strain. Moreover, an efficacy trial of intermittent 
PrEP can be initiated only after the following pharmacological issues are understood. First, the 
overall drug concentrations achieved in humans receiving various intermittent PrEP regimens 
must be understood. Secondly, the intra-individual variability exhibited between proposed 
intermittent PrEP regimens must be carefully considered. Lastly, drug concentrations must then 
be evaluated in light of in vitro and animal model data to estimate the range of concentrations 
that may be necessary to provide preventive concentrations of the relevant antiretroviral drugs.   
1.1.2
Significance of Determining Pharmacology of Intermittent PrEP 
The data generated by the proposed study will be used to develop a comprehensive multi-
compartment pharmacokinetic (PK) model.  Assuming dose proportionality is found, this model 
will be used to estimate extracellular and intracellular drug exposure in multiple compartments 
for any dose and dosing interval combination.  This model will be critical to the following three 
areas: 1) developing an optimal pharmacology-based adherence intervention for future PrEP 
studies, 2) assessing adherence in ongoing studies with currently stored samples, 3) developing a 

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pharmacokinetic-pharmacodynamic (PK/PD) model to determine target intracellular drug 
exposures that prevent HIV infection. 
1.1.3
Need for Accurate Adherence Data 
All large-scale PrEP trials will need to determine medication adherence accurately in order to 
interpret the success or failure of any pharmacologic intervention. Acquisition of HIV infection 
during such trials may result from failure to achieve protective drug concentrations at the site of 
action. This failure may be from either (1) full adherence that did not achieve adequate drug 
concentrations from the prescribed regimen, or (2) poor adherence to the prescribed regimen, 
resulting in concentrations too low to provide protection from HIV infection. Differentiating 
between these two etiologies of PrEP failure are impossible using traditional adherence measures 
[pill counts, medication history and computer chips in the cap of pill containers (e.g., MEMS 
caps)] as none of these actually document pill-taking behavior.  Using a pharmacologic measure 
in combination with a robust PK model will allow interpretation of adherence using stored 
samples from completed studies, and will provide real-time information that could be used to 
guide behavioral interventions.
1.1.4
Drug Concentration as an Adherence Measure 
In clinical trials, measurement of drug concentrations has been proposed as a potential adherence 
measure to assess medication adherence more accurately. In this method, blood and other clinical 
samples of interest are sampled at specified times and the resultant “observed” drug 
concentrations are compared to the “expected” drug concentrations. The proportional difference 
between the expected and observed drug concentrations is taken as a rough estimate of the 
proportion of prescribed doses that were actually taken. Any differences between the observed 
and expected values are attributed primarily to adherence, since the expected value estimates 
(described below) control for adherence, and since other significant sources of variation (e.g., 
assay variability and environmental variables) are reasonably assumed to be stable over time.
1.1.5
Drug Concentration Simulation Models
In computer simulation models, the “expected” drug concentration is based on the PK 
characteristics of the drug and the prescribed dosing regimen of the study.  To facilitate these 
simulations, PK data are available from several reports in which TDF (Viread®) has been 
administered to human research participants. From these studies, one can make some estimates 
of the effective intracellular half-life for tenofovir diphosphate (TFV-DP), the active moiety of 
the drug, which appears to be approximately ~150 hours (Hawkins et al., 2005; Pruvost et al., 
2005; Pruvost et al., 2009). From these studies, however, the inter-individual variability for half-
life and peak concentration (Cmax) for TFV-DP is relatively large (coefficient of variation around 
50% to 65%). Accordingly, the best parameter estimates for such a simulation come from 

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individual study participants. To avoid intensive, frequent PK sampling, which would be 
cumbersome in a large clinical trial, one can employ a combination of (1) population-based 
estimates from the studies like those cited above, and (2) data from a few samples from each 
individual in a prospective study. To remove variability in adherence in estimation of individual 
PK parameter estimates, samples collected from the individuals tested should follow an observed 
dose, thus insuring 100% adherence without variability. With an observed dose, the possibility of 
variable adherence affecting subsequent drug concentrations is removed.   This leaves the 
following variables to consider: inter-individual PK variability, assay variability, and unknown 
sources of noise that may vary with time in the environment. Examples of simulations of 
proposed dosing strategies of TFV-DP in PBMCs (Figure 1-1) and TFV in blood serum (Figure 
1-2) are shown below. 
Figure 1-1.  Simulation of tenofovir diphosphate (TFV-DP) concentration in PBMCs (y-axis) 
versus time (x-axis) anticipated with the four different regimens planned for this study: TDF 300 mg 
daily (blue, QD 300), TDF 600 mg twice weekly (Monday & Thursday, red, BIW 600), TDF 300 mg 
twice weekly (Monday & Thursday, green, BIW 300), TDF 300 mg weekly (Monday, purple, QW 300); 
note all regimens are administered as Truvada® (TDF/FTC). Note the minor increase in Ctau and Cmax
between Day 28 and Day 35 where intra-individual variability will be estimated. Simulation parameters 
are based on data from previous studies (Hawkins et al., 2005; Pruvost et al., 2005; Pruvost et al., 2009). 
Simulations assume dose-proportionality and do not include intra-individual variation.

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Figure 1-2. Simulation of tenofovir concentration in blood serum (y-axis) versus time (x-axis)
anticipated with the four different regimens planned for this proposal. The regimens are described in 
Figure 1-1. Note the near complete overlap in concentration profiles between all regimens on days when 
the dose is taken. This is due to a very low accumulation index (see text below).
To use the computer modeling methods described above, two key facts must be determined with 
regard to the interaction of drug, participant, and regimen: intra-individual variability and dose-
proportionality. Knowledge of these variables is essential for planning future studies that use 
estimated PK parameters based on individual drug concentrations to evaluate adherence.
1.1.6
Importance of Intra-individual Variability 
Knowledge of intra-individual variability in drug concentrations is necessary to determine 
accurate sample size estimates for studies that plan to use drug concentration monitoring as an 
adherence measure. Without this parameter, estimates can be made from inter-individual 
variability from existing unpaired studies. However, the inter-individual approach typically 
results in overestimates of variability and hence a larger than necessary sample size. 
Alternatively, some conservative assumptions can be made for intra-participant correlation, 
though this introduces an additional variable that may cause estimates to be either too high or too 
low. 
1.1.7
Importance of Dose-proportionality
In order for drug concentration to be used as an adherence measure, the dose-proportionality 
(i.e., drug concentration at specified times changes proportionally with change in the frequency 
and amount of drug taken) must be stable at concentrations throughout the adherence range of 
interest: from concentrations associated with 100% adherence down to concentrations associated 
with lower levels of adherence.  While dose-proportionality is closely related to first order PK, 

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there are other conditions that will cause disproportionate kinetics, even for a first order drug.  
Very few drugs exhibit mixed or zero order PK in the clinically relevant concentration range 
(e.g., aspirin, phenytoin, and fluoxetine). For these drugs, a proportional increase in dose results 
in a disproportionately large increase in concentration. For other drugs, a proportional increase in 
dose may result in a disproportionately small increase in concentration. This can be seen in drugs 
with saturable absorption, and with drugs that are anabolized to phosphorylated forms within 
cells, such as TFV, FTC, and zidovudine (AZT). For example, because phosphorylation of AZT-
monophosphate within cells is the rate-limiting step in AZT-triphosphate anabolism, increasing 
the AZT dose can result in disproportionately small or no increase in AZT-triphosphate. For 
either cause of disproportionate PK, the assumption of proportionate change in adherence (as 
with expected and observed drug concentrations) becomes more complex. 
While the dose-proportionality for both TDF and FTC are well-established (Barditch-Crovo et 
al., 2001; Gish et al., 2002), this proportionality has not been determined for the phosphorylated 
moieties of these drugs and must be established for one to use TFV-DP or FTC-TP as measures 
of adherence to a Truvada® (TDF/FTC) regimen. 
Other proportionality factors are dose frequency and the accumulation of drug concentration with 
time – characteristics that are unique to each drug and dosing regimen. Drugs that are dosed at 
intervals more frequent than their half-life accumulate over time and achieve concentrations that 
are higher at steady-state than following a single dose. This accumulation – the so-called 
“accumulation index” or Rac – varies with the ratio of the drug half-life to dose frequency as 
follows: 
Rac = 1/(1 – e-k )   where k is the elimination rate constant [Ln(2)/half-life] and t is the dosing 
interval. 
The greater the drug half-life, the greater the accumulation index. The higher the accumulation 
index, the greater the magnitude by which drug accumulation will exceed sources of noise in 
drug concentration assessment (e.g., dosing time variation, assay variability, assay sensitivity, 
and inter- and intra-individual variability). When measuring adherence with drug concentration, 
one is assessing the magnitude of difference between the expected concentrations of drug at (1) 
prescribed dosing intervals assuming 100% adherence, and (2) actual drug concentrations from 
an unobserved dosing period. The difference is attributed to less than 100% adherence. A 
reduction in adherence has the equivalent effect on concentration as a reduction in the frequency 
of dosing (i.e., the concentration falls). Drugs with a large accumulation index provide a greater 
degree of detectable difference between 100% adherence and lesser adherence. For drugs with 
little accumulation (dose frequency less than their half-life), it may be very difficult to 
discriminate the concentration associated with a single dose (without any prior doses) from 
perfect adherence. Therefore, a participant with poor adherence can appear to a have high level 

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of adherence with a low accumulation index drug and drug regimen combination. In contrast, 
large accumulation index drug and drug regimen combination is very resistant to “white-coat 
adherence”, in which a participant takes one or several doses within the day prior to a scheduled 
research clinic visit. 
Table 1-1. Accumulation index for parent and phosphorylated moieties of TDF and FTC dosed 
either weekly (168 hours) or daily (24 hours). Drug moieties and regimens are ranked by accumulation 
index. An index of 2 indicates a doubling of concentration at steady-state compared to a single dose. An 
index of 1 indicates no accumulation at steady-state. For example, the concentration observed with 100% 
adherence can look the same as no adherence if the non-adherent participant takes a single dose prior to 
the observed blood collection when TFV, FTC or TDF/FTC are used with prescribed weekly dosing 
regimens. By contrast, the concentration expected with 100% adherence to Truvada® will be 9.6 (TFV-
DP) or 2.7 (FTC-TP) times the concentration observed if a participant has taken only a single dose prior 
to the scheduled observation. (Hawkins et al., 2005; Pruvost et al., 2005; Pruvost et al., 2009; Wang et al., 
2004)
Table 1-1. Accumulation index for parent and phosphorylated 
moieties of TDF and FTC dosed either weekly (168 hours) or daily 
(24 hours).
Drug moiety
Half-life 
(hours)
Dosing 
interval 
(hours, tau)
Dosing 
interval / 
half-life
(tau/HL)
Accumulation 
index
(Rac)
TFV-DP
150
24
0.2
9.5
FTC-TP
36
24
0.7
2.7
TFV-DP
150
168
1.1
1.9
TFV
17
24
1.4
1.6
FTC
10
24
2.4
1.2
FTC-TP
36
168
4.7
1.0
TFV
17
168
9.9
1.0
FTC
10
168
16.8
1.0
For this reason, TFV-DP (the intracellular form of TFV) is an excellent candidate for drug 
adherence monitoring, especially with daily dosing (Table 1-1). With an intracellular half-life of 
150 hours, daily to weekly dosing of TDF has a TFV-DP accumulation index ranging from of 9.5 
to 1.9, offering a great range of sensitivity for determining reduced adherence over a large range 
of dose frequencies.  In contrast, TFV concentration (measured in blood serum), when TDF is 
dosed daily, would be expected to perform much less well, with a small difference in drug 
accumulation at steady-state during daily dosing when compared to a single dose (e.g., taken on 
the day of observation). When using TDF on a weekly dosing regimen, there is no anticipated 
difference between drug concentrations of serum TFV compared to taking a single dose of TDF 
without any prior doses. Contrasting Figure 1-1 with Figure 1-2 illustrates this point graphically. 
The different dose regimens are easily distinguishable in Figure 1 where intracellular TFV-DP is 
measured. In contrast, the TFV serum levels are largely overlapping with daily and weekly 
dosing at least one day per week.

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1.2
Rationale
1.2.1
Overall Study Rationale
This PK study is designed to establish the dose-proportionality of TFV and FTC (serum and 
intracellular forms) with daily to weekly dosing. This information is essential to (1) employ drug 
concentration as an adherence measure in future PrEP studies, and (2) to estimate the anticipated 
concentration of parent and active moieties of TFV and FTC in intermittent PrEP regimens 
associated with full adherence to a prescribed regimen. In addition, intra-individual variability 
will be assessed to improve sample size estimates in future PrEP studies that use drug 
concentration as an adherence measure or where PK-PD correlations are planned. The dose-
proportionality of intra-cellular phosphates of the two components of the study product has not 
previously been established. Given the complexity of movement of these drugs between body 
compartments (central compartment, vaginal mucosal tissue, vaginal lumen, gastrointestinal 
mucosal tissue, gastrointestinal lumen), into cells within these compartments, and intra-cellular 
phosphorylation, it is possible that non-dose proportional kinetics, such as mixed or zero-order 
saturable processes may be operating.
1.2.2
Dosing Regimen Rationale 
We propose to study four different dosing regimens of Truvada® (tablets include 300 mg TDF 
and 200 mg FTC): one tablet daily, two tablets twice weekly, one tablet twice weekly, and one 
tablet weekly. To provide adequate data on dose-proportionality, these regimens are selected to 
cover the range of concentrations achieved between daily and weekly dosing, representing a 7-
fold range of anticipated drug concentrations. These regimens will achieve drug concentrations 
anticipated in current PrEP studies that employ daily dosing, as well as drug concentrations 
likely to be achieved with proposed intermittent PrEP regimens with dosing as infrequent as 
weekly or twice weekly dosing. In addition to studying three different dosing frequencies (daily, 
twice weekly, and weekly), we will study two different doses given twice weekly: a twice-
weekly regular dose (one Truvada® tablet), and a twice-weekly double-dose (two Truvada® 
tablets). This will provide data on dose-proportionality for regimens between the weekly and 
daily dosing. Using the weekly one Truvada® tablet as reference, these four dosing regimens 
represent anticipated drug exposures of 1x (weekly), 2x (twice weekly), 4x (double-dose twice 
weekly), and 7x (daily). These proportions will provide sufficiently rich data to assess dose-
proportionality. If dose-proportionality is determined not to hold between weekly and daily 
dosing, the middle dose regimens provide intermediate points to increase the accuracy with 
which we can determine the point of deviation from dose-proportionality. Further, they will 
provide rich drug concentration data to understand these intermediate dosing regimens, which 
may prove useful in future PrEP efficacy studies should the concentrations of drug prove 
insufficiently low with weekly dosing. 

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1.2.3
Dosing Duration Rationale
Dose-proportionality can be assessed at any point of observation following initiation of dosing. 
Intra-individual variability, however, can best be assessed at two sampling times not anticipated 
to vary based on PK principles. Essentially, by definition, this condition is achieved when 
steady-state has been reached. This study requires direct observation of dosing to rule out 
adherence as a cause for variation in drug concentrations. Therefore, we plan to begin sampling 
for intra-individual variability assessments as early as possible after dosing, balancing logistical 
difficulties (up to daily dosing for study staff and research participants) and PK limitations. 
Intracellular TFV-DP concentrations are estimated to achieve between 93% and 97% of steady-
state Cmin (trough) concentrations after 4 half-lives and 5 half-lives, respectively, have elapsed 
after initiation of Truvada® dosing. Based on an estimated 150-hour TFV-DP intracellular half-
life (FTC-TP will achieve steady-state earlier, given its far shorter 36-hour half-life), the 4 and 5 
half-life points coincide with 28 and 35 days of dosing. Laboratory assay variation for all of the 
assays planned for this study are more than twice as large as the very small 4% difference in 
concentrations that we expect to measure between Day 28 and Day 35. Accordingly, we plan to 
dose for 35 days, and to use samples from Day 28 and Day 35 for our intra-individual variability 
estimates. One could argue for longer periods of dosing to further reduce differences between 
doses in the asymptomatic approach to a true steady-state, but we believe that our proposed plan 
provides a reasonable balance between logistical complexity (daily dosing under direct 
observation) and reasonable experimental variation. 
The double-dose (two Truvada® tablets) twice-weekly regimen is only 4/7th (57%) of total 
weekly dosing for the single dose (one tablet) daily regimen. However, the double-dose twice-
weekly regimen includes doses within each dosing day that are twice the dose recommended in 
the package insert for both FTC and TDF, namely 400 mg and 600 mg, respectively. We believe 
that there is ample evidence that this dose will be safe in these healthy volunteers based on 
previous clinical testing and based upon pharmacological principles. First, regarding prior 
clinical studies, as part of their drug development, both TDF and FTC were studied at these 
doses with daily dosing from 2 to 8 weeks without any observation of dose-dependent adverse 
effects (Barditch-Crovo et al., 2001; Rousseau et al., 2003; Rousseau et al., 2001). 
Kearney, et al., demonstrated bioequivalence for TDF and FTC compared to the Truvada® 
combination (Kearney et al., 2004). Accordingly, the data based on dosing single products is 
provided as evidence of each individual component within the combination. 
In the Barditch-Crovo 2001 paper, one cohort of HIV-infected patients received 600 mg TDF for 
35 days – the highest dose administered (Barditch-Crovo et al., 2001).  The drug demonstrated 
dose-proportionality for peak concentration and area under the concentration time curve in blood 
plasma over the range from 75 mg to 600 mg TDF daily. With regard to safety, 1 placebo and 3 

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TFV subjects discontinued the study early due to lab abnormalities. Six subjects had grade 3 or 4 
adverse events (1 placebo, 5 on TDF). None of the 10 subjects receiving TDF 600 mg daily were 
discontinued for either lab abnormalities or experienced a grade 3 or 4 clinical event. 
Accordingly, there was no dose-response noted in adverse effects in this study over the dose 
range from 75 mg to 600 mg daily for 35 days. We will dose for the same 35 day duration, but 
the 600 mg dose will only be administered twice weekly rather than daily as in this study. 
In the Rousseau 2001 paper, one cohort of 9 HIV-infected subjects received 400 mg daily FTC 
for 14 days.  Dose-proportional increases in FTC and FTC-TP concentrations were seen over this 
period. Among all 38 subjects in the study, FTC was well tolerated at all dosing levels. No 
serious or severe adverse events were noted. Moderate events possibly related to FTC were seen 
and were not dose-related. While this study only dosed for 14 days, it is still highly relevant 
since the only dose that is anticipated to exceed the intracellular concentrations of the single 
daily dose cohort is the first dose of the double dose twice weekly regimen. Thereafter, all 
intracellular concentrations will fall below those of the single daily dose cohort.
In contrast, we plan to administer the double-dose only twice-weekly. Accordingly, we believe 
the double-dose twice-weekly regimen will be at least as safe as the daily dosing regimen. 
Based on pharmacological principles, published estimates of PK parameters for both drugs as 
summarized in Table 1-1 above, only the very first dose of the double-dose regimen will give an 
intracellular concentration higher than the concentration following the first dose of the single 
dose daily regimen. At all other times, the intracellular concentration for the double dose twice 
weekly falls below the single dose daily regimen The accumulation index of the fully 
phosphorylated moieties of tenofovir and emtricitabine dosed daily is 9.5 and 2.7, respectively. 
Thus, at steady-state, the intracellular concentration of the fully phosphorylated moieties exceeds 
that of the peak concentration following a single dose by multiples of those values. Accordingly, 
even with a doubling of a single dose, the peak intracellular concentrations falls well below the 
steady-state concentrations with daily dosing. With twice weekly dosing of Truvada® at double 
the recommended dose, the peak intracellular concentrations are still below those of daily 
Truvada® dosing (since the total weekly dose is only 4/7th the daily dose total. After the first 
dose, the double-dose regimen results in concentrations below the daily single dose regimen (see 
Figure 1-1 above). 
The serum TFV concentrations will be higher with double-dosing twice weekly when compared 
to daily dosing of a single tablet (Figure1- 2). This is because the accumulation index for TFV 
and FTC are only 1.6 and 1.2, respectively, indicating very little accumulation. Accordingly, the 
double dose will exceed the concentration of the daily single dose TFV and FTC, but only on the 
2 days of the week when the double dose is taken.  In addition, the use of double doses with a 
twice-weekly frequency provides the daily dose equivalent most closely associated with coitally-
dependent dosing, in which a dose is taken both before and following intercourse. We believe 
these blood plasma concentrations are less relevant for safety issues as the phosphorylated 

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moieties are the ones associated with the desired antiretroviral and undesired (non-viral) effects 
of the drugs.
2.0
STUDY OBJECTIVES AND DESIGN
2.1
Primary Objectives
The primary objectives of this study are to:
·
Assess dose-proportionality of intracellular TFV-DP and FTC-TP from weekly to daily 
dosing (Arms 1-4).
·
Describe intra-individual variability in intracellular TFV-DP and FTC-TP concentrations 
at steady-state (comparison of Day 28 and Day 35).
2.2
Secondary Objectives
The secondary objectives of this study are to:
·
Describe the relationship between pre-dose (Ct) and decaying concentrations of TFV, 
FTC, and their phosphorylated derivatives (TFV-DP and FTC-TP) in blood serum, 
peripheral blood mononuclear cells (PBMCs), CD4+ blood cells, total tissue cells, CD4+ 
tissue cells, tissue homogenate, semen, and luminal fluid at steady-state (Day 35 [pre-
dose] and Day 49 [decaying, greater than one half-life for TFV-DP]).
·
Describe differences in intracellular TFV-DP and FTC-TP steady-state Ct between men 
and women.
·
Characterize the safety profiles of four different TDF/FTC PrEP regimens.
2.3
Study Design
2.3.1
Identification of Study Design
HPTN 066 will be a Phase 1, multi-site, randomized, four-arm, open label study of the 
concentrations of TFV, FTC, and their metabolites in various body compartments following oral 
Truvada® dosing under direct observation by the site staff (directly observed therapy [DOT]) to 
assure 100% adherence to prescribed dosing regimen. The prescribed dosing regimens range 
from weekly to daily FTC/TDF at one of two possible doses. The overall study schedule and 
sample collection summary is provided in Appendix I.  After participant education, informed 
consent, and screening for study eligibility, participants will have baseline procedures performed 
(e.g. phlebotomy) before the first dose. 

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2.3.2
Study Sampling
Sampling will occur over 7 weeks: at enrollment (pre-dose), pre-dose #2 (Day 1 for treatment 
arm 4, Day 3 for treatment arms 2 and 3, and Day 7 for treatment arm 1) and Days 7, 14, 21, 28, 
35 and 49 to support each of the several objectives as follows:
1. Sample blood serum and blood cells (total cells and CD4+ cells) weekly for 5 weeks 
(Enrollment and Days 7, 14, 21, 28, and 35) to assess rate of accumulation and dose-
proportionality in concentrations of extracellular TFV and FTC and intracellular TFV-DP 
and FTC-TP. 
2. Sample blood serum and blood cells at steady-state (Day 28 and Day 35) to assess intra-
individual variability.
3. When participants consent to participate in HPTN 066, they will be offered the 
opportunity to participate in the intensive sampling cohort (to provide mucosal samples). 
The slots in the intensive sampling cohort will be balanced by gender, so that there will 
be two men and two women who will undergo mucosal sampling in each of the four arms 
of the study. Potential research participants who do not choose to participate in or are 
inevaluable for the intensive sampling cohort will be offered participation in the non-
intensive cohort. The study team will sample rectosigmoid, seminal and vaginal fluid, 
cells, and tissue (in addition to blood) at Days 35 and 49 to compare extracellular 
concentrations of TFV and FTC and intracellular concentrations of TFV-DP and FTC-
TP, and to estimate the elimination rate of drug from these compartments. 
The sampling described must occur on the days prescribed given the need to synchronize post-
dose time (7 days) across the four regimens. The blood sampling time should precede the 
established dosing time by no more than one hour. If the participant cannot have tissue sampled 
as prescribed, sampling should take place within the visit window, described in the SSP. In all 
cases, there must be accurate recording of sample collection times. 
Biopsies will not be performed during menstruation. As such, the timing of the first dose will 
need to take into consideration menstrual patterns and will be adjusted accordingly.
2.3.3
Summary of Major Endpoints
2.3.3.1 Primary Endpoints: 
·
Concentrations of TFV and FTC in blood serum
·
Concentrations of TFV-DP and FTC-TP in PBMCs and CD4+ blood cells

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2.3.3.2 Secondary Endpoints:
·
Concentrations of TFV, FTC, TFV-DP and FTC-TP in blood
·
Concentrations of TFV and FTC in vaginal and rectal secretions
·
Concentrations of TFV and FTC in vaginal/cervical and rectal tissue homogenates
·
Concentrations of TFV-DP and FTC-TP in unselected and CD4+ vaginal/cervical and 
rectal tissue cells and tissue homogenates
·
Concentrations of TFV, FTC, TFV-DP and FTC-TP in fluid (vaginal, rectal, semen), 
cells, and tissue homogenate
·
Adverse experiences, including  safety laboratory studies (hematological, renal, hepatic 
function) 
2.3.4
Description of Study Population
The study population will include approximately 32 evaluable (defined Section 7.3), generally 
healthy 18-44 year-old men and women, who are HIV-uninfected and not pregnant or 
breastfeeding. The upper age limit was selected to provide comparability in age for male 
participants with our pre-menopausal female participants and reduce the variability that might be 
introduced by a wider age range that could have a significant impact in this small study.  
2.3.5
Trial Duration
The total duration of participation from the Enrollment Visit to the Termination Visit is 7 weeks.  
Visits may be completed within specified windows around target dates.  The approximate time to 
complete study enrollment is expected to be six months.  The time of total study duration is 
expected to be approximately twelve months, including the study follow-up period. Detailed 
information regarding visit windows will be thoroughly described in the HPTN 066 Study 
Specific Procedures (SSP) Manual.
2.3.6
Expected Duration of Participation
The expected duration of participation for enrolled participants approximately two months.  No 
study data will be collected after the Day 49 final study visit. 
3.0
STUDY POPULATION
Thirty-two HIV-uninfected participants will be included in this study.  Participants will be 
selected for the study according to the criteria in Section 3.1 and 3.2.  They will be recruited as 
described in Section 3.3 and assigned to a study arm as described in Section 7.4. Issues related to 

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participant retention and withdrawal from the study, are described in Sections 3.4 and 3.5, 
respectively.
3.1
Inclusion Criteria
Participants who meet all of the following criteria are eligible for inclusion in this study:
·
18 to 44 years of age, inclusive on the date of screening.
·
Provides informed consent for the study.
·
Non-reactive HIV rapid test results at the screening and enrollment visits.
·
An estimated calculated creatinine clearance (eCcr) at least 70 mL/min by the 
Cockcroft-Gault formula where:
o
eCcr (female) in mL/min = [(140 - age in years) x (weight in kg) x 0.85] / (72 
x serum creatinine in mg/dL).
o
eCcr (male) in mL/min = [(140 - age in years) x (weight in kg)] / (72 x serum 
creatinine in mg/dL).
·
Participants are sexually active, defined as at least one sex (vaginal or anal 
intercourse) act in the 30 days prior to screening.  
·
Participants must agree to use condoms for all coital events during study 
participation.
·
Intensive sampling cohort only:
o
Not using spermicide as a means of birth control (in conjunction with a 
condom or diaphragm)  
·
Women must:
o
Be pre-menopausal
o
Have regular menstrual cycles with at least 21 days between menses (unless 
on contraception that causes amenorrhea or irregular menses) 
o
Have a negative urine pregnancy test at screening and enrollment
o
Be utilizing an alternative method of birth control in addition to condoms 
(hormonal contraceptive, diaphragm or have undergone surgical sterilization) 
or have a vasectomized exclusive male partner.
o
Intensive sampling cohort only:
§
Have a cervix
§
Have documentation of a normal Pap smear within 12 months

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3.2
Exclusion Criteria
·
At screening:
o
Alanine aminotransferase (ALT) or aspartate aminotransferase (AST) greater 
than 1.5 X the site laboratory ULN (upper limit of normal)
o
Hemoglobin less than 10.0 g/dL
o
Platelet count less than 100,000/mm3
o
Serum phosphate level below site laboratory LLN (lower limit of normal)
o
INR or aPTT greater than site laboratory ULN
o
Other safety tests (bicarbonate (HCO3), potassium (K), chloride (Cl), sodium 
(Na), calcium (Ca), fasting glucose) with results outside of the laboratories 
reference range
o
1+ or greater protein on urine dipstick testing
o
1+ or greater glucose on urine dipstick testing
·
Culture-confirmed urinary tract infection
·
Co-enrollment in any other HIV interventional research study (excluding behavioral 
only interventions) or prior enrollment in the active arm of a HIV vaccine trial.
·
Clinically apparent or patient report of active skin disorders including: rash, pruritus, 
maculopapular rash, urticaria, vesiculobullous rash, and pustular rash.
·
Women who are pregnant or breastfeeding.
·
One or more reactive HIV rapid test results at screening or enrollment, even if HIV 
infection is not confirmed.
·
Positive hepatitis B surface antigen (HBsAg) test.
·
Excessive use of alcohol (more than 4 drinks a day on a regular basis).
·
Interleukin therapy; medications with significant nephrotoxic potential, including but 
not limited to amphotericin B, aminoglycosides, cidofovir, foscarnet and systemic 
chemotherapy; and medications that may inhibit or compete for elimination via active 
renal tubular secretion (including but not limited to probenecid).
·
Participants with a history of having a gastrectomy, colostomy, ileostomy, or any 
other procedure altering the gastrointestinal tract or drug absorption. 
·
Intensive sampling cohort only:
○
A positive test for syphilis, gonorrhea, or Chlamydia 
○
A positive test for HSV-2 (individuals with active lesions only)
○
Findings consistent with bacterial vaginosis, vaginal candidiasis, or 
trichomonas (women only)
○
History of STI within 3 months prior to enrollment
○
Medications that prolong clotting time (e.g., warfarin, heparin, clopidogrel 
classes.)
○
Abnormalities of the colorectal mucosa, or significant colorectal symptom(s), 
which in the opinion of the clinician represents a contraindication to biopsy 

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(including but not limited to presence of any unresolved injury, infectious or 
inflammatory condition of the local mucosa, and presence of symptomatic 
external hemorrhoids).
○
Clinically apparent pelvic exam finding (observed by study staff) of genital 
lesions, erythema, edema or any other abnormal physical or pelvic exam 
finding that, in the opinion of the investigator or designee, would 
contraindicate study participation. 
○
Women who have had cervical procedures (conization, LEEP procedure, 
cryosurgery) within the previous 6 months.
○
Spermicide as a method for contraception within last 30 days
·
Any other reason or condition that in the judgment of the investigator, would make 
participation in the study unsafe, complicate interpretation of study outcome data, or 
otherwise interfere with achieving the study objectives.
3.3
Recruitment Process
Participants will be recruited from a variety of venues. Examples of such venues include local 
health clinics, local health departments, and family planning clinics. In order to reach a broader 
audience, college and local newspaper advertisements will be utilized. Fliers will be posted on 
frequently visited local venues. Electronic advertisement will include secure list-serves 
(university) and craigslist.com. All advertising materials must undergo approval by local IRB’s. 
3.4
Participant Retention
Once a participant enrolls in this study, the study site will make every effort to retain him/her for 
7 weeks of the study. Retention goal for the study is 95%. For each participant who is not fully 
evaluable (as defined in Section 7.3), an additional participant will be enrolled. Study site staff 
are responsible for developing and implementing local standard operating procedures (SOPs) to 
target this goal.  Components of such procedures include:
·
Thorough explanation of the study visit schedule and procedural requirements during 
the informed consent process and re-emphasis at each study visit.
·
Thorough explanation of the importance of all study treatment groups to the overall 
success of the study.
·
Collection of detailed locator information at the study Screening Visit, and active 
review and updating of this information at each subsequent visit.
·
Use of appropriate and timely visit reminder mechanisms.

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·
Immediate and multifaceted follow-up on missed visits.
·
Mobilization of trained outreach workers or “tracers” to complete in-person contact 
with participants at their homes and/or other community locations.
3.5
Participant Withdrawal
Regardless of the participant retention methods just described, participants may voluntarily 
withdraw from the study for any reason at any time. The Investigator also may withdraw 
participants from the study in order to protect their safety and/or if they are unwilling or unable 
to comply with required study procedures after consultation with the Protocol Chair, DAIDS 
Medical Officer, SDMC Protocol Statistician, and CORE Protocol Specialist. Study visit and 
dosing requirements for research participant eligibility for safety and PK analyses is described in 
Section 7.2.   
Participants also may be withdrawn if the study sponsor, government or regulatory authorities
(Including the Office for Human Research Protections (OHRP) and FDA), or site IRBs/ECs 
terminate the study prior to its planned end date.  
Every reasonable effort will be made to complete a final evaluation (as described in Section 5.5) 
of participants who terminate from the study prior to the final study visit, and study staff will 
record the reason(s) for all withdrawals from the study in participants’ study records.
4.0
STUDY TREATMENT/PRODUCT/INTERVENTION
4.1
Study Product Formulation/Content/Storage
The study product to be dosed in all treatment arms is Truvada® (FTC/TDF) tablet. Truvada® 
tablets are fixed dose combination tablets containing emtricitabine (FTC) 200 mg and tenofovir 
disoproxil fumarate (TDF) 300 mg in each tablet.  Each bottle contains a silica gel desiccant 
canister that should remain in the original container to protect the study product from humidity.
Study Product Storage
FTC/TDF (Truvada®) study products should be stored at 25°C with excursions permitted to 
15°C - 30°C (59°F-86°F).

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4.2
Study Product Regimen(s)
Study participants who meet the inclusion/exclusion criteria (see Sections 3.1 and 3.2) will be 
randomized to one of four open-label treatment arms with emtricitabine (FTC) 200 mg/tenofovir 
disoproxil fumarate (TDF) 300 mg (Truvada®).
Table 4-1. Study Product Dosing Arms
Treatment 
Arm
Study Product
Number 
of Tablets
Route
Frequency
Duration
1
FTC 200 mg/TDF 300mg
One(1)
Orally
Once Weekly
5 Weeks
2
FTC 200 mg/TDF 300mg
One(1)
Orally
Twice Weekly
5 Weeks
3
FTC 200 mg/TDF 300mg
Two(2)
Orally
Twice Weekly
5 Weeks
4
FTC 200 mg/TDF 300mg
One(1)
Orally
Once Daily
5 Weeks
4.3
Study Product Administration 
FTC/TDF (Truvada®) tablets are administered orally with or without food.
4.3.1
Directly Observed Therapy (DOT) Guidelines
DOT is defined as directly observed therapy. A person identified by the Clinical Research Site 
(CRS) must observe and document the doses of study products taken by the study 
participant. Participants randomized to all arms will have all doses (as per randomized study 
arm) observed and documented. 
The method by which DOT is to occur should be discussed at length with the participant at study 
screening. Participants and coordinators should formulate a well-described plan for DOT prior to 
entry. It is possible that participants may start coming to the clinic for DOT for the first few 
weeks and then switch to an alternate location as CRS study staff and alternative facilities 
permit.
Each CRS will be responsible for determining who can perform DOT for that site. The 
individual(s) could administer the study product or observe the study products being taken, 
depending upon his/her certification and legal requirements of the local jurisdiction. Someone 
who is not medically trained cannot offer advice about the study product.
The DOT observer must be an objective third party arranged by the CRS. Friends or family 
members are not permitted as observers. Individuals who might perform DOT include 
physicians, registered nurses, licensed practical nurses, physician’s assistants, nursing assistants, 
certified home health aides, pharmacists, social workers, outreach workers, or other responsible
individuals designated by the CRS.  

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DOT may be delivered in any location deemed appropriate by the CRS (i.e., in the home, clinic, 
pharmacy, or outreach center). The study staff are responsible for ensuring that confidentiality is 
maintained at the selected locations. Examples of clinic-based DOT used at sites include:
·
Participant presenting daily to the site pharmacy for observed dosing by the site 
pharmacist who then documents on DOT participant log.
·
Participant presenting daily to the entrance of the hospital/clinic and study coordinator 
bring participant’s prepared study product to participant and study coordinator then 
documents on DOT participant log.
·
Participant and study coordinator meet at a designated off-campus location (coffee shop) 
each morning for DOT and study coordinator documents on DOT participant log.
CRSs that plan to use off-site locations for DOT are advised to give careful consideration to the 
following issues:
●
Confidentiality: Sites should consider where study paper work will be stored, how 
participants will be identified, how participant’s prepared study product will be 
transported, and where participants will be observed. Confidentiality should be discussed 
with participants as plans for their DOT are developed. The participant should be 
comfortable with the person who will be observing DOT. Sites must also discuss 
confidentiality with the DOT observer, noting situations where breaches are most likely 
to occur. DOT observers may be required to sign a confidentiality agreement.
The place of DOT administration should be captured on the study treatment record. The primary 
location of DOT administration and any alternate or secondary locations of DOT administration 
should be documented in the participants’ study files as well.  A DOT log will be used as a 
primary source document to record date, time, place, study product administered or taken, 
observer signature, participant signature, and any intervention for missed DOT visit (i.e., 
telephone call). 
Flexibility and creativity are needed as sites explore options for DOT delivery. It is recognized 
that sites will have some variability in the way they choose to administer DOT, but do need to 
report the delivery mechanisms they choose in the participants’ study files. Sites will be asked to 
inform the study team of the methods they are considering before they are used. Sites that have 
questions should contact the HPTN 066 protocol team for guidance.

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4.3.2
Follow-up for Missed Appointments
Participants who miss a DOT visit must be contacted by the clinic. Sites can be flexible (e.g., 
some sites may choose to do a home visit if this is an available option while others may choose 
telephone contact).  A tracking form has been developed for missed DOT visits as part of the 
study treatment record. Missed visits and reasons for missed visits must be recorded on the 
appropriate CRF.
4.3.3
Take-home Supply (for emergencies only) 
Take-home dosing may be provided to participants for emergency purposes.  Additional details 
on the use of the emergency dose will be provided in the SSP manual.
4.4
Product Supply and Accountability
Study Product Supply, Distribution and Accountability
Emtricitabine (FTC) 200 mg/tenofovir disoproxil fumarate (TDF) 300 mg study product tablets 
are manufactured and provided by Gilead Sciences, Inc. under the trade name Truvada®.
Study Product Acquisition
Emtricitabine (FTC) 200 mg/tenofovir disoproxil fumarate (TDF) 300 mg study product will be 
supplied by the NIAID Clinical Research Products Management Center (CRPMC).  The study 
site pharmacist can obtain FTC/TDF study products through the CRPMC by following the 
instructions in the Pharmacy Guidelines and Instructions for DAIDS Clinical Trial Networks, 
and instructions in the SSP Manual.
Study Product Accountability
The site pharmacist is required to maintain records of all FTC/TDF study products received from 
the CRPMC and subsequently dispensed to study participants.  All unused study products are to 
be held until the study is completed, terminated, or otherwise instructed by the sponsor.  Specific 
instructions will be provided by the sponsor for return or destruction of the study products.
4.5
Toxicity Management
Guidelines for clinical management and product hold/discontinuation are outlined in this section.
In general, the site investigator has the discretion to hold study product at any time if s/he feels 
that continued product use would be harmful to the participant, or interfere with treatment 

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deemed clinically necessary according to the judgment of the investigator.  Unless otherwise 
specified below, the investigator should immediately consult the PSRT for further guidance in 
restarting study drug or progressing to permanent discontinuation. 
4.5.1
Grading System
The grading system is located in the Division of AIDS Table for Grading Severity of Adult and 
Pediatric Adverse Events (DAIDS AE Grading Table), Version 1.0, December 2004; 
Clarification August 2009, which can be found on the Regulatory Support Center (RSC) Web 
site: http://rsc.tech-res.com/safetyandpharmacovigilance/. 
4.5.2
Dose Modification 
No dose modifications are permitted.
If absolutely necessary, dosing may be delayed for 2 days in Arms 1-3 during the first 3 weeks 
on study. Thereafter, all doses in week 4 and 5 must be taken as scheduled. All doses for 
research participants in Arm 4 must be taken as scheduled. Failure to comply with these schedule 
requirements will result in termination of the participant from the protocol after a final safety 
evaluation. 
4.5.3
Discontinuation of Study Product in the Presence of Toxicity
Grade 1 or 2
In general, participants who develop a Grade 1 or 2 AE regardless of relatedness to study product, 
and that is not specifically addressed below may continue use of the study product per protocol.  
Grade 3
Participants who develop a Grade 3 AE or toxicity that is not specifically addressed below and is 
judged to be related to study product should have that study product permanently discontinued.  
In general, and unless otherwise decided in consultation with the PSRT, the investigator should 
re-evaluate the participant at least weekly up to 2 weeks to document resolution of toxicity to less 
than Grade 2.  
Grade 4
Participants who develop a Grade 4 AE or toxicity that is not specifically addressed below 
regardless of relationship to study product should have the study product permanently 
discontinued. The participant should be re-evaluated at least weekly up to 2 weeks to show that 
the adverse event resolves to less than Grade 2.

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4.5.4
General Criteria for Discontinuation of Study Product
Participants may voluntarily discontinue the study products for any reason at any time.  Site IoRs 
will permanently discontinue participants from study product per protocol for any of the specific 
criteria below, which may be further clarified in the SSP.  Site IoRs also may permanently 
discontinue participants from study product for use of prohibited medication (per Section 4.9), 
for reasons not shown here or in the SSP (e.g., to protect participants’ safety and/or if 
participants are unable or unwilling to comply with study product use procedures).  In such 
cases, the Site IoRs should first query the PSRT for review.  The PSRT will provide a written 
response to the site indicating whether the PSRT has recommended permanent discontinuation of 
study product based on careful review of all relevant data.
The criteria for permanent discontinuation of study product use for an individual participant are:
·
Study product-related toxicity requiring permanent discontinuation of study product per 
the guidelines above and below
·
Completion of regimen as defined in the protocol
·
Request by participant to terminate study product
·
Clinical reasons determined by the physician 
·
Acquires HIV infection, hepatitis B infection 
·
Becomes pregnant
Any research participant who discontinues participation prematurely for any reason should, prior 
to discontinuation and dismissal from the study, undergo the safety evaluations regularly 
scheduled for the final study visit on Day 49, at the time of discontinuation.  
4.6
Management of Specific Toxicities
Specific guidance related to permanent discontinuation is also noted here as it pertains to the 
clinical management of toxicities.
4.6.1
Nausea and Vomiting
Participants with Grade 1 or 2 nausea or vomiting may be treated symptomatically with 
hydration, oral antiemetic therapies or antiemetic suppositories at the discretion of the site 
investigator.  Participants should be instructed to take oral study product with food.  
Participants with Grade ³ 3 nausea and vomiting for which an alternative etiology is not 
established must permanently discontinue the study product and be treated symptomatically. 

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4.6.2
Diarrhea
Participants with diarrhea of any toxicity grade may be treated symptomatically with permitted 
antimotility agents and rehydration at the discretion of the site investigator.
Participants with new onset Grade ³ 3 diarrhea that is unresponsive to antimotility agents and for 
which an alternative etiology (e.g., infectious diarrhea) is not established must permanently 
discontinue study product and be treated symptomatically.
4.6.3
AST/ALT Elevations
Careful assessments should be done to rule out the use of alcohol, non-study medication-related 
drug toxicity, or viral hepatitis as the cause of elevation in AST or ALT of any grade.  The 
participant must be carefully assessed for any symptoms or signs of hepatotoxicity, including 
fatigue, malaise, anorexia and nausea, jaundice, acholic stools, right upper quadrant pain or 
hepatomegaly.  If the AST/ALT elevation is considered most likely to be due to concomitant 
illness or medication, standard management, including discontinuation of the likely causative 
agent, should be undertaken. If symptoms or signs of clinical hepatitis are present, study 
treatment must be discontinued (see below).
Grade 1
For study participants with less than Grade 1 ALT and AST at study entry, an increase to Grade 
1 ALT or AST even in an asymptomatic participant may be of concern.  
ALT or AST must be repeated as soon as possible (at most within 1 week of the receipt of the 
results) of a new Grade 1 ALT or AST. Study treatment may be continued while repeating ALT 
and AST at the discretion of the investigator provided the participant is asymptomatic.  
Grade 2
Participants should have ALT or AST re-checked as soon as possible (at most within 1 week) 
and then be followed weekly until levels are Grade £ 1. The frequency of follow up may be 
altered at the discretion of the site investigator following consultation with the PSRT. Study 
treatment may continue at the discretion of the investigator provided the participant is 
asymptomatic.
Grade 3/4
Study product should be permanently discontinued for any ALT or AST of Grade 3 or 4. 
Participants should have ALT or AST re-checked as soon as possible (at most within 1 week).  
Participants should then be followed weekly until levels are Grade £ 1. 

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4.6.4
Creatinine Clearance
If the creatinine clearance is <50mL/min, it should be confirmed within 1 week of the receipt of 
the results in consultation with the PSRT.  If the creatinine clearance is confirmed to be 
<50mL/min, the study product must be permanently discontinued.  Participants who fail to have a 
confirmed test will permanently discontinue the study product. 
4.6.5
Hypophosphatemia
Grades 1 and 2.
The phosphate should be repeated within 1 week of the receipt of the results.  Supplemental 
phosphate should be given with phosphate-rich food or fluid with or without neutral phosphate 
solution.  Other causes of phosphate loss should be thoroughly evaluated.
Grade 3/4
The phosphate should be repeated within 48 hours of the receipt of the results.  Supplemental 
phosphate should be given with phosphate-rich food or fluid with or without neutral phosphate 
solution, and other causes of low phosphate should be thoroughly investigated. During the time 
that supplemental phosphate is provided to the participant and the time that testing is repeated, 
sites should follow product hold guidelines described in Section 4.5.  If alternative etiologies for 
Hypophosphatemia cannot be identified, the study team will give consideration to 
discontinuation of study drug. Study medication should be discontinued for resistance to 
supplemental phosphate intervention after 2 days.
4.6.6
Genital Sexually Transmitted Infection/Reproductive Tract Infection
Testing for STIs will be performed at screening for all participants. Persons identified as having 
an STI will be excluded from the intensive cohort and will be referred for management according 
to the local standard of care. Testing will be performed at other study visits only if the participant 
develops signs/symptoms of an STI or expresses a concern about having acquired a STI infection 
after enrollment. If any participant acquires an STI during the study, clinical management will be 
provided in accordance with current CDC guidelines. Probenecid may not be given with study 
medications. If probenecid is necessary for STI treatment, the study dosing will be discontinued.  
Study product need not be held in the event of genital STI/ reproductive tract infection (RTI) 
requiring treatment, unless other product hold guidelines apply. An STI will result in removal 
from the study only if treatment is not complete within one week prior to the tissue sampling 
days in the protocol (intensive sampling cohort only).

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4.6.7
HIV and Hepatitis B
HIV Infection
HIV testing will be performed at screening and enrollment. Individuals with one or more reactive 
HIV rapid test results (regardless of subsequent test results) will be excluded from the study and 
will be referred for management according to the local standard of care. HIV diagnostic testing 
will also be performed at the end of the study, and at any other study visit at which a participant 
has signs/symptoms of acute HIV infection or expresses a concern about having acquired HIV 
infection after enrollment. 
Hepatitis B
Hepatitis B testing (HBsAg) will be performed at screening. Persons with a positive HBsAg test 
will be excluded from the study and will be referred for management according to the local 
standard of care. Hepatitis B testing will be performed at the end of the study and at other study 
visits if the participant expresses a concern about having acquired hepatitis B infection after 
enrollment or as clinically indicated for AST/ALT elevations.  
4.6.8
Management of sampling complications
Tissue and fluids will be obtained in the intensive sampling cohort on Days 35 and 49. All 
subjects will be required to abstain from sexually activity for 72 hours following sample 
collection. Subjects will be contacted by phone 24 hours after sample collection to assess for any 
adverse events. 
Cervicovaginal Fluid
Collection of CVF requires women to be recumbent for 5 minutes to allow fluid to pool in the 
posterior fornix. The vaginal sample aspirator is then gently inserted into the vagina without the 
placement of a speculum. This procedure may cause very slight discomfort, pressure, or irritation 
that should occur only during the procedure (<5 minutes). Any uncomfortable sensations should 
resolve shortly after the procedure.
Subjects will not undergo CVF sampling if evidence of vaginitis is present on initial 
examination.
Cervical Cells
Cervical cells will be collected following the insertion of a speculum into the vagina by inserting 
a cytobrush (two full 360 degree rotations) into the endocervical canal. Minimal discomfort may 
occur with placement of the speculum. Slight bleeding or spotting may occur after the cytobrush 
is removed but no more than that which occurs during a normal pap smear. 

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Seminal Fluid
Semen will be self-collected in a private room in the research unit. Subjects may experience 
psychological distress and local irritation or discomfort. 
Vaginal Biopsies
During this procedure subjects may experience some mild pain when the sample is taken. 
Subjects will receive topical anesthetic prior to the procedure.  Bleeding may also occur at the 
specimen collection site.  Bleeding may be stopped at the biopsy site with application of a local 
anti-coagulant. In very rare cases, a stitch might need to be placed in order to stop bleeding.  
Under such circumstance, female participants may feel slight soreness or discomfort in the 
vagina for one to two days after the vaginal biopsy.  Participants may experience some vaginal 
spotting or bleeding and slight discharge for up to one week after the biopsy.  In rare instances, a 
vaginal biopsy may cause infection or prolonged bleeding. 
Subjects will not undergo tissue sampling if evidence of vaginitis is present on initial 
examination.
Colon Biopsies
Subjects will be requested to follow a low fiber diet for 3 days prior to the procedure and a clear 
liquid diet for 12 hours prior to the procedure. With the procedure, subjects may experience 
abdominal pain, rectal irritation, bloody bowel movements, dehydration, and changes in heart 
rate. There is rare risk of bleeding, perforation and infection with this procedure. In the event that 
other pathology is observed during flexible sigmoidoscopy (anorectal lesions, polyps, 
ulcerations, etc.), the subject will be informed of the findings and a referral for treatment will be 
provided. 
Study procedures may be terminated in the event that any subject experiences any of the 
following:  bleeding from the site that requires aggressive intervention to stop bleeding following 
the sample collection, experiences severe abdominal pain, any increased tenderness or distention 
in the abdominal region, or a change in vital signs from baseline deemed clinically significant by 
the endoscopist/study physician. 
Any subject who experiences excessive bleeding (in the opinion of the local study physician) 
following any biopsy procedure will not have further tissue sampling.
Rectal Luminal Fluid
To collect the rectal luminal fluid, the clinician will gently insert a clear, plastic, lubricated tube 
into the rectum (no more than 4 inches) through which the luminal fluid will be collected with a 
swab or sponge.

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While this is a common medical procedure, it is possible that the participant could experience 
mild discomfort, embarrassment or, rarely, pain (should they have another condition that is 
already causing pain in the area).  If there is any pain experienced during the procedures, the 
study clinician will adapt or stop the procedure.
4.7
Clinical Management of Pregnancy
All study participants are required to use condoms for all coital events during study participation.  
Women should also be utilizing a second form of contraception as indicated in Section 3.1. 
Study staff will provide contraceptive counseling to enrolled participants as needed throughout 
the duration of study participation and will facilitate access to contraceptive services through 
direct service delivery and/or active referrals to local service providers.  Study staff also will 
provide participants with male condoms and counseling on use of condoms during every sex act 
during study participation.  
Pregnancy testing will be performed at screening, enrollment, Day 28, Day 35 (intensive 
sampling cohort only), and Day 49 (pregnancy testing will be done prior to any vaginal 
procedures). Participants will be encouraged to report all signs or symptoms of pregnancy to 
study staff.  The site IoR or designee will counsel any participants who become pregnant 
regarding possible risks to the fetus according to site-specific SOPs.  The IoR or designee also 
will refer the participant to all applicable services; however, sites will not be responsible for 
paying for pregnancy-related care.  
Participants who are pregnant at the termination visit will continue to be followed until the 
pregnancy outcome is ascertained (or, in consultation with the PSRT, it is determined that the 
pregnancy outcome cannot be ascertained).  Pregnancy outcomes will be reported on relevant 
CRFs. Outcomes meeting criteria for expedited adverse event (EAE) reporting also will be 
reported on EAE forms (see Section 6.3.4 below). Follow-up of pregnant participants beyond 
study duration is only intended to capture pregnancy outcomes. On-going or regular study visits 
will not take place. 
NOTE: Participants who become pregnant during the course of the study will permanently 
discontinue study product.  
4.8
Criteria for Early Termination of Study Participation
Participants may voluntarily withdraw from the study for any reason at any time.  Site IoRs may, 
with the approval of the PSRT, withdraw participants before their scheduled termination visit to 
protect their safety, and/or if participants are unable or unwilling to comply with study 
procedures.  Participants also may be withdrawn if the study sponsors, government or regulatory 

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authorities (including the OHRP and FDA), or site IRBs/ECs terminate the study prior to its 
planned end date.  Site investigators are required to consult the Protocol Chair and Protocol 
Biostatistician prior to the termination of any study participant.  Study staff will record the 
reason(s) for all withdrawals in participants’ study records. Study visit and dosing requirements 
for research participant eligibility for safety and PK analyses is described in Section 7.2.
4.9
Concomitant Medications
With the exception of medications listed as prohibited (see below, this section), enrolled study 
participants may use concomitant medications during study participation.  All concomitant 
medications, over-the-counter preparations, vitamins and nutritional supplements, recreational 
drugs, and herbal preparations reported throughout the course of the study will be recorded on 
CRF designated for that purpose.  
All concomitant medications taken or received by participants within the 4 weeks prior to study 
enrollment will be reported on applicable study CRFs.   Medications used for the treatment of 
AEs that occur during study participation also will be recorded on applicable study case report 
forms.
Should participants report use of any of the following medications, they will be required to 
discontinue use of study products and have termination visit evaluations: interleukin therapy; 
medications with significant nephrotoxic potential, including but not limited to amphotericin B, 
aminoglycosides, cidofovir, foscarnet and systemic chemotherapy; and medications that may 
inhibit or compete for elimination via active renal tubular secretion (including but not limited to 
probenecid).
Medications that prolong clotting time (e.g., warfarin, heparin, clopidogrel classes) are 
exclusionary as are the conditions they treat. Occasional use of aspirin or non-steroidal anti-
inflammatory drugs (NSAIDs) for analgesia is allowed with restriction. Aspirin cannot be taken 
within 1 week of scheduled biopsies. NSAID cannot be taken within 24 hours of scheduled 
biopsy. Detailed instructions for preparation for biopsies will be included in the SSP. 
Spermicide should not be used by participants who undergo mucosal sampling during the study.  
Non-study vaginal and rectal products (other than tampons during menstruation, diaphragms and 
female condoms) are also prohibited for individuals undergoing mucosal sampling.  Participants 
who report current use of these products will be counseled regarding the use of alternative 
methods.  Participants are not expected to require gynecologic surgical procedures during 
enrollment period; however, should such a procedure be required, the site IoR or designee will 
consult the PSRT regarding ongoing product use by the participant.

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5.0
STUDY PROCEDURES
An overview of the study visit and procedures schedule is presented in Appendix I. Presented 
below is additional information on visit-specific study procedures. Detailed instructions to guide 
and standardize all study procedures across sites will be provided in the SSP manual. 
In addition to any Interim Visits that may occur in accordance with guidance outlined in Section 
5.6, the following visits should take place for study participants:
·
Screening
·
Enrollment (first dosing day; Day 0)
·
Pre-dose #2, Sample visit for single blood draw (accumulation index)
·
Day 7, Non-Intensive Sampling visit
·
Day 14, Non-Intensive Sampling visit
·
Day 21, Non-Intensive Sampling visit
·
Day 28, Non-Intensive Sampling visit
·
Day 35, Non-Intensive plus Intensive*† Sampling visit
·
Day 49, Non-Intensive plus Intensive*† Sampling visit, Termination Visit
* Note: Sixteen of the total 32 participants will participate in the intensive sampling. The 
intensive sampling cohort will be divided evenly between men and women from each dosing 
cohort.
† Site staff will follow-up with participants via telephone, 24 hours after biopsy to inquire about 
any complications possibly resulting from the biopsy.  
All sampling will occur no more than one hour prior to the scheduled dosing time. 
As dosing will be observed by a study team member, participants will have up to daily contact 
with the study team. These will be termed “dosing visits” to distinguish them from study visits 
listed above. 
Reimbursement for study visits will either take place at the end of each study visit, at the end of 
the study, or in a pro-rated manner, depending on site specific scheduling and IRB approvals.

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5.1
Screening Visit 
After providing written informed consent, potential participants may be screened for eligibility 
over two or more visits if necessary, and eligibility must be confirmed at the enrollment visit (i.e. 
values from screening need to be checked against the eligibility requirements).  For participants 
who do not meet the eligibility criteria, screening will be discontinued when ineligibility is 
determined.
For participants who are found to be presumptively eligible based on the evaluations listed below 
at these visits, final eligibility will be confirmed at the Enrollment Visit (Day 0), scheduled to 
take place within 30 days of the Screening visit.  

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Table 5-1. Screening Visit (up to 30 days prior to Enrollment Visit)
Component
Procedure/Analysis
Administrative
·
Obtain written informed consent for screening/enrollment
·
Verify age
·
Assign Participant ID (PTID)
·
Collect demographic information
·
Collect locator information
·
Assess behavioral eligibility
·
Schedule next visit
Clinical 
·
Collect medical/menstrual history 
·
Collect concomitant medications
·
Perform physical exam  
·
Perform pelvic exam (intensive sampling cohort)  
·
Provide counseling
o
Contraceptive
o
HIV pre- and post-test
o
HIV/STI risk reduction and condom
·
Refer to care for UTI/RTIs/STIs 
·
Provide male condoms
Blood
·
Collect blood samples
o
HIV diagnostic testing (see SSP)
o
Safety testing (see SSP)
o
Coagulation testing (intensive sampling cohort, see SSP)
o
HBsAg (confirmatory testing if positive)
o
Syphilis testing (confirmatory tests if positive)
o
Store plasma
Urine
·
Collect urine sample
o
Pregnancy testing (women only)
o
Dipstick urinalysis (if positive for leukocyte esterase or nitrates, perform 
urine culture)
o
GC/CT testing (intensive cohort only)
Pelvic 
·
Collect pelvic samples (intensive sampling cohort women only)
o
Vaginal fluid for wet mount (BV, candida, trichomonas), vaginal fluid pH 
o
Pap smear* 
Genital
·
HSV-2 testing (if active lesions) (intensive cohort only)
Rectal
·
GC/CT testing (intensive cohort only)
*Not required if a normal PAP smear result was documented within 12 months of screening.

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5.2
Enrollment Visit
Table 5-2. Enrollment Visit (Day 0)
Component
Procedure/Analysis
Administrative 
·
Review/update locator information
·
Confirm behavioral eligibility, required for eligibility assessment
·
Randomization
·
Schedule next study visit
·
Follow procedures for study arm assignment
Clinical 
·
Update medical/menstrual history, required for eligibility assessment 
·
Update concomitant medications, required for eligibility assessment
·
Review pre-existing conditions, required for eligibility assessment
·
Perform physical exam  (See Appendix IB), required for eligibility assessment
·
*Refer for UTI/RTIs/STIs, treat or refer for other findings
·
Provide counseling
o
Contraceptive
o
HIV pre- and post-test
o
HIV/STI risk reduction and condom 
o
Protocol adherence, product use/adherence 
·
Provide male condoms 
Blood
·
HIV diagnostic testing (see SSP)
·
Store plasma 
·
Collect PK blood specimens 
o
Serum 
o
PBMCs 
o
CD4+ blood cells 
Urine
·
Pregnancy testing (women only)
Study Product 
Supply 
·
Administer first dose of study product
·
Provide an emergency dose(s) of participant’s prepared study product
*If indicated (see SSP)

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5.3
Non-Intensive Sampling Visits 
Table 5-3. Non-Intensive Sampling Visits (Days 1‡, 3 ‡, 7, 14, 21, 28, 35, and 49)
Component
Procedure/Analysis
Administrative
·
Review/update locator information
·
Schedule next study visit
Clinical 
·
Collect interval medical/menstrual history 
·
Review/update concomitant medications
·
Record/update adverse events
·
Coagulation tests (Day 28 only, see SSP, intensive sampling cohort only)
·
Perform directed physical exam 
·
Reinforce counseling
o
Contraception
o
HIV/STI risk reduction and male condom 
o
Protocol adherence, product use/adherence 
·
Provide male condoms
Blood
·
Pre-dose #2: single blood draw for accumulation index (Day 1 for Treatment 
Arm 4, Day 3 for Treatment Arms 2 and 3, Day 7 for Treatment Arm 1)†
·
Safety testing (Days 14, 28, 35 and 49 only, see SSP)
·
Hepatitis B testing (Day 49 only, see SSP)
·
Store plasma (Days 28 and 49 only)
·
HIV testing (Day 49 only)
·
Collect blood specimens for PK analysis
o
Serum 
o
PBMCs 
o
CD4+ blood cells 
Urine
·
Collect urine sample (women only, Day 28 and 49 only)
o
Pregnancy testing 
Study Product 
Supply
·
Administer study product (if applicable)
·
Collect unused emergency dose of participant’s prepared study product on Day 
28 Visit for all arms. Provide a new emergency dose of participants’ prepared 
study product on Day 28 Visit in Treatment Arms 2, 3, and 4.  
·
Collect unused emergency dose of participants’ prepared study product on Day 
31 Visit for treatment Arms 2 and 3, and on Day 34 Visit for Treatment Arm 4.
* If indicated (see SSP)
† Sample collection for each cohort should be done no more than 1 hour prior to the second dose for each cohort. 
These will occur on day 7 (Cohort 1), Day 3 (Cohort 2 and 3), and Day 1 (Cohort 4), as indicated.
‡ Day 1 and Day 3 visits are only for the pre-dose #2 blood draw. Only administrative, blood collection, and study 
product supply procedures will take place one these days.  

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5.4
Intensive Sampling Visits
Table 5-4. Intensive Sampling Visit (Days 35 and 49)
Component
Procedure/Analysis
Administrative
· Review/update locator information
· Schedule next study visit
Clinical 
· Collect interval medical/menstrual history 
· Review/update concomitant medications
· Record/update adverse events
· Perform directed physical exam 
· Perform pelvic exam 
· Reinforce counseling
o
Contraceptive
o
HIV/STI risk reduction and male condom 
o
Protocol adherence 
· Male condoms
Blood
· HIV diagnostic testing (Day 49 only, see SSP)
· Hepatitis B testing (Day 49 only, see SSP)
· Safety testing (see SSP) 
· Store plasma (Day 49 only)
· Collect blood specimens for PK testing
o
Serum 
o
PBMCs 
o
CD4+ blood cells 
Urine
· Collect urine sample (women only) 
o
Pregnancy testing 
Vaginal 
sampling†
· Biopsy, cervical cytobrush, and luminal fluid (see SSP)
Rectosigmoid 
Specimens†
· Flexible sigmoidoscopy with biopsies (up to 30) and collection of luminal fluid 
(see SSP)
Semen†
· Collect semen sample (Men only, see SSP)
*If indicated (see SSP)
† Sixteen of the total 32 participants will participate in the intensive sampling which includes all procedures listed 
above. The intensive sampling cohort will be divided evenly between men and women from each dosing cohort. For 
the remaining subjects, that is, the non-intensive cohort, there will be no rectosigmoid, vaginal, or semen sampling 
on Day 35 and 49. 
5.5
Follow up Procedures for Participants who Discontinue Study Product and for 
Participants who Receive a Biopsy on the Last Study Visit.
Participants who permanently discontinue study product for any reason will be encouraged to 
return to the clinic to have exit safety labs drawn (see SSP). This will be their final scheduled 
visit for the study. 

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Participants who are enrolled in the intensive sampling cohort will have biopsies collected on 
Day 49. As an additional safety check, these participants will receive a follow-up phone call the 
day following the biopsy to evaluate for biopsy related adverse events.
5.6
Interim Contacts and Visits
Interim contacts and visits (those between regularly scheduled follow up visits) may be 
performed at participant request or as deemed necessary by the investigator or designee at any 
time during the study. All interim contacts and visits will be documented in participants’ study 
records and on applicable CRFs.  
Some Interim visits may occur for administrative reasons.  For example, the participant may 
have questions for study staff.  Interim visits at which no data is collected are not documented on 
case report forms. Other interim contacts and visits may occur in response to AEs experienced by 
study participants.  When interim contacts or visits are completed in response to participant 
reports of AEs, study staff will assess the reported event clinically, record the event on the case 
report form, and provide or refer the participant to appropriate medical care.  
5.7
Clinical Evaluations and Procedures
See Appendix I for an outline of physical exam and pelvic exam components. For directed 
physical exams (occurring during visits on Day 0 through 49), the extent of the examination is 
determined by the clinical judgment of the clinician based on interim medical history.  
5.8
HIV and STI Counseling
At any time during the study when HIV or STI testing is done on study participants, they will be 
informed of their test results as soon as they become available. Sites should follow their local 
standard of care for providing HIV and STI counseling and should document these procedure in 
site specific SOPs before study initiation.   
5.9
Participant Preparation for Sampling
Preparation and safety evaluations for biopsies
As stated in sections 4.6.8, participants should start a low fiber diet 3 days before each rectal 
biopsy procedure and have a clear liquid diet 12 hours prior to the procedure. They should also 
refrain from meals (or coffee) the day of the procedure. Enemas will be administered before the 
biopsy procedures take place.  Additional details on the dietary preparation and enemas will be 
included in the SSP. 

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Following enrollment, coagulation testing is done on participants in the intensive sampling 
cohort at the Day 28 visit. If test results are a Grade 2 or higher, the intensive tissue sampling 
will be cancelled. If possible, these participants may be transferred into non-intensive sampling 
cohort if there is an available slot. If a slot is not available, they will be terminated from the 
study. 
Semen sampling preparation
Men should refrain from sexual activity 24-48 hours prior to providing semen samples to allow 
for collection of an adequate number of seminal cells. 
6.0
SAFETY MONITORING AND ADVERSE EVENT REPORTING
6.1
Safety Monitoring 
Close cooperation between the Protocol Chair(s), study site Investigator(s), NIAID 
Medical/Program Officer, CORE Protocol Coordinator, SDMC Biostatistician, and other study 
team members will be necessary in order to monitor participant safety and respond to 
occurrences of toxicity in a timely manner. The team will have regularly scheduled conference 
calls during the period of study implementation, and additional ad hoc calls will be convened if 
required.
The study site Investigators are responsible for continuous close monitoring of all AEs that occur 
among study participants, and for alerting the PSRT if unexpected concerns arise. Accrual (but 
not dosing) will be suspended if two or more study participants experience the same grade 3 or 
higher AE (as defined by the DAIDS Standard Toxicity Tables) judged to be related to product 
use. The protocol team then will review all pertinent safety data and determine whether to 
continue accrual and product use.  A decision to stop the trial may be made by the protocol team 
at this time or at any such time that the team agrees that an unacceptable type and/or frequency 
of AEs has been observed.
A sub-group of the Protocol Team, including the Protocol Chair, DAIDS Medical Officer, one or 
more site clinicians, and the SDMC Clinical Affairs safety Associate, will serve as the Protocol 
Safety Review Team (PSRT) to be chaired by the Protocol Chair.  The HPTN SDMC will 
prepare routine safety data reports for review by the PSRT, which will meet via conference call 
approximately once per month or as needed throughout the period of study implementation to 
review safety data, discuss product use management and address any potential safety concerns.  
The content, format and frequency of safety data reports will be agreed upon by the PSRT and 
the SDMC in advance of study implementation.

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Termination Visit. If the participant has an adverse event (AE) at the termination visit, the AE, 
for study purposes, is closed out as “continuing at the end of study participation” but will be 
followed clinically until resolution or stabilization with updates on status provided to the PSRT.  
Participants who have AEs at the Termination Visit that have not resolved or stabilized will be 
followed beyond the Termination Visit until a clinically acceptable resolution of the AE(s) is 
confirmed and documented.  Clinical acceptability of resolution will be determined by the site 
investigator of record (IoR) in consultation with the Protocol Safety Review Team (PSRT).  For 
participants who are pregnant at the Termination Visit, study site staff will make every effort to 
follow the participant until such time that her pregnancy outcome can be ascertained and 
documented.
6.2
Clinical Data Safety Review
A multi-tiered safety review process will be followed for the duration of this study.  The study 
site investigators are responsible for the initial evaluation and reporting of safety information at 
the participant level, and for alerting the PSRT if unexpected concerns arise.  Participant safety is 
also monitored at the Network level through a series of routine reviews conducted by the SDMC 
Clinical Affairs staff, the PSRT and study sponsors.  Additional reviews may be conducted at 
each of these levels as dictated by the occurrence of certain events.  
HPTN SDMC Clinical Affairs staff will review incoming safety data on an ongoing basis. 
Events identified as questionable, inconsistent, or unexplained will be queried for verification.  
Adverse event reports submitted in an expedited manner to the DAIDS Safety Office will be 
forwarded to the DAIDS Medical Officer and SDMC Clinical Affairs staff for review.
The PSRT will meet regularly via conference call to review clinical data reports generated by the 
HPTN SDMC.  The content, format and frequency of the clinical data reports will be agreed 
upon by the PSRT and the SDMC in advance of study implementation. In addition to the routine 
safety data reviews, the PSRT will convene on an ad hoc basis to make decisions regarding the 
handling of any significant safety concerns.  If necessary experts external to the HPTN 
representing expertise in the fields of antiretroviral therapy, biostatistics, HIV transmission and 
medical ethics may be invited to join the PSRT safety review.  A recommendation to stop the 
trial may be made by the PSRT at this time or at any such time that the team agrees that an 
unacceptable type and/or frequency of AEs has been observed.
In the unlikely event that the protocol team or PSRT has serious safety concerns that lead to a 
decision to permanently discontinue the study product for all participants and stop accrual into 
the study, the protocol team or PRST will request a review of the data by the HPTN Study 

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Monitoring Committee (SMC) before recommending that the study be stopped.  If at any time, a 
decision is made to discontinue the study product in all participants, the site investigators of 
record will notify the responsible IRBs/ECs expeditiously.
6.3
Adverse Event Definition and Reporting Requirements
6.3.1
Adverse Events
An AE is defined as any untoward medical occurrence in a clinical research participant 
administered an investigational product and which does not necessarily have a causal 
relationship with the investigational product.  As such, an AE can be an unfavorable or 
unintended sign (including an abnormal laboratory finding, for example), symptom or disease 
temporally associated with the use of an investigational product, whether or not considered 
related to the product.  The term “investigational product” for this study refers to the oral 
medication Truvada® (tenofovir disoproxil fumarate/emtricitabine).
Study participants will be provided instructions for contacting the study site to report any 
untoward medical occurrences they may experience, except for possible life-threatening events, 
for which they will be instructed to seek immediate emergency care.  Where feasible and 
medically appropriate, participants will be encouraged to seek evaluation where a study clinician 
is based, and to request that the clinician be contacted upon their arrival.  With appropriate 
permission of the participant, whenever possible, records from all non-study medical providers 
related to untoward medical occurrences will be obtained and required data elements will be 
recorded on study CRFs.  All participants reporting an untoward medical occurrence will be 
followed clinically until the occurrence resolves (returns to baseline) or stabilizes.
Study site staff will document in source documents and the appropriate AE Log CRF all AEs 
reported by or observed in enrolled study participants regardless of severity and presumed 
relationship to study product.  AE severity will be graded per the DAIDS Table for Grading 
Adult and Pediatric Adverse Events, Version 1.0, December 2004 (Clarification dated August 
2009).
6.3.2
Serious Adverse Event
Serious adverse event (SAE) will be defined per U.S. Code of Federal Regulations (CFR) 312.32
and International Conference on Harmonization (ICH), “Good Clinical Practice: Consolidated 
Guidance” (E6) and “Clinical Safety Data Management: Definitions and Standards for Expedited 
Reporting” (E2A), as AE occurring at any dose that:
·
Results in death 
·
Is life-threatening

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·
Results in persistent or significant disability/incapacity
·
Is a congenital anomaly/birth defect  
·
Requires inpatient hospitalization or prolongation of existing hospitalization 
This includes important medical events that may not be immediately life-threatening or result in 
death, or hospitalization but may jeopardize the patient or may require intervention to prevent 
one of the outcomes listed above.
Per ICH SAE definition, hospitalization itself is not an adverse event, but is an outcome of the 
event. The following types of hospitalization do not require expedited reporting to DAIDS:
·
Any admission unrelated to an AE (e.g. for labor/delivery, cosmetic surgery, 
administrative, or social admission for temporary placement for lack of place to sleep)
·
Protocol-specified admission (e.g. for procedure required by protocol)
·
Admission for diagnosis or therapy of  a condition that existed before receipt of study 
agent(s) and has not increased in severity or frequency as judged by the clinical 
investigator
6.3.3
Adverse Event Relationship to Study Product
Relatedness is an assessment made by a study clinician of whether or not the event is related to 
the study agent.  Degrees of relatedness will be categorized according to current DAIDS-
approved guidelines.
6.3.4
Expedited Adverse Event Reporting
6.3.4.1 Adverse Event Reporting to DAIDS
Requirements, definitions and methods for expedited reporting of Adverse Events (AEs) are 
outlined in Version 2.0 of the DAIDS EAE Manual, which is available on the RSC website at
http://rsc.tech-res.com/safetyandpharmacovigilance/.
The DAIDS Adverse Experience Reporting System (DAERS), an internet-based reporting 
system must be used for expedited AE reporting to DAIDS.  In the event of system outages or 
technical difficulties, expedited AEs may be submitted via the DAIDS EAE Form. For questions 
about DAERS, please contact DAIDS-ES at DAIDS-ESSupport@niaid.nih.gov or from within 
the DAERS application itself.
Sites where DAERS has not been implemented will submit expedited AEs by documenting the 
information on the current DAIDS EAE Form. This form is available on the RSC website: 
http://rsc.tech-res.com/safetyandpharmacovigilance/.  For questions about EAE reporting, please 
contact the RSC (DAIDSRSCSafetyOffice@tech-res.com).

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6.3.4.2 Reporting Requirements for this Study
The SAE Reporting Category, as defined in Version 2.0 of the DAIDS EAE Manual, will be 
used for this study.
The study agent for which expedited reporting is required is: Truvada® (tenofovir disoproxil 
fumarate / emtricitabine (TDF/FTC)) 
6.3.4.3 Grading Severity of Events
The Division of AIDS Table for Grading the Severity of Adult and Pediatric Adverse Events 
(DAIDS AE Grading Table) is should be used to grade adverse events and is available on the 
RSC website at http://rsc.tech-res.com/safetyandpharmacovigilance/.
6.3.4.4   Expedited AE Reporting Period
·
The expedited AE reporting period for this study is as per the EAE manual.
·
After the protocol-defined AE reporting period, unless otherwise noted, only SUSARs as 
defined in Version 2.0 of the EAE Manual will be reported to DAIDS if the study staff 
become aware of the events on a passive basis (from publicly available information).
6.3.5
Pregnancy Outcomes
Pregnancy outcomes will be collected for all study participants by contacting these women soon 
after anticipated parturition. After the participant’s final study contact, any pregnancy outcomes 
that meet criteria for SAE reporting as described above (e.g., congenital anomalies) occurring 
among participants will continue to be expeditiously reported.
6.3.6
Regulatory Requirements
Information on all reported AEs will be included in reports to applicable regulatory authorities.  
The site IoR/designee will submit AE information in accordance with local regulatory agencies’ 
or other local authorities’ requirements.  The site IoR/designee also will submit AE information 
and any other relevant safety information to the IRB in accordance with IRB requirements.
6.4
Social Harms
In order to prevent adverse social events related to study participation, social harms will be 
monitored throughout the study.  Social harms are any untoward social occurrences that happen 

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to a participant as a result of their participation in the study.  Examples include loss of 
employment, harassment by neighbors, shunned by family, rejection by partner, etc.  Although 
social harms due to this study are expected to be negligible, they will be monitored closely 
throughout the study. Information on social harms will be actively solicited from participants at 
follow-up visits and recorded on case report forms and captured in the study database. 
Participants will also be encouraged to report any social harm on an ad hoc basis when it occurs.
In the event that a participant reports social harm, every effort will be made by study staff to 
provide appropriate care and counseling to the participant, and/or referral to appropriate 
resources for the safety of the participant as needed. Social harms that are judged by the 
Investigator of Record to be serious or unexpected will be reported to responsible site’s IRB/EC 
at least annually, or according to their individual requirements. The nature and frequency of 
these social impact reports will be monitored by the protocol team on a regular basis. In addition, 
these data will be reviewed by the HPTN (SMC).
7.0
STATISTICAL CONSIDERATIONS
7.1
Review of Study Design
HPTN 066 is a Phase 1, multi-site, randomized, four-arm, open label study of TFV and FTC 
concentrations in various body compartments following oral Truvada® dosing under direct 
observation by the study team (directly observed therapy [DOT]) to assure 100% adherence to 
prescribed dosing regimen. The prescribed dosing regimens range from weekly to daily 
TDF/FTC (Truvada®) at one of two possible doses for a total duration of 5 weeks. Thirty two 
participants are sampled weekly for blood (serum and cells). At the end of the 5th week and 7th
week, 16 of the 32 participants also undergo seminal fluid, rectosigmoid (tissue and luminal 
fluid) and vaginal sampling (tissue and luminal fluid). Samples are assayed for TFV, FTC and 
phosphorylated moieties of these drugs.
7.2
Endpoints
7.2.1
Primary Endpoints
Consistent with the primary study objective, the following endpoint will be assessed on Day 28 
and Day 35 across all 4 dosing arms:
·
Concentrations of TFV and FTC in blood serum
·
Concentrations of TFV-DP and FTC-TP in PBMCs and CD4+ blood cells

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7.2.2
Secondary Endpoints
Consistent with the secondary study objective, the following endpoints will be assessed on Day 
35 and Day 49 across all 4 dosing arms:
·
Concentrations of TFV, FTC, TFV-DP and FTC-TP in blood
·
Concentrations of TFV and FTC in vaginal and rectal secretions
·
Concentrations of TFV and FTC in vaginal and rectal tissue homogenates
·
Concentrations of TFV-DP and FTC-TP in unselected and CD4+ vaginal and rectal tissue 
cells and tissue homogenates 
·
Concentrations of TFV, FTC, TFV-DP and FTC-TP in fluid (rectal, vaginal, semen), 
cells, and tissue homogenate.
·
Adverse experiences, including safety laboratory studies (hematological, renal, hepatic 
function) 
7.3
Accrual, Follow-up, Retention, and Sample Size
A total of 32 evaluable men and women will be enrolled over a period of approximately 6 
months at multiple sites. An equal number of participants (8 per arm) will be randomized into 
each of the four dosing arms. Each enrolled participant will be followed for 7 weeks. 
Research participants may miss one of the following pre-steady-state study visits when on drug 
(Day 7, Day 14, Day 21) and remain in the study and remain eligible for evaluation in the PK 
outcome analysis of the study. Day 0, Day 28, Day 35, and Day 49 are required for a participant 
to be considered evaluable. Optimally, all study product dosing will occur on the date prescribed. 
However, some forgiveness of strict adherence to the schedule will be tolerated as long as the 
total dose within each week is consistent with the prescribed total. Therefore, for arms 1, 2, and 
3, study drug dosing can be delayed by 2 days, during the first 3 weeks of the study. Thereafter, 
all doses in week 4 and 5 must be taken as scheduled. In Arm 4, all doses for research 
participants must be taken as scheduled. Failure to comply with these scheduled requirements 
will result in discontinuation from the study after a final safety evaluation. 
Evaluable participants are expected to represent 95% of participants enrolled into the study. No 
adjustments are made for retention losses since all discontinued or inevaluable participants will 
be replaced. Each study site will establish participant retention procedures to target lost-to-
follow-up rates of <5% in order to minimize additional enrollments to compensate for 
inevaluable participants.
For the intensive cohort, a subject must have all blood on Days 0, 28, 35 and 49, and vaginal, 
seminal, and rectal samples on Days 35, and 49 to be evaluable. If the subject is inevaluable for 

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the intensive cohort, but has met evaluability criteria for the non-intensive cohort AND there is 
an opening in the non-intensive arm-gender cohort matching the subject, then they may be 
switched in assignment to the non-intensive cohort. If the intensive subject fails to complete all 
required samples OR there is not an opening consistent with their gender/arm, then the 
participant is discontinued after the final safety evaluation. In either case, the subject needs to be 
replaced in the intensive sampling group. 
Typical sample sizes for phase I PK studies generally range between 6 and 12 subjects per arm. 
A number of different approaches have been used to assess dose proportionality in the literature 
(Gough et al., 1995; Hummel et al., 2009; Smith et al., 2000). We consider the sample size using 
the following two approaches: 
1) fitting a simple linear regression model between the PK parameter (y) and dose 
dose
y
+
=
, where the hypothesis that 
0
=
is tested and the lack of fit of the model is 
generally tested by adding a quadratic term; and 
2) using the power model approach to fit 
)
log(
)
log(
dose
y
+
=
, where 90% CI around 
is 
compared to the limits 
)
)
ln(
)
ln(
1,
)
ln(
)
ln(
1(
r
r
U
L
+
+
, where 
)
,
(
U
L
are the pre-defined limits and r
is the ratio of the highest to the lowest dose. The power model and the CI approach has become 
more preferred choice in recent years (Gough et al., 1995; Smith et al., 2000), however, Hummel 
et al. (Hummel et al., 2009) suggested that the choice of bioequivalence default values of 
8.0
=
L
and 
25
.1
=
U
seems to be impractically strict when applied over a dose range that is 
more than doubling apart and proposed a more lenient criterion 
5.0
=
L
and 
2
=
U
.
The empirical power of both approaches are computed using a simulation study with the 
following steps: 
1. Generate random samples of size n from a normal distribution 
)
,
14
(
´
=
´
=
CV
sd
dose
N
, where 14 is the weekly dosing Day 35 C from Figure 
1-1, and dose = 1, 2, 4 and 7 for Arm 1 to 4 respectively.
2. Fit linear regression model 
2
2
1
0
dose
dose
y
´
+
´
+
=
(Smith et al., 2000) and test 
for nonsignificant intercept and quadratic terms (Zhou et al., 2006).
3. Fit linear model 
)
log(
)
log(
dose
y
+
=
and compare 90% CI of 
with the pre-
specified critical interval 
)
36
.1
)
7
ln(
)
2
ln(
1,
64
.0
)
7
ln(
)
5.0
ln(
1(
=
+
=
+
4. Repeat steps 1-3 for 1000 times. The empirical power is calculated as the proportion of 
the replications in which nonsignificant intercept and quadratic term holds for approach 1 
or the CI falls entirely within the critical interval for approach 2.

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There are at least 4 available groups of participants for estimating inter-individual variability to 
be used here for the purposes of estimating sample size. These studies, however, present a broad 
range of CV. In one HIV prevention study in which research participants took a daily oral dose 
of TDF, the inter-individual coefficient of variation in TFV-DP concentrations in PBMCs was 
46%. Hawkins et al. (Hawkins et al., 2005) report range of median concentrations of 85-110 
fmol/M cells (Table 7-1) for 7 subjects evaluated. Imputing values from the extremes of this 
range would yield a CV of approximately 14%. Pruvost et al. (Pruvost et al., 2009) report (Table 
7-3) range of TFV-DP CV% as 28, 36, 35, 54 in a drug interaction study with LPV/r and NVP. 
Pruvost et al. (Pruvost et al., 2005) report (Table 7-1 groups 1 and 3) inter-individual CV% of 
42% (group 1) and 83% (group 3). Note none of the above studies were done in healthy 
volunteers with one single drug dosed under direct observation. Given that the DOT used in this 
study is likely to reduce the variability, a CV estimate of 25-35% seems to be realistic. In the 
simulation study, we explored a range of CV values from 25% to 45%.
Table 7-1. Power for dose proportionality assessment, 
05
.0
=
Sample size (per arm)
CV
Simple linear model and 
hypothesis testing approach
Power model and CI 
approach
N=6
25%
93.3%
98.4%
35%
93.6%
74.9%
45%
93.2%
33.3%
N=8
25%
95.0%
99.7%
35%
95.1%
81.1%
45%
93.8%
41.8%
N=10
25%
95.5%
99.8%
35%
95.0%
85.0%
45%
94.4%
43.5%
N=12
25%
96.1%
99.7%
35%
93.9%
85.2%
45%
94.6%
43.6%
Simulation results suggested that either 6 or 12 participants per arm would yield sufficient power 
for a range of coefficient of variation parameters (25% - 45%) when dose proportionality is 
assessed using hypothesis testing approach based on simple linear regression model. However, 
the empirical power associated with the power model and the CI approach is more heavily 
depend on the value of CV in the relevant range. With CV ≤ 35%, a reasonable degree of 
variation given the argument above, we would have ≥ 81% power to claim dose proportionality 
with 8 participants per arm. For this reason, in addition to cost and operational considerations, 8 
subjects per arm was selected as the minimum size to provide adequate power for the dose-
proportionality assessment, our primary objective. 

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For the secondary objective assessing difference in intracellular TFV-DP and FTC-TP steady 
state Day 35C (pre-dose concentration) between men and women, we assume the mean TFV-DP 
steady state concentration C is 100 fmol/106 cells, and the coefficient of variation between 
people is 35%. Based on the results from Van Belle and Martin (Van Belle and Martin, 1993), 
we would have only 35% power to detect a 1.5 fold difference (i.e., 100 to 150) between men 
and women within each dosing arm if an arm has 4 men and 4 women (i.e., a total of 32 
participants distributed evenly into four arms, 50% men and 50% women). If dose-
proportionality holds, then we would be able to make the comparison between men and women 
across dosing arms. We then would have 89% power to detect a 1. 5 fold difference with a total 
sample size of 32 (16 men and 16 women) selected for this study. Table 7-2 provides power for a 
range of CV, sample size and minimal detectable difference. The power would reduce with 
smaller sample size, smaller minimal detectable difference, and larger CV.
Table 7-2. Power to detect the minimal detectable difference (in fold) for various sample 
sizes and coefficients of variance (CV)
Minimal detectable X-fold difference
CV
N per group 
1.25
1.5
1.75
0.25
3
0.18
0.48
0.73
4
0.24
0.60
0.85
6
0.33
0.78
0.95
12
0.58
0.97
0.99
16
0.70
0.99
0.99
24
0.86
0.99
0.99
0.35
3
0.12
0.28
0.45
4
0.14
0.35
0.57
6
0.19
0.49
0.74
12
0.34
0.78
0.96
16
0.43
0.89
0.99
24
0.59
0.97
0.99
0.45
3
0.09
0.19
0.30
4
0.10
0.23
0.38
6
0.13
0.33
0.53
12
0.22
0.57
0.82
16
0.28
0.69
0.91
24
0.40
0.86
0.98

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7.4
Study Arm Assignment
At the time of consent, subjects will designate whether they will participate in the non-intensive 
or intensive sampling cohort. Assignment prioritization will go to the intensive sampling cohort.  
Within each of these 2 cohorts and each gender stratum, enrolled participants will be assigned at 
random to one of the four study arms in a 1:1:1:1 ratio.  The permuted block randomization 
scheme will not be stratified by site and will be generated and maintained by the HPTN SDMC. 
For each randomized participant who is not fully evaluable (see section 7.3) an additional 
participant of the same gender will be enrolled into the same arm and the same cohort (intensive 
or non-intensive sampling). Details of the randomization system will be in the SSP Manual. 
7.5
Blinding
There will be no blinding of the study. All of the outcome measures are objectively determined 
drug concentrations for which there should be minimal risk for bias. 
7.6
Data Analysis 
7.6.1
Primary Analyses
7.6.1.1 Dose Proportionality 
Dose proportionality will be assessed by a confidence interval approach based on the power 
model, using Day 35 pre-dose concentration (Ct) data from all evaluable participants in Arms 1-
4. Dose-proportionality will be declared if the 90% confidence interval of the slope completely 
falls in the critical interval 
)
36
.1
)
7
ln(
)
2
ln(
1,
64
.0
)
7
ln(
)
5.0
ln(
1(
=
+
=
+
. Analyte concentrations assessed 
for dose-proportionality will include TFV and FTC in fluids and their phosphorylated moieties, 
TFV-DP and FTC-TP, in cells. TFV-DP and FTC-TP will also be assessed in tissue 
homogenates. Weight-based or creatinine clearance-adjusted intracellular PK parameters may 
also be used in evaluating dose-proportionality (as well as secondary outcomes). Biological 
matrices included in the analysis will include: serum, PBMC, CD4+ blood cells, rectosigmoid 
and vaginal luminal fluid, semen, and rectosigmoid and vaginal tissue specimens (total tissue 
cells, CD4+ tissue cells, tissue homogenate) (see Table 7-3).  

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Table 7-3. Biological matrices for analyses
Blood Samples
Vaginal, Rectosigmoid, and Semen Samples
Serum
PBMC
CD4+ 
blood cells
Total tissue 
cells
CD4+ 
tissue cells
Luminal 
fluid & 
semen
Homogenate
TFV-DP
X
X
X
X
X
FTC-TP
X
X
X
X
X
TFV
X
X
X
FTC
X
X
X
Descriptive statistics, such as means, medians, variance, and interquartile ranges, will be used to 
summarize Day 35 pre-dose concentration (Ct) by gender and by dose arm. Simple comparison 
of median concentration level between dose arms will be done with standard non-parametric 
methods.   Gender- or dosing-specific CVs of Day 35 pre-dose concentration (Ct) will be 
estimated. These CV estimates would be informative to the future studies even if dose 
proportionality does not hold. 
7.6.1.2 Intra-Participant Variability
Intra-participant variability will be described by the coefficient of variation of concentrations 
assessed on Day 28 and Day 35, which are assumed to be at steady-state, using data from all 
evaluable participants across dosing arms. We will confirm that steady-state was achieved by 
evaluating drug concentration data from Day 7, 14, and 21. If the dose-proportionality is 
established as described earlier, then dose-adjusted concentrations will be used for a more robust 
assessment.  These variability estimates will be calculated for all analytes in blood and blood 
cells. As supportive evidence, the accumulation data from Day 7, 14, and 21 will also be used to 
estimate the elimination rate constant. Tissue and luminal assessments cannot be calculated due 
to the absence of multiple steady-state samples. 
7.6.2
Secondary Analyses
7.6.2.1 Intracellular Concentration Decay
The apparent elimination rate constant (ke), for intracellular phosphates (TFV-DP and FTC-TP) 
will be determined by estimating the negative slope of the decline in the natural log of 
concentration between Day 35 and Day 49. Half-life will be estimated as the natural log of 2 
divided by the elimination rate constant (ke). These estimates will be performed for each research 
participant and summarized using descriptive statistics for each arm and the study as a whole.  
Biological matrices included in the analysis are the same as shown in Table 7-3.

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7.6.2.2 Differences between Men and Women
Because the genital tracts of men and women differ greatly in terms of anatomy and physiology 
(e.g. pH differences) as well as drug metabolism, it is feasible that there may be gender-specific 
differences in the pharmacology of different doses of tenofovir and emtricitabine in different 
mucosal compartments. Differences in tissue concentrations of these drugs could be relevant in 
understanding if gender-related differences are detected in intermittent PrEP efficacy trials. 
Some gender-related differences may be primarily explained by differences in body mass index 
between men and women, so determining the specific reasons for potential gender-related 
differences may be challenging with relatively small sample sizes. Men and women will be 
recruited into the study in equal proportions for comparison. If dose-proportionality is 
established in the primary objectives, then dose-adjusted Day 35 Ct concentration differences 
(each analyte, location, and matrix) between men and women will be tested pooling all research 
participants using ANOVA. If dose-proportionality is not established in the primary aim, 
intracellular PK parameters will be summarized by standard descriptive statistics (such as means, 
medians, and ranges) for each gender within each dose arm. Non-parametric methods may be 
used to compare Ct between men and women. The lack of statistical power of these comparisons 
due to small sample size is acknowledged.
7.6.2.3 Safety Evaluation
Adverse events will be presented by individual listings and in frequency tables broken down by 
body system.
Laboratory test values will be presented in participant listings and in summaries.  Graded 
laboratory values will be summarized.  Boxplots of local laboratory values will be generated for 
baseline values and for values measured during the course of the study. Each boxplot will show 
the 1st quartile, the median, and the 3rd quartile. Outliers, or values outside the boxplot, will also 
be plotted. If appropriate, horizontal lines representing boundaries for abnormal values will be 
plotted.
8.0
HUMAN SUBJECTS CONSIDERATIONS
8.1
Ethical Review
This protocol and the template informed consent form(s) contained in Appendices II and III —
and any subsequent modifications — will be reviewed and approved by the HPTN Protocol 
Review Committee and DAIDS Prevention Science Review Committee with respect to scientific 
content and compliance with applicable research and human subjects regulations. 

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The protocol, site-specific informed consent form, participant education and recruitment 
materials, and other requested documents — and any subsequent modifications — also will be 
reviewed and approved by the ethical review bodies responsible for oversight of research 
conducted at the study site. 
Subsequent to initial review and approval, the responsible IRBs/ECs will review the protocol at 
least annually.  The Investigator will make safety and progress reports to the IRBs/ECs at least 
annually, and within three months of study termination or completion.  These reports will include 
the total number of participants enrolled in the study, the number of participants who completed 
the study, all changes in the research activity, and all unanticipated problems involving risks to 
human subjects or others. Study sites will submit documentation of continuing review to the 
DAIDS Protocol Registration Office, via the HPTN CORE, in accordance with the current 
DAIDS Protocol Registration Policy and Procedure Manual.
8.2
Informed Consent
Written informed consent will be obtained from each study participant. Each study site is 
responsible for developing a study ICF for local use, based on the template in Appendices II and 
III, which describes the purpose of the study, the procedures to be followed, and the risks and 
benefits of participation, in accordance with all applicable regulations.  The study site is also 
responsible for translating the template form into additional languages (if applicable), and 
verifying the accuracy of the translation by performing an independent back-translation. Literate 
participants will document their provision of informed consent by signing their informed consent 
forms.  Participants will be provided with a copy of their informed consent forms. 
8.3
Risks
Phlebotomy, STI Screening. Phlebotomy may lead to discomfort, feelings of dizziness or 
faintness, and/or bruising, swelling and/or infection.  Pelvic examination may cause mild 
discomfort and/or vaginal bleeding or spotting.  Disclosure of STI status may cause sadness or 
depression in volunteers.  Participation in clinical research includes the risks of loss of 
confidentiality and discomfort with personal nature of questions.  
Confidentiality. Although study sites make every effort to protect participant privacy and 
confidentiality, it is possible that participants' involvement in the study could become known to 
others, and that social harms may result. 
STI Reporting. Participants in sites requiring partner notification in response to diagnosed STI 
or HIV infection could have problems in their relationships with their sexual partners.  
Participants also could have problems in their partner relationships associated with use or 

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attempted use of study products.  In addition, participants could misunderstand the current 
experimental status of the study medication and as a result increase their HIV risk behaviors 
while in the study.
Flexible Sigmoidoscopy with Biopsy. Flexible sigmoidoscopy is a commonly practiced medical 
procedure and the endoscopic procedures done in this trial will not involve any unusual risks or 
discomforts. The risks associated with these procedures include mild discomfort and the feeling 
of having a “bloated stomach”.  Endoscopic biopsies are painless and heal quickly within 3 days. 
On extremely rare occasions, the endoscopic procedure or biopsies may lead to pain, infection 
(sepsis), bleeding or perforation of the gastrointestinal tract. Perforation occurs approximately 
once out of every 100,000 procedures. If this extremely rare complication occurs, antibiotics and 
surgery to repair the tear may be necessary.  Participants will be counseled to refrain from sexual 
intercourse for at least 72 hours following the biopsy. 
Anoscopy and luminal fluid collection. It is possible that the participant may experience mild 
discomfort, embarrassment or, rarely, pain (should they have another condition that is already 
causing pain in the area) during this procedure.  If there is any pain experienced during the 
procedures, the study clinician will adapt or stop the procedure.
Enema. The primary risk from having an enema is temporary discomfort. The procedure may 
cause a bloated or crampy feeling. Some air may be pumped into the rectum as well, causing 
flatulence. Very rarely, the enema tube could make a hole in the rectum causing a bowel 
perforation requiring surgery.
Vaginal Biopsy. Vaginal biopsy carries the risk of discomfort or pain during the procedure and 
for a few hours afterwards.  Participants may have mild vaginal spotting (bleeding) for one or 
two days, and will be instructed to avoid sexual intercourse until bleeding stops.  Some 
temporary discomfort with sexual intercourse may occur if the biopsy areas are still healing.  
There is a small risk of infection and heavier bleeding.  Participants will be instructed to contact 
the clinic if symptoms are bothersome, if heavy bleeding is noted (soaking through a pad or 
tampon in an hour or less) or if the participant develops any abnormal odor or discharge from the 
vagina. Participants will be counseled to refrain from sexual intercourse for at least 72 hours 
following the biopsy.

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Study Product
FTC/TDF Combination Tablet. Truvada® tablets are fixed dose combination tablets 
containing emtricitabine (FTC) 200 mg and tenofovir disoproxil fumarate (TDF) 300 mg in each 
tablet. There is very little experience in healthy volunteers taking Truvada®. No new or 
unexpected side effects are observed with the TDF 300 mg / FTC 200 mg combination tablet 
than those observed when each drug is given separately.  For study Arm 4, the TDF/FTC dosing 
regimens are consistent with package insert recommendations. Participants in Arms 1 and 2 will 
be taking the typical TDF/FTC dose, but at reduced frequency. Participants in Arm 3 will be 
taking twice the recommended dose, but at reduced frequency (twice weekly rather than daily), 
representing 57% of the weekly recommended dose. Individual daily doses of this magnitude 
were studied during development of both TDF and FTC and these were well tolerated. 
FTC. The following side effects have been associated with the use of FTC: headache, dizziness, 
tiredness, inability to sleep, unusual dreams, loose or watery stools, upset stomach (nausea) or 
vomiting, abdominal pain, rash, itching, skin darkening of the palms and/or soles, increased 
cough, runny nose, abnormal liver function tests, increases in pancreatic enzyme, increased 
triglycerides, increased creatine phosphokinase. 
FTC is a pregnancy category B medication.  No controlled human studies of FTC among 
pregnant women have been conducted.  
TDF. The most common side effects associated with oral TDF in patients with HIV infection are 
nausea, headache, diarrhea, vomiting, asthenia, flatulence, abdominal distension/pain and 
anorexia.  Less common side effects of TDF include kidney toxicities and low blood phosphate.  
Other side effects reported in the post-marketing period include weakness, pancreatitis, 
dizziness, shortness of breath, and rash.  TDF has been associated with decreased bone mineral 
density in HIV-infected patients taking TDF tablets for 24-48 weeks. 
TDF is a pregnancy category B medication.  No extensive controlled human studies of TDF 
among pregnant women have been conducted.  
Nucleotide Analogues. Lactic acidosis (elevated lactic acid levels in the blood) and severe 
hepatomegaly (enlarged liver) with steatosis (fatty liver) that may result in liver failure, other 
complications or death have been reported with the use of antiretroviral nucleoside analogues 
alone or in combination.  The liver complications and death have been seen more often in 
women on these drug regimens.  Some nonspecific symptoms that might indicate lactic acidosis 
include: unexplained weight loss, stomach discomfort, nausea, vomiting, fatigue, cramps, muscle 

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pain, weakness, dizziness and shortness of breath. These effects are not expected in this study 
involving a brief exposure to antiretroviral drug.
Antiretroviral Drugs. Redistribution/accumulation of body fat including central obesity, 
dorsocervical fat enlargement (buffalo hump), peripheral wasting, facial wasting, breast 
enlargement, and "cushingoid appearance" have been observed in persons receiving antiretroviral 
drugs. The mechanism and long-term consequences of these events are currently unknown. A 
causal relationship has not been established.  These effects are not expected in this study 
involving a brief exposure to antiretroviral drug.
8.4
Benefits
There will be no direct benefits to participants in this study.  However, participants and others
may benefit in the future from information learned from this study. Specifically, information 
learned in this study may lead to the development of a safe and effective PrEP regimen that 
prevents HIV infection.  
8.5
Incentives
Pending IRB/EC approval, participants will be compensated for their time and effort in this 
study, and/or be reimbursed for travel to study visits and time away from work. Site-specific 
reimbursement amounts will be specified in the study ICFs. 
8.6
Confidentiality
All study-related information will be stored securely at the study site.  All participant 
information will be stored in locked file cabinets in areas with access limited to study staff.  All 
laboratory specimens, reports, study data collection, process, and administrative forms will be 
identified by a coded number only to maintain participant confidentiality.  All records that 
contain names or other personal identifiers, such as locator forms and informed consent forms, 
will be stored separately from study records identified by code number.  All local databases will 
be secured with password-protected access systems.  Forms, lists, logbooks, appointment books, 
and any other listings that link participant ID numbers to other identifying information will be 
stored in a separate, locked file in an area with limited access.
Participant’s study information will not be released without the written permission of the 
participant, except as necessary for monitoring by the NIAID and/or its contractors, Gilead 
Sciences; representatives of the HPTN CORE, SDMC, and/or NL, U.S. FDA, OHRP, and/or site 
IRBs/ECs.  

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The HPTN has obtained a Certificate of Confidentiality from the US Department of Health and 
Human Services that will be applicable for this study.  Sites may register under the Certificate 
through the HPTN CORE once they have obtained local IRB approvals for the study.  This 
Certificate protects study staff from being compelled to disclose study-related.  information by 
any US Federal, State or local civil, criminal, administrative, legislative, or other body.
All protected health information (PHI) will be protected according to the provisions of the 
HIPAA and will only be used or disclosed as allowed by the privacy rule pursuant to relevant 
waivers or authorizations, or as required by federal law.
8.7
Communicable Disease Reporting Requirements
Study staff will comply with all applicable local requirements to report communicable diseases 
identified among study participants to local health authorities.  Participants will be made aware 
of all reporting requirements during the study informed consent process.
8.8
Study Discontinuation
The study may be discontinued at any time by NIAID, site IRBs/ECs, the HPTN, Gilead 
Sciences, OHRP or other governmental agencies as part of their duties to ensure that research 
subjects are protected.  
9.0
LABORATORY SPECIMENS AND BIOHAZARD CONTAINMENT
9.1
Local Laboratory Specimens
As described in Section 5, the following types of specimens will be collected for testing at the 
local laboratory: 
Blood
·
HIV diagnostic testing (see SSP)
o
HIV rapid test
o
HIV Western blot (for HIV confirmatory testing)
o
APTIMA HIV-1 RNA Qualitiative Assay (GenProbe, Inc., GenAptima assay)
·
Safety testing: 
o
CBC
o
AST/ALT
o
Creatine and calculated creatinine clearance
o
phosphate
o
HCO3, Na, Cl, K, Ca, glucose
·
Coagulation testing:

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o
INR, PT and PTT
·
Hepatitis testing (HBsAg)
·
Syphilis testing
·
Plasma storage
·
Blood samples for PK analysis (drug levels)
o
Serum
o
PBMCs
o
CD4+ cells
·
Quantitative HIV RNA assay (for any participant who acquires HIV infection during the 
study).
·
CD4 cell count (for any participant who acquires HIV infection during the study)
·
Genotyping and additional plasma storage (for any participant who acquires HIV 
infection during the study– see Appendix 1D)
Urine
·
Pregnancy testing (women only)
·
CT/GC testing
·
Dipstick for urinalysis. If positive for leukocyte esterase or nitrates, perform urine culture
Pelvic Specimens (women only)
·
Vaginal wet mount (for BV, candida, trichomonas)
·
Vaginal pH (if Amstel criteria are used for diagnosis of BV)
·
Pap smear (if a normal result is not documented within 12 months)
Responsibility of Study Sites
Study sites must adhere to standards of good clinical laboratory practice (GCLP), the HPTN 
Manual of Laboratory Operations, and local Standard Operating Procedures (SOPs) for proper 
collection, processing, labeling, and transport of specimens to the local laboratory. 
Procedures Performed at Local Laboratories
General requirements
Local laboratories must adhere to standards of GCLP, the HPTN Manual of Laboratory 
Operations, and local SOPs for proper specimen receipt, testing, aliquoting, storage, and 
shipment of specimens (see below). Specimen testing and storage must be documented using the 
Laboratory Data Monitoring Systems (LDMS).  Clinical sites in must document that their 
clinical laboratories are CLIA-certified and must meet DAIDS requirements for external quality 
assurance (QA) testing. External Quality Control (QC) of CLIA-waived tests (e.g. HIV rapid 
tests) may be required. HPTN NL staff will conduct period visits to each site to assess the 

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implementation of on-site QC procedures, including proper maintenance of laboratory testing 
equipment and use of appropriate reagents. HPTN NL staff will follow-up directly with site staff 
to resolve any QA or QC problems identified through proficiency testing and/or on-site 
assessments.
HIV diagnostic testing
HIV infection status will be assessed both at screening, enrollment, at the end of the study, and at 
any study visit at which a participant expresses a concern about having acquired HIV infection. 
Diagnostic testing algorithms are provided in the SSP. HIV rapid testing will be used to screen 
participants for HIV infection.  HIV rapid test kits must be FDA-cleared. Preference is given to 
FDA-cleared OraSure Oraquick HIV-1/2, Clearview HIV-1/2 Stat Pak, or Clearview Complete 
HIV-1/2.  Participants with one or more reactive HIV rapid test results will not be eligible for 
enrollment, regardless of subsequent test results. In those cases, HIV infection status will be 
confirmed using local HIV testing guidelines. After enrollment, if HIV infection is suspected or 
documented, the HPTN NL should be contacted. HIV infection must be confirmed for any 
enrolled participant using two independent samples.
Processing of samples for PK analysis (measurement of drug levels)
Blood samples will be processed locally to isolate serum and cells of interest. Detailed 
procedures for processing, freezing, storing, and shipping tissue samples are provided in the SSP 
Manual. 
Shipping
Sites must have the capability for shipping samples to the HPTN NL for Pharmacology, 
Virology, QA, and other testing. All specimens must be shipped in accordance with International 
Air Transportation Association (IATA) specimen shipping regulations and must be documented
in the LDMS. 
Sample storage
Specimens will be stored at study sites and shipped to the HPTN NL upon request. 
9.2
Network Laboratory Specimens
Testing Performed at the HPTN NL
Virology
The HPTN NL will perform testing to determine HIV infection status in selected cases and will 
perform HIV diagnostic testing for QC. The HPTN NL may also perform specialized assays to 
characterize HIV viruses and the immune response to HIV infection in any participant who 
becomes HIV-infected during the study. HIV resistance testing will be performed at the time of 
HIV diagnosis (see Appendix ID). HIV resistance test results from that study visit (time of HIV 

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diagnosis) will be provided to study sites at study closure. Results from that testing will be 
provided to study sites prior to study closure with approval of the HPTN NL and the Protocol 
Chair. That testing may be performed at a commercial laboratory (e.g., Monogram Biosciences). 
Additional testing may be performed for research purposes, either at the HPTN NL, or at an 
outside laboratory designated by the HPTN NL. This may include testing with assays used for 
analysis of HIV incidence testing to evaluate the fading of resistant variants (including minority 
variants assays) HIV subtyping, and other testing to characterize HIV viruses and/or the host 
response to HIV infection. Results from those tests will not be returned to study sites or study 
participants. 
STI Testing
The HPTN NL will perform HSV-2 PCR testing and GC/CT nucleic acid amplification testing 
(NAAT). Results from these tests will be returned to study participants.
Pharmacology
The HPTN NL will perform all drug testing for this protocol. Drugs will be measured in serum, 
PBMCs, CD4+ blood cells, total tissue cells, CD4+ tissue cells, tissue homogenate, and luminal 
(vaginal and rectosigmoid) fluid using assays that have an assay validation report (AVR) and 
SOP approved by the CPQA. Results from these tests will not be returned to study participants.
As described in Section 5, the following types of specimens will be collected for testing at the 
HPTN NL: 
Blood
·
Serum for drug levels
·
PBMC for drug levels
·
CD4+ blood cells for drug levels
·
Plasma for storage (for QC of HIV testing, and for viral characterization in any 
participant who acquires HIV infection during the study)
Rectosigmoid Sampling
·
Tissue biopsy intact (for homogenate)
·
Tissue total cells lysate
·
Tissue CD4+ cells lysate
·
Luminal fluid
Vaginal Sampling
·
Tissue biopsy intact (for homogenate)
·
Tissue total cells lysate
·
Tissue CD4+ cells lysate
·
Luminal fluid

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·
Cervical cells (from cytobrush)
Note: Vaginal tissues will be processed in the priority listed above and will be dependent upon 
the amount of tissue collected during the biopsy. Sites will collect up to 5 biopsies; however, the 
upper limits will be dependent upon site specific IRB approvals.
Genital swabs
·
For HSV-2 testing (only if active lesions are present)
Seminal Fluid
·
Drug levels
Rectal swabs
·
For GC/CT testing
All specimen collection and specimen processing procedures will be outlined in the study 
specific procedures manual. 
Each study site will adhere to standards of good laboratory practice and the HPTN Network 
Laboratory Manual for proper collection, processing, labeling, and transport of specimens for the 
NL.  All specimens will be shipped in accordance with IATA specimen shipping regulations. All 
shipments will be documented using the HPTN LDMS as described in the SSP manual.
9.3
Quality Control and Quality Assurance Procedures
The clinical sites will document that their clinical laboratories are CLIA-certified and/or 
participate in DAIDS sponsored EQA programs. NL staff will conduct periodic visits to each site 
to assess the implementation of on-site laboratory QC procedures, including proper maintenance 
of laboratory testing equipment and use of appropriate reagents. NL staff will follow-up directly 
with site staff to resolve any QC or QA problems identified through proficiency testing and/or 
on-site assessments. Throughout the course of the study, the HPTN NL will select a random 
sample of stored specimens to test for QA purposes. NL staff will follow-up directly with site 
staff to resolve any QA problems identified through this process.
QC for HIV Diagnostic Testing
Before performing HIV diagnostic testing, all sites must validate their testing algorithm, and the 
validation study must be approved by the HPTN NL. Local laboratories will perform testing for 
HIV diagnosis at screening, enrollment, and other scheduled visits. Algorithms for HIV 
diagnostic testing are provided in the SSP. 

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QC for HIV RNA Monitoring 
Quantitative HIV RNA (viral load) testing will be performed at local laboratories for any 
participant with confirmed HIV infection. Note that this is distinct from use of qualitative HIV 
RNA testing that may be performed to determine HIV infection status in selected cases. Local 
laboratories must participate in the DAIDS Virology QA program, with EQA results that are 
deemed satisfactory by the HPTN NL.
QC for CD4 Cell Count Determination
CD4 cell count testing will be performed at local laboratories for any participant with confirmed 
HIV infection. U.S. laboratories performing CD4 cell count testing must be CLIA-certified; 
enrollment in the DAIDS Immunology QA (IQA) program is preferred. 
9.4
Specimen Storage and Possible Future Research Testing
Study site staff will store all plasma specimens collected in this study until all protocol-related 
testing has been completed, including QC testing and other testing performed at or coordinated 
by the HPTN NL. Study staff will also store all samples collected for drug testing as directed by 
the NL. The study site will be informed by SCHARP when shipments to the NL are required, 
and will be instructed which samples to ship. In addition, study participants will be asked to 
provide written informed consent for their specimens to be stored after the end of the study for 
possible future testing.  The specimens of participants who do not consent to long-term storage 
and additional testing will be destroyed at the end of the study.
9.5
Biohazard Containment
As the transmission of HIV and other blood-borne pathogens can occur through contact with 
contaminated needles, blood, and blood products, appropriate blood and secretion precautions 
will be employed by all personnel in the drawing of blood and shipping and handling of all 
specimens for this study, as currently recommended by the U.S. Centers for Disease Control and 
Prevention.  All infectious specimens will be transported in accordance with U.S. regulations (42 
CFR 72).
10.0
ADMINISTRATIVE PROCEDURES
10.1
Study Activation
Prior to implementation of this protocol, and any subsequent full version amendments, each site 
must have the protocol and the protocol consent form(s) approved, as appropriate, by their local 
institutional review board (IRB)/ethics committee (EC) and any other applicable regulatory 
entity (RE). Upon receiving final approval, sites will submit all required protocol registration 
documents to the DAIDS Protocol Registration Office (DAIDS PRO) at the Regulatory Support 
Center (RSC).  The DAIDS PRO will review the submitted protocol registration packet to ensure 
that all of the required documents have been received.

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Site-specific informed consent forms (ICFs) WILL be reviewed and approved by the DAIDS 
PRO and sites will receive an Initial Registration Notification from the DAIDS PRO that 
indicates successful completion of the protocol registration process. A copy of the Initial 
Registration Notification should be retained in the site's regulatory files.
Upon receiving final IRB/EC and any other applicable RE approval(s) for an amendment, sites 
should implement the amendment immediately.  Sites are required to submit an amendment 
registration packet to the DAIDS PRO at the RSC.  The DAIDS PRO will review the submitted 
protocol registration packet to ensure that all the required documents have been received. Site-
specific ICF(s) WILL NOT be reviewed and approved by the DAIDS PRO and sites will receive 
an Amendment Registration Notification when the DAIDS PRO receives a complete registration 
packet. A copy of the Amendment Registration Notification should be retained in the site's 
regulatory files.
For additional information on the protocol registration process and specific documents required 
for initial and amendment registrations, refer to the current version of the DAIDS Protocol 
Registration Manual.
Pending successful protocol registration and submission of all required documents, CORE staff 
will “activate” the site to begin study operations.  Study implementation may not be initiated 
until a study activation notice is provided to the site.
10.2
Study Coordination
The United States Food and Drug Administration (U.S. FDA) has been informed of this study
and has provided an IND-exemption.
Assignment of all sponsor responsibilities for this study will be specified in a Clinical Trials 
Agreement executed by DAIDS and Gilead Sciences.
Study implementation will be directed by this protocol as well as the SSP manual. The SSP 
manual — which will contain reference copies of the Requirements for Source Documentation in 
DAIDS Funded and/or Sponsored Clinical Trials, as well as the DAIDS Manual for Expedited 
Reporting of Adverse Events to DAIDS, Version 2.0, dates January 2010 and the DAIDS 
Toxicity Tables — will outline procedures for conducting study visits; data and forms 
processing; AE assessment, management and reporting; dispensing study products and 
documenting product accountability; and other study operations.  
Study CRFs will be developed by the study team and HPTN SDMC.  Data will be transferred to 
the HPTN SDMC, entered, and cleaned using the SDMC DataFax data management system. 

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Quality control reports and queries routinely will be generated and distributed to the study sites 
for verification and resolution.   
Close coordination between protocol team members will be necessary to track study progress, 
respond to queries about proper study implementation, and address other issues in a timely 
manner.  Rates of accrual, adherence, follow-up, and AE incidence will be monitored closely by 
the team as well as the HPTN Study Monitoring Committee.  The Protocol Chair, DAIDS 
Medical Officer, Protocol Biostatistician, and CORE Protocol Specialist will address issues 
related to study eligibility and AE management and reporting as needed to assure consistent case 
management, documentation, and information-sharing across sites. 
10.3
Study Monitoring
On-site study monitoring may be performed in accordance with DAIDS policies.  Study monitors 
will visit the site to:
·
Verify compliance with human participants and other research regulations and guidelines; 
·
Assess adherence to the study protocol, study-specific procedures manual, and local 
counseling practices; and
·
Confirm the quality and accuracy of information collected at the study site and entered 
into the study database.  
Site investigators will allow study monitors to inspect study facilities and documentation (e.g., 
informed consent forms, clinic and laboratory records, other source documents, case report 
forms), as well as observe the performance of study procedures. Investigators also will allow 
inspection of all study-related documentation by authorized representatives of the HPTN CORE, 
SDMC, NL, NIAID, Gilead Sciences, FDA and other U.S. government and regulatory 
authorities. A site visit log will be maintained at the study site to document all visits.
10.4
Protocol Compliance
The study will be conducted in full compliance with the protocol.  The protocol will not be 
amended without prior written approval by the Protocol Chair and NIAID Medical Officer.  All 
protocol amendments must be submitted to and approved by the relevant IRB(s)/EC(s) and the 
DAIDS Regulatory Support Center (RSC) prior to implementing the amendment.

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10.5
Investigator's Records
The study site investigator will maintain, and store in a secure manner, complete, accurate and 
current study records throughout the study.  The investigator will retain all study records for at 
least three years after the completion of the study, unless directed otherwise by DAIDS.  Study 
records include administrative documentation, including site registration documents and all 
reports and correspondence related to the study, as well as documentation related to each 
participant screened and/or enrolled in the study, including informed consent forms, locator 
forms, case report forms, notations of all contacts with the participant, and all other source 
documents.
10.6
Use of Information and Publications
Publication of the results of this study will be governed by HPTN policies.  Any presentation, 
abstract, or manuscript will be submitted by the Investigator to the HPTN Manuscript Review 
Committee, DAIDS, and Gilead Sciences for review prior to submission.

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11.0
REFERENCES
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R.L. Coleman, P.D. Lamy, J.O. Kahn, I. McGowan, and P.S. Lietman. 2001. Phase i/ii 
trial of the pharmacokinetics, safety, and antiretroviral activity of tenofovir disoproxil 
fumarate in human immunodeficiency virus-infected adults. Antimicrob Agents 
Chemother. 45:2733-9.
Cardo, D.M., D.H. Culver, C.A. Ciesielski, P.U. Srivastava, R. Marcus, D. Abiteboul, J. 
Heptonstall, G. Ippolito, F. Lot, P.S. McKibben, and D.M. Bell. 1997. A case-control 
study of HIV seroconversion in health care workers after percutaneous exposure. Centers 
for Disease Control and Prevention Needlestick Surveillance Group. N Engl J Med. 
337:1485-90.
Garcia-Lerma, G., M.-E. Cong, and J. Mitchell. 2009. Prevention of Rectal Simian HIV 
Transmission in Macaques by Intermittent Pre-exposure Prophylaxis with Oral Truvada 
(Abstract 47). In 16th Conference on Retroviruses and Opportunistic Infections (CROI 
2009), Montreal, Canada.
Garcia-Lerma, J.G., R.A. Otten, S.H. Qari, E. Jackson, M.E. Cong, S. Masciotra, W. Luo, C. 
Kim, D.R. Adams, M. Monsour, J. Lipscomb, J.A. Johnson, D. Delinsky, R.F. Schinazi, 
R. Janssen, T.M. Folks, and W. Heneine. 2008. Prevention of rectal SHIV transmission in 
macaques by daily or intermittent prophylaxis with emtricitabine and tenofovir. PLoS 
Med. 5:e28.
Gish, R.G., N.W. Leung, T.L. Wright, H. Trinh, W. Lang, H.A. Kessler, L. Fang, L.H. Wang, J. 
Delehanty, A. Rigney, E. Mondou, A. Snow, and F. Rousseau. 2002. Dose range study of 
pharmacokinetics, safety, and preliminary antiviral activity of emtricitabine in adults with 
hepatitis B virus infection. Antimicrob Agents Chemother. 46:1734-40.
Gough, K., M. Hutchinson, and O. Keene. 1995. Assessment of dose proportionality: report from 
the pharmaceutical industry. Drug Information Journal. 29:1039-1048.
Hawkins, T., W. Veikley, R.L. St Claire, 3rd, B. Guyer, N. Clark, and B.P. Kearney. 2005. 
Intracellular pharmacokinetics of tenofovir diphosphate, carbovir triphosphate, and 
lamivudine triphosphate in patients receiving triple-nucleoside regimens. J Acquir 
Immune Defic Syndr. 39:406-11.
Hummel, J., S. McKendrick, C. Brindley, and R. French. 2009. Exploratory assessment of dose 
proportionality: review of current approaches and proposal for a practical criterion. 
Pharm Stat. 8:38-49.
Kearney, B.P., J. Zong, J. Begley, and J. Shah. 2004. Bioequivalence of combination tenofovir 
DF/emtricitabine tablets for one-pill once daily administration [poster no. 7.3]. In 5th 
International Workshop on Clinical Pharmacology of HIV Therapy, Rome, Italy.
Pruvost, A., E. Negredo, H. Benech, F. Theodoro, J. Puig, E. Grau, E. Garcia, J. Molto, J. Grassi, 
and B. Clotet. 2005. Measurement of intracellular didanosine and tenofovir 
phosphorylated metabolites and possible interaction of the two drugs in human 
immunodeficiency virus-infected patients. Antimicrob Agents Chemother. 49:1907-14.
Pruvost, A., E. Negredo, F. Theodoro, J. Puig, M. Levi, R. Ayen, J. Grassi, and B. Clotet. 2009. 
Pilot pharmacokinetic study of human immunodeficiency virus-infected patients 
receiving tenofovir disoproxil fumarate (TDF): investigation of systemic and intracellular 
interactions between TDF and abacavir, lamivudine, or lopinavir-ritonavir. Antimicrob 
Agents Chemother. 53:1937-43.

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Rousseau, C.M., R.W. Nduati, B.A. Richardson, M.S. Steele, G.C. John-Stewart, D.A. Mbori-
Ngacha, J.K. Kreiss, and J. Overbaugh. 2003. Longitudinal analysis of human 
immunodeficiency virus type 1 RNA in breast milk and of its relationship to infant 
infection and maternal disease. J Infect Dis. 187:741-7.
Rousseau, F.S., J.O. Kahn, M. Thompson, D. Mildvan, D. Shepp, J.P. Sommadossi, J. 
Delehanty, J.N. Simpson, L.H. Wang, J.B. Quinn, C. Wakeford, and C. van der Horst. 
2001. Prototype trial design for rapid dose selection of antiretroviral drugs: an example 
using emtricitabine (Coviracil). J Antimicrob Chemother. 48:507-13.
Schechter, M., R.F. do Lago, A.B. Mendelsohn, R.I. Moreira, L.H. Moulton, and L.H. Harrison. 
2004. Behavioral impact, acceptability, and HIV incidence among homosexual men with 
access to postexposure chemoprophylaxis for HIV. J Acquir Immune Defic Syndr. 
35:519-25.
Smith, B.P., F.R. Vandenhende, K.A. DeSante, N.A. Farid, P.A. Welch, J.T. Callaghan, and S.T. 
Forgue. 2000. Confidence interval criteria for assessment of dose proportionality. Pharm 
Res. 17:1278-83.
Subbarao, S., A. Ramos, C. Kim, D. Adams, M. Monsour, S. Butera, T. Folks, and R.A. Otten. 
2007. Direct stringency comparison of two macaque models (single-high vs. repeat-low) 
for mucosal HIV transmission using an identical anti-HIV chemoprophylaxis 
intervention. J Med Primatol. 36:238-43.
UNAIDS. 2008. Report on the Global AIDS Epidemic.
Van Belle, G., and D.C. Martin. 1993. Sample Size as a Function of Coefficient of Variation and 
Ratio of Means. American Statistician. 47:165-167.
Veazey, R.S., M.S. Springer, P.A. Marx, J. Dufour, P.J. Klasse, and J.P. Moore. 2005. Protection 
of macaques from vaginal SHIV challenge by an orally delivered CCR5 inhibitor. Nat 
Med. 11:1293-4.
Wang, L.H., J. Begley, R.L. St Claire, 3rd, J. Harris, C. Wakeford, and F.S. Rousseau. 2004. 
Pharmacokinetic and pharmacodynamic characteristics of emtricitabine support its once 
daily dosing for the treatment of HIV infection. AIDS Res Hum Retroviruses. 20:1173-82.
Zhou, J., J. Li, and B. Coate. 2006. Empirical Power Estimation for Phase I Dose Proportionality 
Studies Based on Power-Law Model Using Confidence Interval Criteria. In SUGI 31, San 
Francisco, California.

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APPENDICES

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APPENDIX I: SCHEDULE OF EVALUATIONS AND PROCEDURES

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APPENDIX IA: SCHEDULE OF EVALUATIONS AND PROCEDURES - GENERAL PROCEDURES
Phase
Screening
Enrollment
Accumulation
Steady-State
Decay
Study Week
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Study Day
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
Treatment Arm and 
Regimen
1. FTC 200 mg/TDF 
300 mg one tablet orally 
once weekly for 5 
weeks
X
X
X
X
X
2. FTC 200 mg/TDF 
300 mg  one tablet 
orally twice weekly for 
5 weeks
X
X
X
X
X
X
X
X
X
X
3. FTC 200 mg/TDF 
300 mg  two tablets 
orally twice weekly for 
5 weeks
X
X
X
X
X
X
X
X
X
X
4. FTC 200 mg/TDF 
300 mg C one tablet 
once daily for 5 weeks
X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X
Administrative and 
Behavioral Procedures
Informed Consent
X
Locator Information
X X
X
X
X
X
X
X
Demographic 
Information
X
Medical History 
Questionnaire
X

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APPENDIX IB: SCHEDULE OF EVALUATIONS AND PROCEDURES - CLINICAL PROCEDURES AND LABORATORY 
TESTS
Phase
SCR
ENR
Accumulation
Steady-State
Decay
Study Week
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Study Day
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
Clinical 
Procedures and 
Laboratory 
Tests
Blood 
(Serum/Plasma)
HIV Diagnostic 
Testinga
X X
X
Safety testingb
X
X
X
X
X
Coagulation 
testingc
X
X
HBsAg*
X
X
Syphilis Testingk, *
X
Plasma storage
X X
X
X
Urine
Pregnancy testj 
(women)
X X
X
X
X
Dipstick 
urinalysisd, *
X
GC/CT testing k, *
X

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Phase
SCR
ENR
Accumulation
Steady-State
Decay
Study Week
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Study Day
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
Pelvic (women) 
(intensive cohort
only)
Wet mounte
X
Vaginal fluid pHf
X
Pap Smearg
X
Genital lesions 
(if present;
(intensive cohort 
only)
HSV-2 testingh, *
X
Rectal (intensive 
cohort only)
GC/CT testingi, *
X

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FOOTNOTES FOR APPENDIX IB
a Perform testing as described in the SSP. Participants with one or more HIV rapid test results are not eligible for enrollment, even if HIV infection is
not confirmed. HIV diagnostic testing should be repeated at any study visit if a participant expresses a concern that he/she may have acquired HIV 
infection. 
b Safety testing includes CBC, ALT/AST, phosphate, creatinine (for estimated creatinine clearance), electrolytes (HCO3, K, Cl, Na), Ca, glucose. 
c Coagulation testing includes INR, PT and PTT. Intensive sampling cohort only.
d If urine dipstick is positive for leukocyte esterase or nitrates, perform urinalysis along with urine culture.
e Wet mount includes testing for BV, candida, trichomonas.
f Vaginal pH testing is only required if Amstel criteria are used for diagnosis of BV.
g A Pap smear will be performed if a normal test result cannot be documented within the previous 12 months.
h A PCR-based test for HSV-2 will be performed at the HPTN NL.
i Rectal GC/CT NAAT testing will be performed at the HPTN NL.
j Pregnancy testing will be done before any vaginal study procedures are performed. On Day 35, pregnancy testing will only be performed on women 
in the intensive sampling cohort. 
k These tests are only for participants in the intensive sampling cohort
* Individuals with a positive result for any of these tests are not eligible for enrollment, and will be referred for care. Confirmatory testing should be 
performed as needed according to local testing guidelines. Testing should be repeated at any study visit if a participant expresses a concern that he/she 
may have acquired hepatitis B or an STI.

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APPENDIX IC: SCHEDULE OF EVALUATIONS AND PROCEDURES - SAMPLE COLLECTION FOR PK EVALUATION 
(DRUG TESTING)
Phase
SCR
ENR
Accumulation
Steady-State
Decay
Study Week
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Study Day
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
PK Sampling
Blood serum
X 4
2
3
X
1
X
X
X
X
X
Blood PBMC
X 4
2
3
X
1
X
X
X
X
X
Blood CD4+ cells
X 4
2
3
X
1
X
X
X
X
X
Rectosigmoid 
tissue homogenate
Xa
Xa
Rectosigmoid
tissue total cells
Xa
Xa
Rectosigmoid
tissue CD4+ cells
Xa
Xa
Rectosigmoid 
luminal fluid
Xa
Xa
Vaginal tissue 
homogenate
Xa
Xa
Vaginal tissue 
total cells
Xa
Xa
Vaginal tissue 
CD4+ cells
Xa
Xa
Vaginal fluid
Xa
Xa
Semen
Xa
Xa

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In rows for Blood Serum, Blood PBMC, and Blood CD4 cells, days 1-7, the number indicates a sample collection for each cohort (indicated by cohort 
number, as listed in the protocol schema) no more than one hour before the second dose for each cohort. These will occur on day 7 (Cohort 1), Day 3
(Cohort 2 and 3), and Day 1 (Cohort 4).
a Sixteen participants who agree to intensive tissue sampling will be enrolled in a balanced fashion across dosing cohorts such that two men and two 
women per cohort are enrolled.  For other participants, they will only have blood and blood cells collected on Day 35 and 49. 

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APPENDIX 1D: ADDITIONAL PROCEDURES FROM STUDY PARTICIPANTS WITH 
CONFIRMED HIV INFECTION
1 All stored samples from HIV seroconverters will be centralized at the HPTN NL. 
2 The HPTN NL will coordinate HIV resistance testing using samples shipped to the HPTN NL. HIV 
resistance test results from this study visit (time of HIV diagnosis) will be provided to study sites at study 
closure. Results from this testing may be provided to study sites prior to study closure with approval of 
the HPTN NL and the Protocol Chair. 
3 Additional testing may be performed at the HPTN NL or at another laboratory designated by the HPTN 
NL for research purposes; those results will not be returned to study sites or study participants. This 
testing may include HIV subtyping, minority variants assays, other assays to characterize HIV viruses 
and/or the host response to HIV infection. Additional safety testing will also be performed on all 
participants who seroconvert during the study. 
4 Participants who seroconvert during study participation will have a final safety visit (visit may be split if 
necessary) after confirmation of their HIV status.
Time of diagnosis
All subsequent visits4
CD4 cell count testing
X
HIV viral load testing
X
Shipment of samples to the HPTN NL1
X
X
HIV resistance testing 2
X
Other testing 3
X
X
Additional plasma storage
X
X

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APPENDIX II: SCREENING/ENROLLMENT SAMPLE INFORMED CONSENT 
FORM (Intensive Sampling Cohort)

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SAMPLE INFORMED CONSENT DOCUMENT (Intensive Sampling Cohort)
HPTN 066
DOSE-PROPORTIONALITY AND INTRA-INDIVIDUAL VARIABILITY OF INTRACELLULAR 
TENOFOVIR DIPHOSPHATE AND EMTRICITABINE TRIPHOSPHATE IN HEALTHY VOLUNTEERS
29 NOVEMBER 2010
VERSION 2.0
PRINCIPAL INVESTIGATOR: [Insert Name]
PHONE:  [Insert Number]
GENERAL OVERVIEW 
You are being asked to take part in a research study. Joining this study is voluntary. 
You may refuse to join, or you may withdraw your consent to be in the study, for any reason.
Research studies are designed to obtain new knowledge that may help other people in the future.  
You may not receive any direct benefit from being in the research study. There also may be risks 
to being in research studies.
Details about this study are discussed below.  It is important that you understand this information 
so that you can make an informed choice about being in this research study.  You will be given a 
copy of this consent form.  You should ask the researchers named above, or staff members who 
may assist them, any questions you have about this study at any time.
YOUR PARTICIPATION IS VOLUNTARY
This consent form gives information about the study that will be discussed with you. Once you 
understand the study, and if you agree to take part, you will be asked to sign your name on this 
form. You will be given a copy of this form to keep. 
Before you learn about the study, it is important that you know the following:
·
Your participation is voluntary.  You do not have to take part in the screening tests or in 
the study.
·
You may decide not to take part in the study or to leave the study at any time without 
losing your regular medical care.
·
If you decide not to take part in the study, you can still join another study at a later time if 
there is one available and you qualify.    
·
You cannot join this study if you are taking part in another study of drugs, medical 
devices or vaginal/rectal products.  You are asked to tell the study staff about any other 
studies you are taking part in or thinking of taking part in.  This is very important for 
your safety.  

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PURPOSE OF THE STUDY
This study is being conducted to look at how the body handles the drug Truvada® when it is 
given in different amounts and at different time schedules. This study will also look at the 
difference of medication amount in men and women.
The study will look at levels of Truvada® in the blood, reproductive tract and gastrointestinal 
tract of female and male volunteers.  The aim is to establish how much of the drug (taken by 
mouth) reaches the reproductive and gastrointestinal tracts when given at different doses and at 
different dose schedules. It is believed that the presence of this drug in the reproductive and 
gastrointestinal tracts may be beneficial in the prevention of the transmission of the AIDS virus.  
About 32 participants will be enrolled in the study at 2 different sites in the United States.  About 
16 people will be enrolled in the study here at [study site].  The study should take approximately 
12 months to complete. Each participant will be followed for approximately 2 months.
The study will take samples of blood once a week for the first 5 weeks after taking Truvada® 
according to the dose you are assigned to. After that time, blood samples and tissue samples from 
the gastrointestinal tract (from men and women) will be collected. Samples will also be collected 
from the vagina in women. Men will be asked to donate a semen sample. All of these samples 
are to look at drug concentrations.  You will then have a second set of blood sample and tissue 
biopsies after you have stopped taking the study drug for 2 weeks. The study will also collect 
information on the safety of the medication.
Only 16 of the 32 participants will provide tissue/semen/vaginal samples as part of study 
participation. The remaining participants will only provide blood samples. Potential participants 
will decide with study staff which group they will enroll in before they are consented for the 
study. 
Truvada® is a tablet that is a combination of two medications—emtricitabine and tenofovir. This 
medication is used to treat people who have HIV.  The tablets are generally safe when used as 
treatment for HIV.  These tablets are not a cure for HIV/AIDS, but they are very effective for 
improving the health of people who have HIV/AIDS.  Although these tablets improve the health 
of people with HIV, we do not know if they work to protect against HIV. The results of this 
study may be used in the future to determine if this medication can be taken by healthy people to 
prevent them from getting HIV.
The United States Food and Drug Administration (U.S. FDA) has been informed of this study. 
Truvada has not been approved by the U.S. FDA for the prevention of HIV. 
Before you decide whether to take part in the study, we would like to explain the purpose of the 
study, the risks and benefits, what is expected of you, and what to expect from us.  This consent 
form might contain some words that are unfamiliar.  Please ask questions about anything you do 
not understand or want to learn more about.

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STUDY GROUPS
There are four study groups.  Each group will have 8 people.  There will be 4 women and 4 men 
in each group.  If you decide to take part in the study, you will be randomly assigned to one of 
the four groups, like flipping a coin.  Once you are assigned to a group, you cannot change to 
another group.  The four groups will each be given the same medication, but it will be provided 
in different doses.
·
One group will take one Truvada® tablet one time each week for five weeks
·
One group will take one Truvada® tablet two times each week for five weeks
·
One group will take two Truvada® tablets two times each week for five weeks
·
One group will take one Truvada® tablet each day for five weeks
All four groups are very important to the study.  It is very important that each person takes 
his/her medication as provided.  Because of this, all participants will be observed by a study 
employee when taking the medication so that we can be sure that the participant has taken the 
medication on the right day and at the right time.  This means that some people will be required 
to come to the clinic every day (including Saturday and Sunday) to get their medication.
CRITERIA FOR STUDY PARTICIPATION
In order to participate in the study, you must meet all of the following criteria:
·
Be 18 to 44 years of age, inclusive on the date of screening.
·
Provide informed consent for the study.
·
Provide information on how to contact and locate you
·
Be willing to be tested for HIV infection and receive your test results.
·
Be HIV negative.
·
Have good kidney function.
·
Be sexually active (at least one sex act - vaginal or anal intercourse - in the previous 30 
days). 
·
Agree to use condoms.
·
Women must:
o
Be pre-menopausal
o
Have regular menstrual cycles with at least 21 days between menses (unless on 
contraception that causes lack of or irregular menses) 
o
Have a cervix
o
Have a negative urine pregnancy test
o
Have documentation of a normal Pap smear within 12 months
o
Be utilizing an alternative method of birth control in addition to condoms 
(hormonal contraceptive, diaphragm or have undergone surgical sterilization) or 
have a vasectomized exclusive male partner.
o
Not using spermicide as a means of birth control (with a condom or a diaphragm)

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REASONS WHY YOU WOULD NOT BE ABLE TO PARTICIPATE 
You will not be able to participate in the study if any of the following things apply to you:
·
Blood tests that show the following:
○
Poor liver function
○
Anemia (low blood counts)
○
Abnormal laboratory values
·
Co-enrollment in any other HIV research study that involves an experimental drug or 
prior enrollment in the active arm of a HIV vaccine trial.
·
Active skin disorders.
·
Excessive use of alcohol (more than 4 drinks a day on a regular basis).
·
Any procedure that has altered the stomach or intestines or drug absorption.
·
Abnormalities that will not allow the biopsies to be taken. 
·
Pelvic exam abnormalities. 
·
Women who have had cervical procedures within the previous 6 months.
·
A positive test for syphilis, gonorrhea, chlamydia, hepatitis B, or HSV-2 (individuals 
with active lesions only).
·
Bacterial vaginosis, vaginal candidiasis, or trichomonas (women only).
·
Sexually transmitted infection within the last 3 months
·
Women who are pregnant or breastfeeding.
·
A reactive or positive HIV test result at screening or enrollment, even if you are found to 
be HIV-uninfected based on further testing.
·
Medications that prolong blood clotting time.
·
Medications that affect the kidney or liver. 
·
Anything else in the judgment of the study clinician that would make participation in the 
study unsafe, complicate interpretation of study outcome data, or otherwise interfere with 
achieving the study objectives.
STUDY PROCEDURES
Screening and Enrollment Procedures:
If you agree to have the screening tests, the screening visits will be performed in one to two 
visits, but may be done in more than two if necessary. However, if a person agrees to be enrolled, 
the enrollment visit must take place within 30 days of the very first screening visit.  
Screening Visit:
Your screening visit will happen after you read, discuss, understand, and sign this form. Study 
staff will help you understand the form and answer your questions before you sign this form. The 
procedures done at this visit will take about 2-3 hours.   

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The study staff will: 
·
Ask you where you live and other questions about you, your medical health, and your 
sexual practices.  
·
Discuss the details of “directly observed therapy” and identify how you will be able to 
come to the clinic (or other identified designation) to receive your medication. You will 
need to understand that you have a 1 in 4 chance of having to come to the clinic every 
day to receive your medication.
·
If these questions show that you may be eligible for the study and you are a woman, you 
will give urine for a pregnancy test.   If you are pregnant, you are not eligible for the 
study.  The study staff will refer you to available sources of medical care and other 
services you may need.  If the study is still open after your pregnancy, you can come 
back here to find out if you are eligible then.
●
If you are a woman who is not pregnant or breastfeeding or a man, you will talk with 
study staff about HIV, HIV testing, and ways to avoid HIV and other infections passed 
through sex. You will be asked to give a blood sample (up to 20 mL or 4 teaspoons) for 
HIV testing. You will be told your result as soon as it is available. You will talk with the 
study staff about the meaning of your results and how you feel about them. Sometimes 
HIV tests are not clearly positive, but also not clearly negative. In that case, we will do 
more tests until we know your status for sure. You must receive your HIV test results to 
be in the study. If the test shows that you have HIV, you will not be eligible for the study. 
The study staff will tell you about other studies you may be eligible for, if any. They will 
refer you to available sources of medical care and other services you may need. Your 
partner(s) may also have access to free HIV counseling and testing if needed.
●
Measure your weight.
●
Perform a complete physical examination including your genital area. 
●
Examine inside your vagina (if you are a woman).
o
The study staff will collect fluid from your vagina to test for infections. These 
infections are called trichomoniasis, candidiasis, and bacterial vaginosis (BV). 
●
Talk with you about contraception and ways to prevent sexually transmitted diseases.
●
Test your urine for:
o
Chlamydia and gonorrhea.  These are sexually transmitted infections.  
o
Common urine infections and the health of your kidneys
●
Collect 40 mL (about 8 teaspoons) of blood for :
o
Syphilis testing.  This is a sexually transmitted infection.  
o
To check the health of your blood, liver and kidneys.
o
Hepatitis B testing.  This is an infection of the liver that can be passed from mother 
to baby, through sex, or through body fluids infected with hepatitis B.  If the tests 
show that you have hepatitis B that is active in your liver, you will not be eligible 
for the study.  If the tests show that you have had hepatitis B in the past, but it is no 
longer active in your liver, or that you are immune to hepatitis B because of a prior 
immunization, you may be eligible for the study.
o
Plasma storage for study related testing and long-term storage (if you provide 
consent)
●
Give you condoms.

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The results of the tests listed above will be available within 1-2 weeks after your visit.  You will 
be contacted about the results of your tests when they are available.  A small amount of plasma 
will be stored from this visit. No other samples collected at the time of screening will be kept or 
used for any other tests other than those listed above.
Confirmation of Eligibility:
Once all the results of the screening tests are known, the study staff will:
●
Tell you your test results from your screening visit and what they mean.  If the results 
show that you might have some health problems, you may not be eligible for the study.  
Study staff will refer you to available sources of medical care and other services you may 
need.  Later, if these problems resolve, you can come back to find out if you are eligible 
at that time.
●
Refer you for treatment of a urinary tract infections and sexually transmitted infections, if 
you need it.  If you need this treatment, you will not be eligible for the study.
●
If you have a reactive or positive HIV test or a test that shows that you have active 
hepatitis B infection, you will not be eligible for the study, but you will be referred to the 
appropriate clinic or other medical resource.
●
Give you referrals for other health services if you need them. 
●
Identify a clear plan for directly observed therapy. 
Enrollment Visit:
If you are eligible for this study and decide to take part in the study, you will be asked to return 
for the enrollment visit. At that time, the study staff will: 
●
Perform a urine pregnancy test (women only)
●
Collect up to 50 mL (about 10 teaspoons) of blood for:
o
HIV diagnostic testing 
o
To check the amount of medication (Truvada®) in your blood.
●
Review and update your contact details.
●
Review and update your medical history.
●
Talk with you again about HIV and other infections passed through sex and how to avoid 
these. 
●
Identify your study group.
●
Give you condoms.
●
Administer your first dose of study medication to you and review the procedures for all 
future doses to be observed by study personnel.
Before your second dose:
Before you take your second dose of study medication, you will have blood drawn (up to 40 mL 
or about 8 teaspoons).
At Visits on Days 7, 14, 21, and 28, you will:
●
Tell study staff if you had any health problems since your last visit.
●
Tell study staff about any medications, herbal treatments and supplements you are taking.
●
Tell study staff any new information on where you live and how to keep in contact with 
you. They will use this information to remind you of visits. If you miss a visit, the study 

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staff will try to contact you by [site-specific methods]. They also may visit your home. 
They will try to reach you through the contact people that you list.  If they talk to these 
people, they will not say why they want to reach you.  
●
Talk with study staff about ways to avoid HIV and other infections passed through sex.  
●
Talk to study staff about adherence to the directly observed therapy regimen (if 
necessary).
●
Talk with study staff about contraception.
·
Give a urine sample for a pregnancy test (women only, Day 28 only).
·
Give blood (up to 65 mL or about 13 teaspoons): 
o
To check the amount of medication (Truvada®) in your blood. 
o
To confirm that the medication is not causing problems for certain parts of your 
body such as the liver and kidneys and to check the health of your blood (Days 14 
and 28 only). 
o
Plasma storage for study related testing and long-term storage (if you provide 
consent; Days 28).
●
Get condoms.
The Enrollment and Days 7, 14, 21, and 28 visits will take 1-2 hours each.
At Visits on Days 35 and 49, you will:
●
Participate in the same activities as the previous visits.  
●
Have an exam of the rectum and colon and small samples of your rectal tissue will be 
taken to measure the amount of the medications in your rectal tissue. For this, you will be 
taken to the endoscopy area where a doctor will perform a flexible sigmoidoscopy (also 
known as an endoscopy). To do this, the doctor will use an endoscope to look inside 
your rectum. An endoscope is a long, thin tube with a camera in it.  The doctor will use a 
long wire with a tiny pincher to take up to 30 small biopsies of your intestinal lining. 
Each biopsy is a small piece of tissue measuring smaller than the surface of a pencil 
eraser and about the thickness of a dime.
●
Have a clear, plastic, lubricated tube gently inserted into the rectum (no more than 4 
inches) in order to collect the rectal luminal fluid with a swab.
●
Have a vaginal exam (women only), and vaginal fluids, vaginal cells and small samples 
of vaginal tissue will be collected to measure the medication levels in your vaginal fluids, 
cells and tissue. (These samples will not be taken during menstruation so the timing of 
your menstrual cycle will have to be considered before you take the first study 
medication dose. Pregnancy tests will be performed just prior to any vaginal sampling 
and confirmed to be negative prior to any vaginal procedure.)  
●
Provide a semen sample (men only). 
●
Provide 50 mL (about 10 teaspoons) of blood for: 
o
Tests to confirm that the medication is not causing problems for certain parts of 
your body such as the liver and kidneys,
o
HIV test and Hepatitis B test and (Day 49 only)
o
Plasma storage for study related testing and long-term storage (if you provide 
consent; Day 49 only).
●
Urine for a pregnancy test (women only).

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The Days 35 and 49 visits may take 2-4 hours each. In some cases, due to scheduling limitations, 
these visits may require visiting the clinic on 2 different days.
Men will be asked to abstain from sex for 24-48 hours before semen collection. 
All participants should start a low fiber diet 3 days before each rectal biopsy procedure and have 
a clear liquid diet 12 hours prior to the procedure. They should also refrain from meals (or 
coffee) the day of the procedure. Enemas will be administered before the biopsy procedures take 
place. An enema is a procedure that introduces liquids into the rectum and colon through the 
anus to cleanse the lower intestinal tract.
One day after the biopsies, a study team member will contact you by phone to see how you’re 
doing.
AT ANY TIME IN THE STUDY
If you have health problems that may be caused by sexually transmitted infections, you will:
·
Have an exam of your genital area and inside your vagina.
·
Give blood, urine, and/or vaginal fluid to test for infections passed through sex or other 
vaginal infections
·
Be referred for treatment of any sexually transmitted infection.
Any time HIV or STI testing is done during the study, study staff will contact you to provide you 
with the test results as soon as they become available. Study staff will also provide you with 
information related to your health that is discovered at any study visit.
If you have to leave the study for any reason, you will have one additional follow-up visit where 
you will have the same activities as at the Day 7 through Day 21 visits as well as blood tests to 
confirm the drug has not caused any problems in certain parts of your body such as the liver and 
kidneys. You will also have an HIV and hepatitis b test and a urine pregnancy test (women only).
Study staff will review and assess any medical conditions that you have during the study. Study 
staff may contact you after the study has ended to be sure that you do not have any continuing 
medical problems that require medical attention.
POSSIBLE FUTURE TESTS
Some of the blood that you give during this study may be left over after all of the study tests are 
completed.  The study staff also would like to keep your leftover blood. Future tests on stored 
blood may be unrelated to this study. You will be asked to sign at the end of this consent form to 
give permission for that. Even if you do not give permission to store your blood after the study, 
you can still be in this study. You may also withdraw your consent for specimen storage at any 
time. 
RISKS AND/OR DISCOMFORTS
Blood Draws and Sexually Transmitted Infection Testing
Whenever your blood is drawn, you may:

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·
Feel discomfort or pain. 
·
Feel dizzy or faint. 
·
Have a bruise, swelling, small clot, or infection where the needle goes in your arm or 
finger.
When you have genital exams, you may:
·
Feel discomfort in your genital area and/or inside your vagina.
·
Women may have a small amount of vaginal bleeding which will stop shortly after the 
exam.
Rectal Exam and Tissue Samples
Whenever you have a rectal exam and tissue samples are taken, you may:
·
Have mild discomfort 
·
Have the feeling of a “bloated stomach” 
On extremely rare occasions, you may have: 
·
Pain 
·
Infection
·
Bleeding or 
·
Perforation of the gastrointestinal tract (occurs about once out of every 100,000 
procedures and may require hospitalization and surgical management) 
When rectal fluid is collected via insertion of the plastic tube into the anus, on rare occasion you 
may:
·
Experience mild discomfort 
·
Have pain (should you have another condition that is already causing pain in the area) 
When you have an enema, you may experience:
·
Temporary discomfort caused by a bloated or cramping feeling  
·
Some air may be pumped into the rectum causing gas  
·
On extremely rare occasion, the enema tube could make a hole in the rectum requiring 
surgery
You should not have anal sexual intercourse or insert anything into your rectum for at least 72 
hours after the tissue sampling.
Vaginal Exam and Tissue Samples
Whenever you have a vaginal exam and tissue samples are taken, you may:
·
Have discomfort or pain during the procedure and for a few hours after  
·
Have mild vaginal spotting (bleeding) for one or two days
·
Have temporary discomfort with sexual intercourse  
There is a very small risk that you may:
·
Get an infection 

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·
Have heavy bleeding  
You should not have vaginal sexual intercourse or insert anything into our vagina for at least 72 
hours after the tissue sampling.
Study Medications
About 5 out of 100 people with HIV taking Truvada® tablets have these occasional side effects 
which usually go away after stopping the drug:
·
Upset stomach, vomiting, gas, loose or watery stools  
·
Dizziness or headache 
·
Abdominal pain 
·
Lack of energy/general body weakness
·
Mild problems of kidney function that are only detected by laboratory tests
·
Shortness of breath or cough 
·
Rash, including allergic reaction 
·
Anxiety
·
Joint pain, muscle pain, or other pain syndrome
·
Fever
Fewer than 5 out of 100 people with HIV taking Truvada® tablets have:
·
Skin discoloration/darkening of the palms and/or soles of the feet
Potentially serious side effects are rare, but include:
·
Liver function problems
·
Serious kidney damage or failure
·
Low phosphate levels (a chemical in the blood) or protein or sugar in the urine
·
Inflammation or swelling and possible damage to the pancreas
·
Bone pain and bone changes such as thinning and softening which may increase the risk 
of  breakage
·
Allergic reaction
·
Lactic acidosis and severe hepatomegaly (enlarged liver) with steatosis (fatty liver) that 
may result in liver failure, other complications and death have been reported with the use 
of antiretroviral nucleoside analogues alone or in combination.  The liver complications 
and death have been seen more often in women on these drug regimens. Some 
nonspecific symptoms that might indicate lactic acidosis include: unexplained weight 
loss, stomach discomfort, nausea, vomiting, fatigue, weakness, and shortness of breath.
·
If you are infected with both Hepatitis B and HIV, you should be aware that your liver 
function tests may increase, and symptoms associated with hepatitis (an acute 
inflammation of the liver) may worsen if emtricitabine or tenofovir is stopped.
Some people taking emtricitabine, one of the ingredients in Truvada®, experienced the following 
side effects:
·
Inability to sleep, unusual dreams
·
Runny nose

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·
Rash, itching, which sometimes can be a sign of an allergic reaction
·
Increases in pancreatic enzyme (substances in the blood), which could mean a problem 
with the pancreas 
·
Increased triglycerides 
·
Increased creatine phosphokinase (CPK), which could mean muscle damage
Some people taking tenofovir, one of the ingredients in Truvada®, experienced the following 
side effects:
·
Depression
·
Inflammation or swelling and possible damage to the pancreas and liver
·
Allergic reaction: symptoms may include fever, rash, upset stomach, vomiting, loose or 
watery stools, abdominal pain, achiness, shortness of breath, a general feeling of illness 
or a potentially serious swelling of the face, lips, and/or tongue
You could have these side effects or other side effects that we do not know about.
Other Possible Risks
We do not know if there are other risks if you use herbal treatments or supplements while you 
are using the tablets.  Please tell study staff if you are using any herbal treatments or 
supplements.  
We will perform an HIV test, which is routinely done before HIV drugs are tested in non-HIV 
subjects. You will be counseled before and after this test is done. In accordance with some state 
laws, all positive tests must be reported to the state health department. In the event that you test 
positive for HIV, you will be notified and provided with counseling. If you do test positive, the 
test results will become part of public health records.  A positive result will be placed in your 
hospital chart in order to provide you with the best of care.  You are not required to make these 
results available to your insurance company or employer.
We will perform screening for common sexually transmitted diseases, including bacterial 
vaginosis, syphilis, gonorrhea, Chlamydia, HSV-2 and trichomonas.  This is being done because 
infections could affect the measurements of drug concentrations in cervicovaginal fluid or 
vaginal tissue that will be taken in this study.  Syphilis, gonorrhea, and Chlamydia are reportable 
sexually transmitted infections in some states, and positive results must be reported with your 
name to the local health department.  With any positive result for any of these tests, you will be 
referred to an infectious diseases provider for confirmation (if needed) and treatment.   
In addition, there may be uncommon or previously unknown risks that might occur. You should 
report any problems to the researchers immediately.
There may also be some social risks to participating in this study. You may feel embarrassed or 
uncomfortable with some of the questions you will be asked, some of the procedures that will be 
done, or some of the test results that you will receive. You may also experience stigma as a result 
of being involved in an HIV study because people may assume that you are HIV-infected. 

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Pregnancy and Breastfeeding
If you become pregnant during the study, or within 28 days after completing the study, you 
should notify the study personnel right away for safety monitoring. If you become pregnant 
during the study, you will stop using the tablets right away. The study staff will refer you to 
available sources of medical care and other services you or your baby may need. The study does 
not pay for this care.  
Please tell your obstetrician about your study participation. Your study doctor will ask that you, 
or your obstetrician, provide updates on the progress of your pregnancy and its outcome.  The 
study doctor will make this information available to the study sponsor for safety monitoring 
follow-up.  
We do not know if Truvada® tablets have any effect on pregnancy, on the fetuses of women 
who use tablets when pregnant, or on the babies of women who use tablets when breastfeeding. 
Because of this, pregnant women and women who are breastfeeding will not be allowed to 
participate in this study. Women who join the study must use condoms in addition to an effective 
contraception and method must have pregnancy tests at the screening visit, before starting the 
study, and at visits 4, 5, and 6 while in the study.  Effective contraception includes hormonal 
methods (such as the birth control pill or shot), intrauterine contraceptive device (IUCD); and 
sterilization.  You should not use spermicides as a method of contraception while participating in 
this study.  
BENEFITS
You will not receive any direct benefit from being in this study. You or others may benefit in the 
future from information learned in this study. 
NEW INFORMATION
You will be told any new information learned during this study that might affect your 
willingness to stay in the study. For example, if information becomes available that shows that 
the medication may be causing bad effects, you will be told about this. You will also be told 
when the results of the study may be available, and how to learn about them.  
WHY YOU MAY BE WITHDRAWN FROM THE STUDY WITHOUT YOUR 
CONSENT
You may be withdrawn from the study without your consent if any of the following occur: 
·
Become pregnant.
·
Are breastfeeding.  
·
Are taking certain medications that affect your kidneys.
·
Are unable or unwilling to follow all of the study procedures or instructions.  
·
Could be harmed by continuing to take tablets.
·
The study is stopped or canceled.
·
The study staff feels that staying in the study would be harmful to you.
·
You are not able to attend clinic visits or complete all of the study procedures.
·
You are not able to have directly observed therapy according to study instructions.

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·
Other reasons, as decided by the study staff.
·
You become HIV positive
If you become HIV positive during the course of the study or are withdrawn for any reason, you 
will have a final study visit for safety testing before you are withdrawn from study participation. 
ALTERNATIVES TO PARTICIPATION
[Sites to include/amend the following if applicable: There may be other studies going on here or
in the community that you may be eligible for. If you wish, we will tell you about other studies
that we know about. There also may be other places where you can go for HIV counseling and
testing and contraception. We will tell you about those places if you wish.]
COSTS TO YOU
There is no cost to you for being in this study. 
REIMBURSEMENT
[Sites to insert information about local reimbursement amount and schedule:]
You will receive [$xx] for your time, effort, and travel to and from the clinic. 
CONFIDENTIALITY
We will do everything we can to keep your personal information confidential.  To help us keep 
your personal information confidential, we have obtained a Certificate of Confidentiality from 
the US Federal Government.  This certificate protects researchers from being forced to tell 
people who are not connected with this study, such as the court system, about your participation.  
However, it is not possible to guarantee confidentiality. Your personal information may be 
disclosed if required by law. 
Even with the Certificate of Confidentiality, if the study staff learns of possible child abuse 
and/or neglect or a risk of harm to yourself or others, we will be required to tell the proper 
authorities.
The study staff will also use your personal information, if needed, to verify that you are not 
taking part in any other research studies. This includes other studies conducted by [site name] 
and studies conducted by other researchers that study staff know about. Any publication of this 
study will not use your name or identify you personally.
Your records may be reviewed by:
●
the United States Food and Drug Administration (FDA)
●
the United States National Institutes of Health (NIH)
●
the United States Department of Heath and Human Services (DHHS), Office of Human 
Research Protection (OHRP)
●
[insert names of applicable IRBs/ECs]
●
study staff
●
study monitors
●
the company that makes Truvada® tablets (Gilead Sciences)

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[Sites to include/amend the following if applicable:] [Local/state/national] regulations require 
study staff to report the names of people who test positive for [HIV and other infections] passed 
during sex to the [local health authority]. Outreach workers from the [health authority] may then 
contact you about informing your partners, since they also should be tested. If you do not want to 
inform your partners yourself, the outreach workers will contact them, according to the 
confidentiality guidelines of the [health authority].
RESEARCH-RELATED INJURY
[Sites to specify institutional policy:] It is unlikely that you will be injured as a result of study 
participation. If you are injured, the [institution] will give you immediate necessary treatment for 
your injuries. You [will/will not] have to pay for this treatment. You will be told where you can 
get additional treatment for your injuries. There is no program to pay money or give other forms 
of compensation for such injuries either through this institution or the NIH. You do not give up 
any legal rights by signing this consent form.
PROBLEMS OR QUESTIONS
If you ever have any questions about the study, or if you have a research-related injury, you 
should contact [insert name of the investigator or other study staff] at [insert telephone number 
and/or physical address].
If you have questions about your rights as a research participant, you should contact [insert name 
or title of person on the IRB/EC or other organization appropriate for the site] at [insert physical 
address and telephone number].
If you have questions about whom to contact at the research site, you should contact [insert name 
of the investigator or community educator or CAB member] at [insert physical address and 
telephone number].

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SIGNATURES
[Insert signature blocks as required by the local IRB/EC:] If you have read this consent form, or 
had it read and explained to you, and you understand the information, and you voluntarily agree 
to join the study, please sign your name or make your mark below. Also, please indicate if you 
are willing to allow your leftover samples to be stored for future testing. 
Specimens Stored for Future Testing
_____ My initials indicate that any left-over samples may be stored for future testing:
_____ I do not agree to allow leftover samples to be saved for long term storage and future 
testing after the study
____________________ 
________________________ 
______________
Participant Name (print)
Participant Signature
Date
____________________ 
________________________
______________
Study Staff Conducting 
Study Staff Signature
Date
Consent Discussion (print)
____________________ 
________________________ 
______________
Witness Name (print)
Witness Signature
Date

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APPENDIX III: SCREENING/ENROLLMENT SAMPLE INFORMED CONSENT 
FORM (Non-Intensive Sampling Cohort)

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SAMPLE INFORMED CONSENT DOCUMENT (NON-Intensive Sampling Cohort)
HPTN 066
DOSE-PROPORTIONALITY AND INTRA-INDIVIDUAL VARIABILITY OF INTRACELLULAR 
TENOFOVIR DIPHOSPHATE AND EMTRICITABINE TRIPHOSPHATE IN HEALTHY VOLUNTEERS
29 NOVEMBER 2010
VERSION 2.0
PRINCIPAL INVESTIGATOR: [Insert Name]
PHONE:  [Insert Number]
GENERAL OVERVIEW 
You are being asked to take part in a research study. Joining this study is voluntary. 
You may refuse to join, or you may withdraw your consent to be in the study, for any reason.
Research studies are designed to obtain new knowledge that may help other people in the future.  
You may not receive any direct benefit from being in the research study. There also may be risks 
to being in research studies.
Details about this study are discussed below.  It is important that you understand this information 
so that you can make an informed choice about being in this research study.  You will be given a 
copy of this consent form.  You should ask the researchers named above, or staff members who 
may assist them, any questions you have about this study at any time.
YOUR PARTICIPATION IS VOLUNTARY
This consent form gives information about the study that will be discussed with you. Once you 
understand the study, and if you agree to take part, you will be asked to sign your name on this 
form. You will be given a copy of this form to keep. 
Before you learn about the study, it is important that you know the following:
·
Your participation is voluntary.  You do not have to take part in the screening tests or in 
the study.
·
You may decide not to take part in the study or to leave the study at any time without 
losing your regular medical care.
·
If you decide not to take part in the study, you can still join another study at a later time if 
there is one available and you qualify.    
·
You cannot join this study if you are taking part in another study of drugs, medical 
devices or vaginal/rectal products.  You are asked to tell the study staff about any other 
studies you are taking part in or thinking of taking part in.  This is very important for 
your safety.  

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PURPOSE OF THE STUDY
This study is being conducted to look at how the body handles the drug Truvada® when it is 
given in different amounts and at different time schedules. This study will also look at the 
difference of medication amount in men and women.
The study will look at levels of Truvada® in the blood, reproductive tract and gastrointestinal 
tract of female and male volunteers.  The aim is to establish how much of the drug (taken by 
mouth) reaches the reproductive and gastrointestinal tracts when given at different doses and at 
different dose schedules. It is believed that the presence of this drug in the reproductive and 
gastrointestinal tracts may be beneficial in the prevention of the transmission of the AIDS virus. 
However, half of the study participants will only participate in the blood portion of the study. 
That is the group that you are being evaluated for. Potential participants will decide with study 
staff which group they will enroll in before they are consented for the study.
The study will take samples of blood once a week for the first 5 weeks after taking Truvada® 
according to the dose you are assigned to. All of these samples are to look at drug 
concentrations.  The study will also collect information on the safety of the medication.
Truvada® is a tablet that is a combination of two medications—emtricitabine and tenofovir. This 
medication is used to treat people who have HIV.  The tablets are generally safe when used as 
treatment for HIV.  These tablets are not a cure for HIV/AIDS, but they are very effective for 
improving the health of people who have HIV/AIDS.  Although these tablets improve the health 
of people with HIV, we do not know if they work to protect against HIV. The results of this 
study may be used in the future to determine if this medication can be taken by healthy people to 
prevent them from getting HIV.
The United States Food and Drug Administration (U.S. FDA) has been informed of this study. 
Truvada has not been approved by the U.S. FDA for the prevention of HIV. 
About 32 participants will be enrolled in the study at 2 different sites in the United States.  About 
16 people will be enrolled in the study here at [study site].  The study should take approximately 
12 months to complete. Each participant will be followed for approximately 2 months.
Before you decide whether to take part in the study, we would like to explain the purpose of the 
study, the risks and benefits, what is expected of you, and what to expect from us.  This consent 
form might contain some words that are unfamiliar.  Please ask questions about anything you do 
not understand or want to learn more about.
STUDY GROUPS
There are four study groups.  Each group will have 8 people.  There will be 4 women and 4 men 
in each group.  If you decide to take part in the study, you will be randomly assigned to one of 
the four groups, like flipping a coin.  Once you are assigned to a group, you cannot change to 
another group.  The four groups will each be given the same medication, but it will be provided 
in different doses.

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·
One group will take one Truvada® tablet one time each week for five weeks
·
One group will take one Truvada® tablet two times each week for five weeks
·
One group will take two Truvada® tablets two times each week for five weeks
·
One group will take one Truvada® tablet each day for five weeks
All four groups are very important to the study.  It is very important that each person takes 
his/her medication as provided.  Because of this, all participants will be observed by a study 
employee when taking the medication so that we can be sure that the participant has taken the 
medication on the right day and at the right time.  This means that some people will be required 
to come to the clinic every day (including Saturday and Sunday) to get their medication.
CRITERIA FOR STUDY PARTICIPATION
In order to participate in the study, you must meet all of the following criteria:
·
Be 18 to 44 years of age, inclusive on the date of screening.
·
Provide informed consent for the study.
·
Provide information on how to contact and locate you
·
Be willing to be tested for HIV infection and receive your test results.
·
Be HIV negative.
·
Have good kidney function.
·
Be sexually active (at least one sex act - vaginal or anal intercourse - in the previous 30 
days).
·
Agree to use condoms.
·
Women must:
o
Be pre-menopausal
o
Have regular menstrual cycles with at least 21 days between menses (unless on 
contraception that causes lack of or irregular menses) 
o
Have a negative urine pregnancy test
o
Be utilizing an alternative method of birth control in addition to condoms 
(hormonal contraceptive, diaphragm or have undergone surgical sterilization) or 
have a vasectomized exclusive male partner.
REASONS WHY YOU WOULD NOT BE ABLE TO PARTICIPATE 
You will not be able to participate in the study if any of the following things apply to you:
·
Blood tests that show the following:
○
Poor liver function
○
Anemia (low blood counts)
○
Abnormal laboratory values
·
Co-enrollment in any other HIV research study that involves an experimental drug or 
prior enrollment in the active arm of a HIV vaccine trial.

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·
Active skin disorders.
·
Positive test for Hepatitis B
·
Excessive use of alcohol (more than 4 drinks a day on a regular basis).
·
Women who are pregnant or breastfeeding.
·
A reactive or positive HIV test result at screening or enrollment, even if you are found to 
be HIV-uninfected based on further testing.
·
Medications that affect the kidney or liver. 
·
Anything else in the judgment of the study clinician that would make participation in the 
study unsafe, complicate interpretation of study outcome data, or otherwise interfere with 
achieving the study objectives.
STUDY PROCEDURES
Screening and Enrollment Procedures:
If you agree to have the screening tests, the screening visits will be performed in one to two 
visits, but may be done in more than two if necessary. However, if a person agrees to be enrolled, 
the enrollment visit must take place within 30 days of the very first screening visit.  
Screening Visit:
Your screening visit will happen after you read, discuss, understand, and sign this form. Study 
staff will help you understand the form and answer your questions before you sign this form. The 
procedures done at this visit will take about 2-3 hours.   
The study staff will: 
·
Ask you where you live and other questions about you, your medical health, and your 
sexual practices.  
·
Discuss the details of “directly observed therapy” and identify how you will be able to 
come to the clinic (or other identified designation) to receive your medication. You will 
need to understand that you have a 1 in 4 chance of having to come to the clinic every 
day to receive your medication.
·
If these questions show that you may be eligible for the study and you are a woman, you 
will give urine for a pregnancy test.   If you are pregnant, you are not eligible for the 
study.  The study staff will refer you to available sources of medical care and other 
services you may need.  If the study is still open after your pregnancy, you can come 
back here to find out if you are eligible then.
●
If you are a woman who is not pregnant or breastfeeding or a man, you will talk with 
study staff about HIV, HIV testing, and ways to avoid HIV and other infections passed 
through sex. You will be asked to give a blood sample (up to 20 mL or 4 teaspoons) for 
HIV testing. You will be told your result as soon as it is available. You will talk with the 
study staff about the meaning of your results and how you feel about them. Sometimes 
HIV tests are not clearly positive, but also not clearly negative. In that case, we will do 
more tests until we know your status for sure. You must receive your HIV test results to 
be in the study. If the test shows that you have HIV, you will not be eligible for the study. 

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The study staff will tell you about other studies you may be eligible for, if any. They will 
refer you to available sources of medical care and other services you may need. Your 
partner(s) may also have access to free HIV counseling and testing if needed.
●
Measure your weight.
●
Perform a complete physical examination. 
●
Talk with you about contraception and ways to prevent sexually transmitted diseases.
●
Test your urine for:
o
The health of your kidneys
●
Collect 35 mL (about 7 teaspoons) of blood:
o
To check the health of your blood, liver and kidneys.
o
Hepatitis B testing.  This is an infection of the liver that can be passed from mother 
to baby, through sex, or through body fluids infected with hepatitis B.  If the tests 
show that you have hepatitis B that is active in your liver, you will not be eligible 
for the study.  If the tests show that you have had hepatitis B in the past, but it is no 
longer active in your liver, or that you are immune to hepatitis B because of a prior 
immunization, you may be eligible for the study. 
o
Plasma storage for study related testing and long-term storage (if you provide 
consent)
●
Give you condoms.
The results of the tests listed above will be available within 1-2 weeks after your visit.  You will 
be contacted about the results of your tests when they are available.  A small amount of plasma 
will be stored from this visit. No other samples collected at the time of screening will be kept or 
used for any other tests other than those listed above.
Confirmation of Eligibility:
Once all the results of the screening tests are known, the study staff will:
●
Tell you your test results from your screening visit and what they mean.  If the results 
show that you might have some health problems, you may not be eligible for the study.  
Study staff will refer you to available sources of medical care and other services you may 
need.  Later, if these problems resolve, you can come back to find out if you are eligible 
at that time.
●
Refer you for treatment of a urinary tract infections and sexually transmitted infections, if 
you need it.  If you need this treatment, you will not be eligible for the study.
●
If you have a reactive or positive HIV test or a test that shows that you have active 
hepatitis B infection, you will not be eligible for the study, but you will be referred to the 
appropriate clinic or other medical resource.
●
Give you referrals for other health services if you need them. 
●
Identify a clear plan for directly observed therapy. 
Enrollment Visit:
If you are eligible for this study and decide to take part in the study, you will be asked to return 
for the enrollment visit. At that time, the study staff will: 
●
Perform a urine pregnancy test (women only)
●
Collect up to 50 mL of blood (about 10 teaspoons) for:

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o
HIV diagnostic testing and
o
To check the amount of medication (Truvada®) in your blood.
●
Review and update your contact details.
●
Review and update your medical history.
●
Talk with you again about HIV and other infections passed through sex and how to avoid 
these. 
●
Identify your study group.
●
Give you condoms.
●
Administer your first dose of study medication to you and review the procedures for all 
future doses to be observed by study personnel.
Before your second dose:
Before you take your second dose of study medication, you will have blood drawn (40 mL or 
about 8 teaspoons).
At Visits on Days 7, 14, 21, 28, 35, and 49 you will:
●
Tell study staff if you had any health problems since your last visit.
●
Tell study staff about any medications, herbal treatments and supplements you are taking.
●
Tell study staff any new information on where you live and how to keep in contact with 
you. They will use this information to remind you of visits. If you miss a visit, the study 
staff will try to contact you by [site-specific methods]. They also may visit your home. 
They will try to reach you through the contact people that you list.  If they talk to these 
people, they will not say why they want to reach you.  
●
Talk with study staff about ways to avoid HIV and other infections passed through sex.  
●
Talk to study staff about adherence to the directly observed therapy regimen (if 
necessary).
●
Talk with study staff about contraception.
●
Give blood (up to 65 mL or about 13 teaspoons): 
o
To check the health of your blood, liver and kidneys.  
o
To check the amount of medication (Truvada®) in your blood.
o
To confirm that the medication is not causing problems for certain parts of your 
body such as the liver and kidneys (Day 14, 28, 35, and 49 only). 
o
For an HIV test and Hepatitis test (Day 49 only).
o
Plasma storage for study related testing and long-term storage (if you provide 
consent; Days 28 and 49 only).
●
Get condoms.
●
Give a urine sample for a pregnancy test (Women only; Day 28 and 49 only).  
The Enrollment and all subsequent study visits will take 1-2 hours each.

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AT ANY TIME IN THE STUDY
If you have health problems that may be caused by sexually transmitted infections, you will:
·
Have an exam of your genital area and inside your vagina.
·
Give blood, urine, and/or vaginal fluid to test for infections passed through sex or other 
vaginal infections
·
Be referred for treatment of any sexually transmitted infection.
Any time HIV or STI testing is done during the study, study staff will contact you to provide you 
with the test results as soon as they become available. Study staff will also provide you with 
information related to your health that is discovered at any study visit. 
If you have to leave the study for any reason, you will have one additional follow-up visit where 
you will have the same activities as at the Day 7 through Day 21 visits as well as blood tests to 
confirm the drug has not caused any problems in certain parts of your body such as the liver and 
kidneys. You will also have an HIV and hepatitis b test and a urine pregnancy test (women only).
Study staff will review and assess any medical conditions that you have during the study. Study 
staff may contact you after the study has ended to be sure that you do not have any continuing 
medical problems that require medical attention.
POSSIBLE FUTURE TESTS
Some of the blood that you give during this study may be left over after all of the study tests are 
completed.  The study staff also would like to keep your leftover blood. Future tests on stored 
blood may be unrelated to this study. You will be asked to sign at the end of this consent form to 
give permission for that. Even if you do not give permission to store your blood after the study, 
you can still be in this study. You may also withdraw your consent for specimen storage at any 
time.
RISKS AND/OR DISCOMFORTS
Blood Draws and Sexually Transmitted Infection Testing
Whenever your blood is drawn, you may:
·
Feel discomfort or pain. 
·
Feel dizzy or faint. 
·
Have a bruise, swelling, small clot, or infection where the needle goes in your arm or 
finger.
When you have genital exams, you may:
·
Feel discomfort in your genital area and/or inside your vagina.
·
Women may have a small amount of vaginal bleeding which will stop shortly after the 
exam.
Study Medications
About 5 out of 100 people with HIV taking Truvada® tablets have these occasional side effects 
which usually go away after stopping the drug:
·
Upset stomach, vomiting, gas, loose or watery stools  

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·
Dizziness or headache 
·
Abdominal pain 
·
Lack of energy/general body weakness
·
Mild problems of kidney function that are only detected by laboratory tests
·
Shortness of breath or cough 
·
Rash, including allergic reaction 
·
Anxiety
·
Joint pain, muscle pain, or other pain syndrome
·
Fever
Fewer than 5 out of 100 people with HIV taking Truvada® tablets have:
·
Skin discoloration/darkening of the palms and/or soles of the feet
Potentially serious side effects are rare, but include:
·
Liver function problems
·
Serious kidney damage or failure
·
Low phosphate levels (a chemical in the blood) or protein or sugar in the urine
·
Inflammation or swelling and possible damage to the pancreas
·
Bone pain and bone changes such as thinning and softening which may increase the risk 
of  breakage
·
Allergic reaction
·
Lactic acidosis and severe hepatomegaly (enlarged liver) with steatosis (fatty liver) that 
may result in liver failure, other complications and death have been reported with the use 
of antiretroviral nucleoside analogues alone or in combination.  The liver complications 
and death have been seen more often in women on these drug regimens. Some 
nonspecific symptoms that might indicate lactic acidosis include: unexplained weight 
loss, stomach discomfort, nausea, vomiting, fatigue, weakness, and shortness of breath.
·
If you are infected with both Hepatitis B and HIV, you should be aware that your liver 
function tests may increase, and symptoms associated with hepatitis (an acute 
inflammation of the liver) may worsen if emtricitabine or tenofovir is stopped.
Some people taking emtricitabine, one of the ingredients in Truvada®, experienced the following 
side effects:
·
Inability to sleep, unusual dreams
·
Runny nose
·
Rash, itching, which sometimes can be a sign of an allergic reaction
·
Increases in pancreatic enzyme (substances in the blood), which could mean a problem 
with the pancreas 
·
Increased triglycerides 
·
Increased creatine phosphokinase (CPK), which could mean muscle damage
Some people taking tenofovir, one of the ingredients in Truvada®, experienced the following 
side effects:
·
Depression
·
Inflammation or swelling and possible damage to the pancreas and liver

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·
Allergic reaction: symptoms may include fever, rash, upset stomach, vomiting, loose or 
watery stools, abdominal pain, achiness, shortness of breath, a general feeling of illness 
or a potentially serious swelling of the face, lips, and/or tongue
You could have these side effects or other side effects that we do not know about.
Other Possible Risks
We do not know if there are other risks if you use herbal treatments or supplements while you 
are using the tablets.  Please tell study staff if you are using any herbal treatments or 
supplements.  
We will perform an HIV test, which is routinely done before HIV drugs are tested in non-HIV 
subjects. You will be counseled before and after this test is done. In accordance with some state 
laws, all positive tests must be reported to the state health department. In the event that you test 
positive for HIV, you will be notified and provided with counseling. If you do test positive, the 
test results will become part of public health records.  A positive result will be placed in your 
hospital chart in order to provide you with the best of care.  You are not required to make these 
results available to your insurance company or employer.
In addition, there may be uncommon or previously unknown risks that might occur.  You should 
report any problems to the researchers immediately.
There may also be some social risks to participating in this study. You may feel embarrassed or 
uncomfortable with some of the questions you will be asked, some of the procedures that will be 
done, or some of the test results that you will receive. You may also experience stigma as a result 
of being involved in an HIV study because people may assume that you are HIV-infected. 
Pregnancy and Breastfeeding
If you become pregnant during the study, or within 28 days after completing the study, you 
should notify the study personnel right away for safety monitoring. If you become pregnant 
during the study, you will stop using the tablets right away. The study staff will refer you to 
available sources of medical care and other services you or your baby may need. The study does 
not pay for this care.  
Please tell your obstetrician about your study participation. Your study doctor will ask that you, 
or your obstetrician, provide updates on the progress of your pregnancy and its outcome.  The 
study doctor will make this information available to the study sponsor for safety monitoring 
follow-up.  

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We do not know if Truvada® tablets have any effect on pregnancy, on the fetuses of women who 
use tablets when pregnant, or on the babies of women who use tablets when breastfeeding. 
Because of this, pregnant women and women who are breastfeeding will not be allowed to 
participate in this study. Women who join the study must use condoms in addition to an effective 
contraception method and must have pregnancy tests at the screening visit, before starting the 
study, and at visit 4 while in the study.  Effective contraception includes hormonal methods 
(such as the birth control pill or shot), intrauterine contraceptive device (IUCD); and 
sterilization.  You should not use spermicides as a method of contraception while participating in 
this study.  
BENEFITS
You will not receive any direct benefit from being in this study. You or others may benefit in the 
future from information learned in this study. 
NEW INFORMATION
You will be told any new information learned during this study that might affect your 
willingness to stay in the study. For example, if information becomes available that shows that 
the medication may be causing bad effects, you will be told about this. You will also be told 
when the results of the study may be available, and how to learn about them.  
WHY YOU MAY BE WITHDRAWN FROM THE STUDY WITHOUT YOUR 
CONSENT
You may be withdrawn from the study without your consent if any of the following occur: 
·
Become pregnant.
·
Are breastfeeding.  
·
Are taking certain medications that affect your kidneys.
·
Are unable or unwilling to follow all of the study procedures or instructions.  
·
Could be harmed by continuing to take tablets.
·
The study is stopped or canceled.
·
The study staff feels that staying in the study would be harmful to you.
·
You are not able to attend clinic visits or complete all of the study procedures.
·
You are not able to have directly observed therapy according to study instructions.
·
Other reasons, as decided by the study staff. 
·
You become HIV positive
If you become HIV positive during the course of the study or are withdrawn for any reason, you 
will have a final study visit for safety testing before you are withdrawn from study participation.
ALTERNATIVES TO PARTICIPATION
[Sites to include/amend the following if applicable: There may be other studies going on here or
in the community that you may be eligible for. If you wish, we will tell you about other studies
that we know about. There also may be other places where you can go for HIV counseling and
testing and contraception. We will tell you about those places if you wish.]

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COSTS TO YOU
There is no cost to you for being in this study. 
REIMBURSEMENT
[Sites to insert information about local reimbursement amount and schedule:]
You will receive [$xx] for your time, effort, and travel to and from the clinic.
CONFIDENTIALITY
We will do everything we can to keep your personal information confidential.  To help us keep 
your personal information confidential, we have obtained a Certificate of Confidentiality from 
the US Federal Government.  This certificate protects researchers from being forced to tell 
people who are not connected with this study, such as the court system, about your participation.  
However, it is not possible to guarantee confidentiality. Your personal information may be 
disclosed if required by law. 
Even with the Certificate of Confidentiality, if the study staff learns of possible child abuse 
and/or neglect or a risk of harm to yourself or others, we will be required to tell the proper 
authorities.
The study staff will also use your personal information, if needed, to verify that you are not 
taking part in any other research studies. This includes other studies conducted by [site name] 
and studies conducted by other researchers that study staff know about. Any publication of this 
study will not use your name or identify you personally.
Your records may be reviewed by:
●
the United States Food and Drug Administration (FDA)
●
the United States National Institutes of Health (NIH)
●
the United States Department of Heath and Human Services (DHHS), Office of Human 
Research Protection (OHRP)
●
[insert names of applicable IRBs/ECs]
●
study staff
●
study monitors
●
the company that makes Truvada® tablets (Gilead Sciences)
[Sites to include/amend the following if applicable: ] [Local/state/national] regulations require 
study staff to report the names of people who test positive for [HIV and other infections] passed 
during sex to the [local health authority]. Outreach workers from the [health authority] may then 
contact you about informing your partners, since they also should be tested. If you do not want to 
inform your partners yourself, the outreach workers will contact them, according to the 
confidentiality guidelines of the [health authority].
RESEARCH-RELATED INJURY
[Sites to specify institutional policy:] It is unlikely that you will be injured as a result of study 
participation. If you are injured, the [institution] will give you immediate necessary treatment for 
your injuries. You [will/will not] have to pay for this treatment. You will be told where you can 

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get additional treatment for your injuries. There is no program to pay money or give other forms 
of compensation for such injuries either through this institution or the NIH. You do not give up 
any legal rights by signing this consent form.
PROBLEMS OR QUESTIONS
If you ever have any questions about the study, or if you have a research-related injury, you 
should contact [insert name of the investigator or other study staff] at [insert telephone number 
and/or physical address].
If you have questions about your rights as a research participant, you should contact [insert name 
or title of person on the IRB/EC or other organization appropriate for the site] at [insert physical 
address and telephone number].
If you have questions about whom to contact at the research site, you should contact [insert name 
of the investigator or community educator or CAB member] at [insert physical address and 
telephone number].

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SIGNATURES
[Insert signature blocks as required by the local IRB/EC:] If you have read this consent form, or 
had it read and explained to you, and you understand the information, and you voluntarily agree 
to join the study, please sign your name or make your mark below. Also, please indicate if you 
are willing to allow your leftover samples to be stored for future testing.
Specimens Stored for Future Testing
_____ My initials indicate that any left-over samples may be stored for future testing:
_____ I do not agree to allow leftover samples to be saved for long term storage and future 
testing after the study
____________________ 
________________________ 
______________
Participant Name (print)
Participant Signature
Date
____________________ 
________________________
______________
Study Staff Conducting 
Study Staff Signature
Date
Consent Discussion (print)
____________________ 
________________________ 
______________
Witness Name (print)
Witness Signature
Date